headings in a research paper

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Formatting Research Paper Headings and Subheadings

Different style guides have different rules regarding the formatting of headings and subheadings in a paper, but what information you should actually put into your subheadings is a different question and often up to personal taste. Here we quickly summarize general guidelines, different approaches, and what not to do when choosing headings for a research paper.

Does it matter how I name my sections and subsections?

The main sections of a research paper have general headers and are often journal-specific, but some (e.g., the methods and discussion section) can really benefit from subsections with clear and informative headers. The things to keep in mind are thus the general style your paper is supposed to follow (e.g., APA, MLA), the specific guidelines the journal you want to submit to lists in their author instructions , and your personal style (e.g., how much information you want the reader to get from just reading your subsection headers). 

Table of Contents:

• Style Guides: Rules on Headings and Subheadings
• What Sections and Subsections Do You Need? 
• How Should You Name Your Sections and Subsections?
• Avoid These Common Mistakes

Style Guides: Research Paper Heading and Subheading Format

Headers identify the content within the different sections of your paper and should be as descriptive and concise as possible. That is why the main sections of research articles always have the same or very similar headers ( Introduction, Methods, Results, Discussion ), with no or only small differences between journals. However, you also need to divide the content of some of these sections (e.g., the method section) into smaller subsections (e.g., Participants, Experimental Design, and Statistical Analysis ), and make sure you follow specific journal formatting styles when doing so. 

If the journal you submit to follows APA style , for example, you are allowed to use up to five levels of headings, depending on the length of your paper, the complexity of your work, and your personal preference. To clearly indicate how each subsection fits into the rest of the text, every header level has a different format – but note that headers are usually not numbered because the different formatting already reflects the text hierarchy.

APA style headings example structure

Level 1 Centered, Bold, Title Case

Text begins as a new paragraph.

Level 2 Left-aligned, Bold, Title Case

Level 3 Left-aligned, Bold Italic, Title Case

Level 4     Indented, Bold, Title Case, Period . Text begins on the same                    

                                 line and continues as a regular paragraph.

Level 5     Indented, Bold Italic, Title Case, Period. Text begins on the                           

                                 same line and continues as a regular paragraph.

If you only need one section header (e.g. Methods ) and one level of subsection headers (e.g., Participants, Experimental Design, and Statistical Analysis ), use Level 1 and Level 2 headers. If you need three levels of headings, use Levels 1, 2, and 3 (and so on). Do not skip levels or combine them in a different way. 

If you write a paper in Chicago style or MLA style , then you don’t need to follow such exact rules for headings and subheadings. Your structure just has to be consistent with the general formatting guidelines of both styles (12-pts Times New Roman font, double-spaced text, 0.5-inch indentation for every new paragraph) and consistent throughout your paper. Make sure the different formatting levels indicate a hierarchy (e.g., boldface for level 1 and italics for level 2, or a larger font size for level 1 and smaller font size for level 2). The main specifics regarding Chicago and MLA headings and subheadings are that they should be written in title case (major words capitalized, most minor words lowercase) and not end in a period. Both styles allow you, however, to number your sections and subsections, for example with an Arabic number and a period, followed by a space and then the section name. 

MLA paper headings example structure

1. Introduction

2. Material and Methods

2.1 Subject Recruitment

2.2 Experimental Procedure

2.3 Statistical Analysis

3.1 Experiment 1

3.2 Experiment 2

4. Discussion

5. Conclusion

What research paper headings do you need?

Your paper obviously needs to contain the main sections ( Introduction, Methods, Results, Discussion, and maybe Conclusion ) and you need to make sure that you name them according to the target journal style (have a look at the author guidelines if you are unsure what the journal style is). The differences between journals are subtle, but some want you to combine the results and discussion sections, for example, while others don’t want you to have a separate conclusion section. You also need to check whether the target journal has specific rules on subsections (or no subsections) within these main sections. The introduction section should usually not be subdivided (but some journals do not mind), while the method section, for example, always needs to have clear subsections.

How to Name Your Sections and Subsections

The method section subheadings should be short and descriptive, but how you subdivide this section depends on the structure you choose to present your work – which can be chronological (e.g., Experiment 1, Experiment 2 ) or follow your main topics (e.g., Visual Experiment, Behavioral Experiment, Questionnaire ). Have a look at this article on how to write the methods for a research paper if you need input on what the best structure for your work is. The method subheadings should only be keywords that tell the reader what information is following, not summaries or conclusions. That means that “ Subject Recruitment ” is a good methods section subheading, but “ Subjects Were Screened Using Questionnaires ” is not.  

The subheadings for the result section should then follow the general structure of your method section, but here you can choose what information you want to put in every subheading. Some authors keep it simple and just subdivide their result section into experiments or measures like the method section, but others use the headings to summarize their findings so that the reader is prepared for the details that follow. You could, for example, simply name your subsections “ Anxiety Levels ” and “ Social Behavior ,” if those are the measures you studied and explained in the method section. 

Or, you could provide the reader with a glimpse into the results of the analyses you are going to describe, and instead name these subsections “ Anxiety-Like Behaviors in Mutant Mice ” and “ Normal Social Behaviors in Mutant Mice .” While keeping headings short and simple is always a good idea, such mini-summaries can make your result section much clearer and easier to follow. Just make sure that the target journal you want to submit to does not have a rule against that. 

Common Heading and Subheading Mistakes 

Subheadings are not sentences.

If your heading reads like a full sentence, then you can most probably omit the verb or generally rephrase to shorten it. That also means a heading should not contain punctuation except maybe colons or question marks – definitely don’t put a period at the end, except when you have reached heading level 4 in the APA formatting style (see above) and the rules say so.  

Be consistent

Always check your numbering, for example for spaces and periods before and after numbers (e.g., 3.2. vs 3.2 ), because readability depends on such features. But also make sure that your headings are consistent in structure and content: Switching between short keyword headings (e.g., “ Experiment 2 ”) and summary headings (e.g., “ Mice Do not Recognize People ”) is confusing and never a good idea. Ideally, subheadings within a section all have a similar structure. If your first subsection is called “ Mice Do not Recognize People ,” then “ People Do not Recognize Mice” is a better subheader for the next subsection than “Do People Recognize Mice? ”, because consistency is more important in a research paper than creativity. 

Don’t overdo it

Not every paragraph or every argument needs a subheading. Only use subheadings within a bigger section if you have more than one point to make per heading level, and if subdividing the section really makes the structure clearer overall.

Before submitting your journal manuscript to academic publishers, be sure to get English editing services , including manuscript editing or paper editing from a trusted source. And receive instant proofreading and paraphrasing with Wordvice AI, our AI online text editor , which provides unlimited editing while drafting your research work.

Want to create or adapt books like this? Learn more about how Pressbooks supports open publishing practices.

13.1 Formatting a Research Paper

Learning objectives.

• Identify the major components of a research paper written using American Psychological Association (APA) style.
• Apply general APA style and formatting conventions in a research paper.

In this chapter, you will learn how to use APA style , the documentation and formatting style followed by the American Psychological Association, as well as MLA style , from the Modern Language Association. There are a few major formatting styles used in academic texts, including AMA, Chicago, and Turabian:

• AMA (American Medical Association) for medicine, health, and biological sciences
• APA (American Psychological Association) for education, psychology, and the social sciences
• Chicago—a common style used in everyday publications like magazines, newspapers, and books
• MLA (Modern Language Association) for English, literature, arts, and humanities
• Turabian—another common style designed for its universal application across all subjects and disciplines

While all the formatting and citation styles have their own use and applications, in this chapter we focus our attention on the two styles you are most likely to use in your academic studies: APA and MLA.

If you find that the rules of proper source documentation are difficult to keep straight, you are not alone. Writing a good research paper is, in and of itself, a major intellectual challenge. Having to follow detailed citation and formatting guidelines as well may seem like just one more task to add to an already-too-long list of requirements.

Following these guidelines, however, serves several important purposes. First, it signals to your readers that your paper should be taken seriously as a student’s contribution to a given academic or professional field; it is the literary equivalent of wearing a tailored suit to a job interview. Second, it shows that you respect other people’s work enough to give them proper credit for it. Finally, it helps your reader find additional materials if he or she wishes to learn more about your topic.

Furthermore, producing a letter-perfect APA-style paper need not be burdensome. Yes, it requires careful attention to detail. However, you can simplify the process if you keep these broad guidelines in mind:

• Work ahead whenever you can. Chapter 11 “Writing from Research: What Will I Learn?” includes tips for keeping track of your sources early in the research process, which will save time later on.
• Get it right the first time. Apply APA guidelines as you write, so you will not have much to correct during the editing stage. Again, putting in a little extra time early on can save time later.
• Use the resources available to you. In addition to the guidelines provided in this chapter, you may wish to consult the APA website at http://www.apa.org or the Purdue University Online Writing lab at http://owl.english.purdue.edu , which regularly updates its online style guidelines.

General Formatting Guidelines

This chapter provides detailed guidelines for using the citation and formatting conventions developed by the American Psychological Association, or APA. Writers in disciplines as diverse as astrophysics, biology, psychology, and education follow APA style. The major components of a paper written in APA style are listed in the following box.

These are the major components of an APA-style paper:

Body, which includes the following:

• Headings and, if necessary, subheadings to organize the content
• In-text citations of research sources
• References page

All these components must be saved in one document, not as separate documents.

The title page of your paper includes the following information:

• Title of the paper
• Author’s name
• Name of the institution with which the author is affiliated
• Header at the top of the page with the paper title (in capital letters) and the page number (If the title is lengthy, you may use a shortened form of it in the header.)

List the first three elements in the order given in the previous list, centered about one third of the way down from the top of the page. Use the headers and footers tool of your word-processing program to add the header, with the title text at the left and the page number in the upper-right corner. Your title page should look like the following example.

The next page of your paper provides an abstract , or brief summary of your findings. An abstract does not need to be provided in every paper, but an abstract should be used in papers that include a hypothesis. A good abstract is concise—about one hundred fifty to two hundred fifty words—and is written in an objective, impersonal style. Your writing voice will not be as apparent here as in the body of your paper. When writing the abstract, take a just-the-facts approach, and summarize your research question and your findings in a few sentences.

In Chapter 12 “Writing a Research Paper” , you read a paper written by a student named Jorge, who researched the effectiveness of low-carbohydrate diets. Read Jorge’s abstract. Note how it sums up the major ideas in his paper without going into excessive detail.

Write an abstract summarizing your paper. Briefly introduce the topic, state your findings, and sum up what conclusions you can draw from your research. Use the word count feature of your word-processing program to make sure your abstract does not exceed one hundred fifty words.

Depending on your field of study, you may sometimes write research papers that present extensive primary research, such as your own experiment or survey. In your abstract, summarize your research question and your findings, and briefly indicate how your study relates to prior research in the field.

Margins, Pagination, and Headings

APA style requirements also address specific formatting concerns, such as margins, pagination, and heading styles, within the body of the paper. Review the following APA guidelines.

Use these general guidelines to format the paper:

• Set the top, bottom, and side margins of your paper at 1 inch.
• Use double-spaced text throughout your paper.
• Use a standard font, such as Times New Roman or Arial, in a legible size (10- to 12-point).
• Use continuous pagination throughout the paper, including the title page and the references section. Page numbers appear flush right within your header.
• Section headings and subsection headings within the body of your paper use different types of formatting depending on the level of information you are presenting. Additional details from Jorge’s paper are provided.

Begin formatting the final draft of your paper according to APA guidelines. You may work with an existing document or set up a new document if you choose. Include the following:

• Your title page
• The abstract you created in Note 13.8 “Exercise 1”
• Correct headers and page numbers for your title page and abstract

APA style uses section headings to organize information, making it easy for the reader to follow the writer’s train of thought and to know immediately what major topics are covered. Depending on the length and complexity of the paper, its major sections may also be divided into subsections, sub-subsections, and so on. These smaller sections, in turn, use different heading styles to indicate different levels of information. In essence, you are using headings to create a hierarchy of information.

The following heading styles used in APA formatting are listed in order of greatest to least importance:

• Section headings use centered, boldface type. Headings use title case, with important words in the heading capitalized.
• Subsection headings use left-aligned, boldface type. Headings use title case.
• The third level uses left-aligned, indented, boldface type. Headings use a capital letter only for the first word, and they end in a period.
• The fourth level follows the same style used for the previous level, but the headings are boldfaced and italicized.
• The fifth level follows the same style used for the previous level, but the headings are italicized and not boldfaced.

Visually, the hierarchy of information is organized as indicated in Table 13.1 “Section Headings” .

Table 13.1 Section Headings

A college research paper may not use all the heading levels shown in Table 13.1 “Section Headings” , but you are likely to encounter them in academic journal articles that use APA style. For a brief paper, you may find that level 1 headings suffice. Longer or more complex papers may need level 2 headings or other lower-level headings to organize information clearly. Use your outline to craft your major section headings and determine whether any subtopics are substantial enough to require additional levels of headings.

Working with the document you developed in Note 13.11 “Exercise 2” , begin setting up the heading structure of the final draft of your research paper according to APA guidelines. Include your title and at least two to three major section headings, and follow the formatting guidelines provided above. If your major sections should be broken into subsections, add those headings as well. Use your outline to help you.

Because Jorge used only level 1 headings, his Exercise 3 would look like the following:

Citation Guidelines

In-text citations.

Throughout the body of your paper, include a citation whenever you quote or paraphrase material from your research sources. As you learned in Chapter 11 “Writing from Research: What Will I Learn?” , the purpose of citations is twofold: to give credit to others for their ideas and to allow your reader to follow up and learn more about the topic if desired. Your in-text citations provide basic information about your source; each source you cite will have a longer entry in the references section that provides more detailed information.

In-text citations must provide the name of the author or authors and the year the source was published. (When a given source does not list an individual author, you may provide the source title or the name of the organization that published the material instead.) When directly quoting a source, it is also required that you include the page number where the quote appears in your citation.

This information may be included within the sentence or in a parenthetical reference at the end of the sentence, as in these examples.

Epstein (2010) points out that “junk food cannot be considered addictive in the same way that we think of psychoactive drugs as addictive” (p. 137).

Here, the writer names the source author when introducing the quote and provides the publication date in parentheses after the author’s name. The page number appears in parentheses after the closing quotation marks and before the period that ends the sentence.

Addiction researchers caution that “junk food cannot be considered addictive in the same way that we think of psychoactive drugs as addictive” (Epstein, 2010, p. 137).

Here, the writer provides a parenthetical citation at the end of the sentence that includes the author’s name, the year of publication, and the page number separated by commas. Again, the parenthetical citation is placed after the closing quotation marks and before the period at the end of the sentence.

As noted in the book Junk Food, Junk Science (Epstein, 2010, p. 137), “junk food cannot be considered addictive in the same way that we think of psychoactive drugs as addictive.”

Here, the writer chose to mention the source title in the sentence (an optional piece of information to include) and followed the title with a parenthetical citation. Note that the parenthetical citation is placed before the comma that signals the end of the introductory phrase.

David Epstein’s book Junk Food, Junk Science (2010) pointed out that “junk food cannot be considered addictive in the same way that we think of psychoactive drugs as addictive” (p. 137).

Another variation is to introduce the author and the source title in your sentence and include the publication date and page number in parentheses within the sentence or at the end of the sentence. As long as you have included the essential information, you can choose the option that works best for that particular sentence and source.

Citing a book with a single author is usually a straightforward task. Of course, your research may require that you cite many other types of sources, such as books or articles with more than one author or sources with no individual author listed. You may also need to cite sources available in both print and online and nonprint sources, such as websites and personal interviews. Chapter 13 “APA and MLA Documentation and Formatting” , Section 13.2 “Citing and Referencing Techniques” and Section 13.3 “Creating a References Section” provide extensive guidelines for citing a variety of source types.

Writing at Work

APA is just one of several different styles with its own guidelines for documentation, formatting, and language usage. Depending on your field of interest, you may be exposed to additional styles, such as the following:

• MLA style. Determined by the Modern Languages Association and used for papers in literature, languages, and other disciplines in the humanities.
• Chicago style. Outlined in the Chicago Manual of Style and sometimes used for papers in the humanities and the sciences; many professional organizations use this style for publications as well.
• Associated Press (AP) style. Used by professional journalists.

References List

The brief citations included in the body of your paper correspond to the more detailed citations provided at the end of the paper in the references section. In-text citations provide basic information—the author’s name, the publication date, and the page number if necessary—while the references section provides more extensive bibliographical information. Again, this information allows your reader to follow up on the sources you cited and do additional reading about the topic if desired.

The specific format of entries in the list of references varies slightly for different source types, but the entries generally include the following information:

• The name(s) of the author(s) or institution that wrote the source
• The year of publication and, where applicable, the exact date of publication
• The full title of the source
• For books, the city of publication
• For articles or essays, the name of the periodical or book in which the article or essay appears
• For magazine and journal articles, the volume number, issue number, and pages where the article appears
• For sources on the web, the URL where the source is located

The references page is double spaced and lists entries in alphabetical order by the author’s last name. If an entry continues for more than one line, the second line and each subsequent line are indented five spaces. Review the following example. ( Chapter 13 “APA and MLA Documentation and Formatting” , Section 13.3 “Creating a References Section” provides extensive guidelines for formatting reference entries for different types of sources.)

In APA style, book and article titles are formatted in sentence case, not title case. Sentence case means that only the first word is capitalized, along with any proper nouns.

Key Takeaways

• Following proper citation and formatting guidelines helps writers ensure that their work will be taken seriously, give proper credit to other authors for their work, and provide valuable information to readers.
• Working ahead and taking care to cite sources correctly the first time are ways writers can save time during the editing stage of writing a research paper.
• APA papers usually include an abstract that concisely summarizes the paper.
• APA papers use a specific headings structure to provide a clear hierarchy of information.
• In APA papers, in-text citations usually include the name(s) of the author(s) and the year of publication.
• In-text citations correspond to entries in the references section, which provide detailed bibliographical information about a source.

