哈克尼西棉细胞质雄性不育相关线粒体基因多态性分析

Polymorphism Analysis of Mitochondrial Genes Associated Cytoplasmic Male Sterility in Cotton(Gossypium harknessii Brandegee)

【目的】研究棉花细胞质雄性不育的相关线粒体基因。【方法】利用线粒体基因atp A、atp6、atp9、cob、coxⅠ、coxⅡ、coxⅢ、nad3、nad6、nad9探针,对哈克尼西棉细胞质雄性不育系S、保持系F、杂种一代H进行限制性片段长度多态性分析。【结果】atp A、coxⅢ、nad3、nad6在3个材料间具有多态性。atp A、nad6基因探针与所有酶的杂交结果均显示出多态性,且在不育系与杂种一代中表现一致,而在保持系中差异明显。atp A/Eco RⅠ在3个材料中的杂交结果均显示2条带,其中1条2.2 kb的杂交带在3个材料中相同,另一条在不育系和杂种一代中大小为3.2 kb,而在保持系中为4.8 kb;atp A/PstⅠ杂交结果中,在不育系和杂种一代中的条带大小为17.0kb,而在保持系中为10.2 kb。nad6探针与4个酶的杂交结果均为不育系和杂种一代比保持系多1~2条杂交带。coxⅢ/Eco RⅠ在3个材料中均有1条2.5 kb的杂交带,但在保持系与杂种一代中比不育系多1条1.7 kb的弱带。nad3/Bam HⅠ的杂交结果在不育系与保持系中表现一致,而在杂种一代中缺少1条9.5 kb的杂交带。【结论】推测atp A、nad6基因参与调控不育系的形成,而coxⅢ、nad3可能受到恢复系核基因的调控,在不育系育性恢复过程中发挥较为重要的作用。

[Objective] Cytoplasmic male sterility （CMS） is closely associated with the mitochondrial genome. The aim of this study was to identify CMS-related mitochondrial genes in cotton. [Method] Ten mitochondrial gene probes （i.e., atpA, atp6, atp9, cob, coxI, coxII, coxIII, nad3, nad6, and nad9） were used to analyze restriction fragment length polymorphisms in a Gossypium harknessii Brandegee cytoplasmic male sterile line （S）, maintainer line （F）, and hybrid （H）. [Result] Ten probe/enzyme combinations for four probes （i.e., atpA, coxIII, nad3, and nad6） revealed polymorphisms among lines S, F, and H. All enzyme digestions for two probes （i.e., atpA and nad6） displayed the polymorphisms in three lines, with the same patterns for lines S and H. For atpA, the EcoR I digestion revealed two fragments in the three lines. The 2.2 kb fragment was common to all three lines, while the second fragment was 3.2 kb in lines S and H, but 4.8 kb in line F. The atpA/PstI combination produced a 17.0 kb fragment in lines S and H, but a 10.2 kb fragment in line F. For nad6, one or two additional fragments of the same length were detected in lines S and H, while line F had more than two additional fragments. With coxIII as a probe, the EcoR I digestion resulted in a 2.5 kb fragment in three lines, as well as a 1.7 kb fragment in lines F and H, but not in line S. With the nad3 probe, the same patterns were observed for lines S and F, while the pattern for line H differed because of a lack of a 9.5 kb fragment. [Conclusion] Both atpA and nad6 are involved in regulating the development of CMS, while coxIII and nad3 may be regulated by nuclear genes to help restore fertility in cytoplasmic male sterile lines.