An efficient TILLING platform for cultivated tobacco

doi:10.1111/jipb.12784

	An efficient TILLING platform for cultivated tobacco
	Gao, Yu-Long ; Yao, Xue-Feng ; Li, Wen-Zheng ; Song, Zhong-Bang ; Wang, Bing-Wu ; Wu, Yu-Ping ; Shi, Jun-Li ; Liu, Guan-Shan ; Li, Yong-Ping ; Liu, Chun-Ming
	2020
发表期刊	JOURNAL OF INTEGRATIVE PLANT BIOLOGY
ISSN	1672-9072
卷号	62 期号:2 页码:165-180
摘要	Targeting-induced local lesions in genomes (TILLING) is a powerful reverse-genetics tool that enables high-throughput screening of genomic variations in plants. Although TILLING has been developed for many diploid plants, the technology has been used in very few polyploid species due to their genomic complexity. Here, we established an efficient capillary electrophoresis-based TILLING platform for allotetraploid cultivated tobacco (Nicotiana tabacum L.) using an ethyl methanesulfonate (EMS)-mutagenized population of 1,536 individuals. We optimized the procedures for endonuclease preparation, leaf tissue sampling, DNA extraction, normalization, pooling, PCR amplification, heteroduplex formation, and capillary electrophoresis. In a test screen using seven target genes with eight PCR fragments, we obtained 118 mutants. The mutation density was estimated to be approximately one mutation per 106kb on average. Phenotypic analyses showed that mutations in two heavy metal transporter genes, HMA2S and HMA4T, led to reduced accumulation of cadmium and zinc, which was confirmed independently using CRISPR/Cas9 to generate knockout mutants. Our results demonstrate that this powerful TILLING platform (available at ) can be used in tobacco to facilitate functional genomics applications.
学科领域	Biochemistry & Molecular Biology ; Plant Sciences
DOI	10.1111/jipb.12784
收录类别	SCI
语种	英语
WOS关键词	P-TYPE ATPASES ; POINT MUTATIONS ; DISCOVERY ; TRANSLOCATION ; BIOSYNTHESIS ; MUTAGENESIS ; RESOURCE ; NICOTINE ; MUTANTS ; CADMIUM
WOS研究方向	Biochemistry & Molecular Biology ; Plant Sciences
WOS记录号	WOS:000511460800002
出版者	WILEY
文献子类	Article
出版地	HOBOKEN
EISSN	1744-7909
资助机构	Yunnan Province Tobacco funding [2014YN06, 2014YN05, 2016YN25, 2017YN03] Funding Source: Medline ; Bureau of National Tobacco funding [110201601031 [JY-05], 110201401005 [JY-05], 110201401006 [JY-06]] Funding Source: Medline ; National Transgenic Research Projects [2016ZX08009-001-006-002, 2016ZX08009-003-005] Funding Source: Medline
作者邮箱	cmliu@ibcas.ac.cn
引用统计	被引频次：12[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	http://ir.ibcas.ac.cn/handle/2S10CLM1/21929
专题	中科院植物分子生理学重点实验室
作者单位	1.[Gao, Yu-Long 2.Li, Wen-Zheng 3.Song, Zhong-Bang 4.Wang, Bing-Wu 5.Wu, Yu-Ping 6.Shi, Jun-Li 7.Yunnan Acad Tobacco Agr Sci, Natl Tobacco Genet Engn Res Ctr, Key Lab Tobacco Biotechnol Breeding, Kunming 650021, Yunnan, Peoples R China 8.[Yao, Xue-Feng 9.Chinese Acad Sci, Inst Bot, Key Lab Plant Mol Physiol, Beijing 100093, Peoples R China 10.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 11.Chinese Acad Agr Sci, Tobacco Res Inst, Qingdao 266101, Peoples R China 12.Chinese Acad Agr Sci, Inst Crop Sci, Beijing 100081, Peoples R China
推荐引用方式 GB/T 7714	Gao, Yu-Long,Yao, Xue-Feng,Li, Wen-Zheng,et al. An efficient TILLING platform for cultivated tobacco[J]. JOURNAL OF INTEGRATIVE PLANT BIOLOGY,2020,62(2):165-180.
APA	Gao, Yu-Long.,Yao, Xue-Feng.,Li, Wen-Zheng.,Song, Zhong-Bang.,Wang, Bing-Wu.,...&Liu, Chun-Ming.(2020).An efficient TILLING platform for cultivated tobacco.JOURNAL OF INTEGRATIVE PLANT BIOLOGY,62(2),165-180.
MLA	Gao, Yu-Long,et al."An efficient TILLING platform for cultivated tobacco".JOURNAL OF INTEGRATIVE PLANT BIOLOGY 62.2(2020):165-180.

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