浙贝母SAI基因的克隆、生物信息学及表达分析  

Cloning,Bioinformatics and Expression Analysis of SAI Gene in Fritillaria thunbergii

作　　者：范小平 李梓铭 金泽兰 江建铭[2] 王志安[2] 王忠华[1] Fan Xiaoping;Li Ziming;Jin Zelan;Jiang Jianming;Wang Zhian;Wang Zhonghua(Institute of Biotechnology,Zhejiang Wanli University,Ningbo,315100;Zhejiang Institute of Chinese Meteria Medica,Hangzhou,310023)

机构地区：[1]浙江万里学院生物技术研究所,宁波315100 [2]浙江省中药研究所,杭州310023

出　　处：《分子植物育种》2024年第12期3895-3904,共10页Molecular Plant Breeding

基　　金：国家重点研发计划项目(2017YFC1700700);现代农业产业技术体系建设专项(CARS-21);浙江省中药材新品种选育重大科技专项(2016C02058);浙江省重点实验室建设项目(2011E10015);浙江省重点研发计划项目(2017C02011)共同资助。

摘　　要：为研究可溶性酸性转化酶(soluble acid invertase, SAI)基因在浙贝母生物合成中的作用机制,本研究采用RT-PCR技术首次从浙贝母(Fritillaria thunbergii)中克隆得到一个具有完整开放阅读框的可溶性酸性转化酶基因(FtSAI),该基因全长2 095 bp,共编码634个氨基酸,SAI蛋白序列与百合(Lilium davidii)、郁金香(Tulipa gesneriana)、甜蕉(Dessert banana)、枣椰子(Date palm)的相似性在76%以上。序列比对分析显示SAI是具有β-呋喃果糖苷酶活性位点WMN-DPN-G/A box、半胱氨(Cys) WECXDF和Glyco-32保守结构域。浙贝母SAI蛋白与同属百合科的郁金香(T. gesneriana CAA64953.1)、百合(L. davidii AGW23637.1)的SAI蛋白聚为一类。SAI基因在浙贝母各组织中均有表达,在鳞茎中表达量最低。SAI基因在鳞茎不同发育阶段表达也有显著差异,在鳞茎发育前中期SAI表达量处于较高水平,在后期表达量显著降低。该结果为进一步研究浙贝母SAI蛋白功能以及揭示该蛋白在浙贝母鳞茎淀粉-蔗糖合成代谢过程中发挥的作用提供了一定基础。To study the mechanism of soluble acid invertase(SAD)gene in the biosynthesis of Fritillaria thunbergi,a soluble acid invertase gene(FtSAI)with open reading frame was cloned from F.thunbergi by RT-PCR in this study.The full-length cDNA of SAI gene was 2095 bp,encoded 634 amino acids.The SAI protein sequence of F.thunbergi had more than 76%similarty with the L.davidi,T.gesneriana,dessert banana and date palm.Sequence alignment analysis showed that SAI had a β-furan fruit glycosidase active site WMN-DPN-G/A box,Cysteone(Cys)WECXDF and Glyco-32 conserved domains.The SAI protein of F.thunbergi was grouped together with that of T.gesneriana(CAA64953.1)and L.davidi(AGW23637.1),which belong to the same family of Liliaceae.The SAI gene was expressed in all tissues of F.thunbergi,and the expression level was the lowest in bulb.The expression of SAI gene was also significantly different at different developmental stages of bulb,and the expression level of SAI gene was at a high level in the early and middle stages of bulb development,while the expression level was significantly reduced in the late stage.The results lay a foundation for further study on the function of SAI protein in F.thunbergi and reveal the role of starch-sucrose synthesis in the scale of F.thunbergi.

关 键 词：浙贝母 可溶性酸性转化酶 生物信息学分析 表达分析 

分 类 号：S567.231[农业科学—中草药栽培]