腰果AoCDPK26基因的克隆、亚细胞定位及表达特性分析  

Cloning,subcellular localization and expression characteristics analysis of AoCDPK26 gene in Anacardium occidentale

作　　者：秦灏 黄海杰[2] 黄伟坚[2] 张中润[2] 夏益华 赵丽 QIN Hao;HUANG Haijie;HUANG Weijian;ZHANG Zhongrun;XIA Yihua;ZHAO Li(College of Tropical Agriculture and Forestry/College of Agriculture and Rural Affairs/School of Rural Revitalization,Hainan University,Danzhou,Hainan,571737,China;Tropical Crops Genetic Resources Institute,Chinese Academy of Tropical Agricultural Sciences,Haikou,Hainan,571101,China;College of Science,Qiongtai Normal University/Tropical Biodiversity and Biotic Resource Utilization Laboratory,Qiongtai Normal University,Haikou,Hainan,571129,China)

机构地区：[1]海南大学热带农林学院/农业农村学院/乡村振兴学院,海南儋州571737 [2]中国热带农业科学院热带作物品种资源研究所,海口571101 [3]琼台师范学院理学院/热带生物多样性和生物资源利用实验室,海口571129

出　　处：《中国南方果树》2024年第6期47-56,67,共11页South China Fruits

基　　金：琼台师范学院高层次人才科研启动经费(30030007076);2024年农业农村部乐东腰果种质资源圃运行费(102125221630030009052)资助。

摘　　要：CDPK基因家族在植物非生物胁迫中发挥着重要作用,为了探究腰果Anacardium occidentale L.中AoCDPK26基因的功能和表达特性,以腰果叶片为试材,通过RT-PCR扩增技术,获得AoCDPK26基因序列,利用生物信息学分析方法对其理化性质、氨基酸序列、蛋白质结构进行初步分析预测,利用RT-qPCR技术测定其在腰果不同组织部位、低温和不同植物生长调节剂处理后的表达特性。在烟草中构建pEGAD-35S::AoCDPK26-GFP重组载体对AoCDPK26基因进行亚细胞定位。结果表明,腰果AoCDPK26基因编码区序列含有1692 bp的开放阅读框1个,编码氨基酸564个,系统进化树分析发现该序列与开心果具有较高的亲缘关系。AoCDPK26基因在腰果的根、茎、叶、花、果梨和果仁中均有表达,其中叶片中的表达量最高。在低温胁迫下,处理5 d后叶片AoCDPK26基因的表达量达到最高,是对照表达量的38倍;用茉莉酸甲酯(MeJA)处理腰果幼苗24 h后,叶片AoCDPK26基因表达量增幅最大。在烟草组织亚细胞定位发现,AoCDPK26定位于细胞膜和细胞核中。说明腰果AoCDPK26基因可能参与低温胁迫和内源激素响应的调控。The CDPK gene family plays an important role in anti-abiotic stress of plants.To investigate the function and expression characteristics of AoCDPK26 gene in cashew(A.occidentale),the leaves of cashew were used as experimental material,and the AoCDPK26 gene sequence was obtained by RT-PCR.The physicochemical properties,amino acid sequence and protein structure were preliminary predicted by bioinformatic analysis.The expression characteristics of AoCDPK26 gene in different tissues of cashew seedling,under different treatments with low temperature stress and different plant hormones were analyzed by Real-time fluorescence quantitative PCR.In addition,the recombinant vector PEGAD-35S::AOCDPK26-GFP was constructed and subcellular localization of AoCDPK26 gene was carried out in tobacco.The results indicated that the coding region of AoCDPK26 gene had one open reading frame of 1,692 bp nucleotides,encoding 564 amino acids,which shared a high similarity with Pistacia vera.The AoCDPK26 gene was expressed in the roots,stems,leaves,flowers,cashew apple and cashew nuts,with the highest expression in leaves.The expression of AoCDPK26 gene reached peak at 5 days after low temperature treatment,increased by 38 times compared to the control.The greatest increase in AoCDPK26 gene expression was observed in cashew leaves after 24 h treated with methyl jasmonate(MeJA).Subcellular localization analysis demonstrated that AoCDPK26 was localized in cell membrane and nucleus.It illustrated that the AoCDPK26 gene may be involved in the regulation of low temperature stress and hormone response in A.occidentale.

关 键 词：腰果 AoCDPK26 基因克隆 植物生长调节剂 表达量 亚细胞定位 

分 类 号：S667.9[农业科学—果树学]