Construction of a genomewide RNAi mutant library in rice

doi:10.1111/pbi.12093

CORC > 中国农业科学院 > 作物科学研究所

	Construction of a genomewide RNAi mutant library in rice
	Wang, Lei 1; Zheng, Jie 1; Luo, Yanzhong 1; Xu, Tao 1; Zhang, Qiuxue 1; Zhang, Lan 1; Xu, Miaoyun 1; Wan, Jianmin 2; Wang, Ming-Bo 3; Zhang, Chunyi 1
刊名	PLANT BIOTECHNOLOGY JOURNAL
	2013
卷号	11 期号:8 页码:997-1005
关键词	Rice RNAi gene silencing siRNA library hairpin RNA
ISSN号	1467-7644
DOI	10.1111/pbi.12093
通讯作者	Wang, Lei
英文摘要	Long hairpin RNA (hpRNA) transgenes are a powerful tool for gene function studies in plants, but a genomewide RNAi mutant library using hpRNA transgenes has not been reported for plants. Here, we report the construction of a hpRNA library for the genomewide identification of gene function in rice using an improved rolling circle amplification-mediated hpRNA (RMHR) method. Transformation of rice with the library resulted in thousands of transgenic lines containing hpRNAs targeting genes of various function. The target mRNA was down-regulated in the hpRNA lines, and this was correlated with the accumulation of siRNAs corresponding to the double-stranded arms of the hpRNA. Multiple members of a gene family were simultaneously silenced by hpRNAs derived from a single member, but the degree of such cross-silencing depended on the level of sequence homology between the members as well as the abundance of matching siRNAs. The silencing of key genes tended to cause a severe phenotype, but these transgenic lines usually survived in the field long enough for phenotypic and molecular analyses to be conducted. Deep sequencing analysis of small RNAs showed that the hpRNA-derived siRNAs were characteristic of Argonaute-binding small RNAs. Our results indicate that RNAi mutant library is a high-efficient approach for genomewide gene identification in plants.
学科主题	Biotechnology & Applied Microbiology ; Plant Sciences ; BIOTECHNOLOGY & APPLIED MICROBIOLOGY ; PLANT SCIENCES
语种	英语
出版者	WILEY-BLACKWELL
WOS记录号	WOS:000324639600009
内容类型	期刊论文
源URL	[http://111.203.20.206/handle/2HMLN22E/5787]
专题	作物科学研究所
作者单位	1.Chinese Acad Agr Sci, Natl Key Facil Crop Gene Resources & Genet Improv, Biotechnol Res Inst, Beijing 100193, Peoples R China 2.Chinese Acad Agr Sci, Crop Sci Inst, Natl Key Facil Crop Gene Resources & Genet Improv, Beijing 100193, Peoples R China 3.CSIRO Plant Ind, Canberra, NSW, Australia
推荐引用方式 GB/T 7714	Wang, Lei,Zheng, Jie,Luo, Yanzhong,et al. Construction of a genomewide RNAi mutant library in rice[J]. PLANT BIOTECHNOLOGY JOURNAL,2013,11(8):997-1005.
APA	Wang, Lei.,Zheng, Jie.,Luo, Yanzhong.,Xu, Tao.,Zhang, Qiuxue.,...&Fan, Yunliu.(2013).Construction of a genomewide RNAi mutant library in rice.PLANT BIOTECHNOLOGY JOURNAL,11(8),997-1005.
MLA	Wang, Lei,et al."Construction of a genomewide RNAi mutant library in rice".PLANT BIOTECHNOLOGY JOURNAL 11.8(2013):997-1005.