Writing for Success Copyright © 2015 by University of Minnesota is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License , except where otherwise noted.

APA Style 7th Edition: Citing Your Sources

• Basics of APA Formatting
• In Text Quick View
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• Conference Presentations
• Social Media
• Legal References
• Reports and Gray Literature

What are headings?

Levels of headings.

• Academic Integrity and Plagiarism
• Additional Resources
• Reference Page

Headings are used to effectively organize ideas within a study or manuscript.  It can also highlight important items, themes or topics within sections.  By creating concise headings, the reader can anticipate key points and track the development of your argument.  The heading levels establish the hierarchy of each section and are designated by their formatting.

Adapted from American Psychological Association. (2009). Format for Five Levels of Heading in APA Journals. Publication manual of the American psychological association (6th ed., p. 62) Washington, D.C.: American Psychological Association.

• If you need to use subsections in any given section, use a least two, otherwise omit their use.
• Do not label headings with numbers or letters
• Use of title case : Use of both upper and lower case letters, all major words are capitalized
• Paragraph headings are immediately followed by text for that subsection, rather than starting on a new line.  The heading sits at the start of the first paragraph for that section.
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• Last Updated: Aug 8, 2024 10:42 AM
• URL: https://libguides.usc.edu/APA7th

APA Style 7th Edition

• Student Paper
• Professional Paper
• In-Text Citations
• Reference Citations
• Additional Resources
• Citing Artificial Intelligence

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• Last Updated: Aug 2, 2024 1:56 PM
• URL: https://libguides.unco.edu/apa

• The Complete Guide to APA Format in 2020

APA Headings and Subheadings

• Headings and Subheadings
• Discussion Section
• Websites and Online Sources
• Journals and Periodicals
• Other Print Sources
• Other Non-Print Sources
• In-text Citations
• Footnotes and Endnotes
• Using MyBib Responsibly
• Miscellaneous Questions

There are five levels of headings available to use in an APA formatted paper. These headings indicate the hierchy of the information within the paper.

For example, if your paper contains the standard main sections Method , Results , and Discussion , these should all use the first-level heading.

• Method (level one)
• Results (level one)
• Discussion (level one)

If you want to include Participants and Instruments within the Method section, you would make these a level two heading.

• Participants (level two)
• Instruments (level two)

A subsection of Participants , like Age or Race , would be level three headings. This pattern continues down to level five headings.

• Age (level three)
• Race (level three)

Styling the Different Headings

Each level has specific formatting requirements to make it visibly obvious which level it is. As with all other text in an APA formatted paper, the font size should be 12pt. The differences between each level are as follows:

Level One Heading

Level one headings are centered and bolded, with all major words capitalized (articles and conjunctions are not captalized).

Level Two Heading

Level two headings are similar to level one headings, except are aligned on the left.

Level Three Heading. Level three headings are similar to levels one and two, except they're indented by 5 spaces, followed by a period, and the text following the heading does not start on a new line.

Level Four Heading. The level four heading is identical to level three, except it is also italicized.

Level Five Heading. Finally, the level five heading is identical to level four, except it is no longer bold.

Further Learning

The following video by the University of Maryland is excellent at visually explaining everything on this page.

Encyclopedia

Writing with artificial intelligence, apa headings and subheadings.

• © 2023 by Jennifer Janechek - IBM Quantum
• headings when there are at least two subsections within a larger section.
• using subheadings only when the paper has at least two subsections within a larger section

Table of Contents

APA headings and subheadings refers to the rules for formatting sections of documents in the 7th Edition of the American Psychological Association’s Publication Manual .

A research paper written in APA style should be organized into sections and subsections using the five levels of APA headings. 

Related Concepts:

Notice how sections contain at least two smaller subsections in the example below:

Design  

Participants. , demographics..

Characteristics.

Limitations

Starting with the first level of heading, the subsections of the paper should progressively use the next level(s) of heading without skipping any levels. Major sections of the paper’s main body, including the Method, Results, and Discussion sections, should always be formatted with the first level of heading. However, keep in mind that the Introduction section, which is preceded by the full title of the paper, should be presented in plain type. Any subsections that fall under the major sections are formatted with the next level of heading.

Note that all paragraphs of the main body, including those that fall under subsections of a larger section, still maintain the pattern of indentation, use Times New Roman font, 12 pt., and are double-spaced. There are no extra lines or spaces between paragraphs and headings.

How are the five levels of APA-style headings formatted?

Format each of the five levels of APA-style headings as demonstrated in the example below. Note that while the example features headings titled “First Level,” “Second Level,” and so on, each heading in your paper should be named according to the section it describes. 

First level

The first level of heading is bolded and centered, and the first letter of each word in the heading is capitalized. The paragraph text should be typed on the following line and indented five spaces from the left.

Second level

The second level of heading is bolded and situated flush left, and the first letter of each word in the heading is capitalized. The paragraph text should be typed on the following line and indented five spaces from the left.

Third level

The third level of heading is bolded, indented five spaces from the left, and followed by a period. Capitalize only the first letter of the first word in the heading and of proper nouns. The first paragraph following this heading should be typed on the same line as the heading.

Fourth level

The fourth level of heading is bolded, italicized, indented five spaces from the left, and followed by a period. Capitalize only the first letter of the first word in the heading and of proper nouns. The first paragraph following this heading should be typed on the same line as the heading.

Fifth level

The fifth level of heading is italicized, indented five spaces from the left, and followed by a period. Capitalize only the first letter of the first word in the heading and of proper nouns. The first paragraph following this heading should be typed on the same line as the heading.

Method  (1st level)

Design (2nd level)

        Participants (3rd level)

        Demographics. (4th level)

              Age Group. (5th level)

Limitations  (2nd level)

• For student papers, include only the page number in the top right margin of every page of the paper
• For professional papers, place the page number in the top right margin and the running head in the top left margin of every page of the paper. Running heads should be a shortened version of the paper title.

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APA 7th Edition Style Guide: Headings in APA

• About In-text Citations
• In-Text Examples
• What to Include
• Volume/Issue
• Bracketed Descriptions
• URLs and DOIs
• Book with Editor(s)
• Book with No Author
• Book with Organization as Author
• Book with Personal Author(s)
• Chapters and Parts of Books
• Classical Works
• Course Materials
• Journal Article
• Magazine Article
• Multi-Volume Works
• Newspaper Article
• Patents & Laws
• Personal Communication
• Physicians' Desk Reference
• Social Media
• Unpublished Manuscripts/Informal Publications (i.e. course packets and dissertations)
• Formatting Your Paper
• Formatting Your References
• Annotated Bibliography
• Headings in APA
• APA Quick Guide
• Submit your Paper for APA Review

What are headings?

Headings, sections, subsections, or levels of subordination are a style of dividing your research paper into major parts, then minor subsections. Most college papers do not need headings, especially if you are only producing two to five pages. However, if your professor requests you use headings or your are writing an especially long or detailed paper, then use headings to help readers navigate your text. Follow the APA style rules for creating the correct level of heading. Always start with a level one heading and drill down to the last subsection possible (five) in order as seen below. Instructions and examples for headings are available on p. 47- 49 of the new APA 7th Edition manual.

Levels of Headings

Additional Headings Resources

• APA Style: Headings This page of the APA Style Blog provides more details about styling paper section headings in APA style.
• Heading Levels Template: Student Paper APA Style 7th Edition This example student paper clearly illustrates how to style section headings including the paper title and the Introduction section (which should not be labeled Introduction as APA assumes all papers begin with an introduction section).

Proper Title Case vs. lowercase paragraph heading

Proper title case is using both uppercase and lowercase letters in a title. It calls for the major words to be capitalized while any small conjunctions are made smaller, i.e., 

The Title of this Paper is Lengthy

Lowercase paragraph heading calls for the first word to be capitalized along with any proper nouns contained within the heading, i.e., 

        The title of this heading is much shorter and all lowercase except for the first word.

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• Last Updated: Sep 3, 2024 11:32 AM
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Home / Guides / Citation Guides / APA Format

APA Format for Students & Researchers

In this guide, students and researchers can learn the basics of creating a properly formatted research paper according to APA guidelines.

It includes information on how to conceptualize, outline, and format the basic structure of your paper, as well as practical tips on spelling, abbreviation, punctuation, and more. The guide concludes with a complete sample paper as well as a final checklist that writers can use to prepare their work for submission.

APA Paper Formatting Basics

• All text should be double-spaced
• Use one-inch margins on all sides
• All paragraphs in the body are indented
• Make sure that the title is centered on the page with your name and school/institution underneath
• Use 12-point font throughout
• All pages should be numbered in the upper right hand corner
• The manual recommends using one space after most punctuation marks
• A shortened version of the title (“running head”) should be placed in the upper left hand corner

Table of Contents

Here’s a quick rundown of the contents of this guide on how to do APA format.

Information related to writing and organizing your paper:

• Paper and essay categories

General paper length

• Margin sizes
• Title pages
• Running Heads
• APA Outline
• APA Abstract
• The body of papers
• APA headings and subheadings
• Use of graphics (tables and figures)

Writing style tips:

Proper tone.

• Reducing bias and labels
• Abbreviation do’s and don’ts
• Punctuation
• Number rules

Citing Your Sources:

• Citing Sources
• In-text Citations
• Reference Page

Proofing Your Paper:

• Final checklist
• Submitting your project

APA Information:

• What is APA
• APA 7 Updates

What you won’t find in this guide: This guide provides information related to the formatting of your paper, as in guidelines related to spacing, margins, word choice, etc. While it provides a general overview of APA references, it does not provide instructions for how to cite in APA format.

For step-by-step instructions for citing books, journals, how to cite a website in APA format, information on an APA format bibliography, and more, refer to these other EasyBib guides:

• APA citation (general reference guide)
• APA In-text citation
• APA article citation
• APA book citation
• APA citation website

Or, you can use our automatic generator. Our APA formatter helps to build your references for you. Yep, you read that correctly.

Writing and Organizing Your APA Paper in an Effective Way

This section of our guide focuses on proper paper length, how to format headings, spacing, and more! This information can be found in Chapter 2 of the official manual (American Psychological Association, 2020, pp. 29-67).

Categories of papers

Before getting into the nitty-gritty details related to APA research paper format, first determine the type of paper you’re about to embark on creating:

Empirical studies

Empirical studies take data from observations and experiments to generate research reports. It is different from other types of studies in that it isn’t based on theories or ideas, but on actual data.

Literature reviews

These papers analyze another individual’s work or a group of works. The purpose is to gather information about a current issue or problem and to communicate where we are today. It sheds light on issues and attempts to fill those gaps with suggestions for future research and methods.

Theoretical articles

These papers are somewhat similar to a literature reviews in that the author collects, examines, and shares information about a current issue or problem, by using others’ research. It is different from literature reviews in that it attempts to explain or solve a problem by coming up with a new theory. This theory is justified with valid evidence.

Methodological articles

These articles showcase new advances, or modifications to an existing practice, in a scientific method or procedure. The author has data or documentation to prove that their new method, or improvement to a method, is valid. Plenty of evidence is included in this type of article. In addition, the author explains the current method being used in addition to their own findings, in order to allow the reader to understand and modify their own current practices.

Case studies

Case studies present information related an individual, group, or larger set of individuals. These subjects are analyzed for a specific reason and the author reports on the method and conclusions from their study. The author may also make suggestions for future research, create possible theories, and/or determine a solution to a problem.

Since APA style format is used often in science fields, the belief is “less is more.” Make sure you’re able to get your points across in a clear and brief way. Be direct, clear, and professional. Try not to add fluff and unnecessary details into your paper or writing.  This will keep the paper length shorter and more concise.

Margin sizes in APA Format

When it comes to margins, keep them consistent across the left, right, top, and bottom of the page. All four sides should be the same distance from the edge of the paper. It’s recommended to use at least one-inch margins around each side. It’s acceptable to use larger margins, but the margins should never be smaller than an inch.

Title pages in APA Format

The title page, or APA format cover page, is the first page of a paper or essay. Some teachers and professors do not require a title page, but some do. If you’re not sure if you should include one or not, ask your teacher. Some appreciate the page, which clearly displays the writer’s name and the title of the paper.

The APA format title page for student papers includes six main components:

• the title of the APA format paper
• names of all authors
• institutional affiliation
• course number and title
• instructor’s name

Title pages for professional papers  also require a running head; student papers do not.

Some instructors and professional publications also ask for an author’s note. If you’re required or would like to include an author’s note, place it below the institutional affiliation. Examples of information included in an author’s note include an ORCID iD number, a disclosure, and an acknowledgement.

Here are key guidelines to developing your title page:

• The title of the paper should capture the main idea of the essay, but should not contain abbreviations or words that serve no purpose. For example, instead of using the title “A Look at Amphibians From the Past,” title the paper “Amphibians From the Past.” Delete the unnecessary fluff!
• Center the title on the page and place it about 3-4 lines from the top.
• The title should be bolded, in title case, and the same font size as your other page text. Do not underline or italicize the title. Other text on the page should be plain (not bolded , underlined, or italicized ). 
• All text on the title page should be double-spaced. The APA format examples paper below displays proper spacing, so go take a look!
• Do not include any titles in the author’s name such as Dr. or Ms. In contrast, for your instructor’s name, use the form they prefer (e.g., Sagar Parekh, PhD; Dr. Minako Asato; Professor Nathan Ian Brown; etc.).
• The institutional affiliation is the school the author attends or the location where the author conducted the research.

In a hurry? Try the  EasyBib title page maker to easily create a title page for free.

Sample of an APA format title page for a student paper:

Sample of title page for a professional paper:

Running heads in APA Format

The 7th edition of the American Psychological Association Publication Manual (p. 37) states that running heads are not required for student papers unless requested by the instructor. Student papers still need a page number included in the upper right-hand corner of every page. The 6th edition required a running head for student papers, so be sure to confirm with your instructor which edition you should follow. Of note, this guide follows the 7th edition.

Running heads are required for professional papers (e.g., manuscripts submitted for publication). Read on for instructions on how to create them.

Are you wondering what is a “running head”? It’s basically a page header at the top of every page. To make this process easier, set your word processor to automatically add these components onto each page. You may want to look for “Header” in the features.

A running head/page header includes two pieces:

• the title of the paper
• page numbers.

Insert page numbers justified to the right-hand side of the APA format paper (do not put p. or pg. in front of the page numbers).

For all pages of the paper, including the APA format title page, include the “TITLE OF YOUR PAPER” justified to the left in capital letters (i.e., the running head). If your full title is long (over 50 characters), the running head title should be a shortened version.

Preparing outlines in APA Format

Outlines are extremely beneficial as they help writers stay organized, determine the scope of the research that needs to be included, and establish headings and subheadings.

There isn’t an official or recommended “APA format for outline” structure. It is up to the writer (if they choose to make use of an outline) to determine how to organize it and the characters to include. Some writers use a mix of roman numerals, numbers, and uppercase and lowercase letters.

Even though there isn’t a required or recommended APA format for an outline, we encourage writers to make use of one. Who wouldn’t want to put together a rough outline of their project? We promise you, an outline will help you stay on track.

Here’s our version of how APA format for outlines could look:

Don’t forget, if you’re looking for information on APA citation format and other related topics, check out our other comprehensive guides.

How to form an abstract in APA

An APA format abstract (p. 38) is a summary of a scholarly article or scientific study. Scholarly articles and studies are rather lengthy documents, and abstracts allow readers to first determine if they’d like to read an article in its entirety or not.

You may come across abstracts while researching a topic. Many databases display abstracts in the search results and often display them before showing the full text of an article or scientific study. It is important to create a high quality abstract that accurately communicates the purpose and goal of your paper, as readers will determine if it is worthy to continue reading or not.

Are you wondering if you need to create an abstract for your assignment? Usually, student papers do not require an abstract. Abstracts are not typically seen in class assignments, and are usually only included when submitting a paper for publication. Unless your teacher or professor asked for it, you probably don’t need to have one for your class assignment.

If you’re planning on submitting your paper to a journal for publication, first check the journal’s website to learn about abstract and APA paper format requirements.

Here are some helpful suggestions to create a dynamic abstract:

• Abstracts are found on their own page, directly after the title or cover page.
• Professional papers only (not student papers): Include the running head on the top of the page.
• On the first line of the page, center the word “Abstract” (but do not include quotation marks).
• On the following line, write a summary of the key points of your research. Your abstract summary is a way to introduce readers to your research topic, the questions that will be answered, the process you took, and any findings or conclusions you drew. Use concise, brief, informative language. You only have a few sentences to share the summary of your entire document, so be direct with your wording.
• This summary should not be indented, but should be double-spaced and less than 250 words.
• If applicable, help researchers find your work in databases by listing keywords from your paper after your summary. To do this, indent and type Keywords : in italics.  Then list your keywords that stand out in your research. You can also include keyword strings that you think readers will type into the search box.
• Active voice: The subjects reacted to the medication.
• Passive voice: There was a reaction from the subjects taking the medication.
• Instead of evaluating your project in the abstract, simply report what it contains.
• If a large portion of your work includes the extension of someone else’s research, share this in the abstract and include the author’s last name and the year their work was released.

APA format example page:

Here’s an example of an abstract:

Visual design is a critical aspect of any web page or user interface, and its impact on a user’s experience has been studied extensively. Research has shown a positive correlation between a user’s perceived usability and a user’s assessment of visual design. Additionally, perceived web quality, which encompasses visual design, has a positive relationship with both initial and continued consumer purchase intention. However, visual design is often assessed using self-report scale, which are vulnerable to a few pitfalls. Because self-report questionnaires are often reliant on introspection and honesty, it is difficult to confidently rely on self-report questionnaires to make important decisions. This study aims to ensure the validity of a visual design assessment instrument (Visual Aesthetics of Websites Inventory: Short version) by examining its relationship with biometric (variables), like galvanic skin response, pupillometry, and fixation information. Our study looked at participants assessment of a webpage’s visual design, and compared it to their biometric responses while viewing the webpage. Overall, we found that both average fixation duration and pupil dilation differed when participants viewed web pages with lower visual design ratings compared to web pages with a higher visual design rating.

