克隆普通小麦3个基因组供体种的穗发芽抗性相关基因AIP2(ABI3interacting protein 2),预测、分析其功能,阐明其保守区域在不同材料间的变异,为其进化研究提供理论依据,同时为普通小麦AIP2基因的研究以及利用该基因改良穗发芽性状奠定基...克隆普通小麦3个基因组供体种的穗发芽抗性相关基因AIP2(ABI3interacting protein 2),预测、分析其功能,阐明其保守区域在不同材料间的变异,为其进化研究提供理论依据,同时为普通小麦AIP2基因的研究以及利用该基因改良穗发芽性状奠定基础。以野生一粒小麦(Triticum boeoticum,AA,2n=14)、拟斯卑尔脱山羊草(Aegilops speltoides,SS,2n=14)、节节麦(Aegilops tauschii,DD,2n=14)为材料,采用同源克隆的方法克隆AIP2基因,利用生物信息学软件分析其基因结构及功能,并进行遗传进化分析。结果表明,从小麦3个供体种材料中成功克隆AIP2基因,该基因开放阅读框非常保守,均为969bp,编码323个氨基酸残基。3个供体材料间AIP2氨基酸序列仅存在7个氨基酸残基的差异,结构域预测结果显示,AIP2蛋白含有锌指环家族典型的锌指环结构域,进化分析发现,AIP2基因在植物中非常保守,推测该基因在禾谷类作物中可能具有相似的功能。展开更多
Pre‐harvest sprouting（PHS） seriously affects wheat yield and quality of the grain. ABI3 is a key factor in the activation of seed development and repression of germination in Arabidopsis. An ABI3‐interacting prot...Pre‐harvest sprouting（PHS） seriously affects wheat yield and quality of the grain. ABI3 is a key factor in the activation of seed development and repression of germination in Arabidopsis. An ABI3‐interacting protein（AIP2） could polyubiquitinate ABI3, impair seed dormancy and promote seed germination in Arabidopsis. In this study,two wheat AIP2 genes, TaAIP2A and TaAIP2B, were isolated.Subcellular localization assay and yeast two‐hybrid analysis revealed that TaAIP2A and TaAIP2B may function through interaction with wheat Viviporous‐1（TaVp1）. The transcripts TaAIP2A and TaAIP2B were more abundant in wheat PHS susceptible cultivars than that of resistant ones, and decreased gradually following seed development. Expression of TaAIP2A and TaAIP2B in Arabidopsis aip2‐1 mutant lines resulted in earlier flowering, promotion of seed germination,and reduced ABA sensitivity, respectively, somehow mimicking the phenotype of the wild type, with TaAIP2B having a stronger role in these aspects. Furthermore, the expression ofupstream genes ABI1 and ABI2 were upregulated, whereas that of downstream genes ABI3 and ABI5 were downregulated in both TaAIP2A and TaAIP2B complemented lines upon ABA treatment. These results suggested that wheat AIP2s could negatively regulate the ABA signaling pathway and play important roles in seed germination, and thus wheat PHS resistance finally.展开更多
基金partly funded by the China National Basic Research Program supported by the Chinese Ministry of Science and Technology(2009CB118300)
文摘Pre‐harvest sprouting（PHS） seriously affects wheat yield and quality of the grain. ABI3 is a key factor in the activation of seed development and repression of germination in Arabidopsis. An ABI3‐interacting protein（AIP2） could polyubiquitinate ABI3, impair seed dormancy and promote seed germination in Arabidopsis. In this study,two wheat AIP2 genes, TaAIP2A and TaAIP2B, were isolated.Subcellular localization assay and yeast two‐hybrid analysis revealed that TaAIP2A and TaAIP2B may function through interaction with wheat Viviporous‐1（TaVp1）. The transcripts TaAIP2A and TaAIP2B were more abundant in wheat PHS susceptible cultivars than that of resistant ones, and decreased gradually following seed development. Expression of TaAIP2A and TaAIP2B in Arabidopsis aip2‐1 mutant lines resulted in earlier flowering, promotion of seed germination,and reduced ABA sensitivity, respectively, somehow mimicking the phenotype of the wild type, with TaAIP2B having a stronger role in these aspects. Furthermore, the expression ofupstream genes ABI1 and ABI2 were upregulated, whereas that of downstream genes ABI3 and ABI5 were downregulated in both TaAIP2A and TaAIP2B complemented lines upon ABA treatment. These results suggested that wheat AIP2s could negatively regulate the ABA signaling pathway and play important roles in seed germination, and thus wheat PHS resistance finally.
