摘要
DNA甲基化对植物的生长发育至关重要,DNA甲基转移酶建立和维持着DNA甲基化,其中结构域重排甲基转移酶DRM2催化非对称位点的从头甲基化。为了初步探究马铃薯StDRM2基因的功能,本研究利用同源克隆基因技术从马铃薯‘青薯9号’中克隆了StDRM2基因,利用Gateway技术构建过表达载体,农杆菌遗传转化马铃薯‘大西洋’,获得5株转基因马铃薯苗。利用qRT-PCR技术测定StDRM2基因在转基因马铃薯及其不同组织(根,茎,叶)中的表达量。结果表明,StDRM2基因CDS序列全长1812 bp,编码603个蛋白质氨基酸。系统发育进化树分析表明,StDRM2与茄科作物的番茄亲缘关系最近;qRT-PCR表明转基因马铃薯中的StDRM2基因表达量显著上调,最高相对表达量是野生型马铃薯的6.77倍。StDRM2基因在转基因马铃薯根、茎、叶中均有表达,其表达量不同且差异显著(P<0.05),其中根中的表达量最高,茎中的表达量最低。本研究中转基因马铃薯的获得为研究StDRM2功能提供试验材料,组织表达模式分析为进一步了解StDRM2基因功能和作用机制提供理论依据。
DNA methylation is essential for plant growth and development,and DNA methyltransferase establishes and maintains DNA methylation.The domain rearrangement of the methyltransferase DRM2 catalyzes de novo methylation of asymmetric sites.In order to preliminarily explore the function of the potato StDRM2 gene.In this study,using the homologous gene cloning technology to clone the StDRM2 gene from potato’Qingshu9’,and the overexpression vector was constructed by using Gateway technology.The transformation of potato’Atlantic’by Agrobacterium tumefaciens,which obtained 5 transgenic potato plants.The qRT-PCR technique was used to determine expression of StDRM2 gene in transgenic potato and its different tissues(root,stem,leaf).The results showed that the full-length CDS sequence of StDRM2 gene was 1812 bp,encoding 603 protein amino acid residues.Phylogenetic tree analysis showed that StDRM2 was closely related to tomato of Solanaceae crops,qRT-PCR showed that the expression of StDRM2 gene was significantly up-regulated of transgenic potatoes,and the highest relative expression was 6.77 times that of wild-type potatoes.And StDRM2 gene expressed in roots,stems and leaves of transgenic potato,which were expressed in a different expression level and there was significant difference(P<0.05),with the expression of root was the highest and stem was the lowest.The acquisition of transgenic potatoes in this study provides experimental materials for studying the function of StDRM2,and the analysis of tissues expression patterns provides a theoretical basis on further understanding of the function and mechanism of StDRM2 gene.
作者
高彦妮
王日寰
王芳
王舰
Gao Yanni;Wang Rihuan;Wang Fang;Wang Jian(Qinghai University,Xining,810016;Qinghai Academy of Agriculture and Forestry Sciences,Xining,810016;Key Laboratory of Potato Breeding of Qinghai,Qinghai Academy of Agriculture and Forestry Sciences,Xining,810016;Key Laboratory of Qinghai-Tibetan Plateau Biotechnology of Ministry of Education,Qinghai Universit,Xining,810016;State Key Laboratory of Plateau Ecology and Agriculture,Qinghai Universiy,Xining,810016)
出处
《分子植物育种》
CAS
北大核心
2021年第6期1778-1786,共9页
Molecular Plant Breeding
基金
现代农业产业技术体系(CARS-9)
专用型马铃薯产业高质量发展关键技术研发与示范(2019-NK-A1)共同资助。
