| 水稻黄绿叶突变体ygl13的鉴定及候选基因分析 |
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| 引用本文： | 王亚琴,施军琼,张婷,李燕,张天泉,张小龙,桑贤春,凌英华,何光华.水稻黄绿叶突变体ygl13的鉴定及候选基因分析[J].中国农业科学,2015,48(21):4197-4208. |
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| 作者姓名： | 王亚琴 施军琼 张婷 李燕 张天泉 张小龙 桑贤春 凌英华 何光华 |
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| 基金项目： | 重庆市自然科学基金（cstc2012jjA80011）、中央高校基本科研业务费（XDJK2014C147）、重庆市攻关项目（cstc2013yykfA80013，cstc2012ggB80004） |
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| 摘 要： | 目的]对水稻黄绿叶突变体ygl13(yellow-green leaf 13)进行表型鉴定和候选基因检测,以便了解水稻叶色形成和调控的分子机制。方法]经甲基磺酸乙酯(EMS)诱变籼稻恢复系缙恢10号(Jinhui 10),从中筛选出1份遗传稳定的黄绿叶突变体命名为ygl13,对突变体的表型进行系统观察,调查其成熟期的主要农艺性状,分别测定野生型和突变体苗期和孕穗期的叶片光合色素含量,同时利用透射电镜观察野生型和突变体ygl13的叶肉细胞及叶绿体结构。将表型正常的不育系西农1A与突变体ygl13杂交,根据F1和F2群体的性状表现与分离情况,分析该突变性状的遗传行为,并以F2作为基因定位群体,对突变体ygl13进行候选基因遴选和突变位点测序验证。结果]突变体ygl13的植株叶片在整个生育期均呈现黄绿色,与野生型缙恢10号相比,突变体ygl13苗期和孕穗期叶片叶绿素a、叶绿素b和类胡萝卜素含量均极显著降低。透射电镜观察结果显示,与野生型相比,突变体ygl13叶绿体结构异常,基质片层减少退化,类囊体片层减少,不规则的散乱分布。农艺性状调查结果表明,突变体ygl13穗总粒数增加了26.06%,株高和结实率分别降低了12.33%和18.82%,但穗长、有效穗、穗实粒数和千粒重无显著差异。F2群体正常叶色的植株数与黄绿叶植株数分离比经χ2测验符合3﹕1分离比例(χ2=2.35χ20.05=3.84),表明ygl13的黄绿叶性状由1对隐性核基因控制。YGL13被定位于第8染色体短臂In Del标记ID43和ID69之间,遗传距离分别为4.0和0.5 c M,区间物理距离约为318 kb,共有52个基因。经测序比对分析发现,ygl13突变体在Os SIG1编码区的第1 005个碱基G突变为碱基A(位于第三外显子),造成编码色氨酸(Trp或W)的密码子突变为终止密码子,导致蛋白翻译提前终止,则该基因编码520个氨基酸的蛋白质突变为334个氨基酸的截短蛋白。q RT-PCR结果表明,突变体ygl13部分光合色素代谢途径和光系统相关基因表达紊乱。结论]水稻突变体ygl13的黄绿叶性状由1对隐性核基因控制,该基因与已报道的水稻质体σ因子Os SIG1为等位基因。
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| 关 键 词： | 水稻（Oryza sativa L.） 黄绿叶突变体 OsSIG1 遗传分析 候选基因分析 /> |
| 收稿时间： | 2015-05-26 |
Characterization and Candidate Gene Analysis of Yellow-Green Leaf Mutant ygl13 in Rice (Oryza sativa) |
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| WANG Ya-qin,SHI Jun-qiong,ZHANG Ting,LI Yan,ZHANG Tian-quan,ZHANG Xiao-long,SANG Xian-chun,LING Ying-hua,HE Guang-hua.Characterization and Candidate Gene Analysis of Yellow-Green Leaf Mutant ygl13 in Rice (Oryza sativa)[J].Scientia Agricultura Sinica,2015,48(21):4197-4208. |
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| Authors: | WANG Ya-qin SHI Jun-qiong ZHANG Ting LI Yan ZHANG Tian-quan ZHANG Xiao-long SANG Xian-chun LING Ying-hua HE Guang-hua |
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| Affiliation: | Rice Research Institute of Southwest University/Chongqing Key Laboratory of Application and Safety Control of Genetically Modified Crops, Chongqing 400715 |
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| Abstract: | 【Objective】The current study was conducted aiming at phenotypic characterization and candidate gene analysis of the yellow-green mutant ygl13, so as to add to our knowledge of the formation and regulation of the molecular mechanisms responsible for leaf-color mutations in rice.【Method】A new rice mutant exhibiting stable inheritance was identified as derived from ethyl methane sulfonate (EMS)-treated restorer line Jinhui10 (Oryza sativa), tentatively named as yellow-green leaf 13 (ygl13). Morphological characteristics, the photosynthetic pigment contents and the agronomic traits were measured systematically. Transmission electron microscopy was conducted to analyze the ultrastructure of the mesophyll cells and chloroplasts in the ygl13 mutant and wide-type plants. The ygl13 was crossed with indica sterile line Xinong1A whose plant and leaves were normally green, and the morphological phenotype and segregation ratio of F1 and F2 were used for genetic analysis, F2 for gene mapping, and putative genes in the fine mapped region were analyzed, and the candidate genes in the mutant and the wild type were sequenced, respectively.【Result】The ygl13 leaves displayed yellow-green compared with the wild type. And the photosynthetic pigment contents of chlorophyll a, chlorophyll b, and carotenoid decreased significantly at the seedling and the booting stages. The results from transmission electron microscope demonstrated that the structure of the chloroplast in the mutant ygl13 developed abnormally with poor thylakoids, less grana stacks and scattered distribution when compared with the wide type. According to the performance of agronomic traits, compared with the wild type Jinhui 10, the grain number per panicle increased by 26.06%, and the pant height and seed setting rate decreased by 12.33% and 18.82%. As for the panicle length, effective panicles per plant, filled grain number per panicle and 1000-grain weight, there was no significant difference between the wild type and ygl13. Genetic analysis demonstrated that the mutant trait was controlled by a single recessive gene as the number of green seedlings verses that of yellow-green seedlings approached 3:1（χ2=2.35＜χ20.05=3.84). Genetic mapping of the mutant gene was conducted using 602 recessive individuals from the F2 segregation population. Finally, YGL13 was mapped on the short arm of chromosome 8 between InDel marker ID43 and ID69, and with an interval of 318 kb. There were 52 genes in this region and the sequencing analysis of these candidate genes between the mutant and its wild type revealed a single base change (G1005A) of the OsSIG1 gene (LOC_Os08g06630) in the encoded product resulted in a premature stop codon and protein truncation with 334 residues not the primary protein with 520 residues in the mutant ygl13. The qRT-PCR results showed that, the expression level of genes associated with pigment metabolism and photosynthesis is in disorder. 【Conclusion】 The mutant ygl13 was controlled by a single recessive gene. The gene YGL13 was allelic to OsSIG1 which was documented as a plastid sigma factor previously in rice. |
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| Keywords: | rice (Oryza sativa L ) yellow-green leaf mutant OsSIG1 genetic analysis candidate gene analysis
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