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为揭示YS型小麦温敏雄性不育的育性转换基础,以该类型不育系A3017不育幼穗和可育幼穗为材料构建正、反杂交SSH-cDNA文库,从可育文库中筛选出一个与G3BP(Ras-GTPase activating protein SH3 domain-binding protein)基因同源的EST序列(GenBank登录号为DY543200)。以该EST序列的同源性比对和拼接结果为依据设计引物,在可育幼穗中扩增出一条1230bp的cDNA序列。该片段含有与G3BP相似的由409个氨基酸组成的结构域,与水稻G3BP的氨基酸序列同源性为79%,被命名为TaG3BP(GenBank登录号为GU475149)。利用Real-time PCR检测TaG3BP基因在YS型小麦温敏雄性不育系花药发育各时期中的表达模式,发现该基因在育性转换的关键时期上调表达,且可育条件下的表达量高于同时期不育条件下的表达量。进一步对TaG3BP在3种不同类型的小麦K型雄性不育材料的不育系及其保持系的幼穗中的表达模式进行半定量RT-PCR分析,结果该基因在保持系幼穗中表达量较高,在不育系中表达量较低。表明该基因可能在其育性转换中具有重要作用。
To reveal the basis of fertility transformation in thermo-sensitive male-sterile wheat (YS) -type wheat, SSH-cDNA libraries of both positive and negative hybrids were constructed from male sterile spikelets and fertile young spikes of this type of male sterile line and screened from fertile An EST sequence homologous to the G3BP (GenBank accession number DY543200) was generated. Primers were designed based on the homology alignment and splicing results of the EST sequences, and a 1230 bp cDNA sequence was amplified from fertile young spikes. The fragment contains a 409 amino acid domain similar to G3BP. The fragment has 79% amino acid sequence identity with rice G3BP and is named TaG3BP (GenBank accession number GU475149). Real-time PCR was used to detect the expression pattern of TaG3BP gene in different stages of anther development in YS-type thermo-sensitive male-sterile wheat. It was found that the expression level of TaG3BP gene was up-regulated during the critical period of fertility transformation, The same period of infertility expression. The semi-quantitative RT-PCR analysis of the expression pattern of TaG3BP in the young panicles of three different types of male sterile genotypes of wheat K-type male sterile lines and their maintainer lines showed that the expression level of TaG3BP in young panicles of maintainer lines Higher, lower in CMS lines. Suggesting that this gene may play an important role in its fertility transformation.