棉花雄性不育细胞质对ATP和H2O2含量的影响分析

Analysis the effects of cotton male sterile cytoplasm on ATP and H2O2 contents

【目的】探究哈克尼西棉不育胞质(CMS-D2)和三裂棉不育胞质(CMS-D8)对ATP和H_(2)O_(2)含量的影响。【方法】选用CMS-D2和CMS-D8两套“三系”(不育系、保持系及其恢复系)材料,测定其不同发育时期的叶片和花蕾中ATP和H_(2)O_(2)含量;并结合已有研究测定候选不育基因orf610a超表达拟南芥植株中ATP和H_(2)O_(2)含量;通过实时荧光定量聚合酶链式反应分析ATP合成相关基因在不育系、保持系以及转基因拟南芥花蕾中的表达量。【结果】与其保持系和恢复系相比,CMS-D2不育系叶片在蕾期和铃期均表现为ATP含量极显著降低、H_(2)O_(2)含量极显著升高;CMS-D8不育系叶片在蕾期表现为ATP含量极显著降低、H_(2)O_(2)含量极显著升高,在铃期表现为ATP含量极显著升高;CMS-D2不育系3 mm、4 mm花蕾中ATP含量极显著降低,4 mm花蕾中H_(2)O_(2)含量极显著上升;CMS-D8不育系5 mm、6 mm花蕾中ATP含量极显著降低。与其保持系相比,CMS-D2和CMS-D8不育系花蕾中,多个ATP合成相关基因的表达量极显著降低。与野生型材料相比,orf610a超表达拟南芥角果和花蕾中的H_(2)O_(2)含量显著升高,ATP含量极显著降低,且大多数ATP合成相关基因均极显著下调表达。【结论】不育胞质会对棉花不同发育时期的叶片和花蕾中ATP和H_(2)O_(2)含量产生一定的影响,二者动态平衡的破坏可能会影响花粉的正常发育。

[Objective]This research aims to investigate the effects of male sterile cytoplasm of Gossypium harknessii(CMS-D2)and Gossypium trilobum(CMS-D8)on ATP and H_(2)O_(2) contents.[Method]The contents of ATP and H_(2)O_(2) in leaves and flower buds of CMS-D2 and CMS-D8"three lines"(sterile line,maintainer line,and restorer line)at different developmental stages were determined,and combined with the subject of existing research,ATP and H_(2)O_(2) contents in Arabidopsis plants overexpressing the sterile candidate gene orf610a were determined.The expression levels of ATP synthesis-related genes in the flower buds of sterile lines,maintainer lines and transgenic Arabidopsis were analyzed by quantitative real-time polymerase chain reaction(qRT-PCR).[Result]During the development of cotton,compared with its maintainer line and restorer line,the leaves of CMS-D2 sterile line showed an extremely significant decrease in ATP content and an extremely significant increase in H_(2)O_(2) content at both the bud stage and boll stage;the leaves of CMS-D8 sterile line showed an extremely significant decrease in ATP content and an extremely significant increase in H_(2)O_(2) content at the bud stage,and an extremely significant increase in ATP content at the boll stage.The ATP content in 3 mm and 4 mm flower buds of CMS-D2 sterile line was extremely significantly decreased,and the H_(2)O_(2) content in 4 mm flower buds was extremely significantly increased;the ATP contents in 5 mm and 6 mm flower buds of CMS-D8 sterile line were extremely significantly decreased.Compared with the maintainer lines,the expression levels of several ATP synthesis-related genes in the flower buds of the CMS-D2 and CMS-D8 sterile lines were extremely significantly reduced.Compared with wild-type Arabidopsis,the H_(2)O_(2) contents in siliques and flower buds of orf610a-overexpressed Arabidopsis were significantly increased,and the ATP content were extremely significantly decreased,and most ATP synthesis-related genes were extremely significantly down-regulated.[Conclusion]The male sterile cytoplasm have certain effects on the contents of ATP and H_(2)O_(2) in leaves and flower buds at different developmental stages of cotton,and the disruption of the dynamic balance may affect the development of pollen.