Keywords : usability, visual design, websites, eye tracking, pupillometry, self-report, VisAWI

The body of an APA paper

On the page after the title page (if a student paper) or the abstract (if a professional paper), begin with the body of the paper.

Most papers follow this format:

• At the top of the page, add the page number in the upper right corner of all pages, including the title page.
• On the next line write the title in bold font and center it. Do not underline or italicize it.
• Begin with the introduction and indent the first line of the paragraph. All paragraphs in the body are indented.

Sample body for a student paper:

Most scientific or professional papers have additional sections and guidelines:

• Start with the running head (title + page number). The heading title should be in capital letters. The abstract page should be page 2.
• The introduction presents the problem and premise upon which the research was based. It goes into more detail about this problem than the abstract.
• Begin a new section with the Method and use this word as the subtitle. Bold and center this subtitle. The Method section shows how the study was run and conducted. Be sure to describe the methods through which data was collected.
• Begin a new section with the Results . Bold and center this subtitle. The Results section summarizes your data. Use charts and graphs to display this data.
• Draw conclusions and support how your data led to these conclusions.
• Discuss whether or not your hypothesis was confirmed or not supported by your results.
• Determine the limitations of the study and next steps to improve research for future studies.

Sample body for a professional paper:

Keep in mind, APA citation format is much easier than you think, thanks to EasyBib.com. Try our automatic generator and watch how we create APA citation format references for you in just a few clicks. While you’re at it, take a peek at our other helpful guides, such as our APA reference page guide, to make sure you’re on track with your research papers.

Proper usage of headings & subheadings in APA Format

Headings (p. 47) serve an important purpose in research papers — they organize your paper and make it simple to locate different pieces of information. In addition, headings provide readers with a glimpse to the main idea, or content, they are about to read.

In APA format, there are five levels of headings, each with a different formatting:

• This is the title of your paper
• The title should be centered in the middle of the page
• The title should be bolded
• Use uppercase and lowercase letters where necessary (called title capitalization)
• Place this heading against the left margin
• Use bold letters
• Use uppercase and lowercase letters where necessary
• Place this heading against the left side margin
• End the heading with a period
• Indented in from the left margin

Following general formatting rules, all headings are double spaced and there are no extra lines or spaces between sections.

Here is a visual APA format template for levels of headings:

Use of graphics (tables and figures) in APA Format

If you’re looking to jazz up your project with any charts, tables, drawings, or images, there are certain APA format rules (pp. 195-250) to follow.

First and foremost, the only reason why any graphics should be added is to provide the reader with an easier way to see or read information, rather than typing it all out in the text.

Lots of numbers to discuss? Try organizing your information into a chart or table. Pie charts, bar graphs, coordinate planes, and line graphs are just a few ways to show numerical data, relationships between numbers, and many other types of information.

Instead of typing out long, drawn out descriptions, create a drawing or image. Many visual learners would appreciate the ability to look at an image to make sense of information.

Before you go ahead and place that graphic in your paper, here are a few key guidelines:

• Follow them in the appropriate numerical order in which they appear in the text of your paper. Example : Figure 1, Figure 2, Table 1, Figure 3.
• Example: Figure 1, Figure 2, Table 1, Figure 3
• Only use graphics if they will supplement the material in your text. If they reinstate what you already have in your text, then it is not necessary to include a graphic.
• Include enough wording in the graphic so that the reader is able to understand its meaning, even if it is isolated from the corresponding text. However, do not go overboard with adding a ton of wording in your graphic.
• Left align tables and figures

In our APA format sample paper , you’ll find examples of tables after the references. You may also place tables and figures within the text just after it is mentioned.

Is there anything better than seeing a neatly organized data table? We think not! If you have tons of numbers or data to share, consider creating a table instead of typing out a wordy paragraph. Tables are pretty easy to whip up on Google Docs or Microsoft Word.

General format of a table should be:

• Table number
• Choose to type out your data OR create a table. As stated above, in APA format, you shouldn’t have the information typed out in your paper and also have a table showing the same exact information. Choose one or the other.
• If you choose to create a table, discuss it very briefly in the text. Say something along the lines of, “Table 1 displays the amount of money used towards fighting Malaria.” Or, “Stomach cancer rates are displayed in Table 4.”
• If you’re submitting your project for a class, place your table close to the text where it’s mentioned. If you’re submitting it to be published in a journal, most publishers prefer tables to be placed in the back. If you’re unsure where to place your tables, ask!
• Include the table number first and at the top. Table 1 is the first table discussed in the paper. Table 2 is the next table mentioned, and so on. This should be in bold.
• Add a title under the number. Create a brief, descriptive title. Capitalize the first letter for each important word. Italicize the title and place it under the table number.
• Only use horizontal lines.
• Limit use of cell shading.
• Keep the font at 12-point size and use single or double spacing. If you use single spacing in one table, make sure all of the others use single spaces as well. Keep it consistent.
• All headings should be centered.
• In the first column (called the stub), center the heading, left-align the information underneath it (indent 0.15 inches if info is more than one line).
• Information in other columns should be centered.
• General . Information about the whole table.
• Specific . Information targeted for a specific column, row, or cell.
• Probability . Explains what certain table symbols mean. For example, asterisks,  p values, etc.

Here’s an APA format example of a table:

We know putting together a table is pretty tricky. That’s why we’ve included not one, but a few tables on this page. Scroll down and look at the additional tables in the essay in APA format example found below.

Figures represent information in a visual way. They differ from tables in that they are visually appealing. Sure, tables, like the one above, can be visually appealing, but it’s the color, circles, arrows, boxes, or icons included that make a figure a “figure.”

There are many commonly used figures in papers. Examples APA Format:

• Photographs
• Hierarchy charts

General format of a figure is the same as tables. This means each should include:

• Figure number

Use the same formatting tables use for the number, title, and note.

Here are some pointers to keep in mind when it comes to APA format for figures:

• Only include a figure if it adds value to your paper. If it will truly help with understanding, include it!
• Either include a figure OR write it all out in the text. Do not include the same information twice.
• If a note is added, it should clearly explain the content of the figure. Include any reference information if it’s reproduced or adapted.

APA format sample of a figure:

Photographs:

We live in a world where we have tons of photographs available at our fingertips.

Photographs found through Google Images, social media, stock photos made available from subscription sites, and tons of other various online sources make obtaining photographs a breeze. We can even pull out our cell phones, and in just a few seconds, take pictures with our cameras.

Photographs are simple to find, and because of this, many students enjoy using them in their papers.

If you have a photograph you would like to include in your project, here are some guidelines from the American Psychological Association.

• Create a reference for the photograph. Follow the guidelines under the table and figure sections above.
• Do not use color photos. It is recommended to use black and white. Colors can change depending on the reader’s screen resolution. Using black and white ensures the reader will be able to view the image clearly. The only time it is recommended to use color photos is if you’re writing about color-specific things. For example, if you’re discussing the various shades of leaf coloration, you may want to include a few photographs of colorful leaves.
• If there are sections of the photograph that are not related to your work, it is acceptable to crop them out. Cropping is also beneficial in that it helps the reader focus on the main item you’re discussing.
• If you choose to include an image of a person you know, it would be respectful if you ask their permission before automatically including their photo in your paper.  Some schools and universities post research papers online and some people prefer that their photos and information stay off the Internet.

B. Writing Style Tips

Writing a paper for scientific topics is much different than writing for English, literature, and other composition classes. Science papers are much more direct, clear, and concise. This section includes key suggestions, explains how to write in APA format, and includes other tidbits to keep in mind while formulating your research paper.

Verb usage in APA

Research experiments and observations rely on the creation and analysis of data to test hypotheses and come to conclusions. While sharing and explaining the methods and results of studies, science writers often use verbs.

When using verbs in writing, make sure that you continue to use them in the same tense throughout the section you’re writing. Further details are in the publication manual (p. 117).

Here’s an APA format example:

We tested the solution to identify the possible contaminants.

It wouldn’t make sense to add this sentence after the one above:

We tested the solution to identify the possible contaminants. Researchers often test solutions by placing them under a microscope.

Notice that the first sentence is in the past tense while the second sentence is in the present tense. This can be confusing for readers.

For verbs in scientific papers, the APA manual recommends using:

• Past tense or present perfect tense for the explantation of the procedure
• Past tense for the explanation of the results
• Present tense for the explanation of the conclusion and future implications

If this is all a bit much, and you’re simply looking for help with your references, try the EasyBib.com APA format generator . Our APA formatter creates your references in just a few clicks. APA citation format is easier than you think thanks to our innovative, automatic tool.

Even though your writing will not have the same fluff and detail as other forms of writing, it should not be boring or dull to read. The Publication Manual suggests thinking about who will be the main reader of your work and to write in a way that educates them.

How to reduce bias & labels

The American Psychological Association strongly objects to any bias towards gender, racial groups, ages of individuals or subjects, disabilities, and sexual orientation (pp. 131-149). If you’re unsure whether your writing is free of bias and labels or not, have a few individuals read your work to determine if it’s acceptable.

Here are a few guidelines that the American Psychological Association suggests :

• Only include information about an individual’s orientation or characteristic if it is important to the topic or study. Do not include information about individuals or labels if it is not necessary.
• If writing about an individual’s characteristic or orientation, for essay APA format, make sure to put the person first. Instead of saying, “Diabetic patients,” say, “Patients who are diabetic.”
• Instead of using narrow terms such as, “adolescents,” or “the elderly,” try to use broader terms such as, “participants,” and “subjects.”
• “They” or “their” are acceptable gender-neutral pronouns to use.
• Be mindful when using terms that end with “man” or “men” if they involve subjects who are female. For example, instead of using “Firemen,” use the term, “Firefighter.” In general, avoid ambiguity.
• When referring to someone’s racial or ethnic identity, use the census category terms and capitalize the first letter. Also, avoid using the word, “minority,” as it can be interpreted as meaning less than or deficient. Instead, say “people of color” or “underrepresented groups.”
• When describing subjects in APA format, use the words “girls” and “boys” for children who are under the age of 12. The terms, “young woman,” “young man,” “female adolescent,” and “male adolescent” are appropriate for subjects between 13-17 years old; “Men,” and “women,” for those older than 18. Use the term, “older adults.” for individuals who are older. “Elderly,” and “senior,” are not acceptable if used only as nouns. It is acceptable to use these terms if they’re used as adjectives.

Read through our example essay in APA format, found in section D, to see how we’ve reduced bias and labels.

Spelling in APA Format

• In APA formatting, use the same spelling as words found in Merriam-Webster’s Collegiate Dictionary (American English) (p. 161).
• If the word you’re trying to spell is not found in Webster’s Collegiate Dictionary, a second resource is Webster’s Third New International Dictionary .
• If attempting to properly spell words in the psychology field, consult the American Psychological Association’s Dictionary of Psychology

Thanks to helpful tools and features, such as the spell checker, in word processing programs, most of us think we have everything we need right in our document. However, quite a few helpful features are found elsewhere.

Where can you find a full grammar editor? Right here, on EasyBib.com. The EasyBib Plus paper checker scans your paper for spelling, but also for any conjunction , determiner, or adverb out of place. Try it out and unlock the magic of an edited paper.

Abbreviation do’s and don’ts in APA Format

Abbreviations can be tricky. You may be asking yourself, “Do I include periods between the letters?” “Are all letters capitalized?” “Do I need to write out the full name each and every time?” Not to worry, we’re breaking down the publication manual’s abbreviations (p. 172) for you here.

First and foremost, use abbreviations sparingly.

Too many and you’re left with a paper littered with capital letters mashed together. Plus, they don’t lend themselves to smooth and easy reading. Readers need to pause and comprehend the meaning of abbreviations and quite often stumble over them.

• If the abbreviation is used less than three times in the paper, type it out each time. It would be pretty difficult to remember what an abbreviation or acronym stands for if you’re writing a lengthy paper.
• If you decide to sprinkle in abbreviations,  it is not necessary to include periods between the letters.
• Example: While it may not affect a patient’s short-term memory (STM), it may affect their ability to comprehend new terms. Patients who experience STM loss while using the medication should discuss it with their doctor.
• Example : AIDS
• The weight in pounds exceeded what we previously thought.

Punctuation in APA Format

One space after most punctuation marks.

The manual recommends using one space after most punctuation marks, including punctuation at the end of a sentence (p. 154). It doesn’t hurt to double check with your teacher or professor to ask their preference since this rule was changed recently (in 2020).

The official APA format book was primarily created to aid individuals with submitting their paper for publication in a professional journal. Many schools adopt certain parts of the handbook and modify sections to match their preference. To see an example of an APA format research paper, with the spacing we believe is most commonly and acceptable to use, scroll down and see section D.

For more information related to the handbook, including frequently asked questions, and more, here’s further reading on the style

It’s often a heated debate among writers whether or not to use an Oxford comma (p. 155), but for this style, always use an Oxford comma. This type of comma is placed before the words AND and OR or in a series of three items.

Example of APA format for commas: The medication caused drowsiness, upset stomach, and fatigue.

Here’s another example: The subjects chose between cold, room temperature, or warm water.

Apostrophes

When writing a possessive singular noun, you should place the apostrophe before the s. For possessive plural nouns, the apostrophe is placed after the s.

• Singular : Linda Morris’s jacket
• Plural : The Morris’ house

Em dashes (long dash) are used to bring focus to a particular point or an aside. There are no spaces after these dashes (p. 157).

Use en dashes (short dash) in compound adjectives. Do not place a space before or after the dash. Here are a few examples:

• custom-built
• 12-year-old

Number rules in APA Format

Science papers often include the use of numbers, usually displayed in data, tables, and experiment information. The golden rule to keep in mind is that numbers less than 10 are written out in text. If the number is more than 10, use numerals.

APA format examples:

• 14 kilograms
• seven individuals
• 83 years old
• Fourth grade

The golden rule for numbers has exceptions.

In APA formatting, use numerals if you are:

• Showing numbers in a table or graph
• 4 divided by 2
• 6-month-olds

Use numbers written out as words if you are:

• Ninety-two percent of teachers feel as though….
• Hundred Years’ War
• One-sixth of the students

Other APA formatting number rules to keep in mind:

• World War II
• Super Bowl LII
• It’s 1980s, not 1980’s!

Additional number rules can be found in the publication manual (p. 178)

Need help with other writing topics? Our plagiarism checker is a great resource for anyone looking for writing help. Say goodbye to an out of place noun , preposition , or adjective, and hello to a fully edited paper.

Overview of APA references

While writing a research paper, it is always important to give credit and cite your sources; this lets you acknowledge others’ ideas and research you’ve used in your own work. Not doing so can be considered plagiarism , possibly leading to a failed grade or loss of a job.

APA style is one of the most commonly used citation styles used to prevent plagiarism. Here’s more on crediting sources . Let’s get this statement out of the way before you become confused: An APA format reference and an APA format citation are two different things! We understand that many teachers and professors use the terms as if they’re synonyms, but according to this specific style, they are two separate things, with different purposes, and styled differently.

A reference displays all of the information about the source — the title, the author’s name, the year it was published, the URL, all of it! References are placed on the final page of a research project.

Here’s an example of a reference:

Wynne-Jones, T. (2015). The emperor of any place . Candlewick Press.

An APA format citation is an APA format in-text citation. These are found within your paper, anytime a quote or paraphrase is included. They usually only include the name of the author and the date the source was published.

Here’s an example of one:

Hypertrophic cardiomyopathy is even discussed in the book, The Emperor of Any Place . The main character, Evan, finds a mysterious diary on his father’s desk (the same desk his father died on, after suffering from a hypertrophic cardiomyopathy attack). Evan unlocks the truth to his father and grandfather’s past (Wynne-Jones, 2015).

Both of the ways to credit another individual’s work — in the text of a paper and also on the final page — are key to preventing plagiarism. A writer must use both types in a paper. If you cite something in the text, it must have a full reference on the final page of the project. Where there is one, there must be the other!

Now that you understand that, here’s some basic info regarding APA format references (pp. 281-309).

• Each reference is organized, or structured, differently. It all depends on the source type. A book reference is structured one way, an APA journal is structured a different way, a newspaper article is another way. Yes, it’s probably frustrating that not all references are created equal and set up the same way. MLA works cited pages are unique in that every source type is formatted the same way. Unfortunately, this style is quite different.
• Most references follow this general format:

Author’s Last name, First initial. Middle initial. (Year published). Title of source . URL.

Again, as stated in the above paragraph, you must look up the specific source type you’re using to find out the placement of the title, author’s name, year published, etc.

For more information on APA format for sources and how to reference specific types of sources, use the other guides on EasyBib.com. Here’s another useful site .

Looking for a full visual of a page of references? Scroll down and take a peek at our APA format essay example towards the bottom of this page. You’ll see a list of references and you can gain a sense of how they look.

Bonus: here’s a link to more about the fundamentals related to this particular style. If you want to brush up or catch up on the Modern Language Association’s style, here’s a great resource on how to cite websites in MLA .

In-text APA citation format

Did you find the perfect quote or piece of information to include in your project? Way to go! It’s always a nice feeling when we find that magical piece of data or info to include in our writing. You probably already know that you can’t just copy and paste it into your project, or type it in, without also providing credit to the original author.

Displaying where the original information came from is much easier than you think.Directly next to the quote or information you included, place the author’s name and the year nearby. This allows the reader of your work to see where the information originated.

APA allows for the use of two different forms of in-text citation, parenthetical and narrative Both forms of citation require two elements:

• author’s name
• year of publication

The only difference is the way that this information is presented to the reader.

Parenthetical citations are the more commonly seen form of in-text citations for academic work, in which both required reference elements are presented at the end of the sentence in parentheses. Example:

Harlem had many artists and musicians in the late 1920s (Belafonte, 2008).

Narrative citations allow the author to present one or both of the required reference elements inside of the running sentence, which prevents the text from being too repetitive or burdensome. When only one of the two reference elements is included in the sentence, the other is provided parenthetically. Example:

According to Belafonte (2008), Harlem was full of artists and musicians in the late 1920s.

