| 摘要: |
| 在T-DNA插入突变体Salk_059463株系的群体中,筛选到两株雄性不育突变体,对T-DNA序列上的一对引物进行PCR鉴定,结果表明:其基因组中没有T-DNA插入.遗传分析表明这两株雄性不育突变体由同一单个隐性基因控制,引起不育的主要原因是从花药发育的第8期开始,小孢子细胞质内容物逐渐减少直至消失,到花药发育的第12期,药室内的小孢子只剩下一个花粉壁空壳,故该突变体命名为opw(only pollen wall).利用图位克隆的方法对OPW基因进行了定位,结果表明OPW基因位于第二条染色体上分子标记T28M21和T3G21之间的12 kb区间内,该区间内一共有21个基因注释.通过克隆区间内的基因并测序发现opw-1突变体基因组中At2g40140基因编码序列的外显子在第289和第290个碱基之间插入了一个A碱基,而opw-2突变体基因组中At2g40140基因编码序列的外显子在第412和第413个碱基之间插入了一个T碱基,造成的编码序列移码使第424至第426碱基成为终止密码子,故At2g40140是编码OPW的候选基因. |
| 关键词: 拟南芥 雄性不育 花粉壁空壳 图位克隆 |
| DOI： |
| 分类号: |
| 基金项目:国家自然科学基金(30970268);上海师范大学大学生创新项目 |
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| Cloning of Gene of male sterile mutant opw(only pollen wall) in Arabidopsis thaliana |
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PANG Chaoting, ZHANG Ting, LI Na, WU Wenyan, GAO Jufang
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College of Life and Environmental Sciences, Shanghai Normal University
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| Abstract: |
| Two male sterile mutant lines with normal vegetative and flora development but no seed yields were screened out from Salk_059463 line,a population of Arabidopsis ecotype Columbia (Col) mutagenized by T-DNA insertion (provided by ARBC).The identification of T-DNA insertion site showed that there were no T-DNA sequences in the genome of these two mutants.Genetic analysis indicated that the mutants were allele and controlled by the same single recessive nuclear gene named only pollen wall,of which the cytoplasm of microspores becomes vanished from anther development stage 8,and there are only pollen wall of pollen grains in the anther at the anther development stage 12.The OPW gene was mapped to a region of 12 kb between the molecular makers T28M21 and T3G21 on chromosome 2 using map-based cloning technique.There are 21 genes annotations in that region.Cloning and sequencing the At2g40140 of the mutants,we found that there was a single A base insertion between 289 to 290 bases in the opw-1 and another single T base insertion between 412 to 413 bases in the opw-2,therefore caused the end code appear in 424-426 bases.So At2g40140 is considered the candidate gene of the OPW. |
| Key words: Arabidopsis thaliana male sterile only pollen wall map-based cloning |
