小麦光温敏雄性不育相关基因的DDRT-PCR分析及功能预测  

Analysis of Photoperiod-temperature Sensitive Male Sterility Related Genes in Wheat with DDRT-PCR

作　　者：赵昌平[1] 张立平[1] 李云伏[1] 马荣才[2] 单福华[1] 张风廷[1] 叶志杰[1] 秦娜[1] 

机构地区：[1]北京市农林科学院北京杂交小麦工程技术研究中心,北京100097 [2]北京市农林科学院北京生物工程技术研究中心,北京100097

出　　处：《中国生物化学与分子生物学报》2007年第1期56-62,共7页Chinese Journal of Biochemistry and Molecular Biology

基　　金：北京市自然科学基金(No.5010001);国家高技术研究发展计划(863计划)(No.2002AA207004);北京农业育种基础研究创新平台项目(No.YZPT01-04)资助~~

摘　　要：以小麦光温敏雄性不育系BS210为试材,分别在北京顺义(可育环境)和安徽阜阳(不育环境)两种诱导育性表现的生态环境下种植,并于雌雄蕊分化期、药隔形成期、花粉母细胞形成期、四分体期和单核期5个时期提取小穗组织的RNA,利用DDRT-PCR方法分离不育系在光、温因子诱导下的差异表达mRNA,并通过反向Northern进行验证.对获得的20条差异表达的EST进行了测序和BLAST分析,得到了4个候选基因的片段,对其进行5′Race扩增、序列分析及功能预测.结果表明它们分别与水稻的DNA修复重组蛋白基因rad50、小麦穗部表达的LRR重复序列型类受体激酶基因、玉米叶片坏死斑点L1s1基因的序列相似性分别为89%、89%和88%,另外还筛选到1个未知功能的新基因片段,它和rad50分别在可育和不育环境下表达的mRNA具有相同的5′端编码区,但3′端非编码区序列出现长度差异.研究结果为深入探讨小麦光温敏雄性不育机理提供了有意义的理论依据.The wheat BS210, a line identified as photoperiod-temperature sensitive genie male sterility, was grown in different ecological environment （Shunyi district in Beijing for fertile, and Fuyang in Anhui province for sterile ）. The total RNAs were isolated from the young fruiting spikes of plants in different developing period （stamens/pistil differentiation, anther chamber formation, pollen mother cell formation, tetrad phase, and mono-nucleate phase）, and the differential expression of the genes were analyzed with mRNA different display technique. 20 differential expression DNA bands were identified with reverse Northern hybridization, and the positive clones were sequenced. The 4 differential expression genes were selected as candidate genes through the BLAST research in GenBank, and therefore the 5′RACEs were conducted. The further homology research indicated that candidate genes A3, B2 and A2 were highly homologieal to Oryza sativa DNA repairrecombination protein （radSO）, Triticum aestivum receptor-like kinase with LRR repeats and Zea mays lethal leaf-spot 1 with 89% ,89% and 88% identity, respectively, while the candidate gene B1 was a novel gene. Interestingly, compared with themselves between the sterile and fertile conditions, high homology was observed in the 5′ region of mRNAs of B1 and A3, while the lengths of 3′ region of mRNAs are different. This study provided some useful information for further understanding the mechanism of photoperiod-temperature sensitive genie male sterility of wheat.

关 键 词：小麦 光温敏雄性不育 DDRT-PCR分析 不育相关基因片段 

分 类 号：Q55[生物学—生物化学]