Reassessment of the Four Yield-related Genes Gn1a, DEP1, GS3, and IPA1 in Rice Using a CRISPR/Cas9 System
Identifieur interne : 000048 ( Pmc/Checkpoint ); précédent : 000047; suivant : 000049Reassessment of the Four Yield-related Genes Gn1a, DEP1, GS3, and IPA1 in Rice Using a CRISPR/Cas9 System
Auteurs : Meiru Li [République populaire de Chine] ; Xiaoxia Li [République populaire de Chine] ; Zejiao Zhou [République populaire de Chine] ; Pingzhi Wu [République populaire de Chine] ; Maichun Fang [République populaire de Chine] ; Xiaoping Pan [République populaire de Chine] ; Qiupeng Lin [République populaire de Chine] ; Wanbin Luo [République populaire de Chine] ; Guojiang Wu [République populaire de Chine] ; Hongqing Li [République populaire de Chine]Source :
- Frontiers in Plant Science [ 1664-462X ] ; 2016.
Abstract
Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated (Cas) systems have been successfully used as efficient tools for genome editing in a variety of species. We used the CRISPR/Cas9 system to mutate the
Url:
DOI: 10.3389/fpls.2016.00377
PubMed: 27066031
PubMed Central: 4811884
Affiliations:
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, <italic>GS3</italic>
, and <italic>IPA1</italic>
in Rice Using a CRISPR/Cas9 System</title>
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<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Reassessment of the Four Yield-related Genes <italic>Gn1a</italic>
, <italic>DEP1</italic>
, <italic>GS3</italic>
, and <italic>IPA1</italic>
in Rice Using a CRISPR/Cas9 System</title>
<author><name sortKey="Li, Meiru" sort="Li, Meiru" uniqKey="Li M" first="Meiru" last="Li">Meiru Li</name>
<affiliation wicri:level="1"><nlm:aff id="aff1"><institution>Key Laboratory of Plant Resources Conservation and Sustainable Utilization, South China Botanical Garden, Chinese Academy of Sciences</institution>
<country>Guangzhou, China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea></wicri:regionArea>
</affiliation>
<affiliation wicri:level="1"><nlm:aff id="aff2"><institution>Guangdong Provincial Key Laboratory of Applied Botany, South China Botanical Garden, Chinese Academy of Sciences</institution>
<country>Guangzhou, China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea></wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Li, Xiaoxia" sort="Li, Xiaoxia" uniqKey="Li X" first="Xiaoxia" last="Li">Xiaoxia Li</name>
<affiliation wicri:level="1"><nlm:aff id="aff3"><institution>Guangdong Provincial Key Lab of Biotechnology for Plant Development, South China Normal University</institution>
<country>Guangzhou, China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea></wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Zhou, Zejiao" sort="Zhou, Zejiao" uniqKey="Zhou Z" first="Zejiao" last="Zhou">Zejiao Zhou</name>
<affiliation wicri:level="1"><nlm:aff id="aff3"><institution>Guangdong Provincial Key Lab of Biotechnology for Plant Development, South China Normal University</institution>
<country>Guangzhou, China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea></wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Wu, Pingzhi" sort="Wu, Pingzhi" uniqKey="Wu P" first="Pingzhi" last="Wu">Pingzhi Wu</name>
<affiliation wicri:level="1"><nlm:aff id="aff1"><institution>Key Laboratory of Plant Resources Conservation and Sustainable Utilization, South China Botanical Garden, Chinese Academy of Sciences</institution>
<country>Guangzhou, China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea></wicri:regionArea>
</affiliation>
<affiliation wicri:level="1"><nlm:aff id="aff2"><institution>Guangdong Provincial Key Laboratory of Applied Botany, South China Botanical Garden, Chinese Academy of Sciences</institution>
<country>Guangzhou, China</country>
</nlm:aff>
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<author><name sortKey="Fang, Maichun" sort="Fang, Maichun" uniqKey="Fang M" first="Maichun" last="Fang">Maichun Fang</name>
