茄子miR171b在抵抗黄萎病菌侵染中的功能  

Function of Eggplant(Solanum melongena)miR171b in Resistance to Verticillium dahliae Infection

作　　者：陈敏[1] 林元秘 朱文姣 杨清[1] CHEN Min;LIN Yuan-Mi;ZHU Wen-Jiao;YANG Qing(College of Life Science,Nanjing Agricultural University,Nanjing 210095,China)

机构地区：[1]南京农业大学生命科学学院,南京210095

出　　处：《农业生物技术学报》2022年第7期1268-1278,共11页Journal of Agricultural Biotechnology

基　　金：江苏省自然科学基金(BK20180519);江苏高校优势学科建设工程资助项目(130809001)。

摘　　要：microRNA171b(miR171b)是miR171家族的成员,在植物的生长发育、应答非生物胁迫和生物胁迫中发挥重要作用。为了揭示miR171b在茄子(Solanum melongena)应答黄萎病菌大丽轮枝菌(Verticillium dahliae)侵染过程中的功能,本研究以茄子栽培品种’苏琦1号’为实验材料,克隆pri-miR171b基因并构建表达载体,通过根癌农杆菌(Agrobacterium tumefaciens)介导的遗传转化系统构建miR171b转基因茄子植株,并对其抗黄萎病能力进行评估分析。抗病性分析表明,与对照相比,miR171b过表达株系对大丽轮枝菌侵染具有较强抗性,病情指数为18.5,大约是对照的1/3~1/2;而miR171b反义抑制株系对大丽轮枝菌侵染较为敏感,病情指数分别为68.4和72.6,是对照的1.5~1.6倍。病原菌内部转录间隔区(internal transcribed spacer,ITS)定量分析显示,miR171b过表达株系维管束组织中大丽轮枝菌的含量较对照显著降低,而miR171b反义抑制株系的菌含量显著增加(P<0.05)。抗氧化酶活性分析显示,miR171b过表达株系中超氧化物歧化酶(superoxide dismutase,SOD)、过氧化物酶(peroxidase,POD)和过氧化氢酶(catalase,CAT)酶活性高于对照株系,而反义抑制株系则低于对照。上述结果表明miR171b参与了茄子对黄萎病的防卫过程,并起着正调控作用。本研究为茄子抗黄萎病育种提供了理论依据。microRNA171b(miR171b)is a member of miR171 family and plays an important role in plant growth and development,response to abiotic and biological stresses.To reveal the function of miR171b in eggplant(Solanum melongena)response to Verticillium dahliae,using eggplant cultivar’Suqi No.1’as the test material in this study,the pri-miR171b gene was cloned and its expression vector was constructed.The miR171b transgenic eggplant plants were regenerated through Agrobacterium tumefaciens-mediated genetic transformation and analyzed their resistance to Verticillium wilt.The disease resistance analysis showed that the miR171b overexpression line was more resistant to the infection of V.dahliae with disease index of 18.5,which was about 1/3 to 1/2 of the control,while miR171b antisense inhibition lines were more sensible with disease index of 68.4 and 72.6,which were 1.5~1.6 times of the control.The quantitative analysis of pathogen internal transcribed spacer(ITS)showed that the content of V.dahliae in the vascular tissues of miR171b overexpression line was significantly lower than that in the control(P<0.05),and its content in miR171b antisense lines was significantly increased(P<0.05).Antioxidant enzyme activity analysis showed that the activities of superoxide dismutase(SOD),peroxidase(POD)and catalase(CAT)enzyme in miR171b overexpression line were higher than those in the control,while their activities were lower in the antisense lines than in the control.The above results showed that miR171b was involved in the defense process of eggplant against Verticillium wilt,and played a positive regulatory role.This study provides a theoretical basis for the breeding of eggplant cultivars with the Verticillium wilt resistance.

关 键 词：茄子 miR171b 黄萎病 功能分析 

分 类 号：S532[农业科学—作物学]