铁皮石斛DoglgC基因克隆及其功能分析  

Cloning of DoglgC from Dendrobium officinale and Functional Analysis

作　　者：杨婷舒 文国松[1] 赵明富[2] Yang Tingshu;Wen Guosong;Zhao Mingfu(Institute of Chinese Medicinal Materials,Yunnan Agricultural University,Kunming,650201;College of Plant Protection,Yunnan Agricultural University,Kunming,650201)

机构地区：[1]云南农业大学中药材研究所,昆明650201 [2]云南农业大学植物保护学院,昆明650201

出　　处：《分子植物育种》2022年第21期7046-7052,共7页Molecular Plant Breeding

基　　金：国家自然科学基金项目(81360611)资助。

摘　　要：本研究通过RT-PCR技术克隆铁皮石斛DoglgC基因以及构建表达载体,研究DoglgC对淀粉、蔗糖等合成的影响。结果表明,获得了1476 bp的开放阅读框;将铁皮石斛DoglgC与拟南芥大亚基ATglgC、水稻小亚基OTglgC和大肠杆菌ECglgC序列进行比对,结果表明序列高度保守。通过系统发育树分析发现DoglgC氨基酸序列和‘小兰屿蝴蝶兰’亲缘关系最近,属于类可逆糖基化多肽分支。利用DNA同源重组技术构建了表达载体p BI121s-DoglgC,用蘸花法将glgC基因转入野生拟南芥中,通过拟南芥植株表型观察发现,转DoglgC的拟南芥生长慢,叶片比野生型拟南芥的小。多糖含量的测定结果表明转DoglgC基因拟南芥多糖含量高于野生型拟南芥(WT),说明DoglgC基因能增加铁皮石斛多糖的合成。本研究验证了DoglgC基因对铁皮石斛多糖含量的影响,利于培育具有更高营养价值的作物。In this study,the DoglgC of Dendrobium officinale was cloned by RT-PCR and constructed the expression vector to study the effect of DoglgC on the synthesis of starch and sucrose.The results showed that the open reading frame of 1476 bp was obtained.The sequence of DoglgC of D.officinale was compared with that of ATglgC of Arabidopsis thaliana,OTglgC of rice small subunit and EcglgC of E.coli,and the results showed that the sequence is highly conserved.Phylogenetic tree analysis showed that the amino acid sequence of DoglgC is closely related to’Xiaolanyu’Phalaenopsis and belonged to the branch of reversibly glycosylated peptides.The expression vector pBI121s-DoglgC was constructed using DNA homologous recombination technology,and the glgC gene was transferred into the wild A.thaliana by the dipping flower method through the phenotype observation of Arabidopsis plants,it is found that the transformed DoglgC Arabidopsis grows slowly and the leaves are smaller than those of wild-type Arabidopsis.The results of determination of polysaccharide content showed that the polysaccharide content of transgenic A.thaliana was higher than that of wild-type Arabidopsis(WT),indicating that DoglgC can increase the synthesis of D.officinale polysaccharides.This study verified the effect of DoglgC on the polysaccharide content of D.officinale,help cultivating crops with higher nutritional value.

关 键 词：铁皮石斛 DoglgC 多糖 拟南芥 

分 类 号：S567.239[农业科学—中草药栽培]