An efficient TILLING platform for cultivated tobacco
文献类型：期刊论文
| 作者 | 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
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| 刊名 | JOURNAL OF INTEGRATIVE PLANT BIOLOGY
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| 出版日期 | 2020 |
| 卷号 | 62期号:2页码:165-180 |
| ISSN号 | 1672-9072 |
| DOI | 10.1111/jipb.12784 |
| 文献子类 | Article |
| 英文摘要 | 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. |
| 学科主题 | Biochemistry & Molecular Biology ; Plant Sciences |
| 出版地 | HOBOKEN |
| 电子版国际标准刊号 | 1744-7909 |
| 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 |
| 语种 | 英语 |
| WOS记录号 | WOS:000511460800002 |
| 出版者 | WILEY |
| 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 |
| 源URL | [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.Chinese Acad Sci, Inst Bot, Key Lab Plant Mol Physiol, Beijing 100093, Peoples R China 2.Chinese Acad Agr Sci, Inst Crop Sci, Beijing 100081, Peoples R China 3.Chinese Acad Agr Sci, Tobacco Res Inst, Qingdao 266101, Peoples R China 4.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 5.[Yao, Xue-Feng 6.Yunnan Acad Tobacco Agr Sci, Natl Tobacco Genet Engn Res Ctr, Key Lab Tobacco Biotechnol Breeding, Kunming 650021, Yunnan, Peoples R China 7.Wu, Yu-Ping 8.Wang, Bing-Wu 9.Song, Zhong-Bang 10.Li, Wen-Zheng |
| 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. |
入库方式： OAI收割
来源：植物研究所
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