StWIP2基因调控马铃薯对晚疫病的抗性  

StWIP2 Regulates the Resistance to Phytophthora infestans in Potato(Solanum tuberosum)

作　　者：朱迪 周云[1,2,3] 杨生龙 孙海宏 贺苗苗[1,2,3] ZHU DiZHOU Yun;YANG Sheng-Long;SUN Hai-Hong;HE Miao-Miao(Academy of Agriculture and Forestry Sciences,Qinghai University,Xining 810016,China;Key Laboratory of Qinghai-Tibetan Plateau Biotechnology(Qinghai University),Ministry of Education,Xining 810016,China;Key Laboratory of Potato Breeding,Qinghai Province/Ministry of Education,Engineering Research Center of Potato in Northwest Region,Xining 810016,China)

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

出　　处：《农业生物技术学报》2024年第11期2484-2497,共14页Journal of Agricultural Biotechnology

基　　金：青海省科技厅基础研究计划项目(2022-ZJ-712);国家马铃薯产业技术体系(CARS-09)。

摘　　要：伤害诱导蛋白(wound-induced proteins,WIPs)在植物应答病原菌侵染的防御反应中发挥了重要作用。本研究从马铃薯(Solanum tuberosum)'青薯9号'中克隆了StWIP2基因(GenBank No.NM_001288699.1),并对StWIP2基因进行生物信息学分析,同时构建StWIP2基因过表达载体和沉默载体,运用反向遗传学手段对StWIP2基因在与晚疫病互作中的功能进行鉴定。生物信息学分析发现,StWIP2基因的CDS全长为636 bp,共编码211个氨基酸。qPCR分析发现,StWIP2基因在'春薯5号'中表达量最高,在二倍体马铃薯DM中表达量最低。在水杨酸(salicylic acid,SA)、脱落酸(abscisic acid,ABA)、茉莉酸甲酯(methyl jasmonate,MeJA)和晚疫病菌胁迫处理下,StWIP2基因均受到调控。过表达StWIP2能够提高植株H_(2)O_(2)和丙二醛(malondialdehyde,MDA)的含量、抗氧化酶的活性及相关防卫基因病程相关蛋白1(pathogenesis related protein 1,StPR1)、StPR5和乙烯响应因子(ethylene responsive factor 1,StERF1)的表达水平来推迟植株发病的时间,提高植株对晚疫病的抗性水平。沉默StWIP2基因后,植株H_(2)O_(2)和MDA的含量降低,抗氧化酶活性降低;防卫基因植物防御素基因1.2(plant defensin gene 1.2,PDF1.2)、StERF1、过氧化物酶基因(peroxidase,StPOD)和苯丙氨酸解氨酶(phenylalanine ammonialyase,StPAL)的表达量降低,StPR1和StPR5的表达受到抑制,植株对晚疫病的抗性水平下降。上述结果表明,StWIP2基因在与晚疫病互作的过程中起到正调控的作用,StWIP2基因可以作为马铃薯抗病育种的候选基因资源加以利用。Wound-induced proteins(WIPs)play an essential role in plants defence responses to pathogen infestation.In this study,the StWIP2 gene(GenBank No.NM_001288699.1)was cloned from potato(Solanum tuberosum)'Qingshu 9',and bioinformatics analyses of the StWIP2 gene were carried out,while overexpression vectors and silencing vectors were constructed,and reverse genetics was used to characterise the function of the StWIP2 gene in the interaction with Phytophthora infestans.Bioinformatics revealed that the StWIP2 gene had a full CDS length of 636 bp and encoded a total of 211 amino acids.qPCR analysis revealed that the StWIP2 gene had the highest expression in'Chunshu 5'and the lowest expression in diploid potato DM.The StWIP2 gene was responsive and differentially expressed under treatments of P.infestans,salicylic acid(SA),abscisic acid(ABA),and methyl jasmonate(MeJA).Overexpression of StWIP2 was able to increase the content of H_(2)O_(2)and malondialdehyde(MDA),the activity of antioxidant enzymes,and the expression levels of pathogenesis related protein 1(StPR1),StPR5 and ethylene responsive factor 1(StERF1),to delay the onset of the disease in the plants and to increase the level of plant resistance to P.infestans.After silencing the StWIP2 gene,the content of H_(2)O_(2)and MDA in plants decreased,and the activity of antioxidant enzymes decreased;The expression levels of plant defense gene 1.2(PDF1.2),StERF1,peroxidase gene(StPOD),and phenylalanine ammonia lyase(StPAL)were reduced,while the expression of StPR1 and StPR5 were inhibited,resulting in a decrease in plant resistance to late blight.The above results indicate that the StWIP2 gene plays a positive regulatory role in the interaction with P.infestans,and the StWIP2 gene can be utilized as a candidate genetic resource for disease resistance breeding in potatoes.

关 键 词：马铃薯 致病疫霉 伤害诱导蛋白基因(StWIP2) 抗病性鉴定 抗氧化酶 

分 类 号：S532[农业科学—作物学]