Wang Dawei, Wang Shaomei, Chao Jiangtao, Wu Xinru, Sun Yuhe, Li Fengxia, Lv Jing, Gao Xiaoming, Liu Guanshan, Wang Yuanying
Tobacco Research Institute, Chinese Academy of Agricultural Sciences, No. 11 Keyuanjingsi Road, Laoshan District, Qingdao, 266101, China.
Key Laboratory for Tobacco Gene Resources, State Tobacco Monopoly Administration, Qingdao, 266101, China.
Planta. 2017 Jul;246(1):149-163. doi: 10.1007/s00425-017-2690-z. Epub 2017 Apr 11.
A novel tobacco mutant library was constructed, screened, and characterized as a crucial genetic resource for functional genomics and applied research. A comprehensive mutant library is a fundamental resource for investigating gene functions, especially after the completion of genome sequencing. A new tobacco mutant population induced by ethyl methane sulfonate mutagenesis was developed for functional genomics applications. We isolated 1607 mutant lines and 8610 mutant plants with altered morphological phenotypes from 5513 independent M2 families that consisted of 69,531 M2 plants. The 2196 mutations of abnormal phenotypes in the M2 putative mutants were classified into four groups with 17 major categories and 51 subcategories. More than 60% of the abnormal phenotypes observed fell within the five major categories including plant height, leaf shape, leaf surface, leaf color, and flowering time. The 465 M2 mutants exhibited multiple phenotypes, and 1054 of the 2196 mutations were pleiotropic. Verification of the phenotypes in advanced generations indicated that 70.63% of the M3 lines, 84.87% of the M4 lines, and 95.75% of the M5 lines could transmit original mutant phenotypes of the corresponding M2, M3, and M4 mutant plants. Along with the increased generation of mutants, the ratios of lines inheriting OMPs increased and lines with emerging novel mutant phenotypes decreased. Genetic analyses of 18 stably heritable mutants showed that two mutants were double recessive, five were monogenic recessive, eight presented monogenic dominant inheritance, and three presented semi-dominant inheritance. The pleiotropy pattern, saturability evaluation, research prospects of genome, and phenome of the mutant populations were also discussed. Simultaneously, this novel mutant library provided a fundamental resource for investigating gene functions in tobacco.
构建、筛选并鉴定了一个新型烟草突变体文库,将其作为功能基因组学和应用研究的重要遗传资源。全面的突变体文库是研究基因功能的基础资源,尤其是在基因组测序完成之后。为功能基因组学应用开发了一个由甲基磺酸乙酯诱变诱导的新型烟草突变群体。我们从由69531株M2植株组成的5513个独立M2家系中分离出1607个突变系和8610株形态表型改变的突变植株。M2假定突变体中2196个异常表型的突变被分为4组,有17个主要类别和51个子类别。观察到的超过60%的异常表型属于包括株高、叶形、叶表面、叶色和开花时间在内的五个主要类别。465个M2突变体表现出多种表型,2196个突变中有1054个是多效性的。对后代的表型验证表明,70.63%的M3株系、84.87%的M4株系和95.75%的M5株系能够传递相应M2、M3和M4突变植株的原始突变表型。随着突变体世代的增加,继承原始突变表型的株系比例增加,出现新突变表型的株系减少。对18个稳定遗传突变体的遗传分析表明,两个突变体是双隐性的,五个是单基因隐性的,八个呈现单基因显性遗传,三个呈现半显性遗传。还讨论了突变群体的多效性模式、饱和度评估、基因组和表型组的研究前景。同时,这个新型突变体文库为研究烟草基因功能提供了基础资源。