<affiliation wicri:level="1"><nlm:aff id="aff1"><institution>Key Laboratory of Plant Resources Conservation and Sustainable Utilization, South China Botanical Garden, Chinese Academy of Sciences</institution>
<country>Guangzhou, China</country>
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<country>Guangzhou, China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea></wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Pan, Xiaoping" sort="Pan, Xiaoping" uniqKey="Pan X" first="Xiaoping" last="Pan">Xiaoping Pan</name>
<affiliation wicri:level="1"><nlm:aff id="aff1"><institution>Key Laboratory of Plant Resources Conservation and Sustainable Utilization, South China Botanical Garden, Chinese Academy of Sciences</institution>
<country>Guangzhou, China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea></wicri:regionArea>
</affiliation>
<affiliation wicri:level="1"><nlm:aff id="aff2"><institution>Guangdong Provincial Key Laboratory of Applied Botany, South China Botanical Garden, Chinese Academy of Sciences</institution>
<country>Guangzhou, China</country>
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<author><name sortKey="Lin, Qiupeng" sort="Lin, Qiupeng" uniqKey="Lin Q" first="Qiupeng" last="Lin">Qiupeng Lin</name>
<affiliation wicri:level="1"><nlm:aff id="aff3"><institution>Guangdong Provincial Key Lab of Biotechnology for Plant Development, South China Normal University</institution>
<country>Guangzhou, China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea></wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Luo, Wanbin" sort="Luo, Wanbin" uniqKey="Luo W" first="Wanbin" last="Luo">Wanbin Luo</name>
<affiliation wicri:level="1"><nlm:aff id="aff3"><institution>Guangdong Provincial Key Lab of Biotechnology for Plant Development, South China Normal University</institution>
<country>Guangzhou, China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea></wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Wu, Guojiang" sort="Wu, Guojiang" uniqKey="Wu G" first="Guojiang" last="Wu">Guojiang Wu</name>
<affiliation wicri:level="1"><nlm:aff id="aff1"><institution>Key Laboratory of Plant Resources Conservation and Sustainable Utilization, South China Botanical Garden, Chinese Academy of Sciences</institution>
<country>Guangzhou, China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea></wicri:regionArea>
</affiliation>
<affiliation wicri:level="1"><nlm:aff id="aff2"><institution>Guangdong Provincial Key Laboratory of Applied Botany, South China Botanical Garden, Chinese Academy of Sciences</institution>
<country>Guangzhou, China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea></wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Li, Hongqing" sort="Li, Hongqing" uniqKey="Li H" first="Hongqing" last="Li">Hongqing Li</name>
<affiliation wicri:level="1"><nlm:aff id="aff3"><institution>Guangdong Provincial Key Lab of Biotechnology for Plant Development, South China Normal University</institution>
<country>Guangzhou, China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea></wicri:regionArea>
</affiliation>
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<series><title level="j">Frontiers in Plant Science</title>
<idno type="eISSN">1664-462X</idno>
<imprint><date when="2016">2016</date>
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<front><div type="abstract" xml:lang="en"><p>Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated (Cas) systems have been successfully used as efficient tools for genome editing in a variety of species. We used the CRISPR/Cas9 system to mutate the <italic>Gn1a</italic>
(Os01g0197700), <italic>DEP1</italic>
(Os09g0441900), <italic>GS3</italic>
(Os03g0407400), and <italic>IPA1</italic>