If there are two authors listed in the source entry, then the parenthetical reference must list them both:

(Smith & Belafonte, 2008)

If there are three or more authors listed in the source entry, then the parenthetical reference can abbreviate with “et al.”, the latin abbreviation for “and others”:

(Smith et al., 2008)

The author’s names are structured differently if there is more than one author. Things will also look different if there isn’t an author at all (which is sometimes the case with website pages). For more information on APA citation format, check out this page on the topic: APA parenthetical citation and APA in-text citation . There is also more information in the official manual in chapter 8.

If it’s MLA in-text and parenthetical citations you’re looking for, we’ve got your covered there too! You might want to also check out his guide on parenthetical citing .

Would you benefit from having a tool that helps you easily generate citations that are in the text? Check out EasyBib Plus!

References page in APA Format

An APA format reference page is easier to create than you probably think. We go into detail on how to create this page on our APA reference page . We also have a guide for how to create an annotated bibliography in APA . But, if you’re simply looking for a brief overview of the reference page, we’ve got you covered here.

Here are some pointers to keep in mind when it comes to the references page in APA format:

• This VIP page has its very own page. Start on a fresh, clean document (p. 303).
• Center and bold the title “References” (do not include quotation marks, underline, or italicize this title).
• Alphabetize and double-space ALL entries.
• Use a readable font, such as Times New Roman, Arial, Calibri, or Lucida (p. 44).
• Every quote or piece of outside information included in the paper should be referenced and have an entry.
• Even though it’s called a “reference page,” it can be longer than one page. If your references flow onto the next page, then that’s a-okay.
• Only include the running head if it is required by your teacher or you’re writing a professional paper.

Sample reference page for a student paper:

Here’s another friendly reminder to use the EasyBib APA format generator (that comes with EasyBib Plus) to quickly and easily develop every single one of your references for you. Try it out! Our APA formatter is easy to use and ready to use 24/7.

Final APA Format Checklist

Prior to submitting your paper, check to make sure you have everything you need and everything in its place:

• Did you credit all of the information and quotes you used in the body of your paper and show a matching full reference at the end of the paper? Remember, you need both! Need more information on how to credit other authors and sources? Check out our other guides, or use the EasyBib APA format generator to credit your sources quickly and easily. EasyBib.com also has more styles than just the one this page focuses on.
• 12-pt. Times New Roman
• 11-pt. Calibri, Arial, Georgia
• 10-pt. Lucida, Sans Unicode, Computer Modern
• If you created an abstract, is it directly after the title page? Some teachers and professors do not require an abstract, so before you go ahead and include it, make sure it’s something he or she is expecting.
• Professional paper — Did you include a running head on every single page of your project?
• Student paper — Did you include page numbers in the upper right-hand corner of all your pages?
• Are all headings, as in section or chapter titles, properly formatted? If you’re not sure, check section number 9.
• Are all tables and figures aligned properly? Did you include notes and other important information directly below the table or figure? Include any information that will help the reader completely understand everything in the table or figure if it were to stand alone.
• Are abbreviations used sparingly? Did you format them properly?
• Is the entire document double spaced?
• Are all numbers formatted properly? Check section 17, which is APA writing format for numbers.
• Did you glance at the sample paper? Is your assignment structured similarly? Are all of the margins uniform?

Submitting Your APA Paper

Congratulations for making it this far! You’ve put a lot of effort into writing your paper and making sure the t’s are crossed and the i’s are dotted. If you’re planning to submit your paper for a school assignment, make sure you review your teacher or professor’s procedures.

If you’re submitting your paper to a journal, you probably need to include a cover letter.

Most cover letters ask you to include:

• The author’s contact information.
• A statement to the editor that the paper is original.
• If a similar paper exists elsewhere, notify the editor in the cover letter.

Once again, review the specific journal’s website for exact specifications for submission.

Okay, so you’re probably thinking you’re ready to hit send or print and submit your assignment. Can we offer one last suggestion? We promise it will only take a minute.

Consider running your paper through our handy dandy paper checker. It’s pretty simple.

Copy and paste or upload your paper into our checker. Within a minute, we’ll provide feedback on your spelling and grammar. If there’s a pronoun , interjection , or verb out of place, we’ll highlight it and offer suggestions for improvement. We’ll even take it a step further and point out any instances of possible plagiarism.

If it sounds too good to be true, then head on over to our innovative tool and give it a whirl. We promise you won’t be disappointed.

What is APA Format?

APA stands for the American Psychological Association . In this guide, you’ll find information related to “What is APA format?” in relation to writing and organizing your paper according to the American Psychological Association’s standards. Information on how to cite sources can be found on our APA citation page. The official American Psychological Association handbook was used as a reference for our guide and we’ve included page numbers from the manual throughout. However, this page is not associated with the association.

You’ll most likely use APA format if your paper is on a scientific topic. Many behavioral and social sciences use this organization’s standards and guidelines.

What are behavioral sciences? Behavioral sciences study human and animal behavior. They can include:

• Cognitive Science
• Neuroscience

What are social sciences? Social sciences focus on one specific aspect of human behavior, specifically social and cultural relationships. Social sciences can include:

• Anthropology
• Political Science
• Human Geography
• Archaeology
• Linguistics

What’s New in the 7th Edition?

This citation style was created by the American Psychological Association. Its rules and guidelines can be found in the Publication Manual of the American Psychological Association . The information provided in the guide above follows the 6th edition (2009) of the manual. The 7th edition was published in 2020 and is the most recent version.

The 7th edition of the Publication Manual is in full color and includes 12 sections (compared to 8 sections in the 6th edition). In general, this new edition differentiates between professional and student papers, includes guidance with accessibility in mind, provides new examples to follow, and has updated guidelines.We’ve selected a few notable updates below, but for a full view of all of the 7th edition changes visit the style’s website linked here .

• Paper title
• Student name
• Affiliation (e.g., school, department, etc.)
• Course number and title
• Course instructor
• 6th edition – Running head: SMARTPHONE EFFECTS ON ADOLESCENT SOCIALIZATION
• 7th edition – SMARTPHONE EFFECTS ON ADOLESCENT SOCIALIZATION
• Pronouns . “They” can be used as a gender-neutral pronoun.
• Bias-free language guidelines . There are updated and new sections on guidelines for this section. New sections address participation in research, socioeconomic status, and intersectionality.
• Spacing after sentences. Add only a single space after end punctuation.
• Tables and figures . The citing format is now streamlined so that both tables and figures should include a name and number above the table/figure, and a note underneath the table/figure.
• 6th ed. – (Ikemoto, Richardson, Murphy, Yoshida 2016)
• 7th ed. – (Ikemoto et al., 2016)
• Citing books. The location of the publisher can be omitted. Also, e-books no longer need to mention the format (e.g., Kindle, etc.)
• Example: https://doi.org/10.1038/s42255-019-0153-5
• Using URLs. URLs no longer need to be prefaced by the words “Retrieved from.”

New citing information . There is new guidance on citing classroom or intranet resources, and oral traditions or traditional knowledge of indigenous peoples.

Visit our EasyBib Twitter feed to discover more citing tips, fun grammar facts, and the latest product updates.

American Psychological Association. (2020). Publication manual of the American Psychological Association (7th ed.) (2020). American Psychological Association. https://doi.org/10.1037/0000165-000

Published October 31, 2011. Updated May 14, 2020.

Written and edited by Michele Kirschenbaum and Elise Barbeau. Michele Kirschenbaum is a school library media specialist and the in-house librarian at EasyBib.com. Elise Barbeau is the Citation Specialist at Chegg. She has worked in digital marketing, libraries, and publishing.

APA Formatting Guide

APA Formatting

• Annotated Bibliography
• Block Quotes
• et al Usage
• Multiple Authors
• Paraphrasing
• Page Numbers
• Parenthetical Citations
• Sample Paper
• View APA Guide

Citation Examples

• Book Chapter
• Journal Article
• Magazine Article
• Newspaper Article
• Website (no author)
• View all APA Examples

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We should not use “et al.” in APA reference list entries. If the number of authors in the source is up to and including 20, list all author names and use an ampersand (&) before the final author’s name. If the number of authors is more than 20, list the first 19 authors’ names followed by an ellipsis (but no ampersand), and then add the final author’s name. An example of author names in a reference entry having more than 20 authors is given below:

Author Surname1, F. M., Author Surname2, F. M., Author Surname3, F. M., Author Surname4, F. M., Author Surname5, F. M., Author Surname6, F. M., Author Surname7, F. M., Author Surname8, F. M., Author Surname9, F. M., Author Surname10, F. M., Author Surname11, F. M., Author Surname12, F. M., Author Surname13, F. M., Author Surname14, F. M., Author Surname15, F. M., Author Surname16, F. M., Author Surname17, F. M., Author Surname18, F. M., Author Surname19, F. M., . . .  Last Author Surname, F. M. (Publication Year).

Alvarez, L. D., Peach, J. L., Rodriguez, J. F., Donald, L., Thomas, M., Aruck, A., Samy, K., Anthony, K., Ajey, M., Rodriguez, K. L., Katherine, K., Vincent, A., Pater, F., Somu, P., Pander, L., Berd, R., Fox, L., Anders, A., Kamala, W., . . . Nicole Jones, K. (2019).

Note that, unlike references with 2 to 20 author names, the symbol “&” is not used here before the last author’s name.

APA 7, released in October 2019, has some new updates. Here is a brief description of the updates made in APA 7.

Different types of papers and best practices are given in detail in Chapter 1.

How to format a student title page is explained in Chapter 2. Examples of a professional paper and a student paper are included.

Chapter 3 provides additional information on qualitative and mixed methods of research.

An update on writing style is included in Chapter 4.

In chapter 5, some best practices for writing with bias-free language are included.

Chapter 6 gives some updates on style elements including using a single space after a period, including a citation with an abbreviation, the treatment of numbers in abstracts, treatment for different types of lists, and the formatting of gene and protein names.

In Chapter 7, additional examples are given for tables and figures for different types of publications.

In Chapter 8, how to format quotations and how to paraphrase text are covered with additional examples. A simplified version of in-text citations is clearly illustrated.

Chapter 9 has many updates: listing all author names up to 20 authors, standardizing DOIs and URLs, and the formatting of an annotated bibliography.

Chapter 10 includes many examples with templates for all reference types. New rules covering the inclusion of the issue number for journals and the omission of publisher location from book references are provided. Explanations of how to cite YouTube videos, power point slides, and TED talks are included.

Chapter 11 includes many legal references for easy understanding.

Chapter 12 provides advice for authors on how to promote their papers.

For more information on some of the changes found in APA 7, check out this EasyBib article .

APA Citation Examples

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Examples: Edited Papers

Need editing and proofreading services, apa headings & subheadings | formatting guidelines & examples.

• Tags: APA , APA Style , Formatting Guidelines

APA headings and subheadings provide a clear structure to your document and make it easy to navigate through. This is especially beneficial when it comes to longer documents. 

The APA format makes use of headings and subheadings to indicate a hierarchy. But what is an APA paper heading and how do you customize it? We will answer all these questions with the help of concrete examples and templates.

Let’s start off with understanding the basics of how to customize these APA format headings and subheadings. 

APA heading format

There are five APA heading levels that can be used as per the latest APA guidelines. You can decide which level of heading to use, depending on the length and complexity of your document.

• The level 1 heading is the main APA heading for your paper, which is used to indicate all the major sections (excluding the introduction). 
• The level 2 heading is included under the level 1 heading and is used to indicate its different aspects.
• The level 3 heading is used to indicate the subpoints to be included under the level 2 heading and so on.

Here are the formatting guidelines for the APA heading formats for the 7th edition:

• Avoid creating too many subheadings. This makes your document shabby and difficult to understand. 
• Do NOT number the APA-style headings and subheadings. The APA paper format does not make use of Roman and Arabic letters and numerals to number the headings.
• Include a double space between the content and the heading for the first 3 levels.
• Use the title case for all your headings.
• Avoid creating a single APA subheading under the parent heading. 

Let’s take a look at how to format each of these APA heading levels in more detail.

APA heading levels

As stated above, the APA Style guide consists of 5 levels of headings. Each level heading should be bolded and in some cases italicized . Here are the guidelines for the five APA format headings:

APA level 1 heading

The level 1 heading for an APA paper is the main heading in an APA-style paper. It is used for all the significant sections of the paper such as “Methods”, “Results”, and “Discussion”. 

Do NOT include the Introduction in the level 1 heading. The heading of your paper present in the APA title page serves as a de facto heading for your introduction. 

Here are the formatting guidelines for an APA level 1 heading:

• Centrally align and bold the heading.
• Leave a double space between the heading and the text under it. 
• Begin writing the text on a new line, leaving an indent 0.5 inches from the left margin. 

Here is the APA heading format example:

APA Level 1 Heading

The text is indented and begins on a new line.

A section label is used to indicate specific parts of your paper including. It is similar to a level 1 heading but is formatted slightly differently. It is bolded and centrally aligned but is placed in a separate line at the top of a new page. 

APA level 2 heading

A level 2 heading is a subsection that is included under the level 1 heading. For instance, subheadings such as “Test Sample” are included under the main heading “Method”. Make sure that there are two or more subheadings present under each main heading. 

The level 2 APA subheading format is as follows:

• Left align and bold the heading.
• Leave a double space between your heading and the text pertaining to it.
• Leave an indent and begin writing the text on a new line.

Here is a level 2 APA heading example:

APA Level 2 Heading

APA level 3 heading

Level 3 APA subheadings are used to further categorize characteristics included under the level 2 subheading. For instance, the level 2 subheading “Test Sample” may further be divided into sample groups such as “Preteens”, “Adolescents”, and “Adults” . 

Here are the formatting guidelines for an APA level 3 heading:

• Left align, bold, and italicize the heading.
• Leave space between the heading and the text included below it.
• Leave a 0.5-inch indent and begin writing the text on a new line.

Here is a level 3 APA heading example:

APA Level 3 Heading

APA level 4 heading

The level 4 APA subheadings are rarely used when it comes to school papers. They’re more often used in more complex and detailed papers such as postgraduate or professional research papers. It is used to further categorize information included in the level 3 heading. For instance, the sample group of “Preteens” can further be divided into categories such as “Children (6–8 years)” and “Children (9–12 years)”.  

Following are the formatting guidelines for an APA level 4 heading:

• Left align, indent, and bold the heading.
• End the heading with a period.
• Begin writing the text after the period in the same line.

Here is a level 4 APA heading example:

APA Level 4 Heading. The text begins in the same line.

APA level 5 heading.

The level 5 heading is the final heading included under the APA formatting guidelines. It is used to further categorize information included under the level 4 heading. For instance, the category “Children (6–8 years)” can be further divided into “Boys” and “Girls”.

Following are the formatting guidelines for an APA level 5 heading:

• Left align, indent, bold, and italicize the heading.
• End your heading with a period.

Here is a level 5 APA heading example:

APA Level 5 Heading. The text begins in the same line.

When you put all of the headings together, it should look something like this:

APA Level 5 Heading . The text begins in the same line.

APA heading format for Microsoft Word and Google Docs

Things get much easier when it comes to formatting APA headings in MS Word and Google Docs. Instead of individually inserting each heading, you can simply make use of the “styles” feature. This allows you to select a heading of your choice with a single click.

Make sure to edit your heading as per the APA formatting guidelines, since these default settings are not available in MS Word or Google Docs.  

APA heading example

To understand how the APA levels are structured, let’s take a look at an APA format for headings. The following research paper outline highlights the methods used to collect the effect of coffee on preteens and young adults. 

Here’s how the headings and text should be structured in the following example.

If you think formatting is not your strong suit and you want to make things easier, consider working with professional editing services . These professionals not only correct structural errors and content flow but also rectify all proofreading errors. 

Need help with formatting your paper? Here are a few more articles you can refer to:

• MLA Format: 9th Edition Paper Formatting Guidelines
• APA Header Format: 5 Steps & Running Head Examples
• Chicago Style Citation: Quick Guide & Examples
• Chicago Title, Cover Page & Body | Paper Format Guidelines
• How to Create In-Text Citations and Reference Page in APA 7

Frequently Asked Questions

When should i use each level of heading, how are headings numbered in apa style, can i use more than five levels of headings in apa style.

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Title Case Capitalization

APA Style uses two types of capitalization for titles of works (such as paper titles ) and headings within works : title case and sentence case .

In title case, major words are capitalized, and most minor words are lowercase. In sentence case, most major and minor words are lowercase ( proper nouns are an exception in that they are always capitalized).

• major words: Nouns, verbs (including linking verbs), adjectives, adverbs, pronouns, and all words of four letters or more are considered major words.
• minor words: Short (i.e., three letters or fewer) conjunctions, short prepositions, and all articles are considered minor words.

Title case capitalization is covered in the seventh edition APA Style manuals in the Publication Manual Section 6.17 and the Concise Guide Section 5.7

How to implement title case

In title case, capitalize the following words in a title or heading:

• the first word of the title or heading, even if it is a minor word such as “The” or “A”
• the first word of a subtitle
• the first word after a colon, em dash, or end punctuation in a heading
• major words, including the second part of hyphenated major words (e.g., “Self-Report,” not “Self-report”)
• words of four letters or more (e.g., “With,” “Between,” “From”)

Lowercase only minor words that are three letters or fewer in a title or heading (except the first word in a title or subtitle or the first word after a colon, em dash, or end punctuation in a heading):

• short conjunctions (e.g., “and,” “as,” “but,” “for,” “if,” “nor,” “or,” “so,” “yet”)
• articles (“a,” “an,” “the”)
• short prepositions (e.g., “as,” “at,” “by,” “for,” “in,” “of,” “off,” “on,” “per,” “to,” “up,” “via”)

When to use title case

Use title case for the following:

• titles of articles, books, reports, and other works appearing in text

In the book Train Your Mind for Peak Performance: A Science-Based Approach for Achieving Your Goals

In the article “Turning Frowns (and Smiles) Upside Down: A Multilevel Examination of Surface Acting Positive and Negative Emotions on Well-Being”

• titles of tests or measures, including subscales

Beck Depression Inventory–II

• all headings within a work (Levels 1–5; these are also bold or bold italic)
• the title of your own paper and of named sections and subsections within it

the Results section

• titles of periodicals (these are also italicized)

Journal of Latinx Psychology

Chicago Tribune

• table titles (these are also italicized)
• figure titles (these are also italicized), axis labels, and legends

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College Assignment Heading: A Simple Guide for Formatting

Crafting an effective college assignment heading is key to making a strong first impression on professors. The title serves as the face of your paper and reflects your focus on precision and adherence to academic standards. Curious about how to create a proper title? This guide provides clear instructions on creating an impressive college paper heading, guiding you to comprehend what a header in writing is, and the appropriate way to format it. By applying these tips and guidelines, be confident your prep aligns with the required scholarly standards. 

