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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。
In order to further reveal the molecular mechanism of thermo-sensitive genic male sterility in wheat (Triticum aestivum L.), wheat Bainong sterility (BNS) sterile lines and fertile anthers were used as materials. Chips technique was used to analyze the expression profiles of tetrad and dinuclear anther genes in sterile and fertile lines. Semi-quantitative RT-PCR was used to validate some fertile candidate genes with significant difference. The results showed that there were 418 effective transcripts differentially expressed between the two materials in the nucleus at 61 127 probes, of which 142 were significantly down-regulated and 276 were significantly up-regulated. The functional classification shows that these genes are mainly involved in important life processes such as metabolism, cytology, biological regulation and stimulus response. In order to verify the authenticity of the chip data, 10 differentially expressed leucine-rich repeats (Ta.17228.1), Ta.37113.2 (unknown), leucine-rich repeats ATP-binding cassette transporter (Ta.21809.1), peptidoglycan binding protein (Ta.22570.1), cytochrome P450 (Ta.19531.1), phosphoribulokinase phosphoribulose kinase, Ta. 18368.1), transcriptional regulator (Ta. 9239.1), peptidase C1A subfamily (Ta.37113.1), thionin super family (Ta.21983.1), V Type proton ATPase subunit D (Ta.18091.1). The results showed that the expression of the differentially expressed genes was consistent with that of the gene chip. Conserved domain analysis of these differentially expressed genes revealed that Ta.18091.1, Ta.21809.1 and Ta.19531.1 were most likely to be the key genes responsible for pollen abortion in BNS. The results provide important theoretical basis for cloning and verification of related genes, and provide the basic information for finding the key genes that cause the BNS abortion in wheat.