(Os08g0509600) genes of rice cultivar Zhonghua 11, genes which have been reported to function as regulators of grain number, panicle architecture, grain size and plant architecture, respectively. Analysis of the phenotypes and frequencies of edited genes in the first generation of transformed plants (T0) showed that the CRISPR/Cas9 system was highly efficient in inducing targeted gene editing, with the desired genes being edited in 42.5% (<italic>Gn1a</italic>
), 67.5% (<italic>DEP1</italic>
), 57.5% (<italic>GS3</italic>
), and 27.5% (<italic>IPA1</italic>
) of the transformed plants. The T2 generation of the <italic>gn1a</italic>
, <italic>dep1</italic>
, and <italic>gs3</italic>
mutants featured enhanced grain number, dense erect panicles, and larger grain size, respectively. Furthermore, semi-dwarf, and grain with long awn, phenotypes were observed in <italic>dep1</italic>
and <italic>gs3</italic>
mutants, respectively. The <italic>ipa1</italic>
mutants showed two contrasting phenotypes, having either fewer tillers or more tillers, depending on the changes induced in the OsmiR156 target region. In addition, we found that mutants with deletions occurred more frequently than previous reports had indicated and that off-targeting had taken place in highly similar target sequences. These results proved that multiple regulators of important traits can be modified in a single cultivar by CRISPR/Cas9, and thus facilitate the dissection of complex gene regulatory networks in the same genomic background and the stacking of important traits in cultivated varieties.</p>
</div>
</front>
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</TEI>
<pmc article-type="research-article"><pmc-dir>properties open_access</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-ta">Front Plant Sci</journal-id>
<journal-id journal-id-type="iso-abbrev">Front Plant Sci</journal-id>
<journal-id journal-id-type="publisher-id">Front. Plant Sci.</journal-id>
<journal-title-group><journal-title>Frontiers in Plant Science</journal-title>
</journal-title-group>
<issn pub-type="epub">1664-462X</issn>
<publisher><publisher-name>Frontiers Media S.A.</publisher-name>
</publisher>
</journal-meta>
<article-meta><article-id pub-id-type="pmid">27066031</article-id>
<article-id pub-id-type="pmc">4811884</article-id>
<article-id pub-id-type="doi">10.3389/fpls.2016.00377</article-id>
<article-categories><subj-group subj-group-type="heading"><subject>Plant Science</subject>
<subj-group><subject>Original Research</subject>
</subj-group>
</subj-group>
</article-categories>
<title-group><article-title>Reassessment of the Four Yield-related Genes <italic>Gn1a</italic>
, <italic>DEP1</italic>
, <italic>GS3</italic>
, and <italic>IPA1</italic>
in Rice Using a CRISPR/Cas9 System</article-title>
</title-group>
<contrib-group><contrib contrib-type="author"><name><surname>Li</surname>
<given-names>Meiru</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>1</sup>
</xref>
<xref ref-type="aff" rid="aff2"><sup>2</sup>
</xref>
<xref ref-type="author-notes" rid="fn002"><sup>†</sup>
</xref>
<uri xlink:type="simple" xlink:href="http://loop.frontiersin.org/people/319344/overview"></uri>
</contrib>
<contrib contrib-type="author"><name><surname>Li</surname>
<given-names>Xiaoxia</given-names>
</name>
<xref ref-type="aff" rid="aff3"><sup>3</sup>
</xref>
<xref ref-type="author-notes" rid="fn002"><sup>†</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Zhou</surname>
<given-names>Zejiao</given-names>
</name>
<xref ref-type="aff" rid="aff3"><sup>3</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Wu</surname>
<given-names>Pingzhi</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>1</sup>
</xref>
<xref ref-type="aff" rid="aff2"><sup>2</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Fang</surname>
<given-names>Maichun</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>1</sup>
</xref>
<xref ref-type="aff" rid="aff2"><sup>2</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Pan</surname>
