摘要
该实验采用定量逆转录PCR(Quantitative reverse transcription-PCR)方法分析三七[Panax notoginseng(Burk)F.H.Chen]病程相关蛋白(pathogensis related protein,PR)基因PnPR1的表达谱,并构建PnPR1的植物超表达载体,通过根癌农杆菌介导转入烟草(Nicotiana tabacum)中过量表达。结果表明:(1)外源茉莉酸甲酯预处理三七根部,能显著提高茄腐镰刀菌(Fusarium solani)侵染过程中PnPR1基因的表达量。(2)4种信号分子茉莉酸甲酯、水杨酸、过氧化氢和乙烯利处理均能够不同程度地提高PnPR1基因在三七根中的表达水平,但3种信号分子抑制剂处理三七根部后PnPR1的表达量下调。(3)PnPR1在T_(2)代转基因烟草中稳定表达,并且转基因烟草株系对茄腐镰刀菌的抗性显著提高。研究认为,PnPR1基因在转录水平上响应茄腐镰刀菌的侵染,并受茉莉酸甲酯等信号分子的诱导,在烟草中过表达PnPR1基因可增强对茄腐镰刀菌的抗性,表明PnPR1是三七应对根腐病菌防卫反应中的抗病基因。
In this study,we used quantitative reverse transcription-PCR to analyze the expression level of PnPR1,a pathogenesis-related protein gene of Panax notoginseng.In addition,the overexpression vector of pCAMBIA2300s-PnPR1 was constructed and then introduced into tobacco(Nicotiana tabacum)with Agrobacterium tumefaciens mediated method.The results showed that:(1)the methyl jasmonate pretreatment of P.notoginseng roots greatly up-regulated the PnPR1 expression during Fusarium solani infection.(2)The expression level of PnPR1 gene was induced by the treatment of four signal molecules(methyl jasmonate,ethephon,salicylic acid,and hydrogen peroxide)to varying degrees.The expression of PnPR1 was down-regulated by three kinds of signal molecule inhibitors.(3)PnPR1 was stably expressed in T_(2) transgenic tobacco,and the resistance of transgenic tobacco lines to F.solani was significantly improved.In conclusion,the PnPR1 gene responds to the infection of F.solani at transcription level and was induced by signal molecules such as methyl jasmonate.Overexpression of PnPR1 in tobacco lines enhanced the resistance to F.solani,indicating that PnPR1 is a disease-resistance gene involved in the defense responses of P.notoginseng to F.solani.
作者
张应鹏
陈虹均
郑锂蕾
王自娥
崔秀明
葛锋
刘迪秋
ZHANG Yingpeng;CHEN Hongjun;ZHENG Lilei;WANG Zi'e;CUI Xiuming;GE Feng;LIU Diqiu(Kunming University of Science and Technology, Faculty of Life Science and Technology, Kunming 650500, China;2Key Laboratory of Panax notoginseng Resources Sustainable Development and Utilization, Kunming 650500, China)
出处
《西北植物学报》
CAS
CSCD
北大核心
2020年第12期2000-2007,共8页
Acta Botanica Boreali-Occidentalia Sinica
基金
国家自然科学基金(82060693)
云南省“万人计划”青年拔尖人才项目。
关键词
三七
病程相关蛋白
过表达
抗性
茄腐镰刀菌
Panax notoginseng(Burk)F.H.Chen
pathogensis-related protein
overexpression
resistance
Fusarium solani