Basic Rules for an Assignment Header

A well-organized college assignment heading is crucial for clearly identifying your assignment and presenting it professionally. Adhering to the basic rules will help you craft a carefully designed title:

•  Include Crucial Information: A typical heading generally features the student and teacher’s name and surname, the due date, and the current course name. Clearly display and align the project’s title to the document’s left margin. 
•  Use the Correct Font and Size: Standard academic papers require a legible font like Times New Roman or sometimes Arial and 12-point size. Avoid choosing decorative fonts that may undermine the professional quality of your composition. 
•  Follow Proper Spacing Guidelines: Your prep’s heading needs to be double-spaced, and coherent with the remainder of the writing. This maintains uniformity and readability within the document. 
•  Add Page Numbers (if indicated): Some teachers might ask for page numbers to be positioned in the header section. So, format them as per the given instructions. 

A well-crafted essay title guarantees your assignment recognition for clarity and compliance with academic standards, paving the way for successful submission.

How to Head an Assignment?

Understanding what is a heading in an essay and grasping how to head a paper correctly are foundational skills in scholarly writing. A proper essay header normally contains the instructor’s and student’s details, the course code or title, the current date, and the prep’s title. Most colleges follow APA, MLA, or Chicago-style guidelines, so familiarize yourself with these standards. Your header needs to be aligned correctly and placed where it doesn’t interfere with your work’s content. Adhering to these guidelines ensures your college paper heading is both succinct and polished.

Formatting Rules for a Heading

Following these guidelines guarantees your heading format meets academic expectations:

•  Ensure Consistent Alignment: Align all components of your header close to the left margin, maintaining uniformity in the paper. 
•  Use Double-Spacing: Keep double-spacing between the lines of your prep’s heading. 
•  Avoid Unnecessary Information: Keep the college heading concise, avoiding any extra text or embellishments. 
•  Capitalization and Punctuation: Properly capitalize and punctuate your essay heading consistent with style guidelines, ensuring consistency across the prep. 
•  Consistency: Use identical size, font, and style for all components in your header. 

Adhering to these rules will give your composition a polished look. This simplifies it for the lecturer to move through your composition.

Why Is a Good College Assignment Heading Important?

A skillfully designed paper heading gives your assignment a sophisticated look and helps to clearly organize the essentials for your readers. It signals to your lecturer that you have carefully followed educational standards and put thought into every aspect of your project. In addition, a proper heading for an essay guarantees that all relevant data is easily accessible. That’s specifically useful in larger classes where preps are handled by multiple lecturers or TAs. By concentrating on a succinct and accurate essay heading, you set a strong foundation for the rest of your home tasks.

Tips for Refining Your College Paper Heading

Refining your college paper heading is vital to guarantee it meets academic criteria and effectively represents your paper. By carefully focusing on the particulars, you can elevate the expertise and coherence of your home assignment. See some practical tips to consider:

•  Review institutional guidelines: Always verify your institution's detailed formatting instructions or templates to ensure adherence. 
•  Keep it simple: Steer clear of irrelevant information in your heading; a clean, straightforward format is best. 
•  Maintain consistency: Make sure that the font, size, and alignment are uniform across your header. 
•  Double-check accuracy: Verify that all components in your title, comprising your name and surname, course name, and submission date, are correct. 
•  Align with your prep's style: Confirm the header complements the overall tone and style of your written work. 

By applying these suggestions, you can develop an assignment title that fulfills academic standards and improves the overall appearance of your prep.

 Recap

A well-formatted college paper heading is a minor but vital part of academic composition. From understanding what a title is to mastering how to head a text, focusing on these details can make a notable difference in the overall presentation of your composition. Always stick to the fundamental formatting standards and each time confirm that you follow the heading format specified by your educational institution's guidelines. To further enhance your writing, consider utilizing our tools at AI Essay Detector and College Essay Generator to help you create outstanding essays with ease.

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Speaker 1: Today we will discuss what is perhaps the most single important element of your research paper, the title. The title is the first thing that journal editors and reviewers see when they look at your paper. It is also the only piece of information that fellow researchers will see in a database or search engine query. Therefore, you want to make sure the title captures all of the relevant aspects of your study but does so in a way that is accessible and captivating to readers. Follow these steps to create a perfect title for your paper. First, ask yourself some questions about what your paper seeks to answer and what it accomplishes. What is my paper about? My paper studies how program volume affects outcomes for liver transplant patients on waiting lists. What methods or techniques did I use to perform my study? I employed a case study. What or who was the subject of my study? I studied 60 liver transplant patients on a waiting list in the U.S. aged 20 to 50 years. What were the results? Positive correlation between waiting list volume and poor outcome of transplant procedure. After answering these questions, move on to the second step, which is to identify and list key words and phrases from these responses. Program volume, outcomes, liver transplant patients, waiting lists, case study, 60 liver transplant patients, age 20 to 50 years, positive correlation. These keywords will form the foundation of your title. Once you have identified and listed these keywords, use them to create one long sentence. This study used a case study of 60 liver transplant patients around the U.S. aged 20 to 50 years to assess how the waiting list volume affects the outcome of liver transplantation in patients. These indicate a positive correlation between increased waiting list volume and a negative prognosis after transplant procedure. Next create a working title. Remove elements that make it a complete sentence, but keep everything that is most important to what the study is about. Delete all unnecessary or redundant words. Now let's shift some words around and rephrase it a bit to shorten the length and make it leaner and yet more natural. What you are left with is a case study of 60 liver transplant patients around the U.S. aged 20 to 50 years assessing the impact of waiting list volume on outcome of a transplantation and showing a positive correlation between increased waiting list volume and negative prognosis. This is getting closer to what we want in a title, which is just the most important information. But note that the word count for this working title is still 38 words, whereas the average published journal article title is 16 words or fewer. Therefore we need to eliminate some words and phrases that are not essential to the title. In step 5 you will delete all extra words and phrases and put key words at the beginning and end of your title. Since the number of patients studied and the exact outcome are not the most essential elements of this paper, remove these elements first. In addition, the methods used in a study are not usually the most searched for keywords in databases and represent additional details that you may want to remove to make your title a little leaner. So we are left with assessing the impact of waiting list volume on outcome and prognosis in liver transplantation patients. In this final version you can immediately see how much clearer the title is and what the study is about and what it aims to achieve. And note that the important terms are written at the beginning and the end of the title. Assessing, which is the main action of the study, is at the beginning and liver transplantation patients, the specific subject of the study, is given at the end. This will help a lot with search engine and database queries, meaning that a lot more researchers will find your article once it is published. And if you want to add a subtitle to give more detail about methodology, you can do this by putting this information after a colon. A case study of US adult patients ages 20 to 25. We abide strictly by our word count rule, this may be unnecessary, but every journal has its own standard formatting and style guidelines for titles, so it's a good idea to be aware of these both while writing your title and writing the study itself. So let's review these steps. First, answer some basic questions about your paper. Next, identify and list keywords and phrases from these responses. Third, turn these keywords into a long sentence. Out of this long sentence you can create a working title, deleting any unnecessary and redundant words. Last, delete any extra elements to meet a suitable word count for your title. Note key terms at the beginning and the end, and again you may add a subtitle if it seems necessary or important to this study. Keep these tips in mind when creating your research paper title. Write the title after you've written your paper in abstract. Include all of your paper's essential terms. Keep it short and to the point, about 16 words or fewer is best. Avoid using jargon or abbreviations that will not be understandable by the general reader. Use keywords that closely relate to the content of your paper, and never use a period at the end of your title. Remember, your title is not a sentence. For more tips on how to improve your writing, visit wordvice.com and check out our resources page where you will find a great many helpful articles and videos. And be sure to subscribe to our YouTube channel and social media pages to stay up to date with more excellent academic writing and journal publications content.

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What a Thesis Paper is and How to Write One

From choosing a topic and conducting research to crafting a strong argument, writing a thesis paper can be a rewarding experience.

It can also be a challenging experience. If you've never written a thesis paper before, you may not know where to start. You may not even be sure exactly what a thesis paper is. But don't worry; the right support and resources can help you navigate this writing process.

What is a Thesis Paper?

A thesis paper is a type of academic essay that you might write as a graduation requirement for certain bachelor's, master's or honors programs. Thesis papers present your own original research or analysis on a specific topic related to your field.

“In some ways, a thesis paper can look a lot like a novella,” said Shana Chartier , director of information literacy at Southern New Hampshire University (SNHU). “It’s too short to be a full-length novel, but with the standard size of 40-60 pages (for a bachelor’s) and 60-100 pages (for a master’s), it is a robust exploration of a topic, explaining one’s understanding of a topic based on personal research.”

Chartier has worked in academia for over 13 years and at SNHU for nearly eight. In her role as an instructor and director, Chartier has helped to guide students through the writing process, like editing and providing resources.

Chartier has written and published academic papers such as "Augmented Reality Gamifies the Library: A Ride Through the Technological Frontier" and "Going Beyond the One-Shot: Spiraling Information Literacy Across Four Years." Both of these academic papers required Chartier to have hands-on experience with the subject matter. Like a thesis paper, they also involved hypothesizing and doing original research to come to a conclusion.

“When writing a thesis paper, the importance of staying organized cannot be overstated,” said Chartier. “Mapping out each step of the way, making firm and soft deadlines... and having other pairs of eyes on your work to ensure academic accuracy and clean editing are crucial to writing a successful paper.”

How Do I Choose a Topic For My Thesis Paper?

What your thesis paper is for will determine some of the specific requirements and steps you might take, but the first step is usually the same: Choosing a topic.

“Choosing a topic can be daunting," said Rochelle Attari , a peer tutor at SNHU. "But if (you) stick with a subject (you're) interested in... choosing a topic is much more manageable.”

Similar to a thesis, Attari recently finished the capstone  for her bachelor’s in psychology . Her bachelor’s concentration is in forensics, and her capstone focused on the topic of using a combined therapy model for inmates who experience substance abuse issues to reduce recidivism.

“The hardest part was deciding what I wanted to focus on,” Attari said. “But once I nailed down my topic, each milestone was more straightforward.”

In her own writing experience, Attari said brainstorming was an important step when choosing her topic. She recommends writing down different ideas on a piece of paper and doing some preliminary research on what’s already been written on your topic.

By doing this exercise, you can narrow or broaden your ideas until you’ve found a topic you’re excited about. " Brainstorming is essential when writing a paper and is not a last-minute activity,” Attari said.

How Do I Structure My Thesis Paper?

Thesis papers tend to have a standard format with common sections as the building blocks.

While the structure Attari describes below will work for many theses, it’s important to double-check with your program to see if there are any specific requirements. Writing a thesis for a Master of Fine Arts, for example, might actually look more like a fiction novel.

According to Attari, a thesis paper is often structured with the following major sections:

Introduction

• Literature review
• Methods, results

Now, let’s take a closer look at what each different section should include.

Your introduction is your opportunity to present the topic of your thesis paper. In this section, you can explain why that topic is important. The introduction is also the place to include your thesis statement, which shows your stance in the paper.

Attari said that writing an introduction can be tricky, especially when you're trying to capture your reader’s attention and state your argument.

“I have found that starting with a statement of truth about a topic that pertains to an issue I am writing about typically does the trick,” Attari said. She demonstrated this advice in an example introduction she wrote for a paper on the effects of daylight in Alaska:

In the continental United States, we can always count on the sun rising and setting around the same time each day, but in Alaska, during certain times of the year, the sun rises and does not set for weeks. Research has shown that the sun provides vitamin D and is an essential part of our health, but little is known about how daylight twenty-four hours a day affects the circadian rhythm and sleep.

In the example Attari wrote, she introduces the topic and informs the reader what the paper will cover. Somewhere in her intro, she said she would also include her thesis statement, which might be:

Twenty-four hours of daylight over an extended period does not affect sleep patterns in humans and is not the cause of daytime fatigue in northern Alaska .

Literature Review

In the literature review, you'll look at what information is already out there about your topic. “This is where scholarly articles  about your topic are essential,” said Attari. “These articles will help you find the gap in research that you have identified and will also support your thesis statement."

Telling your reader what research has already been done will help them see how your research fits into the larger conversation. Most university libraries offer databases of scholarly/peer-reviewed articles that can be helpful in your search.

In the methods section of your thesis paper, you get to explain how you learned what you learned. This might include what experiment you conducted as a part of your independent research.

“For instance,” Attari said, “if you are a psychology major and have identified a gap in research on which therapies are effective for anxiety, your methods section would consist of the number of participants, the type of experiment and any other particulars you would use for that experiment.”

In this section, you'll explain the results of your study. For example, building on the psychology example Attari outlined, you might share self-reported anxiety levels for participants trying different kinds of therapies. To help you communicate your results clearly, you might include data, charts, tables or other visualizations.

The discussion section of your thesis paper is where you will analyze and interpret the results you presented in the previous section. This is where you can discuss what your findings really mean or compare them to the research you found in your literature review.

The discussion section is your chance to show why the data you collected matters and how it fits into bigger conversations in your field.

The conclusion of your thesis paper is your opportunity to sum up your argument and leave your reader thinking about why your research matters.

Attari breaks the conclusion down into simple parts. “You restate the original issue and thesis statement, explain the experiment's results and discuss possible next steps for further research,” she said.

Find Your Program

Resources to help write your thesis paper.

While your thesis paper may be based on your independent research, writing it doesn’t have to be a solitary process. Asking for help and using the resources that are available to you can make the process easier.

If you're writing a thesis paper, some resources Chartier encourages you to use are:

• Citation Handbooks: An online citation guide or handbook can help you ensure your citations are correct. APA , MLA and Chicago styles have all published their own guides.
• Citation Generators: There are many citation generator tools that help you to create citations. Some — like RefWorks — even let you directly import citations from library databases as you research.
• Your Library's Website: Many academic and public libraries allow patrons to access resources like databases or FAQs. Some FAQs at the SNHU library that might be helpful in your thesis writing process include “ How do I read a scholarly article? ” or “ What is a research question and how do I develop one? ”

It can also be helpful to check out what coaching or tutoring options are available through your school. At SNHU, for example, the Academic Support Center offers writing and grammar workshops , and students can access 24/7 tutoring and 1:1 sessions with peer tutors, like Attari.

"Students can even submit their papers and receive written feedback... like revisions and editing suggestions," she said.

If you are writing a thesis paper, there are many resources available to you. It's a long paper, but with the right mindset and support, you can successfully navigate the process.

“Pace yourself,” said Chartier. “This is a marathon, not a sprint. Setting smaller goals to get to the big finish line can make the process seem less daunting, and remember to be proud of yourself and celebrate your accomplishment once you’re done. Writing a thesis is no small task, and it’s important work for the scholarly community.”

A degree can change your life. Choose your program  from 200+ SNHU degrees that can take you where you want to go.

Meg Palmer ’18 is a writer and scholar by trade who loves reading, riding her bike and singing in a barbershop quartet. She earned her bachelor’s degree in English, language and literature at Southern New Hampshire University (SNHU) and her master’s degree in writing, rhetoric and discourse at DePaul University (’20). While attending SNHU, she served as the editor-in-chief of the campus student newspaper, The Penmen Press, where she deepened her passion for writing. Meg is an adjunct professor at Johnson and Wales University, where she teaches first year writing, honors composition, and public speaking. Connect with her on LinkedIn .

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About southern new hampshire university.

SNHU is a nonprofit, accredited university with a mission to make high-quality education more accessible and affordable for everyone.

Founded in 1932, and online since 1995, we’ve helped countless students reach their goals with flexible, career-focused programs . Our 300-acre campus in Manchester, NH is home to over 3,000 students, and we serve over 135,000 students online. Visit our about SNHU  page to learn more about our mission, accreditations, leadership team, national recognitions and awards.

• Open access
• Published: 30 August 2024

Rice Promoter Editing: An Efficient Genetic Improvement Strategy

• Bowen Wu 1   na1 ,
• Hangfei Luo 1   na1 ,
• Zhongbo Chen 1   na1 ,
• Bakht Amin 1 ,
• Manyu Yang 1 ,
• Zhenghan Li 1 ,
• Shuai Wu 1 ,
• Saleh H. Salmen 3 ,
• Sulaiman Ali Alharbi 3 &
• Zhongming Fang 1 , 2  

Rice volume  17 , Article number:  55 ( 2024 ) Cite this article

Metrics details

Gene expression levels in rice ( Oryza sativa L.) and other plant species are determined by the promoters, which directly control phenotypic characteristics. As essential components of genes, promoters regulate the intensity, location, and timing of gene expression. They contain numerous regulatory elements and serve as binding sites for proteins that modulate transcription, including transcription factors and RNA polymerases. Genome editing can alter promoter sequences, thereby precisely modifying the expression patterns of specific genes, and ultimately affecting the morphology, quality, and resistance of rice. This paper summarizes research on rice promoter editing conducted in recent years, focusing on improvements in yield, heading date, quality, and disease resistance. It is expected to inform the application of promoter editing and encourage further research and development in crop genetic improvement with promote.

