中国南瓜CmoAux/IAA基因的克隆和表达特征分析  

Cloning and expression analysis of CmoAux/IAA gene in Cucurbita moschata

作　　者：任广乾[1] 刘正响 韩圆圆 陈碧华 李新峥 李庆飞 REN Guangqian;LIU Zhengxiang;HAN Yuanyuan;CHEN Bihua;LI Xinzheng;LI Qingfei(Xinxiang City Academy of Agricultural Sciences,Xinxiang 453003,Henan,China;College of Horticulture and Landscape,Henan Institute of Science and Technology,Xinxiang 453003,Henan,China)

机构地区：[1]河南省新乡市农业科学院,河南新乡453003 [2]河南科技学院园艺园林学院,河南新乡453003

出　　处：《中国瓜菜》2024年第10期30-37,共8页China Cucurbits And Vegetables

基　　金：国家自然科学基金(32302559);河南省科技攻关项目(242102110312);河南省高等学校青年骨干教师培养计划(2024GGJS102);河南省重大科技专项(241100110200)。

摘　　要：在前期对中国南瓜败育雌蕊和正常雌蕊进行转录组分析发现,CmoAux/IAA基因可能参与了雌花发育,为进一步探究其生物学功能,利用同源序列克隆法从中国南瓜中克隆得到全长为600 bp的CmoAux/IAA基因,该基因编码199个氨基酸;保守结构域分析结果表明,其属于Aux/IAA家族,包含结构域Ⅰ、Ⅲ、Ⅳ;该蛋白无信号肽和跨膜结构。多序列比对和系统进化树分析结果显示,CmoAux/IAA氨基酸序列与印度南瓜XP_022986212.1的氨基酸序列、美洲南瓜XP_023513171.1的氨基酸序列亲缘关系较近,相似度高达98.01%、96.50%。拟南芥原生质体亚细胞定位结果显示,CmoAux/IAA蛋白定位于细胞核和细胞质。实时荧光定量结果显示,该基因具有组织表达特异性,主要在南瓜植株生长点及雌花柱头中表达,柱头中的相对表达量是雄蕊的137倍,在叶和根中不表达,其可能在植株生长点幼嫩组织分化或雌花发育过程中发挥着重要作用。In the previous study,transcriptomic analysis of abortive pistil and normal pistil of Cucurbita moschata found that CmoAux/IAA gene might be involved in female flower development.In order to further explore its biological function,a 600 bp gene(CmoAux/IAA)was cloned from Cucurbita moschata by homologous sequence cloning method and it encodes 199 amino acids.The results of conserved domain analysis showed that it belongs to the Aux/IAA family and includes domainsⅠ,Ⅲ,andⅣ.The protein has no signal peptide and transmembrane structure.Multiple sequence alignment and phylogenetic tree results showed that the amino acid sequence of CmoAux/IAA were closely related to XP_022986212.1 of Cucurbita maxima and XP_023513171.1 of Cucurbita pepo,and the similarity was 98.01%and 96.50%,respectively.The results of subcellular localization of Arabidopsis protoplasts showed that the protein was located in nucleus and cytoplasm.Real-time fluorescence quantitative results showed that CmoAux/IAA has tissue expression specificity,mainly expressed in the growing points and stigma of female flowers in pumpkin plants,and the expression level of CmoAux/IAA in stigma was 137 fold that of in stamen,but not expressed in leaves and roots and it may play an important role in the differentiation of young tissue or the development of female flowers.

关 键 词：中国南瓜 AUX/IAA 基因克隆 生物信息学分析 亚细胞定位 

分 类 号：S642.1[农业科学—蔬菜学]