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| A reference-guided TILLING by amplicon-sequencing platform supports forward and reverse genetics in barley | |
Jiang, Congcong; Lei, Miaomiao1; Guo, Yu2; Gao, Guangqi; Shi, Lijie1; Jin, Yanlong; Cai, Yu1; Himmelbach, Axel2; Zhou, Shenghui; He, Qiang; Yao, Xuefeng3; Kan, Jinhong; Haberer, Georg4; Duan, Fengying; Li, Lihui; Liu, Jun; Zhang, Jing; Spannagl, Manuel4; Liu, Chunming3 ; Stein, Nils2; Feng, Zongyun1; Mascher, Martin2; Yang, Ping
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| 2022 | |
| 发表期刊 | PLANT COMMUNICATIONS
 |
| ISSN | 2590-3462 |
| 卷号 | 3期号:4 |
| 摘要 | Barley is a diploid species with a genome smaller than those of other members of the Triticeae tribe, making it an attractive model for genetic studies in Triticeae crops. The recent development of barley genomics has created a need for a high-throughput platform to identify genetically uniform mutants for gene function investigations. In this study, we report an ethyl methanesulfonate (EMS)-mutagenized population consisting of 8525M3 lines in the barley landrace Hatiexi'' (HTX), which we complement with a high-quality de novo assembly of a reference genome for this genotype. The mutation rate within the population ranged from 1.51 to 4.09 mutations per megabase, depending on the treatment dosage of EMS and the mutation discrimination platform used for genotype analysis. We implemented a three-dimensional DNA pooling strategy combined with multiplexed amplicon sequencing to create a highly efficient and cost-effective TILLING (targeting induced locus lesion in genomes) platform in barley. Mutations were successfully identified from 72 mixed amplicons within a DNA pool containing 64 individual mutants and from 56 mixed amplicons within a pool containing 144 individuals. We discovered abundant allelic mutants for dozens of genes, including the barley Green Revolution contributor gene Brassinosteroid insensitive 1 (BRI1). As a proof of concept, we rapidly determined the causal gene responsible for a chlorotic mutant by following the MutMap strategy, demonstrating the value of this resource to support forward and reverse genetic studies in barley. |
| 关键词 | barley mutagenesis TILLING amplicon-seq genetics |
| 学科领域 | Biochemistry & Molecular Biology ; Plant Sciences |
| DOI | 10.1016/j.xplc.2022.100317 |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS关键词 | BYMOVIRUS RESISTANCE ; MUTATION MECHANISM ; HIDDEN VARIATION ; GENOME ; IDENTIFICATION ; DISCOVERY ; GENES ; MUTAGENESIS ; POPULATION ; FRAMEWORK |
| WOS研究方向 | Science Citation Index Expanded (SCI-EXPANDED) |
| WOS记录号 | WOS:000844042700007 |
| 出版者 | ELSEVIER |
| 文献子类 | Article |
| 出版地 | AMSTERDAM |
| 资助机构 | National Key Research and Development Program of China [2018YFD1000702/2018YFD1000700] ; Agricultural Science and Technology Innovation Program of the Chinese Academy of Agricultural Sciences (CAAS) , China |
| 作者邮箱 | mascher@ipk-gatersleben.de ; yangping@caas.cn |
| 作品OA属性 | gold, Green Published |
| 引用统计 | |
| 文献类型 | 期刊论文 |
| 条目标识符 | http://ir.ibcas.ac.cn/handle/2S10CLM1/28943 |
| 专题 | 中科院植物分子生理学重点实验室 |
| 作者单位 | 1.Chinese Acad Agr Sci, Inst Crop Sci, Beijing, Peoples R China 2.Sichuan Agr Univ, Coll Agron, Chengdu, Peoples R China 3.Leibniz Inst Plant Genet & Crop Plant Res IPK, Seeland, Germany 4.Chinese Acad Sci, Inst Bot, Beijing, Peoples R China 5.Helmholtz Ctr Munich, German Res Ctr Environm Hlth, Plant Genome & Syst Biol PGSB, Neuherberg, Germany |
| 推荐引用方式 GB/T 7714 | Jiang, Congcong,Lei, Miaomiao,Guo, Yu,et al. A reference-guided TILLING by amplicon-sequencing platform supports forward and reverse genetics in barley[J]. PLANT COMMUNICATIONS,2022,3(4). |
| APA | Jiang, Congcong.,Lei, Miaomiao.,Guo, Yu.,Gao, Guangqi.,Shi, Lijie.,...&Yang, Ping.(2022).A reference-guided TILLING by amplicon-sequencing platform supports forward and reverse genetics in barley.PLANT COMMUNICATIONS,3(4). |
| MLA | Jiang, Congcong,et al."A reference-guided TILLING by amplicon-sequencing platform supports forward and reverse genetics in barley".PLANT COMMUNICATIONS 3.4(2022). |
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| A_reference_guided_T（3848KB） | 期刊论文 | 出版稿 | 开放获取 | CC BY-NC-SA | 浏览 请求全文 | |
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