{"@context":{"@vocab":"https://cir.nii.ac.jp/schema/1.0/","rdfs":"http://www.w3.org/2000/01/rdf-schema#","dc":"http://purl.org/dc/elements/1.1/","dcterms":"http://purl.org/dc/terms/","foaf":"http://xmlns.com/foaf/0.1/","prism":"http://prismstandard.org/namespaces/basic/2.0/","cinii":"http://ci.nii.ac.jp/ns/1.0/","datacite":"https://schema.datacite.org/meta/kernel-4/","ndl":"http://ndl.go.jp/dcndl/terms/","jpcoar":"https://github.com/JPCOAR/schema/blob/master/2.0/"},"@id":"https://cir.nii.ac.jp/crid/1360580240159314560.json","@type":"Article","productIdentifier":[{"identifier":{"@type":"DOI","@value":"10.1111/pbi.13239"}},{"identifier":{"@type":"URI","@value":"https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1111%2Fpbi.13239"}},{"identifier":{"@type":"URI","@value":"https://onlinelibrary.wiley.com/doi/pdf/10.1111/pbi.13239"}},{"identifier":{"@type":"URI","@value":"https://onlinelibrary.wiley.com/doi/full-xml/10.1111/pbi.13239"}}],"dc:title":[{"@value":"Generation of a multiplex mutagenesis population via pooled <scp>CRISPR</scp>‐Cas9 in soya bean"}],"description":[{"type":"abstract","notation":[{"@value":"<jats:title>Summary</jats:title><jats:p>The output of genetic mutant screenings in soya bean [<jats:italic>Glycine max</jats:italic> (L.) Merr.] has been limited by its paleopolypoid genome. <jats:styled-content style=\"fixed-case\">CRISPR</jats:styled-content>‐Cas9 can generate multiplex mutants in crops with complex genomes. Nevertheless, the transformation efficiency of soya bean remains low and, hence, remains the major obstacle in the application of <jats:styled-content style=\"fixed-case\">CRISPR</jats:styled-content>‐Cas9 as a mutant screening tool. Here, we report a pooled <jats:styled-content style=\"fixed-case\">CRISPR</jats:styled-content>‐Cas9 platform to generate soya bean multiplex mutagenesis populations. We optimized the key steps in the screening protocol, including vector construction, sg<jats:styled-content style=\"fixed-case\">RNA</jats:styled-content> assessment, pooled transformation, sg<jats:styled-content style=\"fixed-case\">RNA</jats:styled-content> identification and gene editing verification. We constructed 70 <jats:styled-content style=\"fixed-case\">CRISPR</jats:styled-content>‐Cas9 vectors to target 102 candidate genes and their paralogs which were subjected to pooled transformation in 16 batches. A population consisting of 407 T0 lines was obtained containing all sg<jats:styled-content style=\"fixed-case\">RNA</jats:styled-content>s at an average mutagenesis frequency of 59.2%, including 35.6% lines carrying multiplex mutations. The mutation frequency in the T1 progeny could be increased further despite obtaining a transgenic chimera. In this population, we characterized <jats:italic>gmric1/gmric2</jats:italic> double mutants with increased nodule numbers and <jats:italic>gmrdn1‐1/1‐2/1‐3</jats:italic> triple mutant lines with decreased nodulation. Our study provides an advanced strategy for the generation of a targeted multiplex mutant population to overcome the gene redundancy problem in soya bean as well as in other major crops.</jats:p>"}]}],"creator":[{"@id":"https://cir.nii.ac.jp/crid/1380580240159314569","@type":"Researcher","foaf:name":[{"@value":"Mengyan Bai"}],"jpcoar:affiliationName":[{"@value":"College of Resources and Environment Fujian Provincial Key Laboratory of Haixia Applied Plant Systems Biology Fujian Agriculture and Forestry University Fuzhou China"}]},{"@id":"https://cir.nii.ac.jp/crid/1380580240159314567","@type":"Researcher","foaf:name":[{"@value":"Juehui Yuan"}],"jpcoar:affiliationName":[{"@value":"College of Resources and Environment Fujian Provincial Key Laboratory of Haixia Applied Plant Systems Biology Fujian Agriculture and Forestry University Fuzhou China"}]},{"@id":"https://cir.nii.ac.jp/crid/1380580240159314561","@type":"Researcher","foaf:name":[{"@value":"Suning Li"}],"jpcoar:affiliationName":[{"@value":"FAFU‐UCR Joint Center for Horticultural Biology and Metabolomics Haixia Institute of Science and Technology Fujian Agriculture and Forestry University Fuzhou China"},{"@value":"College of Life Sciences Fujian Agriculture and Forestry University Fuzhou China"}]},{"@id":"https://cir.nii.ac.jp/crid/1380580240159314432","@type":"Researcher","foaf:name":[{"@value":"Huaqin Kuang"}],"jpcoar:affiliationName":[{"@value":"FAFU‐UCR Joint Center for Horticultural Biology and Metabolomics Haixia Institute of Science and Technology Fujian Agriculture and Forestry University Fuzhou China"}]},{"@id":"https://cir.nii.ac.jp/crid/1380580240159314560","@type":"Researcher","foaf:name":[{"@value":"Pingping