<given-names>Xiaoping</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>1</sup>
</xref>
<xref ref-type="aff" rid="aff2"><sup>2</sup>
</xref>
<uri xlink:type="simple" xlink:href="http://loop.frontiersin.org/people/119175/overview"></uri>
</contrib>
<contrib contrib-type="author"><name><surname>Lin</surname>
<given-names>Qiupeng</given-names>
</name>
<xref ref-type="aff" rid="aff3"><sup>3</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Luo</surname>
<given-names>Wanbin</given-names>
</name>
<xref ref-type="aff" rid="aff3"><sup>3</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Wu</surname>
<given-names>Guojiang</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>1</sup>
</xref>
<xref ref-type="aff" rid="aff2"><sup>2</sup>
</xref>
<xref ref-type="author-notes" rid="fn001"><sup>*</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Li</surname>
<given-names>Hongqing</given-names>
</name>
<xref ref-type="aff" rid="aff3"><sup>3</sup>
</xref>
<xref ref-type="author-notes" rid="fn001"><sup>*</sup>
</xref>
<uri xlink:type="simple" xlink:href="http://loop.frontiersin.org/people/310923/overview"></uri>
</contrib>
</contrib-group>
<aff id="aff1"><sup>1</sup>
<institution>Key Laboratory of Plant Resources Conservation and Sustainable Utilization, South China Botanical Garden, Chinese Academy of Sciences</institution>
<country>Guangzhou, China</country>
</aff>
<aff id="aff2"><sup>2</sup>
<institution>Guangdong Provincial Key Laboratory of Applied Botany, South China Botanical Garden, Chinese Academy of Sciences</institution>
<country>Guangzhou, China</country>
</aff>
<aff id="aff3"><sup>3</sup>
<institution>Guangdong Provincial Key Lab of Biotechnology for Plant Development, South China Normal University</institution>
<country>Guangzhou, China</country>
</aff>
<author-notes><fn fn-type="edited-by"><p>Edited by: <italic>Naveen C. Bisht, National Institute of Plant Genome Research, India</italic>
</p>
</fn>
<fn fn-type="edited-by"><p>Reviewed by: <italic>Yuriko Osakabe, The University of Tokushima, Japan; Robert Henry, The University of Queensland, Australia; Rupesh Kailasrao Deshmukh, Laval University, Canada</italic>
</p>
</fn>
<corresp id="fn001">*Correspondence: <italic>Guojiang Wu, <email xlink:type="simple">wugj@scbg.ac.cn</email>
; Hongqing Li, <email xlink:type="simple">hqli@scnu.edu.cn</email>
</italic>
</corresp>
<fn fn-type="other" id="fn002"><p><sup>†</sup>
<italic>These authors have contributed equally to this work.</italic>
</p>
</fn>
<fn fn-type="other" id="fn003"><p>This article was submitted to Plant Genetics and Genomics, a section of the journal Frontiers in Plant Science</p>
</fn>
</author-notes>
<pub-date pub-type="epub"><day>30</day>
<month>3</month>
<year>2016</year>
</pub-date>
<pub-date pub-type="collection"><year>2016</year>
</pub-date>
<volume>7</volume>
<elocation-id>377</elocation-id>
<history><date date-type="received"><day>20</day>
<month>1</month>
<year>2016</year>
</date>
<date date-type="accepted"><day>11</day>
<month>3</month>
<year>2016</year>
</date>
</history>
<permissions><copyright-statement>Copyright © 2016 Li, Li, Zhou, Wu, Fang, Pan, Lin, Luo, Wu and Li.</copyright-statement>
<copyright-year>2016</copyright-year>
<copyright-holder>Li, Li, Zhou, Wu, Fang, Pan, Lin, Luo, Wu and Li</copyright-holder>
<license xlink:href="http://creativecommons.org/licenses/by/4.0/"><license-p>This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.</license-p>
</license>
</permissions>
<abstract><p>Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated (Cas) systems have been successfully used as efficient tools for genome editing in a variety of species. We used the CRISPR/Cas9 system to mutate the <italic>Gn1a</italic>
(Os01g0197700), <italic>DEP1</italic>
(Os09g0441900), <italic>GS3</italic>
(Os03g0407400), and <italic>IPA1</italic>
