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Molecular and genetical dissection ofthe mechanism underlying cytoplasmic male sterility in sugar beet

甜菜细胞质雄性不育机制的分子和遗传解析

基本信息

批准号：
16208001
负责人：
MIKAMI Tetsuo
金额：
$ 32.03万
依托单位：
Hokkaido University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16208001/
关键词：
sugar beet cytoplasmic male sterility restorer-of-fertility gene MPL anther tapetal cells anther-specific lipid transfer protein mitochondria cDNA 非特異的脂質輸送タンパク質 in situハイブリダイゼーション Rf1遺伝子 MPL遺伝子 in situ ハイブリダイゼーションタベート組織四分予期

项目摘要

Dissecting the nature of male-sterility encoding mitochondrial gene(s) and the nuclear genes (named Rf) that suppress male sterility is quite instructive in understanding the mechanism of cytoplasmic male sterility (CMS) in sugar beet. Here we attempted to clone and characterize the Rf1 locus which controls fertility restoration in sugar beet CMS. The results are as follows.1. We were able to restrict the possible location of the Rf1 locus to a minimum of six BAC clones spanning an interval of about 260kb. The nucleotide sequencing of the 260-kb region revealed 28 ORFs.2. Among the 28 ORFs, we found four ORFs which are specifically expressed in flower buds and targeted to mitochondria.3. The four ORFs are adjacent to one another and all code for metalloprotease. We designated the ORFs MPLs. Interestingly, in a nonrestoring (rf1rf1) genotype we identified homologous ORFs that exhibit deletions and are not expressed in flower buds.4. In situ hybridization analysis demonstrated that expression of MPLs is confined to the tapetal cells of anthers at the microspore tetrad and young microspore stages. Taken together, our observations suggest strongly that MPLs5. In an attempt to correlate mitochondrial lesions with cellular metabolic processes in sterile anthers, we found an anther-specific lipid transfer protein gene (named LTP).The gene was revealed to be highly expressed in the tapetal cells of fertile anthers at the young microspore stage but not in sterile anthers throughout flower development. It is most likely that the expression of LTP is down-regulated in the tapetum in response to mitochondrial dysfunction and subsequent physiological changes caused by CMS cytoplasm.

深入研究甜菜细胞质雄性不育的线粒体基因和核基因对甜菜细胞质雄性不育的抑制作用，对深入了解甜菜细胞质雄性不育的机理具有重要的指导意义。在这里，我们试图克隆和表征的Rf 1基因座控制甜菜CMS育性恢复。研究结果如下：1.我们能够限制Rf 1基因座的可能位置，以跨越约260 kb的间隔的最少6个BAC克隆。对260-kb区域进行核苷酸序列测定，共发现28个ORFs.在这28个ORF中，我们发现了4个在花蕾中特异表达的ORF，并且是针对拟南芥的.这四个ORF彼此相邻，都编码金属蛋白酶。我们将ORF命名为MPLs。有趣的是，在一个非恢复（rf 1 rf 1）基因型，我们确定了同源的ORF，表现出缺失，并没有在花蕾中表达。原位杂交分析表明，在小孢子四分体和幼小孢子时期，MPLs仅在花药的绒毡层细胞中表达。总之，我们的观察强烈表明，MPLs 5。为了研究不育花药线粒体损伤与细胞代谢过程的关系，我们发现了一个花药特异的脂质转运蛋白基因（LTP），该基因在小孢子发育时期的可育花药绒毡层细胞中表达量很高，而在整个花发育过程中不表达。这是最有可能的，LTP的表达下调绒毡层响应线粒体功能障碍和随后的生理变化引起的CMS细胞质。