温敏核雄性不育小麦中不育系和可育系花药基因的表达谱分析

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摘要为进一步揭示小麦(Triticum aestivum L.)温敏核雄性不育的分子机理，本研究以小麦百农不育系(Bainong sterility, BNS)不育系和可育系花药为材料，利用Affymetrix 小麦基因芯片技术，分析其不育系和可育系中四分体和二核期花药基因的表达谱，并对部分差异显著的育性候选基因运用半定量RT-PCR技术进行验证。结果表明，在61 127个探针中，两材料间在二核期差异表达的有效转录本有418条，其中显著下调的转录本为142条，显著上调的转录本为276条。功能分类表明，这些基因主要参与代谢、细胞学过程、生物调节和刺激反应等重要生命过程。为验证芯片数据可信性，利用半定量RT-PCR法对10个差异表达显著的基因富含亮氨酸的重复序列基因(Leucine-rich repeats, Ta.17228.1)、Ta.37113.2(未知)、ATP结合盒转运子(ATP-binding cassette transporter, Ta.21809.1)、肽聚糖结合蛋白(peptidoglycan binding protein, Ta.22570.1)、细胞色素P450(cytochrome P450, Ta.19531.1)、磷酸核酮糖激酶(phosphoribulose kinase, Ta.18368.1)、转录调节因子(transcriptional regulator, Ta.9239.1)、肽酶C1A的亚家族(peptidase C1A subfamily, Ta.37113.1)、泛素家族(thionin super family, Ta.21983.1)、V型质子ATP酶亚基D(V-type proton ATPase subunit D, Ta.18091.1)进行验证，结果表明，差异基因的表达情况与基因芯片检测结果一致。对这些差异基因进行保守结构域分析发现，Ta.18091.1、Ta.21809.1和Ta.19531.1最有可能是引起BNS花粉败育的关键基因。该研究结果为相关基因克隆和验证提供重要的理论依据，进而为找到引起小麦BNS败育的关键基因提供基础资料。

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	白羿雄
	宋瑜龙
	牛艳波
	于永昂
	刘宏伟
	王小利
	马守才
	牛娜
	张改生
	马翎健
	张小红
	王军卫

关键词 ：小麦, 温敏核雄性不育, 百农不育系(BNS), 基因芯片, 半定量RT-PCR

Abstract：To reveal the molecular mechanism of the temperature/photoperiod sensitive male sterile lines of wheat (Triticum aestivum L.), the differently expressed genes in anthers at the tetrad and bicellular stage of the sterile and fertile lines of Bainong sterility (BNS) were analyzed using Affymetrix gene chip technology of wheat. The semi-quantitative RT-PCR was utilized to characterize some sterile candidate genes. The results showed that among the 61 127 probes transcripts, there were 418 valid transcripts presenting different expression between the sterile and fertile lines. Among these transcripts, 142 of them were down-regulated and 276 of them were up-regulated in the bicellular stage. Functional classification indicated that these genes participated in a series of significant life processes of metabolism process, cytological process, biological regulation and irritative reaction etc. Moreover, to verify chip data, 10 genes of leucine-rich repeats (Ta.17228.1), ATP-binding cassette transporter (Ta.21809.1), peptidoglycan binding protein (Ta.22570.1), Ta.37113.2(unknown), cytochrome P450 (Ta.19531.1), phosphoribulose kinase (Ta.18368.1), transcriptional regulator (Ta.9239.1), peptidase C1A subfamily (Ta.37113.1), thionin super family (Ta.21983.1) and V-type proton ATPase subunit D (Ta.18091.1), differentially expressed significantly, were identified by cDNA semi-quantitative RT-PCR. Result showed that the expression models of the 10 genes were concordant with the result of gene chip detection. Conservative structure domain analysis showed that Ta.18091.1, Ta.21809.1 and Ta.19531.1 most likely involved in the abortive anthers in the BNS sterile lines. The results provide important theoretical basis for sterility-related genes cloning and verification, and contribute to find the key gene which leads to pollen abortion in thermo-sensitive genic male sterile wheat (BNS).

Key words： Wheat Thermo-sensitive genic male sterility Bainong sterility (BNS) Gene chips Semi-quantitative RT-PCR

收稿日期: 2015-01-13 出版日期: 2015-06-01

基金资助:国家“863”计划重大专项;国家自然科学基金;陕西省“13115”科技创新工程重大科技专项

通讯作者: 王军卫 E-mail: wjw@nwsuaf.edu.cn

引用本文:

白羿雄宋瑜龙牛艳波于永昂刘宏伟王小利马守才牛娜张改生马翎健张小红王军卫. 温敏核雄性不育小麦中不育系和可育系花药基因的表达谱分析[J]. , 2015, 23(8): 1011-1019.

链接本文:

http://journal05.magtech.org.cn/Jwk_ny/CN/ 或 http://journal05.magtech.org.cn/Jwk_ny/CN/Y2015/V23/I8/1011

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