(Os08g0509600) genes of rice cultivar Zhonghua 11, genes which have been reported to function as regulators of grain number, panicle architecture, grain size and plant architecture, respectively. Analysis of the phenotypes and frequencies of edited genes in the first generation of transformed plants (T0) showed that the CRISPR/Cas9 system was highly efficient in inducing targeted gene editing, with the desired genes being edited in 42.5% (<italic>Gn1a</italic>
), 67.5% (<italic>DEP1</italic>
), 57.5% (<italic>GS3</italic>
), and 27.5% (<italic>IPA1</italic>
) of the transformed plants. The T2 generation of the <italic>gn1a</italic>
, <italic>dep1</italic>
, and <italic>gs3</italic>
mutants featured enhanced grain number, dense erect panicles, and larger grain size, respectively. Furthermore, semi-dwarf, and grain with long awn, phenotypes were observed in <italic>dep1</italic>
and <italic>gs3</italic>
mutants, respectively. The <italic>ipa1</italic>
mutants showed two contrasting phenotypes, having either fewer tillers or more tillers, depending on the changes induced in the OsmiR156 target region. In addition, we found that mutants with deletions occurred more frequently than previous reports had indicated and that off-targeting had taken place in highly similar target sequences. These results proved that multiple regulators of important traits can be modified in a single cultivar by CRISPR/Cas9, and thus facilitate the dissection of complex gene regulatory networks in the same genomic background and the stacking of important traits in cultivated varieties.</p>
</abstract>
<kwd-group><kwd>CRISPR/Cas9 system</kwd>
<kwd>gene editing</kwd>
<kwd><italic>Oryza sativa</italic>
L.</kwd>
<kwd>yield-related genes</kwd>
<kwd>yield-related traits</kwd>
</kwd-group>
<counts><fig-count count="5"></fig-count>
<table-count count="3"></table-count>
<equation-count count="0"></equation-count>
<ref-count count="59"></ref-count>
<page-count count="13"></page-count>
<word-count count="0"></word-count>
</counts>
</article-meta>
</front>
</pmc>
<affiliations><list><country><li>République populaire de Chine</li>
</country>
</list>
<tree><country name="République populaire de Chine"><noRegion><name sortKey="Li, Meiru" sort="Li, Meiru" uniqKey="Li M" first="Meiru" last="Li">Meiru Li</name>
</noRegion>
<name sortKey="Fang, Maichun" sort="Fang, Maichun" uniqKey="Fang M" first="Maichun" last="Fang">Maichun Fang</name>
<name sortKey="Fang, Maichun" sort="Fang, Maichun" uniqKey="Fang M" first="Maichun" last="Fang">Maichun Fang</name>
<name sortKey="Li, Hongqing" sort="Li, Hongqing" uniqKey="Li H" first="Hongqing" last="Li">Hongqing Li</name>
<name sortKey="Li, Meiru" sort="Li, Meiru" uniqKey="Li M" first="Meiru" last="Li">Meiru Li</name>
<name sortKey="Li, Xiaoxia" sort="Li, Xiaoxia" uniqKey="Li X" first="Xiaoxia" last="Li">Xiaoxia Li</name>
<name sortKey="Lin, Qiupeng" sort="Lin, Qiupeng" uniqKey="Lin Q" first="Qiupeng" last="Lin">Qiupeng Lin</name>
<name sortKey="Luo, Wanbin" sort="Luo, Wanbin" uniqKey="Luo W" first="Wanbin" last="Luo">Wanbin Luo</name>
<name sortKey="Pan, Xiaoping" sort="Pan, Xiaoping" uniqKey="Pan X" first="Xiaoping" last="Pan">Xiaoping Pan</name>
<name sortKey="Pan, Xiaoping" sort="Pan, Xiaoping" uniqKey="Pan X" first="Xiaoping" last="Pan">Xiaoping Pan</name>
<name sortKey="Wu, Guojiang" sort="Wu, Guojiang" uniqKey="Wu G" first="Guojiang" last="Wu">Guojiang Wu</name>
<name sortKey="Wu, Guojiang" sort="Wu, Guojiang" uniqKey="Wu G" first="Guojiang" last="Wu">Guojiang Wu</name>
<name sortKey="Wu, Pingzhi" sort="Wu, Pingzhi" uniqKey="Wu P" first="Pingzhi" last="Wu">Pingzhi Wu</name>
<name sortKey="Wu, Pingzhi" sort="Wu, Pingzhi" uniqKey="Wu P" first="Pingzhi" last="Wu">Pingzhi Wu</name>
<name sortKey="Zhou, Zejiao" sort="Zhou, Zejiao" uniqKey="Zhou Z" first="Zejiao" last="Zhou">Zejiao Zhou</name>
</country>
</tree>
</affiliations>
</record>
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