| 小麦生理型与遗传型雄性不育相关基因锰超氧化物歧化酶(Mn-SOD)及果糖1,6-二磷酸醛缩酶(FBA)的表达分析 |
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| 引用本文： | 李莉,张改生,张龙雨,李亚鑫.小麦生理型与遗传型雄性不育相关基因锰超氧化物歧化酶(Mn-SOD)及果糖1,6-二磷酸醛缩酶(FBA)的表达分析[J].农业生物技术学报,2012,20(3):225-234. |
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| 作者姓名： | 李莉 张改生 张龙雨 李亚鑫 |
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| 作者单位： | 西北农林科技大学国家杨凌农业生物技术育种中心/国家小麦改良中心杨凌分中心/小麦育种教育部工程研究中心/陕西省作物杂种优势研究与利用重点实验室,杨凌,712100 |
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| 基金项目： | 国家高技术研究发展计划(863)重大专项(No.2011AA10A106);国家自然科学基金(No.31071477,31171611);高等学校博士学科点专项科研基金(No.20090204110024);陕西省"13115"科技创新工程重大科技专项(No.2010ZDKG-68) |
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| 摘 要： | 锰超氧化物歧化酶(Mn-SOD)是一种重要的抗氧化剂,果糖1,6-二磷酸醛缩酶(FBA)是叶绿体光合碳化阶段起重要调节作用的关键酶.本研究以小麦(Triticum aestivum L.)生理型与遗传型等基因雄性不育系及其对应育性正常的保持系为材料,选取花粉小孢子发育各时期的花药及三核期子房,对供试材料花药全蛋白表达差异研究的基础上,对雄性不育相关基因(Mn-SOD)及FBA进行核酸水平的实时荧光定量PCR分析,结果发现,与可育保持系相比,(1)生理型雄性不育系Mn-SOD基因在幼穗期、单核期和三核期表达量显著下调,而酶活力在幼穗期上调,在单核期下调；遗传型雄性不育系Mn-SOD基因在幼穗期和单核期表达量下调,酶活力在幼穗期上调,在二核期下调.(2)生理型雄性不育系和遗传型雄性不育系FBA基因表达量在幼穗期和单核期均下调,而对应同时期的FBA酶活力也下调,而遗传型不育系FBA基因在三核期表达量和FBA酶活力均上调.(3)Mn-SOD和FBA在遗传型雄性不育系三核期子房和花药中表达量均高于生理型雄性不育系和正常可育系,而在生理型雄性不育系花药中,Mn-SOD表达量明显低于对照可育系,在子房中其表达量略高于正常可育系.基因表达具有组织特异性,其蛋白表达较基因表达具有一定滞后性.Mn-SOD基因过量表达(单核至二核期),从而清除花药代谢紊乱产生的过多的活性氧,维持细胞正常功能；而花粉败育(生理型和遗传型雄性不育)导致花药失去活力,从而使花药叶绿体光合能力下降,FBA下调表达.研究结果提示,不同发育时期FBA及Mn-SOD酶活力变化与基因表达水平相一致,且这两个指标的变化直接或间接的影响了小麦花药的育性程度.
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| 关 键 词： | 小麦雄性不育 锰超氧化物歧化酶(Mn-SOD) 果糖1 6-二磷酸醛缩酶(FBA) 实时荧光定量 相对表达量 |
Analysis of Gene Expression for Manganese Superoxide Dismutase (Mn-SOD) and Fructose-l.6-bisphosphate Aldolase (FBA) in Genetic and Physiological Male Sterility Lines of Wheat |
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| LI Li
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ZHANG Gai-Sheng
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ZHANG Long-Yu
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LI Ya-Xin.Analysis of Gene Expression for Manganese Superoxide Dismutase (Mn-SOD) and Fructose-l.6-bisphosphate Aldolase (FBA) in Genetic and Physiological Male Sterility Lines of Wheat[J].Journal of Agricultural Biotechnology,2012,20(3):225-234. |
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| Authors: | LI Li ZHANG Gai-Sheng ZHANG Long-Yu LI Ya-Xin |
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| Institution: | LI Li ZHANG Gai-Sheng* ZHANG Long-Yu LI Ya-Xin Northwest Agriculture and Forestry University, State Yangling Agricultural Biotechnology & Breeding Center /Yangling Branch of State Wheat Improvement Centre/Wheat Breeding Engineering Research Center, Ministry of Education / Shaanxi Key Laboratory of Crop Heterosis, Yangling 712100, China |
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| Abstract: | Manganese superoxide dismutase(Mn-SOD) is an important anti-oxidant enzyme, which bind manganese functions primarily to protect mitochondrial components from superoxide liberated as a normal byproduct of respiration. Fructose-1,6-bisphosphate aldolase(ALD/FBA) plays a critical role in the photosynthesis in plants. The physiological male sterility, induced by chemical hybridize agent, was different with genetic male sterility which caused by it’s own genetic background of sterile cytoplasm. The wheat anthers and ovaries of genetic male sterility, normal fertility and physiological male sterility which induced by chemical hybridize agent SQ-1 were collected as materials for RNA extraction.Based on the differential expressed proteins identified by 2-DE technology, the Mn-SOD and FBA protein were significant and were analyzed by Real-time quantitive PCR. Compared to the normal fertility, the Mn-SOD was distinct down-regulated at young spike stage, uninucleate and trinucleate stages in physiological male sterility , while it’s enzyme activity was especially higher than that of control at young spike stage and significantly decreased at binucleate stage. For the genetic male sterility, Mn-SOD was down-regulated at young spike and uninucleate stages, and the enzyme activity was increased at young spike stage but declined at binucleate stage. The FBA was up-regulated in both genetic and physiological male sterility lines at the young spike and uninucleate stages compared with the control, accordingly, it’s enzyme activity was also fluctuated with the gene expression. Both the two genes were up-regulated in the ovary and anther of two male sterility lines at the trinucleate stage, and highest expressed in the genetic male sterility, but the expression of Mn-SOD in anther of physiological male sterility was lower than that of control. Mn-SOD and FBA genes were tissue-specific expressed in anther and ovary, and the protein abundance was hysteretic to gene expression. The Mn-SOD was over expressed during uninucleate and binucleate stages, which eliminated abundant active oxygen to keep the normal functions of cells. However, the abortion of anthers was also contributed to the decrease of photosynthesis capability, therefore, result in down-regulated expression of FBA.The study of the both enzyme activity and the gene expression level in different development stages among different male sterile lines contributed to the prefound learn about male sterility, and the consistence of this two index indicate a high relationship of male sterile and this two genes. |
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| Keywords: | Male sterility of wheat Real-time PCR Manganese superoxide dismutase(Mn-SOD) Fructose-1 6-bisphosphate aldolase(FBA) Relative expression |
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