水稻ygl98黄绿叶突变基因的精细定位与遗传分析

doi:10.3724/SP.J.1006.2011.00991

摘要/Abstract

摘要： 通过EMS诱变获得一份遗传稳定的水稻黄绿叶突变体ygl98，该突变体整个生育期呈黄绿色。与野生型相比，突变体的叶绿素和类胡萝卜素含量分别下降45.3%和45.6%，有效穗数和结实率分别减少14.4%和10.7%，株高降低7.4%。透射电镜观察表明，ygl98突变体的叶绿体形状不规则，叶绿体中有许多空的囊泡状结构，类囊体数目减少，每个基粒仅由少数几个类囊体垛叠而成。遗传分析表明，ygl98的突变性状由1对隐性核基因控制。利用(ygl98/浙辐802) F₂作为定位群体，将突变基因定位在第3染色体长臂InDel标记I3和I4之间，遗传距离分别为0.07 cM和0.19 cM，两标记之间的物理距离约为44.2 kb，此区间内包含8个预测基因。基因组序列分析发现，ygl98突变体在编码镁离子螯合酶ChlD亚基的OsChlD基因编码区第1 522碱基处(位于第10外显子)，碱基G突变为碱基A，从而造成编码蛋白序列第508位的丙氨酸(Ala)突变成苏氨酸(Thr)。该基因是已报道的水稻黄绿叶基因Chlorina-1的等位基因，但突变体表型有明显区别，Chlorina-1突变体在2~3周龄幼苗时开始出现黄绿叶，且该黄绿叶性状仅在苗期表现，而ygl98突变体整个生育期都表现为黄绿叶，这可能是OsChlD基因组序列的突变位点不同造成的。

关键词: 水稻, 黄绿叶突变体, OsChlD, 遗传分析, 精细定位

Abstract: A yellow-green leaf mutant ygl98 was isolated by chemistry mutagenesis. Its whole plant exhibited yellow-green trait throughout the growing period. Compared with its wild-type parent 10079, the contents of chlorophyll and carotenoid decreased by 45.3% and 45.6%; and at the maturity, the number of productive panicles per plant, seed setting rate and plant height reduced by 14.4%, 10.7%, and 7.4%, respectively. The result of electron microscopic observation revealed that the chloroplasts in the ygl98 mutant were out-of-shape. A lot of cystic structures and poor thylakoids were observed in the chloroplasts of the ygl98 mutant, and grana stacks appeared to be less dense compared to those of wild type. Genetic analysis showed that the yellow-green leaf trait of the ygl98 mutant was controlled by one pair of recessive nuclear genes. Genetic mapping of the mutant gene was conducted by using 771 yellow-green leaf individuals from the F₂ mapping population of ygl98/Zhefu 802. Finally, the mutant gene was mapped between InDel markers I3 and I4 on the long arm of chromosome 3, with genetic distances of 0.07 and 0.19 cM, respectively, and with physical distance of 44.2 kb, in which eight predicted genes had been annotated. Sequencing analysis of these candidate genes between the mutant and its wild-type revealed the single base change (G1522A) of the gene for magnesium-chelatase ChlD subunit resulted in a missense mutation (A508T) in the encoded product. The same gene mutation caused by OsChlD(Chlorina-1) was documented previously. The Chlorina-1 mutant displays a severe yellowish-green leaf phenotype only at the seedling stage, and the abnormal leaf color is ?rst observed on the leaves of 2- to 3-week-old seedlings, while the ygl98 mutant exhibits yellow-green trait throughout the growing period. The different phenotypes of the two mutants may be caused by the different mutational sites of OsChlD genomic sequence.

Key words: Oryza sativa L., Yellow-green leaf mutant, OsChlD, Genetic analysis, Fine-mapping

孙小秋, 王兵, 肖云华, 万春美, 邓晓建, 王平荣. 水稻ygl98黄绿叶突变基因的精细定位与遗传分析[J]. 作物学报, 2011, 37(06): 991-997.

SUN Xiao-Qiu, WANG Bing, XIAO Yun-Hua, MO Chun-Mei, DENG Xiao-Jian, WANG Beng-Rong. Genetic Analysis and Fine-Mapping of the ygl98 Yellow-Green Leaf Gene in Rice[J]. Acta Agron Sin, 2011, 37(06): 991-997.

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