Introduction

The phenotype of rice is influenced by both genetic and environmental factors (Wu et al. 2020 ; Chen et al. 2024 ; Ahmad et al. 2024 ). Genetic modification offers the potential for stable phenotypic performance by altering its genetic information (Zeb et al. 2022 ; Ahmad et al. 2022a , b ). Common strategies include introducing genes from exogenous species, modulating gene expression levels, and completely disrupting gene function (Luo et al. 2024 ; Zeng et al. 2020a , b ). Consequently, a variety of gene editing techniques have been developed and employed to achieve these goals, such as TALENs (Becker and Boch 2021 ), ZFNs (Gupta and Musunuru 2014 ), CRISPR/Cas (Riaz et al. 2022 ; Shaheen et al. 2023 ), Prime Editing (Xu et al. 2020 ; Ahmad et al. 2022a , b ), and Base Editing (Li et al. 2023a , b , c ; Tabassum et al. 2021 ; Monsur et al. 2020 ). The advent of these technologies has revolutionized the precise manipulation of targeted nucleic acids, profoundly impacting modern rice breeding efforts (Lu et al. 2018 ; Ahmad et al. 2021a , b ). For a long period in the past, rice gene editing had concentrated on modification of the coding regions of functional genes. While this approach has yielded some positive results, it has also revealed certain drawbacks. For example, leading to unwanted abnormal phenotypes such as cessation of flowering, reduced yield and increased susceptibility to diseases (Fan et al. 2014 ; Li et al. 2020 ; Tan et al. 2018 ; Zhao et al. 2015 ). Additionally, deletion of entire promoter may result in damage to the three-dimensional genome structure and cause severe phenotypic changes (Spielmann et al. 2018 ). Consequently, researchers have observed that the most effective approach to addressing these limitations may involve mild regulation of gene expression.

In recent years, molecular breeders have shifted their focus from coding sequences to other genomic regions, with particular emphasis on promoters (Jia et al. 2022 ; Liu et al. 2021a , b ; Peng et al. 2017 ). Promoters are DNA sequences comprising of several thousand base pairs (bp) located upstream of the transcribed region of genes. They typically contain multiple transcriptional regulatory elements, known as cis-regulatory elements (CREs), which regulate gene expression by binding to specific proteins (Marand et al. 2023 ). The core promoter is typically defined as the region approximately ± 50 bp on either side of the transcription start site (TSS) (Bernardini et al. 2023 ), which plays a pivotal role in regulating the initiation and direction of transcription (Haberle and Stark 2018 ). The proximal promoter with many primary regulatory elements is found approximately 250 bp upstream from the TSS. It is the binding site for many transcription factors (Thrall et al. 2024 ). The distal promoter, situated further upstream from the proximal promoter, also contains lots of cis-regulatory elements (Hausman 2022 ). Furthermore, upstream of the coding sequence contains the 5′ untranslated region (5′UTR) and the upstream open reading frames (uORFs) located therein. The 5′UTR, which lies between TSS and the first coding base, plays a non-negligible role in transcription regulation (Chung et al. 2006 ; Samadder et al. 2017 ). uORFs, potential open reading frames within the 5′UTR, have been reported to regulate the transcription or translation of downstream main open reading frame (mORF) (Combier et al. 2008 ; Xu et al. 2017 ). The promoter region contains both known and potential factors that regulate gene expression, so it has received attention from researchers in recent years. Consequently, editing this region has been progressively studied in depth. It has been demonstrated that editing promoter sequences can be an effective means for mastering gene expression regulation (Li et al. 2017 ; Zeng et al. 2020a , b ; Wang et al. 2024 ). Editing crop promoters had the potential to generate novel alleles that regulated the expression of target genes on a large range (Rodriguez-Leal et al. 2017 ). Mutations in coding sequences often lead to complete disruption of gene function (Wittkopp and Kalay 2012 ). At some point, it makes more sense to efficiently reduce gene expression without completely disrupting gene function, thereby achieving the desired results with no or only negligible negative effects. Mutations in CREs tends to alter the spatiotemporal expression patterns and intensity of genes (Wu et al. 2021a , b ). Editing specific CREs in promoter region was considered a practical strategy for reducing transcript levels (Haberle and Stark 2018 ). In rice promoter editing, fine-tuning target gene expression through promoter editing had successfully minimized adverse effects on plant growth and development (Wang et al. 2021a , b , c ). Numerous examples demonstrated that regulating gene expression by editing CREs in the promoter regions of target genes is an effective strategy for genetic improvement of rice (Li et al. 2023a , b , c ) and aligns with the theories and practices of modern rice breeding.

As a globally essential food crop, rice serves as a staple for billions, particularly in Asian countries. The focus on enhancing rice traits, such as yield, quality, and resistance, has never been more critical. Although improving rice yield has traditionally been a primary focus of breeders, it is notable that only a few studies have directly addressed yield traits. Instead, the majority of researchers concentrated their efforts on improving rice heading date, eating and cooking qualities (ECQ) and bacterial disease resistance, with only a few studies focusing on other aspects. This trend is somewhat unexpected, but it is worth noting that most existing literatures also considered the impact of promoter region editing on yield when examining other traits. Therefore, promoter gene editing, a powerful tool in genetic improvement, holds vast potential for application in rice production and may serve as a model for breeding other crops.

Moreover, as a model organism among crops, rice offers several advantages for genetic research: it is one of the earliest plants to have its genome sequenced, it is diploid with a relatively small genome, and the costs associated with its well-established genetic transformation are low. These factors make rice uniquely suited for promoter editing studies, providing a fertile ground for the validation and optimization of this technology. Furthermore, rice can rapidly generate large amounts of genetic materials, facilitating the swift validation and refinement of promoter editing techniques. Then, the significance of rice promoter editing extends far beyond this crop; it holds promise for advancing the entire field of plant genetics. In addition, the wealth of wild rice germplasm and regionally cultivated rice varieties, particularly in Asia, adds another layer of complexity and opportunity. The regional specificity and strong local adaptation of commercial rice varieties necessitate tailored promoter editing strategies to accommodate the genetic diversity found in different varieties. This specificity is crucial for the successful improvement of rice varieties. Thus, a comprehensive review of the research progress in rice promoter editing is not only timely but essential.

This paper provides a comprehensive review of recent literature on rice promoter region editing (Table  1 ). As gene editing technology continues to advance with greater precision and efficiency, it is anticipated that an increasing number of studies will focus on promoter region editing in rice and other crops. We believe that this comprehensive review will serve as a valuable reference for future research on promoter editing in other plant species, and will help drive further advancements in crop genetic improvement and agricultural sustainability.

Research Progress on Promoter Region Editing of Rice

In recent years, studies on editing of the promoter region in rice have focused on regulation of heading date, improvement of eating and cooking quality, and enhancement of bacterial disease resistance. In addition, a few studies have explored the improvement of rice yield, fertility restoration of male sterile lines, and enhancement of micronutrient content.

Regulation of Yield-Related Traits

The breeding of high-yielding rice is a topic that is consistently addressed in the field of rice cultivation. Despite the successes of existing high-yielding varieties, breeders continually strive for higher and more consistent yields with unremitting efforts. Rice yield is primarily determined by tillering and panicle architecture traits (Wen et al. 2024 ), including panicle number, grain number per panicle, and grain weight (Amin et al 2024 ; Wang and Li 2005 ). These traits exhibit complex quantitative variation among different rice varieties, as they are typical quantitative traits. To generate a wide range of Quantitative Trait Variants (QTVs) in rice, a CRISPR/Cas12a promoter editing (CAPE) system was developed, which was based on a promoter Key-Region (KR) estimating model and an efficient CRISPR/Cas12a-based multiplexed or singular editing system (Zhou et al. 2023 ). Experimental results with promoter editing of multiple rice genes demonstrated the efficacy of this system in creating QTVs. It was observed that editing KRs with high total scores produced more obvious phenotypes. For instance, promoter-edited OsGBSS1 ( Wx ) and OsGS3 lines demonstrated continuous variation in starch content and seed size (Zhou et al. 2023 ). Furthermore, the expression level of OsGBSS1 correlated strongly with amylose content (AC) (Zhou et al. 2023 ). Additionally, QTVs for seed length and width were also observed (Zhou et al. 2023 ). A comparative analysis of two representative promoter-edited lines revealed no differences in morphology, height, or main panicle length compared to wild-type (WT) (Zhou et al. 2023 ). Promoter-edited OsD2 lines showed a gradual reduction in OsD2 expression, which was accompanied by a decrease in plant height. Nevertheless, main panicle length, seed number, seed size, and 1,000-grain weight were affected to varying extents (Zhou et al. 2023 ). Concurrently, promoter editing of OsD11 yielded outcomes comparable to those of OsD2 (Zhou et al. 2023 ). OsSD1 (also known as GA20ox ) has been successfully employed to generate rice varieties exhibiting semi-dwarfing characteristics while maintaining yield stability (Qiao and Zhao 2011 ). Two promoter-edited OsSD1 lines also exhibited dwarfing traits and showed no differences in grain size or 1000-grain weight compared to WT and null-mutant controls. The promoter-edited OsD18 lines demonstrated a continuous range of variation in plant height, from 64 to 98 cm, with reduced OsD18 expression (Zhou et al. 2023 ). Additionally, main panicle length and grain number exhibited intermediate phenotypes between OsD18 knockout mutants and the WT (Zhou et al. 2023 ). In the field, wild-type plants exhibited a pronounced lodging phenotype, whereas OsD18 promoter-edited lines demonstrated resistance to lodging (Zhou et al. 2023 ). This study highlighted that editing the OsD18 promoter can produce semi-dwarfing QTVs without a significant loss of yield, which is referred to as a "quantitative green revolution trait". The most notable aspect of this research is the development of the robust CAPE system, which effectively introduces QTVs related to grain starch content, grain size, and plant height. This study not only represents a technological innovation in promoter editing but also validates the potential of promoter editing to introduce QTVs in rice. The refinement and widespread adoption of the CAPE system are expected to greatly advance promoter editing research, facilitate the creation of diverse trait variations, and ultimately contribute to the improvement of rice and other crops.

Rice breeding is often constrained by trade-offs among different agronomic traits, including the penalties of yield by plant immunity, dilution effect of plant nutrition, and negative correlations among plant architecture components (Nelson et al. 2018 ; Takatsuji 2017 ; Wang et al. 2021a , b , c ; Wang et al. 2021a , b , c ). These trade-offs, particularly among traits like tiller number, grain number per panicle and grain weight, limit overall rice yield. Many of these trade-offs can be attributed to genetic pleiotropy (Song et al. 2022 ). Pleiotropic genes affecting multiple traits may result in undesirable consequences. When attempting to regulate a specific trait by modifying a pleiotropic gene, there is a possibility that the modification may have an adverse effect on other traits associated with that gene. Successfully breaking the pleiotropy allows for the full utilization of beneficial effects while overcoming its adverse effects on other traits. In a previous study, researchers demonstrated that editing the promoter region of IPA1 effectively reduced its pleiotropy (Song et al. 2022 ). They discovered that certain promoter fragments deletion mutants can influence IPA1 expression in a tissue-specific manner, thereby reducing IPA1 pleiotropy and producing extensive phenotypic variation in tiller number and panicle size (Song et al. 2022 ). One such promoter-edited lines, IPA1-Pro10,which contained a 54 bp deletion, exhibited an increase in both tiller number and panicle size, leading to a greatly improvement in rice yield by overcoming their trade-offs (Song et al. 2022 ). Field experiments showed that the IPA1-Pro10 exhibited a 7.2% increase in tiller number per plant and an 8.5% increase in grain number per panicle compared to ZH11 (Song et al. 2022 ). Additionally, IPA1-Pro10 demonstrated a 15.9% increase in yield while maintaining a similar heading date (Song et al. 2022 ). To elucidate the mechanism underlying this synergistic increase in yield-related traits, researchers conducted further investigations and discovered that the transcription factor An-1 (Luo et al. 2013 ) could directly bind to the GCGCGTGT motif on the promoter of IPA1 , repressing the expression of IPA1 in the spikelet (Song et al. 2022 ). This suggested that An-1 specifically restrains spikelet and secondary meristem of the panicle. The deletion of this region resulted in altered tissue-specific expression of IPA1 and separated the pleiotropy of both IPA1 and An-1 to overcome agronomic trade-offs (Song et al. 2022 ). This study demonstrates that a tiling-deletion screen for cis-regulatory regions (CRRs) holds promise for reducing gene pleiotropy caused by different spatial–temporal functions of the same gene, which is expected to address the breeding difficulty resulting from trade-offs. As many genes affect rice yield (Dong et al. 2022 ), future research should delve deeper into these genes, especially using promoter editing technology.

Regulation of Heading Date

Flowering date is an important trait for rice adaptation to different light conditions in multiple places. Fine-tuning the heading date of desirable rice varieties has received much attention from breeders. There are many genes that can control the heading data of rice (Chen et al. 2022 ). Typically, delaying the heading date in rice is achieved through two main approaches: either elevation of the expression of floral inhibitors, or reduction of the expression of floral inducers (Liu et al. 2021a , b ). Direct editing of the coding region of flowering genes often resulted in null mutations, which can significantly delay or even prevent the heading of rice (Zhao et al. 2015 ). Consequently, mutations caused by editing coding regions often have no practical application value. A more practical goal for rice breeders is to create a series of rice lines with different heading dates by fine-tuning the expression of flowering genes. Rice Flowering Locus T1 ( RFT1 ) was a significant floral regulator under long-day (LD) conditions (Tsuji et al. 2011 ). RFT1 formed a ternary complex (Florigen activation complex, FAC) with 14-3-3 protein and the transcription factor OsFD1, which promoted flowering under LD conditions (Peng et al. 2021 ). Recently, a study successfully created mutants with multiple promoter sequence variations by editing a 0.5 kb region in the promoter of RFT1 (Zhou et al. 2024a , b ). These mutants exhibited delayed heading date ranging from 1.4 to 9.2 days, accompanied by reduced expression of flower-forming genes RFT1 and Heading date 3a ( Hd3a ) (Zhou et al. 2024a , b ). The researchers also assessed the promoter activity of the RFT1 promoter mutant, which showed reduced promoter activity (Zhou et al. 2024a , b ). In addition, their investigations showed that other agronomic traits, such as plant height, panicle number, and 1,000-grain weight, in the mutants were not significantly different from the WT (Zhou et al. 2024a , b ).

Heading date 1 ( Hd1 ), Heading date 7 ( Ghd7 ), and Days to heading 8 ( DTH8 ) were three genes that act mainly upstream of Early heading date 1 ( Ehd1 ), and their combinations largely determined the photoperiod sensitivity and geographic environment adaptation of rice (Zhang et al. 2015a , b ). Hd1 was a photoperiod-sensitive gene that affected heading date and maturity in rice, and both Ghd7 and DTH8 were important flowering suppressors (Xue et al. 2008 ; Wei et al. 2010 ). Using the High-Efficiency Multiplex Promoter-Targeting (HMP) system, the 2.0 kb promoter regions of Hd1 , Ghd7 , and DTH8 were randomly and uniformly edited, and new alleles in their cis-regulatory regions were generated (Zhou et al. 2024a , b ). These mutants exhibited a range of heading dates, for example, the promoter-edited mutants of Hd1 had heading dates ranging from about 73 days like the Hd1 null mutation to about 107 days like the WT. Similar phenotypes were observed in mutant lines of Ghd7 and DTH8 (Zhou et al. 2024a , b ). Notably, several of these mutant lines exhibited precocious heading phenotypes close to the Hd1 null mutation (Zhou et al. 2024a , b ), suggesting that the promoter mutations in these lines are almost equivalent to the null mutation.

The early heading gene Ehd1 promoted rice flowering by activating the expression of flowering genes Hd3a and RFT1 (Doi et al. 2004 ), while Hd2 was a repressor of Ehd1 (Hori et al. 2016 ). In previous study, the heading date of rice was extended by surging the expression of flowering repressors (Liu et al. 2021a , b ). Then, three uORFs in the Hd2 leader sequence were identified and edited, and mutant lines with 4–11 flowering days later than the WT were successfully obtained (Liu et al. 2021a , b ). Quantitative reverse transcription polymerase chain reaction (qRT-PCR) results showed a significant decrease in the expression of Ehd1 , Hd3a and RFT1 (Liu et al. 2021a , b ). Interestingly, the transcript level of Hd2 remained unchanged, but a follow-up study showed that the protein level of Hd2 was actually increased (Liu et al. 2021a , b ), suggesting that the uORF of Hd2 can inhibit the translation without affecting transcription. This study provides new insights into target selection for promoter editing.

Ehd1 not only regulated rice heading date but also affected yield-related agronomic traits (Cho et al. 2016 ; Doi et al. 2004 ). Several Ehd1 promoter mutants with editing the nearby regions of some certain CREs in Ehd1 promoter exhibited varying degrees of reduced expression levels of Ehd1 , Hd3a and RFT1 (Li et al. 2023a , b , c ). This was different from the situation where the Ehd1 null mutation with editing of coding region of Ehd1 led to almost completely suppressed expression of Hd3a and RFT1 (Li et al. 2023a , b , c ). Furthermore, promoter mutants displayed delayed heading dates and enhanced some yield-related traits (Li et al. 2023a , b , c ). The uORF mutants exhibited similar phenotypes (Li et al. 2023a , b , c ), indicating that this uORF positively regulates the function of Ehd1 .