Gong"}],"jpcoar:affiliationName":[{"@value":"FAFU‐UCR Joint Center for Horticultural Biology and Metabolomics Haixia Institute of Science and Technology Fujian Agriculture and Forestry University Fuzhou China"}]},{"@id":"https://cir.nii.ac.jp/crid/1380580240159314564","@type":"Researcher","foaf:name":[{"@value":"Zhihui Zhang"}],"jpcoar:affiliationName":[{"@value":"College of Resources and Environment Fujian Provincial Key Laboratory of Haixia Applied Plant Systems Biology Fujian Agriculture and Forestry University Fuzhou China"},{"@value":"FAFU‐UCR Joint Center for Horticultural Biology and Metabolomics Haixia Institute of Science and Technology Fujian Agriculture and Forestry University Fuzhou China"}]},{"@id":"https://cir.nii.ac.jp/crid/1380580240159314566","@type":"Researcher","foaf:name":[{"@value":"Jiafeng Sun"}],"jpcoar:affiliationName":[{"@value":"College of Resources and Environment Fujian Provincial Key Laboratory of Haixia Applied Plant Systems Biology Fujian Agriculture and Forestry University Fuzhou China"},{"@value":"FAFU‐UCR Joint Center for Horticultural Biology and Metabolomics Haixia Institute of Science and Technology Fujian Agriculture and Forestry University Fuzhou China"}]},{"@id":"https://cir.nii.ac.jp/crid/1380580240159314565","@type":"Researcher","foaf:name":[{"@value":"Bo Liu"}],"jpcoar:affiliationName":[{"@value":"College of Resources and Environment Fujian Provincial Key Laboratory of Haixia Applied Plant Systems Biology Fujian Agriculture and Forestry University Fuzhou China"},{"@value":"FAFU‐UCR Joint Center for Horticultural Biology and Metabolomics Haixia Institute of Science and Technology Fujian Agriculture and Forestry University Fuzhou China"}]},{"@id":"https://cir.nii.ac.jp/crid/1380580240159314562","@type":"Researcher","foaf:name":[{"@value":"Maoxiang Yang"}],"jpcoar:affiliationName":[{"@value":"College of Resources and Environment Fujian Provincial Key Laboratory of Haixia Applied Plant Systems Biology Fujian Agriculture and Forestry University Fuzhou China"},{"@value":"FAFU‐UCR Joint Center for Horticultural Biology and Metabolomics Haixia Institute of Science and Technology Fujian Agriculture and Forestry University Fuzhou China"}]},{"@id":"https://cir.nii.ac.jp/crid/1380580240159314568","@type":"Researcher","foaf:name":[{"@value":"Lan Yang"}],"jpcoar:affiliationName":[{"@value":"Key Laboratory of Molecular Biology and Gene Engineering in Jiangxi Province College of Life Science Nanchang University Jiangxi China"}]},{"@id":"https://cir.nii.ac.jp/crid/1380580240159314434","@type":"Researcher","foaf:name":[{"@value":"Shikui Song"}],"jpcoar:affiliationName":[{"@value":"FAFU‐UCR Joint Center for Horticultural Biology and Metabolomics Haixia Institute of Science and Technology Fujian Agriculture and Forestry University Fuzhou China"}]},{"@id":"https://cir.nii.ac.jp/crid/1380580240159314433","@type":"Researcher","foaf:name":[{"@value":"Dong Wang"}],"jpcoar:affiliationName":[{"@value":"Key Laboratory of Molecular Biology and Gene Engineering in Jiangxi Province College of Life Science Nanchang University Jiangxi China"}]},{"@id":"https://cir.nii.ac.jp/crid/1380580240159314563","@type":"Researcher","foaf:name":[{"@value":"Yuefeng Guan"}],"jpcoar:affiliationName":[{"@value":"FAFU‐UCR Joint Center for Horticultural Biology and Metabolomics Haixia Institute of Science and Technology Fujian Agriculture and Forestry University Fuzhou China"}]}],"publication":{"publicationIdentifier":[{"@type":"PISSN","@value":"14677644"},{"@type":"EISSN","@value":"14677652"}],"prism:publicationName":[{"@value":"Plant Biotechnology Journal"}],"dc:publisher":[{"@value":"Wiley"}],"prism:publicationDate":"2019-09-09","prism:volume":"18","prism:number":"3","prism:startingPage":"721","prism:endingPage":"731"},"reviewed":"false","dc:rights":["http://creativecommons.org/licenses/by-nc/4.0/"],"url":[{"@id":"https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1111%2Fpbi.13239"},{"@id":"https://onlinelibrary.wiley.com/doi/pdf/10.1111/pbi.13239"},{"@id":"https://onlinelibrary.wiley.com/doi/full-xml/10.1111/pbi.13239"}],"createdAt":"2019-08-27","modifiedAt":"2024-11-21","relatedProduct":[{"@id":"https://cir.nii.ac.jp/crid/1360298341434361728","@type":"Article","resourceType":"学術雑誌論文(journal article)","relationType":["isReferencedBy"],"jpcoar:relatedTitle":[{"@value":"Development of Ethyl Methanesulfonate Mutant Edamame Soybean (Glycine max (L.) Merr.) Populations and Forward and Reverse Genetic Screening for Early-Flowering Mutants"}]}],"dataSourceIdentifier":[{"@type":"CROSSREF","@value":"10.1111/pbi.13239"},{"@type":"CROSSREF","@value":"10.3390/plants11141839_references_DOI_6ZPmy2Mib09P8TSPx3xxYgjzDvn"}]}