XIAO Lianjie,HUANG Jie,CAO Penghui,et al.Analysis and gene mapping of rice premature senescence mutant zs[J].Journal of Nanjing Agricultural University,2018,41(5):793-800.[doi:10.7685/jnau.201802016]





Analysis and gene mapping of rice premature senescence mutant zs
肖连杰 黄捷 曹鹏辉 牟昌龄 Thanhliem Nguyen 刘世家 陈亮明 江玲
南京农业大学作物遗传与种质创新国家重点实验室/江苏省植物基因工程技术研究中心, 江苏 南京 210095
XIAO Lianjie HUANG Jie CAO Penghui MOU Changling Thanhliem Nguyen LIU Shijia CHEN Liangming JIANG Ling
State Key Laboratory of Crop Genetics and Germplasm Enhancement/Research Center of Jiangsu Plant Gene Engineering, Nanjing Agricultural University, Nanjing 210095, China
riceleaf premature senescencefine mappingsequence analysiscytochrome P450
[目的]本研究旨在对水稻叶片早衰突变体zs进行遗传分析及基因定位,探索水稻叶片早衰的分子机制。[方法]从甲基磺酸乙酯(EMS)诱变‘宁粳1号’突变体库中筛选鉴定了1个稳定遗传的早衰突变体zs,构建突变体zs与籼稻品种‘N22’的F2群体,对控制突变性状的基因进行定位,利用RT-qPCR对叶绿素合成以及质体发育相关基因的表达水平进行分析。[结果]突变体zs在28℃条件下生长14 d时,zs第2叶叶尖出现黄褐色斑点,并逐渐向叶基部蔓延。突变体zs叶绿素a和叶绿素b含量明显下降。zs叶肉细胞结构异常,叶绿体中类囊体片层结构排列松散。与野生型相比,zs的株高、分蘖数和千粒质量均显著降低。通过构建遗传分离群体,将突变基因定位在第12号染色体短臂上的600 kb的区间内。此区间内含有2个已报道的与叶片衰老相关的基因LOC_Os12g16410LOC_Os12g16720,分别编码异黄酮还原酶和细胞色素单加氧酶(P450)。测序分析发现,仅在LOC_Os12g16720第2外显子上有1个单碱基突变,导致苏氨酸突变为甲硫氨酸。zs叶绿素合成相关基因和质体发育相关基因的表达量显著低于野生型。[结论]鉴定了1个早衰突变体zs,通过精细定位与序列分析,将LOC_Os12g16720作为候选基因,该基因可能参与调控叶绿素合成及质体的发育。
[Objectives]In this study,genetics analysis and gene mapping of zs,a mutant with early senescent leaf in rice,were conducted to explore the molecular mechanism or rice leaf premature senescence.[Methods]A premature senescence mutant zs was isolated from an ethylmethylsulfone-induced ‘Ningjing 1’ mutant library. The F2 population from zs crossed to ‘N22’ (indica cultivar)was used to mapping the gene ZS. The expression level of the genes related to chlorophyll synthesis and plastid development was analyzed by RT-qPCR.[Results]Compared with the wild type,the second leaf tip of the mutant zs displayed yellow brown spots,spreading to the base of the leaf when it grew at 28℃ for 14 days. The content of chlorophyll a and chlorophyll b decreased significantly in zs leaves. The structure of mesophyll cells in zs were abnormal,and the thylakoid lamellae of chloroplasts were loosely arranged. In addition,the plant height,tiller number and 1 000-grain weight of zs decreased significantly. The target gene was finally mapped on the short arm of chromosome 12 near to marker NL12-18 and ZS20 with the physical distance about 600 kb. There were two reported genes related to leaf senescence in this region,LOC_Os12g16410 and LOC_Os12g16720,which encoded isoflavone reductase and cytochrome monooxygenase(P450),respectively. Sequencing analysis found that there was only a single base mutation on the LOC_Os12g16720 gene exon,which caused the substitution of threonine to methionine. Compared with wild type,the expression level of chlorophyll synthesis and plastid development related genes decreased significantly in the mutant zs.[Conclusions]We identified a prematurely senescence mutant zs. The cytochrome monooxygenase gene(LOC_Os12g16720) was the candidate gene by fine mapping and sequencing. ZS may be involved in the regulation of chlorophyll synthesis and plastid development.


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更新日期/Last Update: 1900-01-01