岷江百合类萌发素蛋白基因LrGLP2的克隆及表达特性分析  

Cloning and Expression Analysis of a Germin-Like Protein Gene from Lilium regale Wilson

作　　者：刘亚龙[1] 李红丽[1] 刘迪秋[1] 张南南[1] 何华[1] 葛锋[1] 陈朝银[1] 

机构地区：[1]昆明理工大学生命科学与技术学院,昆明650500

出　　处：《植物生理学报》2013年第10期1063-1070,共8页Plant Physiology Journal

基　　金：国家自然科学基金(31160401)

摘　　要：类萌发素蛋白是植物中普遍存在的一类可溶性糖蛋白,在植物抗逆胁迫中起着重要作用。依据岷江百合编码GLP的EST序列设计引物,采用快速扩增cDNA末端技术,从岷江百合(Lilium regale Wilson)中克隆得到一个新的GLP基因的全长cDNA序列,命名为LrGLP2。LrGLP2全长cDNA为921 bp,含有654 bp的开放阅读框,49 bp 5非编码区以及218 bp 3 UTR,编码217个氨基酸的蛋白质。LrGLP2编码蛋白质与已知植物GLPs家族成员间的同源性和聚类分析表明LrGLP2与来源于水稻(Oryza sativa)、节节麦(Aegilops tauschii)、葡萄(Vitis vinifera)中的GLPs具有较高的相似性。qRT-PCR分析显示,LrGLP2在岷江百合正常生长发育的根中有一定量的表达,而在茎和叶中几乎检测不到表达量。水杨酸、茉莉酸以及H2O2处理均不同程度抑制LrGLP2的转录水平,但乙烯处理能明显诱导LrGLP2的表达。此外,岷江百合接种尖孢镰刀菌(Fusarium oxysporum f.sp.lilii)后,LrGLP2在接种后2 h表达迅速上调,12 h表达量急剧上升,至24 h表达量达到最大值,之后表达量下降,可见LrGLP2参与岷江百合对尖孢镰刀菌的防卫反应。Germin-like proteins （GLPs） are a class of soluble glycoproteins which play important roles in plant defense responses. According to the EST sequence encoded the GLP, the gene-specific primers were designed to clone the full-length cDNA of a novel GLP gene from Lilium regale Wilson with the method of rapid amplification of cDNA ends. This novel gene was named as LrGLP2. LrGLP2 was 921 bp in length and contained an intact open reading frame （ORF） of 654 bp, a 5＇-untranslated region （UTR） of 49 bp, and 3＇-UTR of 218 bp, and the ORF encoded a protein with 217 amino acids. The LrGLP2 were homologous with the GLPs from Oryza sativa, Aegilops tauschii and Vitis vinifera according to the homology and phylogenetic analyses of the relationship of LrGLP2 with some known plant GLPs. qRT-PCR analysis indicated that the LrGLP2 was expressed at relatively high level in root, and almost no expression was detected in the stem and leaf. H202, jasmonic acid, and salicylic acid treatments inhibited the transcription level of LrGLP2 in different degree; however the expression of LrGLP2 was evidently induced by ethylene. Moreover, the expression of LrGLP2 in L. regale was quickly increased after inoculation with Fusarium oxysporum f. sp. lilii, and transcription level was rapidly up-regulated from 12 h to 24 h after inoculation with the highest expression level at 24 h, then its expression was declined after 24 h, which suggested that LrGLP2 was involved in defense response against the F. oxysporum f. sp. lilii.

关 键 词：岷江百合 类萌发素蛋白 基因克隆 尖孢镰刀菌 防卫反应 

分 类 号：Q943[生物学—植物学]