一个调控水稻叶绿素合成基因wl-1的精细定位  

Fine Mapping of wl-1 in Rice,a Gene Regulating the Synthesis of Chlorophyll

作　　者：李潜龙 圣忠华[2] 陈炜 魏祥进[2] 焦桂爱[2] 唐绍清[2] 胡培松[1,2] 王建龙[1] Li Qianlong;Sheng Zhonghua;Chen Wei;Wei Xiangjin;Jiao Guiai;Tang Shaoqing;Hu Peisong;Wang Jianlong(Southem Regional Collaborative Innovation Center for Grain and Oil Crops in China,Agricultural College,Hunan Agricultural University,Changsha, 410128;Key Laboratory of Rice Biology and Genetics Breeding of Ministry of Agriculture,China National Rice Research Institute,Hangzhou,310006)

机构地区：[1]湖南农业大学农学院南方粮油作物协同创新中心,长沙410128 [2]中国水稻研究所农业部水稻生物学与遗传育种重点实验室,310006

出　　处：《分子植物育种》2019年第1期132-139,共8页Molecular Plant Breeding

基　　金：浙江省农业新品种选育重大专项(2016C02050-2);国家重点研发项目(2016YFD0101801;2017YFD0100300);植物转基因专项(2016ZX08001006)共同资助

摘　　要：为改良优异两系不育系株1S (Z1S)的株型特征,利用甲基磺酸乙酯(EMS)诱变技术处理株1S (Z1S),得到一个苗期为白条纹的突变体,命名为wl-1。以该突变体与粳稻品种日本晴杂交,得F1种子,进而获得自交F2群体。遗传分析表明wl-1受单个隐性基因控制。利用图位克隆技术将wl-1初定位于水稻第3号染色体上RM15851和RM15880两个标记之间。进一步扩大遗传定位群体,最终将wl-1精细定位于标记inedl3和indel4之间的122 kb区域内。生物信息学分析表明该区域有12个ORF。对这12个ORF编码区逐个测序分析表明,其第10个ORF (LOC-03g52170),编码4-羟基-3-甲基丁-2-烯基二磷酸还原酶的第2个外显子上的第165位碱基处存在1个碱基A替换为T的突变。基因表达分析表明,在幼苗期与野生型株1S相比,突变体中LOC-03g52170的表达量显著低于相应的野生型;同时突变体中叶绿素合成相关基因、光合作用相关基因以及叶绿体发育相关基因的表达量发生了显著变化。对叶色基因wl-1 (LOC-03g52170)的挖掘,有利于进一步阐释叶绿体和叶绿素合成的生物学机理,同时为水稻高光效育种提供理论和技术支撑。In order to improve the plant type characteristics of excellent two-line male sterile line Zhu1 S(Z1 S),the EMS mutagenesis technology was used as the method to treat Zhu1 s(Z1 S). A mutant with white stripes at seedling stage named wl-1 was obtained. The mutant was hybridized with japonica rice Nipponbare to obtain F1 seed and obtain self-bred F2 population. Genetic analysis showed that the wl-1 was controlled by a single recessive Gene. Using mapping based cloning techniques, the wl-1 was initially located between RM15851 and RM15880 markers on the rice chromosome 3. To further expand the genetic mapping population, WL-1 was finally fine positioned in the 122 kb region between In-Del markers indel3 and Indel4. Bioinformatics analysis showed that there were 12 ORFs in the fine mapping region. Sequencing analysis of the 12 ORFs coding regions one by one showed that the 10 thORF(LOC-03 g52170) encoded 4-hydroxy-3-methylbut-2-enyl diphosphate reductase, which there was a replaced with T at the 165 thbase on the second exon. Gene expression analysis showed that the expression level of LOC-03 g52170 was significantly lower in the mutant than that of wild-type Z1 S at the seedling stage, while the expression of chlorophyll synthesis related genes, photosynthesis related genes and chloroplast development related genes in mutants changed significantly. The excavation of leaf color gene wl-1(LOC-03 g-52170) would helpful to further elucidate the biological mechanism of chloroplast and chlorophyll synthesis, and to provide theoretical and technical support for high photosynthetic efficiency rice breeding.

关 键 词：wl-1 白条纹叶 基因精细定位 水稻 

分 类 号：S511[农业科学—作物学]