马铃薯StWRKY15在转基因烟草的耐寒性分析  

Analysis of Cold Tolerance of Potato StWRKY15 in Transgenic Tobacco

作　　者：冯月娟 杨振红 王芳[1,2,3,4,5] Feng Yuejuan;Yang Zhenhong;Wang Fang(College of Agriculture and Animal Husbandry,Qinghai University,Xining,810016;Academy of Agricultural and Forestry Sciences,Qinghai Uni-versity,Xining,810016;Engineering Research Center of Potato in Northwest Region Ministry of Education,Qinghai University,Xining,810016;Key Laboratory of Biotechnology on Qinghai-Tibet Plateau,Ministry of Education,Qinghai University,Xining,810016;Qinghai Key Laboratory of Potato Breeding,Qinghai Academy of Agriculture and Forestry Sciences,Xining,810016)

机构地区：[1]青海大学农牧学院,西宁810016 [2]青海大学农林科学院,西宁810016 [3]青海大学,西北马铃薯教育部工程研究中心,西宁810016 [4]青海大学,青藏高原生物技术教育部重点实验室,西宁810016 [5]青海省农林科学院,青海省马铃薯育种重点实验室,西宁810016

出　　处：《分子植物育种》2021年第21期7101-7112,共12页Molecular Plant Breeding

基　　金：青海省科技厅重点研发与转化计划项目(2020-HZ-806);财政部和农业农村部:国家现代农业产业技术体系项目(CARS-10)共同资助。

摘　　要：WRKY转录因子在植物的正常生长发育以及逆境生长过程中起着重要作用。本研究以4℃作为低温胁迫水平,对‘青薯9号’马铃薯组培苗进行胁迫处理。通过RT-qPCR技术对马铃薯的5个WRKY转录因子基因进行表达分析,筛选出对低温响应的基因,构建表达载体遗传转化烟草,初步分析基因的功能。结果显示,4℃低温胁迫下,马铃薯中的StWRKY15基因上调显著(P<0.05);STRING预测分析StWRKY15编码蛋白的互作蛋白,主要包括StWRKY72编码蛋白、StGRAS6编码蛋白、c2结构域QUIRKY蛋白;通过双酶切方法成功获得重组质粒PRI101-StWRKY15;冻融法介导转化烟草,过表达StWRKY15基因显著提高了转基因烟草的耐寒性。4℃低温胁迫下,StWRKY15基因的过表达明显增加了脯氨酸、可溶性糖及总蛋白含量,降低了丙二醛含量;胁迫10 d时,转基因烟草的长势优于野生型烟草。综上所述,可推断出StWRKY15对低温胁迫有响应,在逆境胁迫下可减轻植株受到的损伤,在一定范围内增强转基因烟草的耐寒性。本研究为进一步了解马铃薯WRKY转录因子的功能提供了理论依据。WRKY transcription factors play an important role on normal growth and development of plants and the growth process of adversity. In this study, 4 ℃ is employed as the low temperature stress level to stress the potato plantlets in vitro of ’Qingshu 9’. The expression analysis of 5 WRKY transcription factor genes of potato is executed by RT-qPCR technology, and the genes that are responsive to low temperature is detected. The expression vector was constructed and transformed into tobacco to analyze the function of the gene. The results indicated that under 4 ℃ low temperature stress, StWRKY15 gene in potato is up-regulated significantly(P<0.05). Using STRING predicte and analyze the interaction proteins of StWRKY15 encoding protein, mainly including StWRKY72 encoding protein, StGRAS6 encoding protein, and c2 domain QUIRKY protein;the recombinant plasmid PRI101-StWRKY15 was successfully obtained by double enzyme digestion method;The recombinant plasmid is transformed into tobacco by freeze-thaw method and overexpression StWRKY15 genes significantly improves the cold tolerance of transgenic modified tobacco. Under 4 ℃ low temperature stress, the overexpression of StWRKY15 gene significantly increased the content of proline, soluble sugar and total protein, and decreases the content of malondialdehyde;When the stress was 10 days, the growth of the transgenic tobacco is better than wild-type tobacco. In summary, it can be inferred that StWRKY15 responds to low temperature stress, and can reduce plant damage under adversity stress, and enhance the cold tolerance of transgenic tobacco within a certain range. This study provides a theoretical basis for further researching of the function of potato WRKY transcription factor.

关 键 词：马铃薯 WRKY转录因子 生物信息学分析 低温胁迫 

分 类 号：S532[农业科学—作物学]