泸定百合泛素基因(Ls-Ub)的克隆及表达分析  

Cloning and Expression Analysis of the Ubiquitin(Ls-Ub) Gene from Lilium sargentiae

作　　者：马璐琳[1] 崔光芬[1] 贾文杰[1] 王祥宁[1] 段青[1] 杜文文[1] 王继华[1] Ma Lulin;Cui Guangfen;Jia Wenjie;Wang Xiangning;Duan Qing;Du Wenwen;Wang Jihua(Flower Research Institute of Yunnan Academy of Agricultural Sciences,Yunnan Flower Breeding Key Lab,National Engineering Research Center for Ornamental Horticulture,Kunming,65020)

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

出　　处：《分子植物育种》2018年第16期5216-5222,共7页Molecular Plant Breeding

基　　金：云南省科技创新人才计划-科技领军人才(2016HA005);云南省现代农业花卉苗木产业技术体系建设(2017K JTX0010);云南省重大科技专项(农业)(2016ZA005)共同资助

摘　　要：泛素(ubiquitin,Ub)与植物对外界的多种胁迫反应相关。为了研究百合中的泛素基因与百合抗镰刀菌病害的关系,本研究根据百合尖孢镰刀菌(Fusarium oxysporum f.sp.lilii)诱导的泸定百合(Lilium sargentiae Wilson)SSH文库中筛选到的Ub EST序列,通过RACE结合RT-PCR技术从泸定百合中克隆到1个Ub基因全长c DNA序列,该基因全长996 bp,包含一个长690 bp编码229个氨基酸的开放阅读框,命名为Ls-Ub(Gen Bank登录号:KY996308)。序列比对分析结果表明Ls-Ub与其它物种Ub基因编码的氨基酸序列具有高度的相似性。Q-PCR结果显示百合镰刀菌诱导后Ls-Ub强烈上调表达,且明显高于水处理对照的表达水平。推测该基因可能与百合抗镰刀菌病害相关。本研究将为揭示百合抗镰刀菌枯萎病的分子机制提供一定理论依据。Ubiquitin（Ub） is correlated with the response of plant to various stresses outside. To investigate the relationship between the Ub gene and disease resistance to Fusarium wilt in lily, a full-length c DNA sequence of an Ub gene was cloned from Lilium sargentiae by using RACE and RT-PCR, according to the screened Ub EST sequence in the（suppression subtractive hybridization） SSH library of Fusarium sargentiae induced by Fusarium oxysporum f. sp. lilii. The full length of the Ub c DNA sequence was 996 bp with a 690 bp open reading frame（ORF）which encoded 229 amino acids, and the gene was named Ls-Ub（Gen Bank No.： KY996308）. Sequence alignment analysis indicated that the amino acid sequence encoded by Ls-Ub was highly homologous with that by Ub gene of other plant species. Q-PCR analysis showed that the Ls-Ub expression was strongly up-regulated after induced by F. oxysporum which was significantly higher than the expression of the water treatment control. It was speculated that the Ls-Ub gene might play a certain role in the disease resistance to Lily Fusarium wilt. The research might provide some valuable information for further disclosing the molecular mechanism of resistance to Lily Fusarium wilt.

关 键 词：泸定百合 百合尖孢镰刀菌 过氧化氢酶基因 基因表达 

分 类 号：S436.8[农业科学—农业昆虫与害虫防治]