A reference-guided TILLING by amplicon-sequencing platform supports forward and reverse genetics in barley

doi:10.1016/j.xplc.2022.100317

CORC > 植物研究所 > 中国科学院植物研究所 > 中科院植物分子生理学重点实验室

	A reference-guided TILLING by amplicon-sequencing platform supports forward and reverse genetics in barley
	Jiang, Congcong ; Lei, Miaomiao 1; Guo, Yu 2; Gao, Guangqi ; Shi, Lijie 1; Jin, Yanlong ; Cai, Yu 1; Himmelbach, Axel 2; Zhou, Shenghui ; He, Qiang
刊名	PLANT COMMUNICATIONS
	2022
卷号	3 期号:4
关键词	barley mutagenesis TILLING amplicon-seq genetics
ISSN号	2590-3462
DOI	10.1016/j.xplc.2022.100317
文献子类	Article
英文摘要	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.
学科主题	Biochemistry & Molecular Biology ; Plant Sciences
出版地	AMSTERDAM
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)
语种	英语
出版者	ELSEVIER
WOS记录号	WOS:000844042700007
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
内容类型	期刊论文
源URL	[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).