泸定百合丝氨酸/苏氨酸蛋白激酶基因(LsS/TK)的克隆与表达分析  

Cloning and Expression Analysis of a Serine/Threonine Protein Kinase Gene(LsS/TK)in Lilium sargentiae

作　　者：马璐琳[1] 段青[1] 杜文文[1] 张艺萍[1] 崔光芬[1] 贾文杰[1] 吴学尉 王祥宁[1] 王继华[1] Ma Lulin;Duan Qing;Du Wenwen;Zhang Yiping;Cui Guangfen;Jia Wenjie;Wu Xuewei;Wang Xiangning;Wang Jihua(Yunnan Flower Breeding Key Laboratory,National Engineering Research Center for Ornamental Horticulture,Flower Research Institute of Yunnan Academy of Agricultural Sciences,Kunming,650205;School of Agriculture,Yunnan University,Kunming,650091)

机构地区：[1]云南省农业科学院花卉研究所,国家观赏园艺工程技术研究中心,云南省花卉育种重点实验室,昆明650205 [2]云南大学农学院,昆明650091

出　　处：《分子植物育种》2020年第18期5925-5932,共8页Molecular Plant Breeding

基　　金：国家重点研发计划(2019YFD1000400);云南省重点研发计划-农业领域(2018BB010);云南省科技领军人才计划(2016HA005);教育部省合建项目(2018BSHJ0108)共同资助。

摘　　要：本研究根据从之前已构建的百合尖孢镰刀菌(Fusarium oxysporum f.sp.lilii)诱导的泸定百合(Lilium sargentiae Wilson)SSH文库中筛选到的一个S/TK EST序列,通过RACE结合RT-PCR技术克隆到1个泸定百合S/TK基因全长cDNA序列,该基因全长2016 bp,开放阅读框长1440 bp,编码479个氨基酸,命名为LsS/TK。保守域结构分析表明Ls S/TK具有典型的S/TK激酶保守亚结构域。进化树分析结果显示LsS/TK与拟南芥的APTK19的进化关系最近,而与其它植物的S/TK的进化关系都较远。实时定量PCR结果显示百合镰刀菌诱导后LsS/TK表现上调表达,推测该基因可能在一定程度上参与泸定百合抗镰刀菌病害途径。In this study,applying the S/TK EST sequence identified in the(suppression subtractive hybridization)SSH library induced by Fusarium oxysporum f.sp.lilii,we cloned a novel S/TK gene from Lilium argentiae Wilson through RACE and RT-PCR.This S/TK gene was named as LsS/TK with a 2016 bp cDNA sequence,containing a1440 bp complete open reading frame(ORF)and encoding 478 amino acids.The analysis of conserved domain structure showed that LsS/TK protein harbored a typical S/TK kinase conserved subdomain.Phylogenetic analysis indicated that LsS/TK has the highest similarity with APTK19 from Arabidopsis thaliana,but has lower homologous to the S/TK from other plants.The RT-qPCR analysis showed that the LsS/TK was up-regulated after infected by F.oxysporum.It was speculated that the LsS/TK gene might play a certain role in the disease resistance reaction of Lily Fusarium wilt.

关 键 词：泸定百合(Lilium sargentiae Wilson) 丝氨酸/苏氨酸蛋白激酶 LsS/TK 

分 类 号：S682.29[农业科学—观赏园艺]