Chen Qian, Zhao Tingting, Duan Lili, Mo Zejun, Tian Maozhu, Li Zhenhua, Liu Yang, Liu Renxiang
Guizhou University College of Agriculture/College of Tobacco, Guizhou Provincial Key Laboratory of Tobacco Quality Research, Guiyang 550025, China.
Open Life Sci. 2021 Aug 24;16(1):815-826. doi: 10.1515/biol-2021-0087. eCollection 2021.
The sterile line is the basis of crop heterosis utilization. To broaden the sources of male sterility in tobacco, the L. gene was cloned from the tobacco variety K326 by homologous cloning based on the ( L. ) gene sequence of male-sterility genes in pepper. The protein structure and physicochemical properties of the two genes were determined by bioinformatics analysis, and the function of the gene was verified by the CRISPR/Cas9 system. The results showed that the sequences of Ntms1 and Cams1 were 85.25% similar, and plant homeodomains were found in both genes; the physical and chemical properties were also very similar. It is speculated that the gene had the same function as the gene in controlling male sterility. Compared to the wild-type plants, the filaments of the knockout mutant plants were shorter, and the stamen was shorter than the pistil. The anthers did not develop fully and had few viable pollen grains; the tapetum and the anther wall had developed abnormally, and the anther chamber was severely squeezed. The malondialdehyde content in the mutant plants was significantly higher than that in the wild-type plants, while self-fertility was significantly lower in the mutant plants. The results showed that the gene plays an important role in regulating fertility in tobacco.
不育系是作物杂种优势利用的基础。为拓宽烟草雄性不育的来源，基于辣椒雄性不育基因（L.）的基因序列，通过同源克隆从烟草品种K326中克隆了L.基因。通过生物信息学分析确定了这两个基因的蛋白质结构和理化性质，并利用CRISPR/Cas9系统验证了该基因的功能。结果表明，Ntms1和Cams1的序列相似度为85.25%，且两个基因中均发现了植物同源结构域；其理化性质也非常相似。推测该基因在控制雄性不育方面与该基因具有相同功能。与野生型植株相比，该基因敲除突变体植株的花丝较短，雄蕊短于雌蕊。花药发育不完全，有活力的花粉粒较少；绒毡层和花药壁发育异常，药室严重挤压。突变体植株中的丙二醛含量显著高于野生型植株，而突变体植株的自交育性显著较低。结果表明，该基因在调控烟草育性方面发挥重要作用。