Improvement of Quality

As living standards rise, people are paying greater attention to the flavor and appearance of rice quality. Studies have demonstrated that gene editing certain genes can improve quality of rice (Hui et al. 2022 ; Mao et al. 2021 ). Amylose content (AC) is a significant indicator of the eating and cooking quality (ECQ) of rice (Li et al. 2016 ). In general, rice grains with higher AC and lower gel consistency showed poorer taste profile, whereas grains with moderate AC (15–20%) and higher gel consistency (60–80 mm) exhibited superior taste profile for the majority of population (Zeng et al. 2020a , b ). Wx ( OsGBSS1 ) was regarded as the most pivotal gene regulating AC (Sano 1984 ; Zhu et al. 2024 ). The Wx -encoded granule-bound starch synthase 1 (GBSS1) was the primary enzyme responsible for influencing AC (Mikami et al. 2008 ; Zhang et al. 2019 ). Natural variations in Wx gave rise to different levels of AC, as well as variations in ECQ among different rice varieties (Zhang et al. 2019 ). In addition to regulating AC and ECQ, Wx also affected gel consistency, water absorption, and elongation of cooked rice (Kim et al. 2021 ; Tian et al. 2009 ; Zhang et al. 2021 ). This evidence demonstrates that Wx plays a significant role in determining the ECQ of rice. Based on the regulatory role of Wx on AC, a lot of researchers believe that editing Wx to regulate AC for improving rice quality is a viable strategy.

Moreover, it has been demonstrated that moderate regulation of AC could be achieved by precisely regulating the expression of Wx (Huang et al. 2020 ). Wx1764178A identified by Tang et al. ( 2024 ) was an allele of Wx that had an adenine variant SNP (A-SNP) in its promoter region. In previous study, the CRISPR/Cas9 system was employed to target the sequence in the vicinity of the A-SNP in the promoter of this allele for gene editing (Huang et al. 2020 ). Compared to the WT, the promoter-edited lines exhibited reduced Wx expression, with a reduction in apparent amylose content (AAC) of approximately 10% (Huang et al. 2020 ). It is noteworthy that the target is situated 2,500 bp away from the upstream region of the Wx promoter. This suggests that the discovery and utilization of polymorphic sites beyond 2000 bp upstream of the promoter may also have potential for applications.

Reducing rice chalkiness is also a crucial aspect in achieving high-quality rice. Wxa , which was also an allele of Wx , contributed to the production of rice with a high AC (25–30%), which negatively affected the flavor of rice (Wang et al. 1995 ). By modifying the region between 500 and 760 base pairs upstream of the Wxa promoter, edited lines demonstrated a notable decrease in AC (Li et al. 2023a , b , c ). Furthermore, these edited lines were accompanied by an improvement in chalky grain percentage (CGP) and chalk grain grade (CGG), without any adverse effects on other agronomic traits (Li et al. 2023a , b , c ). Other studies have also conducted research on promoter editing in Wxa . Gene editing of several CREs, including Endosperm-box, A-box, CAAT-box and 5’UTR intronic splicing site (5’UISS), in the Wxa promoter region has been conducted (Zeng et al. 2020a , b ). The findings indicated a notable decline in Wxa mRNA levels among a portion of promoter region element mutants, as well as across the entirety of 5′UISS mutants. Rapid visco analysis of starch quality indicated that the viscosity index of the edited lines correlated with their ACs, exhibiting various degrees of variability (Zeng et al. 2020a , b ). Meantime, the primary agronomic characteristics of the mutants, including grain weight, grain length, grain width, and plant height, were not significantly different from those of the WT (Zeng et al. 2020a , b ).

Wxb was a weak allele of Wx that produced moderate AC (15–18%) (Isshiki et al. 1998 ). By editing the region near the TATA-box of the Wxb promoter, mutant lines were successfully generated with reduced the expression level of Wx (Huang et al. 2020 ). Some promoter-edited lines exhibited markedly diminished AC, whereas others demonstrated no discernible alterations (Huang et al. 2020 ). Notably, one edited line demonstrated a moderate reduction in AC under a variety of growth conditions (Huang et al. 2020 ). This particular line was distinguished by the alteration of only the bases situated in the vicinity of the TATA box, without any impact on other sequences, resulting in a significant change in Wx expression (Huang et al. 2020 ). This study demonstrates that targeting the core region of a gene promoter for editing is a viable method for regulating gene expression.

A recent study introduced an engineered CRISPR/FrCas9 system from Faecalibaculum rodentium for efficient plant genome editing, overcoming challenges with TA-rich core promoters (Wang et al. 2024 ). The study showed that the system can efficiently edit the core promoter region of the Wx gene. The transcript abundance of Wx in the promoter edited lines were reduced to a certain extent, accompanied by a decrease in AC (Wang et al. 2024 ). This research highlights the high efficiency of FrCas9-mediated core promoter editing in rice, and its potential for versatile base editing as well as precise gene expression regulation.

In addition to Wx , Glucan , Water-Dikinase 1 ( GWD1 ) was a pivotal gene that regulated both the degradation and synthesis of rice starch (Ritte et al. 2002 ). GWD1 catalyzed the reaction of α-glucan and adenosine triphosphate (ATP) with water to produce α-glucan phosphate monoesters, adenosine monophosphate (AMP), and orthophosphate (Ritte et al. 2002 ). Additionally, GWD1 catalyzed the phosphorylation of starch, regulating both starch degradation and synthesis (Mikkelsen et al. 2005 ; Skeffington et al. 2019 ; Xu et al. 2017 ). Using the CRISPR/Cas9 system, researchers successfully edited the promoter of GWD1 , generating gwd1 weak mutants (Wang et al. 2021a , b , c ). These promoter mutant lines exhibited a notable reduction in chalky grain grade and chalky grain percentage, although there was only a slight alteration in grain shape (Wang et al. 2021a , b , c ). Compared to WT, the transcript levels of GWD1 in the promoter-edited mutants were approximately 50% of those in WT. Additionally, the peak viscosity (PKV), hot paste viscosity (HPV), cool paste viscosity (CPV), and breakdown viscosity (BDV) of the gwd1 mutant rice flour exhibited a slight decrease, whereas the setback viscosity (SBV) demonstrated an increase compared with WT (Wang et al. 2021a , b , c ). In contrast to the significant reduction in grain yield observed in the null mutant of GWD1 (Hirose et al. 2013 ), the authors noted that the mutation caused by promoter editing of GWD1 had relatively mild effects and did not significantly impact yield-related traits (Wang et al. 2021a , b , c ).This implied that the ECQ of the promoter-edited lines was somewhat decreased (Wang et al. 2021a , b , c ). In contrast to the significant reduction in grain yield observed in the null mutant of GWD1 (Hirose et al. 2013 ), the authors indicated that the mutation caused by promoter editing of GWD1 had relatively mild effects and did not affect yield-related traits (Wang et al. 2021a , b , c ).

SLG7 , an allele of GL7 and GW7 , was associated with grain shape and starch quality (Wang et al. 2015 ). AH2 was a MYB-domain protein with a crucial target site AC II element (ACCAATCC) that played a crucial role in the development of grain and hull (Ren et al. 2019 ). Research has shown that AH2 was essential for determining the development of hull epidermis, palea identity, and grain size (Ren et al. 2019 ). Promoter-edited lines of SLG7 were created by targeting several CREs on the SLG7 promoter (Tan et al. 2023 ). The researchers identified the AC II element at one of the mutation sites, suggesting that mutation at this site may affect the interaction between AH2 and SLG7 and thus affecting SLG7 expression (Tan et al. 2023 ). Some SLG7 promoter-edited mutants exhibited elevated SLG7 expression, reduced chalkiness, longer grains, higher amylose content, and decreased gel consistency, however, the yield per plant did not differ from that of WT (Tan et al. 2023 ). Further analysis revealed that the binding ability of AH2 to the mutant of SLG7 promoter was significantly reduced, and the repressive effect of AH2 protein on the mutant promoter was also significantly reduced (Tan et al. 2023 ). This study highlights the significant potential of targeting CREs (e.g., transcription factor binding sites) in promoter regions to create beneficial mutations and improve rice traits.

Enhancement of Resistance to Bacterial Diseases

Rice is susceptible to a range of pathogenic agents, including fungi, bacteria, viruses, and other microorganisms. The most prevalent diseases such as sheath blight, blast, bacterial leaf blight, and tungro cause severe yield losses every year. In order to overcome these rice diseases, gene editing technology provides breeders with an effective way (Ahmad et al. 2020 ). Plant pathogens were able to identify specific signals from the host plant, which then initiated the infection process (Ontoy and Ham 2024 ). Bacteria employed specialized secretion systems to deliver effector molecules into plant cells, thereby modulating host responses and facilitating invasion (Zhang et al. 2022 ). Rice possesses a multitude of genes that confer resistance to specific pathogens, known as resistance genes ( R genes). Conversely, there are susceptibility genes ( S genes) that, when recognized by pathogens, make rice more vulnerable to disease. Two destructive rice bacterial diseases, bacterial leaf blight (BLB) and bacterial leaf streak (BLS), are caused by two different strains of the rice Xanthomonas oryzae : pv. oryzae ( Xoo ) and pv. oryzicola ( Xoc ) respectively. These pathogens are widely distributed and have significant negative impacts on global rice production. BLB was the most destructive disease, accounting for 40% of rice pest and disease losses (Shaheen et al. 2019 ; Syed-Ab-Rahman et al. 2020 ). Xoo and Xoc are two common pathogenic bacteria that promote rice disease by injecting Transcription-Activator-Like Effectors (TALEs) into rice plant cells. These TALEs recognized Effector-Binding Elements (EBEs) in the promoters of homologous host genes, which resulted in the activation of rice S genes (Bezrutczyk et al. 2018 ; White et al. 2009 ). Consequently, many researchers have endeavored to regulate crucial R genes or S genes to develop rice varieties with the intention of obtaining disease resistance. This ultimately aims to breed rice varieties with enhanced resistance to the disease.

Moreover, it indicated that breeding resistant rice lines was the most effective way to tackle BLB caused by Xoo (Eom et al. 2019 ). The Sugars Will Eventually Be Exported Transporter ( SWEET ) gene family encodes sugar-transporting proteins required for normal plant growth and development. However, certain SWEET genes may be co-opted by pathogenic microorganisms to fulfill their nutrient requirements, making rice susceptible to disease (Gupta 2020 ). In rice, OsSWEET11 , OsSWEET13 and OsSWEET14 have been identified as key susceptibility genes for BLB (Chen et al. 2010 ; Yuan and Wang 2013 ). The CRISPR/Cas9 system was employed to edit six EBEs in the promoters of OsSWEET11 , OsSWEET13 , and OsSWEET14 (Oliva et al. 2019 ). This approach conferred robust broad-spectrum resistance to the edited lines without compromising agronomic traits in rice (Oliva et al. 2019 ). Evaluation of plant height, panicle length, tiller number, and fertility demonstrated that the majority of promoter-edited lines exhibited comparable performance to that of the WT parents (Oliva et al. 2019 ).

In a comparable study, genetically editing the EBEs in the promoters of OsSWEET11a (an allele of OsSWEET11 ), OsSWEET13 , and OsSWEET14 resulted in the production of various mutant lines that exhibited complex resistance or susceptibility to diverse bacterial strains (Liu et al. 2024 ). They also found that the deletion of EBE sites from the OsSWEET11a and OsSWEET14 promoters resulted in a reduction in pollen viability and seed setting rate (Liu et al. 2024 ). Furthermore, EBE editing in the OsSWEET13 promoter showed a positive effect on rice yield (Liu et al. 2024 ). Although EBEs editing may still have a negative impact on rice growth and development, it is reasonable to conclude that promoter editing is likely to have a less severe impact compared to null mutations that directly target the coding sequence.

By utilizing CRISPR/Cas9 to target EBEs in four of the SWEET genes ( OsSWEET11 , OsSWEET13 , OsSWEET14 , and OsSWEET15 ), promoter-edited lines were created to make changes in single or multiple EBEs (Eom et al. 2019 ). AXO1947, an African Xoo strain that causes bacterial blight in rice, exhibited a reduction in virulence in OsSWEET14 single-knockout lines, yet was unable to infect the SWEET13; SWEET14 double mutants (Eom et al. 2019 ). Growth and yield evaluations under greenhouse conditions revealed no discernible differences in performance for the OsSWEET13 and OsSWEET14 knockout mutant lines, when compared to the WT (Eom et al. 2019 ). Similarly, field experiments indicated that these lines exhibited comparable outcomes in terms of both yield and growth to the WT (Eom et al. 2019 ). The researchers concluded that EBE-edited lines in which the normal promoter function of OsSWEET13 and OsSWEET14 were affected would not cause yield loss based on this evidence (Eom et al. 2019 ). Although the experiments yielded positive results, they also indicated that the resistance conferred by the SWEET promoter editing variant that prevents binding of the TALEs would be rendered ineffective by the emergence of new pathogenic Xoo strains (Eom et al. 2019 ). Consequently, the SWEET promoter-edited lines may need to be used in combination with other R genes.

TALE Tal2g was a primary virulence factor of Xoc , which induced the expression of the S gene OsSULTR3;6 in rice, thereby making it susceptible to disease (Cernadas et al. 2014 ). EBEs in the promoter regions of Xoo susceptibility genes ( OsSWEET11 and OsSWEET14 ), and Xoc susceptibility gene ( OsSULTR3;6 ) were edited to obtain two promoter-edited rice lines of GT0105 and ZT0918 (Ni et al. 2021 ). These lines demonstrated significantly improved resistance to both Xoo and Xoc (Ni et al. 2021 ). Moreover, morphometric and other agronomic traits of GT0105 and ZT0918 exhibited no notable divergence from those of WT (Ni et al. 2021 ). This indicates that editing the EBEs in the promoters of S genes did not result in any alteration in the original biological functions of these genes. The result of qRT-PCR confirmed that the expression of S genes mentioned above in the edited rice lines was no longer responsive to pathogens carrying the corresponding TALEs (Ni et al. 2021 ). According to the study, this is the first instance in which engineered rice lines resistant to Xoo and Xoc had been produced through promoter editing of S genes. Furthermore, the editing process had not affected other agronomic traits. In a similar study, mutants edited at a 20-bp nucleotide target site overlapping the PthXo3, AvrXa7 and TalF EBEs were obtained in the OsSWEET14 promoter (Duy et al. 2021 ). It is pertinent to note that the frequency of individuals with CRISPR/Cas9-induced mutations in T0 transgenic rice was 90% in this study (Duy et al. 2021 ). This high frequency of mutations serves to demonstrate the effectiveness of the CRISPR/Cas9 system as a tool for gene editing in rice. What’s more, transgene-free plants with superiority of CRISPR/Cas9 technology can be generated from the majority of T1 segregation populations, without the necessity for laborious crossing or backcrossing procedures.

In addition, researchers have turned their attention to R genes, attempting to obtain enhanced resistance by editing the promoter regions of R genes. Two principal molecular strategies have emerged for improving rice resistance. The first strategy involves reducing the susceptibility of S genes to TALEs. The second strategy focuses on insertion of EBEs induced by the causal bacterium into the dysfunctional promoters of R genes. Typically, the transcription of R genes was strictly controlled, but it can be activated by TALEs following pathogen attachment (Zhang et al. 2015a , b ). This characteristic suggests that inserting EBEs into the promoters of less responsive R gene alleles could facilitate their recognition by TALEs, thus enhancing resistance. The R gene Xa23 conferred the widest range of resistance to BLB in rice (Wang et al. 2014 ). The resistance of Xa23 is a consequence of the interaction between Xa23 and the TALE avrXa23 of Xoo . Upon infection of rice by Xoo , the secreted AvrXa23 can be captured by a 28-bp EBE (EBE AvrXa23 ) in the promoter of Xa23 , resulting in the activation of Xa23 expression and the development of strong BLB resistance in rice. Unfortunately, EBE AvrXa23 was present in only a few rice varieties, whereas xa23s , the alleles of Xa23 that do not contain EBE AvrXa23 , were present in most sensitive rice genotypes (Cui et al. 2017 ). This characterization provides an opportunity for molecular breeders to improve BLB resistance in susceptible rice.

The insertion of EBE AvrXa23 into the promoter region of the xa23 allele in Nipponbare was achieved through CRISPR/Cas9-mediated precise homology directed repair (HdR) (Wei et al. 2021 ). This resulted in the transformation of BLB-susceptible rice variety with Nipponbare background into BLB-resistant line without significant effects on nutritional and yield traits (Wei et al. 2021 ). The investigation is groundbreaking, which represents a significant advancement in utilization of the highly efficacious BLB resistance gene Xa23 and the application of genome editing technology in the genetic improvement of rice. It also serves as a paradigm for the study and utilization of pivotal genomic elements in rice. Later studies have confirmed the feasibility of this strategy. An efficient nonhomologous end-joining-mediated targeted insertion method was used to edit the promoter of xa23 (Kumar et al. 2023 ). In this study, the insertion of EBE AvrXa7/PthXo3 and EBE TalC into the xa23 promoter region resulted in the activation of xa23 expression, enabling it to respond to Xoo strains expressing the homologous TALEs and confer resistance to specific strains (Kumar et al. 2023 ). In another study, a modified prime editing (PE) system was used to knock-in EBE AvrXa23 into the xa23 promoter and conferred robust BLB resistance in susceptible rice (Gupta et al. 2023 ). This is the first report of using PE system to edit a non-functional rice R gene and efficiently incorporate a 30 bp cis-regulatory element. As the PE system continues to be refined and popularized, it is expected to provide a valuable tool for the efficient and precise editing of various genes, particularly in the promoter regions.

Other Improvements in Rice

Cytoplasmic male sterility (CMS) is a significant agricultural trait that is characterized by the absence of functional pollen due to abnormal gene expression in mitochondrion. The presence of nuclear-encoded fertility restorer (Rf) gene was able to restore the function of the pollen in male sterile lines (Kazama et al. 2016 ). Retrograde-Regulated Male Sterility ( RMS ) serves as a restorer gene for cytoplasmic male sterile lines of rice. RNA-interference-mediated silencing of RMS had been shown to restore fertility in CMS lines, however, overexpression of RMS in fertility-restoring lines had been observed to induce pollen abortion (Fujii and Toriyama 2009 ). Wild-type rice that normally expresses RMS remained completely sterile (Fujii and Toriyama 2009 ). The CRISPR-edited line created by targeting the RMS promoter between the coding sequence of PPR2 (a Rf gene) and RMS was successfully developed, leading to partial restoration of fertility (Suketomo et al. 2020 ). The seed setting rate recovered to 15.9% in the T0 generation and 12.5% in the T1 generation, accompanied by a decrease in RMS expression (Suketomo et al. 2020 ).

