黄秋葵八氢番茄红素合成酶PSY基因的克隆与分析  

Cloning and Expression Analysis of a Phytoenesynthase Gene in Hibiscus esculentus L.

作　　者：李永平 陈敏氡[1,2,3,4] 朱海生 温庆放[1,2,3,4] 刘建汀 林思锻[5] LI Yongping;CHEN Mindong;ZHU Haisheng;WEN Qingfang;LIU Jianting;LIN Siduan(Fujian Key Laboratory of Vegetable Genetics and Breeding,Fuzhou 350013,China;Crops Research Institute,Fujian Academy of Agricultural Sciences,Fuzhou 350013,China;Vegetable Research Center,Fujian Academy of Agricultural Sciences,Fuzhou 350013,China;Fujian Engineering Research Center for Vegetables,Fuzhou 350013,China;Institute of Agricultural Science,Luojiang District,Quanzhou 362011,China)

机构地区：[1]福建省蔬菜遗传育种重点实验室,福州350013 [2]福建省农业科学院作物研究所,福州350013 [3]福建省农业科学院蔬菜研究中心,福州350013 [4]福建省蔬菜工程技术研究中心,福州350013 [5]泉州市洛江区农业科学研究所,泉州362011

出　　处：《中国细胞生物学学报》2019年第10期1938-1947,共10页Chinese Journal of Cell Biology

基　　金：福建省属公益类科研院所基本科研专项(批准号:2019R1031-4);福建省重大专项专题(批准号:2018NZ0002-3);福建省农业科学院蔬菜科技创新团队(批准号:STIT2017-1-2);国家特色蔬菜产业技术体系项目(批准号:CARS-24-G-07)资助的课题~~

摘　　要：该研究根据黄秋葵(Hibiscus esculentus L.)转录组测序获得的八氢番茄红素合成酶PSY基因序列,设计引物克隆验证得到一条全长为1399 bp的cDNA,开放阅读框(ORF)包含1155个碱基;预测编码384个氨基酸,理论分子量为43.45 kDa,等电点(pI)为8.729;编码的蛋白与亚洲棉(Gossypium arboreum)、陆地棉(Gossypium hirsutum)、雷蒙德氏棉(Gossypium raimondii)、海岛棉(Gossypium barbadense)、可可(Theobroma cacao)同源蛋白的相似性均在90%以上,显示黄秋葵PSY基因具高度的保守性,将基因命名为HePSY,GeneBank登录号为:MG372369。通过荧光定量PCR分析表明,HePSY基因在黄秋葵根、茎、叶、花和果荚中均有表达;叶片生长中以成熟叶中表达最高,果实发育中花后4天表达量高。类胡萝卜素含量以成熟叶的最高,果实以花后4天含量最高,与HePSY基因的表达呈正相关。该研究揭示了PSY基因表达和类胡萝卜素积累的特性,为开展黄秋葵类胡萝卜素分子调控机制研究奠定了基础。In this study,a PSY gene was cloned from Hibiscus esculentus L.by RT-PCR and RACE technique.The full-length cDNA sequence of PSY was 1399 bp,which contained a 1155 bp open reading frame(ORF)that encoded 384 amino acids with a predicted molecular weight of 43.45 kDa and a hypothetical isoelectric point(pI)of 8.729.The encoded protein had more than 90%homology with Gossypium arboretum,Gossypium hirsutum,Gossypium raimondii,Gossypium barbadense,and Theobroma cacao,which indicated that PSY gene of Okra was highly conserved.The gene was named HePSY and the GeneBank accession number was MG372369.Real-time PCR analysis revealed that HePSY could be expressed in different tissues of Hibiscus esculentus L.,including roots,stems,leaves,flowers and fruits,and the level was the highest in mature leaves and 4 days after flowering during the process of leaf and fruit development,respectively.Carotenoid contents was explored.Hibiscus esculentus L.carotenoid was also the highest in mature leaves and 4 days after flowering during the process of leaf and fruit development respectively,which positive correlation with HePSY gene expression.These results revealed the expression of HePSY gene and the accumulation characteristics of carotenoid contents,and provided a base for the study of carotenoid molecular regulation mechanism in Hibiscus esculentus L.

关 键 词：黄秋葵 八氢番茄红素合成酶 类胡萝卜素 

分 类 号：S649[农业科学—蔬菜学]