枣ATP1基因克隆及其在雄性可育和不育品种中的表达分析

以枣树雄性不育材料JMS1及雄性可育品种‘月光’及花粉中度可育品种‘冬枣’为试材,克隆ATP1基因,并比较其在枣雄性可育和不育材料之间的表达差异。枣ATP1基因包含完整开放阅读框为1 530bp,其编码蛋白与苹果和西瓜同源蛋白相似性系数分别达95.88%和98.62%。ATP1基因在3个品种中不存在碱基差异,且确定该基因为线粒体基因。利用实时荧光定量PCR技术检测ATP1基因组织特异表达发现,JMS1的嫩枝、老枝、叶、蕾中ATP1基因的表达量较高,但在花、花药、花萼、子房中,JMS1的ATP1基因表达量均低于‘月光’。在单花发育前期JMS1中ATP1表达量高于‘月光’和‘冬枣’,在单花发育后期其表达量低于‘月光’,但高于‘冬枣’。枣花蕾发育早期ATP1基因异常高表达可能影响了ATP合酶的合成,可能与JMS1花粉完全败育的主要原因有关。

ATP1 gene cloning and its expression in male-sterile and male-fertile cultivars of Chinese jujube(Ziziphus jujuba Mill.)

ATP1 gene was isolated and its expression difference in the male-fertile and male-sterile germplasm was analyzed in this study,using a male-sterile germplasm JMS1,male-fertile cultivar ‘Yueguang’ and ‘Dongzao’ with middle fertility of pollen.ATP1 gene contained 1 530bp open reading frame,with a high degree of homology with ATP1 in Malus domestica (95.88％)and Citrullus lanatus(98.62％).ATP1 was mitochondrial gene,which did not show sequence differences among three cultivars.Tissue-specific expression analysis by qRT-PCR showed that the expression of ATP1 in young branches,old branches,leaves and buds of JMS1 was higher,yet it was lower in flower,anther,calyx,ovary of JMS1 than that of ‘Yueguang’.The expression of ATP1 gene kept higher level in JMS1 than ‘Yueguang’ and ‘Dongzao’ at the early development stage of flower.In contrast,the gene expression at the late development stage of flower in JMS1 was lower than ‘Yueguang’ and still higher than ‘Dongzao’.The severe expression of ATP1 gene influenced ATP synthase production during early anther development stage and was probably associated with complete pollen abortion in JMS1.