Zinc (Zn) is a key micronutrient that plays a role in a multitude of biochemical processes in plants, such as serving as a co-factor for enzymes, participating in the synthesis of nucleotides and some plant hormones. Iron (Fe) is another essential micronutrient crucial for plant growth and development. It is the building block of several enzymes and plays a significant role in maintaining the structure of chloroplast. The OsNAS2 , which encoded nicotinamide synthase in rice, had been demonstrated to enhance bioavailable Fe content with elevated expression level (Lee et al. 2017 ). OsNAS2 showed great potential for Fe/Zn biofortification of rice endosperm (Johnson et al. 2011 ), and was regulated through a B-type OsRR-mediated signaling pathway (Gao et al. 2019 ). Disruption of the binding site of OsRR with the ARR1AT motif may diminish the inhibitory impact of the OsRR on OsNAS2 , resulting in elevated intragrain Zn/Fe concentrations (Gao et al. 2019 ). CRISPR/Cas9 system was employed to edit the ARR1AT motif in the OsNAS2 promoter, resulting in increased Zn/Fe concentrations in the promoter-edited lines (Ludwig et al. 2024 ). The 4 bp deletion-edited line among them exhibited elevated grain number per plant, grain weight per plant, panicle length, plant height, and Zn content relative to WT (Ludwig et al. 2024 ). The expression level of OsNAS2 was found to be significantly elevated in certain promoter-edited lines (Ludwig et al. 2024 ). These results showed that editing the ARR1AT motif in the OsNAS2 promoter enhanced Zn/Fe uptake or translocation in rice (Ludwig et al. 2024 ). This study demonstrates the feasibility of regulating the uptake, translocation, and accumulation of micronutrients in rice through promoter editing. Additionally, it provides insights into fine-tuning the micronutrient content of rice to create more nutritious varieties in the future.

Vacuolar H + translocating pyrophosphatase (V-PPase) affects cytoplasmic pH and is an enzyme activity regulator that degrades starch to sucrose. The research showed that editing the GATA element in the promoter of the V-PPase gene significantly downregulated V-PPase and reduced rice chalkiness (Dharwadker et al. 2022 ; Gann et al. 2023 ), while this downregulation also reduced germination and slowed seedling growth. In addition, the lower germination rate and slower seedling growth were found in edited lines, which would be attributed to the limitation of sucrose formation (Dharwadker et al. 2022 ; Gann et al. 2023 ).

Future Perspectives

Promoter editing allows for precise gene regulation by inserting, deleting, or replacing specific bases or fragments within the CREs of genes. This approach enables the development of rice varieties with high yield, superior quality, and enhanced stress tolerance (Fig.  1 ). Although promoter editing can regulate gene expression in a different way from knocking out the coding region and achieve more subtle regulation of gene expression, it still has a certain degree of blindness and lacks perfect theoretical guidance. We identify several key issues with promoter editing and propose potential solutions to address these challenges.

Adapted from the published paper (Gao 2021 ). Fine editing promoter regulatory elements at the single-base level, inserting large functional fragments and promoter recombination or substitution may be the focus of future research. C Promoter editing has the potential to alter gene spatiotemporal expression patterns. One possible approach is to insert tissue- or time-specific promoter elements. Another is to inserting environmentally responsive promoter elements to induce or inhibit gene expression through exogenous treatments such as light, temperature, and chemical reagents

Current applications and future perspectives for rice genetic improvement based on promoter editing. The future exploration of rice promoter CREs and functional motifs has the potential to broaden the scope of promoter editing and improve editing efficiency. Using advanced gene editing systems can also greatly improve the accuracy and efficiency of editing. In addition, it is possible to achieve precise control of gene spatiotemporal expression through promoter editing. This progress could lead to the development of an ideal rice variety with improved traits, such as higher yield, better quality, and greater stress resistance. A Potential targets for promoter editing, their possible effects on changing gene or protein expression levels or patterns, and future applications. In the future, more promoter regulatory elements should be discovered and elucidated, and advanced promoter editing systems should be used to transform them to ultimately achieve efficient rice improvement. B Modification results achievable by various editing strategies.

Deeper understanding of promoter regulatory elements. Generally speaking, promoter editing needs to target promoter regulatory elements with specific functions, which are usually protein binding or recognition sites, but can also include conserved motifs or natural variation sites. The complexity of promoters includes numerous regulatory elements. While some elements have been elucidated through reliable experiments, many others remain with unknown functions. It is crucial to clarify the roles these elements play in regulating downstream genes and how they function. To achieve effective promoter editing, the targeted site must possess potential functionality rather than being an arbitrary sequence. This necessitates a well-designed, targeted approach rather than a blind one. As researchers gain a deeper understanding of the functions and mechanisms of nearly all regulatory elements, conserved motifs, and natural variant sites within gene promoters, promoter editing is expected to more precisely regulate gene expression. For instance, if a SNP in a gene promoter results in different spatial expression patterns across varieties, editing this SNP could potentially modify the gene's expression site. Similarly, if a conserved motif limits gene expression, inserting this motif into another gene could control its expression duration.

Gene editing systems are crucial tools for precise manipulation of genes. Since the inception of the first-generation gene editing system ZFNs, scientists have continually advanced the development of more refined systems, including TALENs, CRISPR/Cas9, CRISPR/Cas12, CRISPR/Cas13, Base Editing, and Prime Editing systems. While ZFNs and TALENs are now largely obsolete, CRISPR/Cas remains widely utilized. Of above systems, CRISPR/Cas9 has become the most widely used genome editing tool. However, each CRISPR/Cas system has its own features. We provided a comprehensive comparison of these CRISPR/Cas systems (Table  2 ). Although the CRISPR/Cas system has made significant strides, it still has limitations (Hussain & Ahmad 2022 ). Many current CRISPR/Cas systems depend on a sequence known as the protospacer adjacent motif (PAM) to bind or cut the target DNA. Targets lacking PAMs cannot be edited using traditional CRISPR/Cas systems, which significantly limits their flexibility. Many promoter regulatory elements or other desired editing sites may not offer adjacent PAMs, posing a considerable obstacle to precise and flexible site-specific editing. To address these limitations, future efforts should focus on developing more efficient and versatile gene editing systems that either require a relaxed PAM sequence or eliminate the need for PAM altogether. Moreover, it is still not easy to accurately insert large fragments at desired sites and target specific bases for editing. To address these issues, the development of gene editing systems must be advanced simultaneously.

The precise regulation of rice growth and development has long been a challenge for breeders. By modifying rice promoters with advanced gene editing techniques, researchers can regulate gene expression within specific ranges, thus influencing rice traits. Future advancements may include the use of diverse gene editing systems to modify rice promoters, creating binding sites for specific transcription factors or DNA-binding proteins. One promising solution is the use of technologies based on CRISPR system to insert functional sites from identified tissue-specific promoters into the promoters of certain genes that are desired to express in specific parts of the body. This approach allows for the expression of genes in specific tissues without the risk of undesirable effects that can arise from using strong promoters that drive expression throughout the entire plant. By integrating key sites of inducible promoters into specific genes, it is also possible to control gene expression at precise times through exogenous induction. Similarly, replacing the promoters of particular genes with tandem tissue-specific and inducible promoters in vitro enables precise regulation of rice growth and development in targeted tissues and at specific developmental stages. This methodology offers a refined approach to managing gene expression, ensuring that desired traits are achieved without compromising other essential characteristics.

Targeting the coding sequence to generate gene mutations is undoubtedly the most direct way to alter phenotypes. However, even with knockouts, residual translation of non-functional proteins can occur, leading to wasted energy and potentially detrimental effects on rice growth and development, which can negatively impact agricultural production. In contrast, modifying promoters avoids the production of redundant proteins, making the resulting mutant lines safer and more efficient by eliminating the waste of energy on ineffective proteins. This approach also allows for precise regulation of gene expression and enables targeted gene knockdown without the lethal effects sometimes associated with complete gene knockouts or the instability of heterozygous knockouts. Given its distinctive advantages—such as straightforward control, high precision, and minimal disruption to the plant’s genetic structure—promoter region editing is likely to become a mainstream breeding method in the future. This approach offers a promising avenue for rice breeding, providing a more efficient means of achieving desired traits while minimizing unintended effects.

Due to genetic linkage and pleiotropy, crop breeding often faces trade-offs between traits. For instance, in rice, enhancing panicle weight can negatively impact tiller number. This mutually restrictive trade-off is a significant bottleneck in conventional breeding techniques. Overcoming these trade-offs is crucial for developing new varieties that are high-yielding, high-quality, and widely resistant. Natural crop evolution involves limited genetic mutations, which constrains breeders' ability to develop high-yielding, disease-resistant, and high-quality crop seeds. Even when beneficial mutations are identified, traditional breeding methods are time-consuming and require continuous refinement each year. Promoters, which control the timing and level of gene expression, play a key role in the pleiotropy of genes, as gene expression is tightly regulated by these elements. By modifying specific elements of a promoter, it is possible to optimize the expression pattern of a target gene and its associated trade-offs, thereby achieving breeding goals that improve yield, quality, and resistance simultaneously. Promoter gene editing technology allows for fine-tuning of gene expression without deleting genes or diminishing their protein-coding ability. This approach offers greater flexibility in improving quantitative traits of crops. By using gene editing to create and select the most ideal variants, breeders can control gene activity more precisely. This not only accelerates crop improvement but also enhances predictability, enabling breeders to "customize" crops according to planting conditions and select the best variants to maximize yields.

Promoter editing, which falls under the broader category of gene editing, raises important issues related to regulation, safety, and risk. With the advent of gene editing technologies like CRISPR, the term “Genetically Modified Crops (GMC)” has become well-known. However, concerns about GMC persist, primarily due to perceived risks to human health, animals, and the environment. These concerns include potential harm from gene-edited components, the possibility of edited genes producing toxic proteins, the risk of resistance genes leading to resistant strains, and the potential creation of superweeds through genetic drift (Ahmad et al. 2021a , b ). To address these concerns, GMC are subjected to rigorous risk, toxicity, and biosafety assessments. Many countries, especially those with advanced regulatory systems, have implemented laws and regulations to control GMC and ensure that consumers have the right to know if the food they purchase is genetically modified. For instance, China, the United States, and several European Union countries employ an end-product-based regulatory framework, labeling GM-free products accordingly and monitoring GMC before commercialization. Promoter editing, which uses techniques like CRISPR to alter genetic material, also falls within the scope of GMC regulations. Given the relatively recent introduction of promoter-edited crops and their limited practical application, there may be gaps in the current regulations, safety, and risk assessments for these crops. Therefore, in-depth research is needed to develop efficient, GM-free editing strategies (He et al. 2022 ). Policymakers should engage in discussions with researchers to formulate appropriate policies, ensuring that promoter-edited crops can be developed safely and effectively for the benefit of humanity in the future.

Taken together, promoter editing allows precise gene regulation by altering specific bases or fragments, enabling the development of better rice varieties. Unlike coding region knockouts, it offers nuanced gene expression control but requires a deeper understanding of promoter elements. Advanced gene editing systems like CRISPR/Cas have improved precision but still face limitations. Promoter editing avoids redundant protein production, and helps balance genetic trade-offs in breeding. However, promoter-edited crops raise regulatory and safety concerns, necessitating rigorous assessments and effective policies to ensure safe and beneficial applications. With continued research and technological advancements, promoter editing has the potential to revolutionize rice and other crop improvement and sustainable agriculture.

The regulation of gene expression at the molecular level is a crucial aspect of modern molecular breeding strategies, with promoters playing a pivotal role in this regulation. Promoter editing offers several key advantages. First, precise gene regulation: promoter editing allows for the fine-tuning of gene expression, which can enhance important agronomic traits in rice. Second, broad range of regulation: promoter editing can regulate gene expression in a wide range, thus generating a series of continuous QTVs, which is conducive to enhancing the ecological adaptability of rice or meets the requirements of more situations. Third, specificity in response: editing specific promoter regulatory elements can alter the sensitivity of the promoter to specific inducers or repressors, thereby achieving specific effects. Nevertheless, rice promoter editing continues to face significant challenges. Identifying appropriate editing sites for more precise modifications remains a critical issue. Additionally, there is a need for more efficient and accurate gene editing systems. However, the potential for rice promoter editing in genetic improvement is substantial. With an enhanced understanding of rice promoter regulatory elements and advancements in gene editing technology, promoter editing is expected to become a more prominent tool in rice genetic improvement. This progress will likely lead to the development of numerous superior rice varieties through promoter editing.

Data Availability

No datasets were generated or analysed during the current study.

Abbreviations

Apparent amylose content

Amylose content

Bacterial leaf blight

Bacterial leaf streak

CRISPR/Cas12a promoter editing

Cis-regulatory elements

Effector-binding elements

Eating and cooking quality

Genetically modified crops

High-efficiency multiplex promoter-targeting

Key-regions

Protospacer adjacent motif

• Promoter editing

Quantitative trait locus

Single nucleotide polymorphism

Transcription-activator-like effectors

Transcription start site

5′ Untranslated region

Cytoplasmic male sterility

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This work was supported by the following research projects: the National Natural Science Foundation of China under Grant [32260498]; the Guizhou Provincial Excellent Young Talents Project of Science and Technology under Grant [Qiankehepingtairencai-YQK (2023) 002]; the Guizhou Provincial Science and Technology Projects under Grants [Qiankehechengguo (2024) General 116] and [Qiankehejichu-ZK (2022) Key 008]; the Key Laboratory of Molecular Breeding for Grain and Oil Crops in Guizhou Province under Grant [Qiankehezhongyindi (2023) 008]; the Key Laboratory of Functional Agriculture of Guizhou Provincial Department of Education under Grant [Qianjiaoji (2023) 007]; the Qiandongnan Science and Technology Support Project under Grant [Qiandongnan Kehe Support (2023) 06]; Researchers supporting project number [RSP2024R385], King Saud University, Riyadh, Saudi Arabia.

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Bowen Wu, Hangfei Luo and Zhongbo Chen have contributed equally to this work.

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Institute of Rice Industry Technology Research, Key Laboratory of Functional Agriculture of Guizhou Provincial, Department of Education, Key Laboratory of Molecular Breeding for Grain and Oil Crops in Guizhou Province, College of Agricultural Sciences, Guizhou University, Guiyang, 550025, China

Bowen Wu, Hangfei Luo, Zhongbo Chen, Bakht Amin, Manyu Yang, Zhenghan Li, Shuai Wu & Zhongming Fang

Key Laboratory of Plant Resource Conservation and Germplasm Innovation in Mountainous Region (Ministry of Education), Guizhou University, Guiyang, 550025, China

Zhongming Fang

Department of Botany and Microbiology, College of Science, King Saud University, 11451, Riyadh, Saudi Arabia

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Wu, B., Luo, H., Chen, Z. et al. Rice Promoter Editing: An Efficient Genetic Improvement Strategy. Rice 17 , 55 (2024). https://doi.org/10.1186/s12284-024-00735-7

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• Risk of fatality, including statistics
• How measles is spread
• Immunization procedures in different regions
• Different regions, focusing on the arguments from those against immunization
• Immunization figures in affected regions
• High number of cases in non-immunizing regions
• Illnesses that can result from measles virus
• Fatal cases of other illnesses after patient contracted measles
• Summary of arguments of different groups
• Summary of figures and relationship with recent immunization debate
• Which side of the argument appears to be correct?

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Follow these steps to start your research paper outline:

• Decide on the subject of the paper
• Write down all the ideas you want to include or discuss
• Organize related ideas into sub-groups
• Arrange your ideas into a hierarchy: What should the reader learn first? What is most important? Which idea will help end your paper most effectively?
• Create headings and subheadings that are effective
• Format the outline in either alphanumeric, full-sentence or decimal format

There are three different kinds of research paper outline: alphanumeric, full-sentence and decimal outlines. The differences relate to formatting and style of writing.

• Alphanumeric
• Full-sentence

An alphanumeric outline is most commonly used. It uses Roman numerals, capitalized letters, arabic numerals, lowercase letters to organize the flow of information. Text is written with short notes rather than full sentences.

• Sub-point of sub-point 1

Essentially the same as the alphanumeric outline, but with the text written in full sentences rather than short points.

• Additional sub-point to conclude discussion of point of evidence introduced in point A

A decimal outline is similar in format to the alphanumeric outline, but with a different numbering system: 1, 1.1, 1.2, etc. Text is written as short notes rather than full sentences.

• 1.1.1 Sub-point of first point
• 1.1.2 Sub-point of first point
• 1.2 Second point

To write an effective research paper outline, it is important to pay attention to language. This is especially important if it is one you will show to your teacher or be assessed on.

There are four main considerations: parallelism, coordination, subordination and division.

Parallelism: Be consistent with grammatical form

Parallel structure or parallelism is the repetition of a particular grammatical form within a sentence, or in this case, between points and sub-points. This simply means that if the first point is a verb , the sub-point should also be a verb.

Example of parallelism:

• Include different regions, focusing on the different arguments from those against immunization

Coordination: Be aware of each point’s weight

Your chosen subheadings should hold the same significance as each other, as should all first sub-points, secondary sub-points, and so on.

Example of coordination:

• Include immunization figures in affected regions
• Illnesses that can result from the measles virus

Subordination: Work from general to specific

Subordination refers to the separation of general points from specific. Your main headings should be quite general, and each level of sub-point should become more specific.

Example of subordination:

Division: break information into sub-points.

Your headings should be divided into two or more subsections. There is no limit to how many subsections you can include under each heading, but keep in mind that the information will be structured into a paragraph during the writing stage, so you should not go overboard with the number of sub-points.

Ready to start writing or looking for guidance on a different step in the process? Read our step-by-step guide on how to write a research paper .

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Gahan, C. (2023, August 15). How to Create a Structured Research Paper Outline | Example. Scribbr. Retrieved September 3, 2024, from https://www.scribbr.com/research-paper/outline/

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