摘要
RPM1属于CC-NBS-LRR类蛋白激酶,在植物抗病反应中起重要作用,其功能在拟南芥、水稻等植物中有深入研究,但在草莓上还鲜有报道。以草莓为试材,克隆FveRPM1基因的编码区序列,发现其长度为2748bp,编码915个氨基酸,属于膜表达蛋白。进化关系分析显示草莓FveRPM1与苹果MdRPM1的亲缘关系最近,而与乌拉尔图小麦TuRPM1的亲缘关系较远。利用RTqPCR技术检测了FveRPM1基因在草莓不同器官、不同激素浓度及病菌处理下的表达量,结果表明:FveRPM1基因在草莓根中高表达,而在新叶中的表达量最低;不同浓度的MeJA和SA分别处理Ruegen草莓12h后FveRPM1基因的表达水平明显上升,其中1μmol·L^(-1)的MeJA处理12h后FveRPM1基因的表达量最高,为对照的13.2倍,而0.1μmol·L^(-1)的SA处理12h后FveRPM1基因的表达量最高,为对照的14倍;灰霉病菌处理能显著增加FveRPM1基因的表达量,同时也能显著增强POD、CAT和SOD的活性。利用农杆菌介导的草莓果实瞬时转化试验,将构建好的FveRPM1基因的过表达载体和干扰载体分别转入艳丽和月华草莓白果期果实中,待果实转红时接种灰霉病菌后发现,过表达FveRPM1基因能够增强草莓果实对灰霉病的抗性,而沉默该基因则降低了草莓果实对灰霉病的抗性,并且FveRPM1基因过表达果实的SOD酶活性最强,FveRPM1基因沉默果实的SOD酶活性最弱,CAT酶活性表现相同的规律,说明FveRPM1基因参与草莓果实对灰霉病的防御响应。研究结果为利用分子手段进行草莓抗病育种奠定一定的理论基础。
RPM1 belongs to the CC-NBS-LRR class of protein kinases,it plays an important role in plant disease resistance response.Its function has been deeply studied in plants such as Arabidopsis thaliana and rice,but it has rarely been reported on strawberries.In this study,strawberry was used as the test material,the coding region of FveRPM1 gene was cloned.It was found that its length is 2,748bp,encoding 915 amino acids,which belongs to membrane-expressed protein.Evolutionary relationship analysis shows that strawberry FveRPM1 is most closely related to MdRPM1,but farther from TuRPM1.RT-qPCR technology was used to detect the expression of FveRPM1 gene in different organs of strawberry,different hormone concentrations and pathogen treatments,the results showed that the FveRPM1 gene was highly expressed in strawberry roots,while the expression level was the lowest in new leaves.The expression level of FveRPM1 gene increased significantly in Ruegen strawberry after treated with different concentrations of MeJA and SA for 12h,respectively.Among them,the expression of FveRPM1 gene was the highest after 1μmol·L^(-1)MeJA treatment for 12h,it was 13.2-fold higher than that in control.However,it was the highest and 14-fold higher than that in control after 0.1μmol·L^(-1)SA treatment for 12h.Botrytis cinerea treatment can significantly increase the expression of FveRPM1 in strawberry,and it can also significantly enhance the activity of POD,CAT and SOD.The overexpression vector and interference vector of FveRPM1 gene were transferred to the white fruit of Yanli and Yuehua by Agrobacteria transient transformation,respectively.The overexpression of the FveRPM1 gene increased the resistance of strawberry fruits to Botrytis cinerea,while its silence reduced the resistance of strawberry fruits to Botrytis cinerea after inoculating Botrytis cinerea when the berries turned red.The SOD activity in the overexpressed fruit of FveRPM1 gene was the strongest,it was the weakest in the silenced fruit.And CAT activity had the same trend.The results showed that FveRPM1 gene is involved in the defense response of strawberry fruit to Botrytis cinerea.The research results will provide a certain theoretical basis for the use of molecular means for strawberry disease resistance breeding.
作者
关宇涵
王雨沙
李晓明
李贺
GUAN Yu-han;WANG Yu-sha;LI Xiao-ming;LI He(College of Horticulture/Liaoning Key Laboratory of Strawberry Breeding and Cultivation,Shenyang Agricultural University,Shenyang 110161,China)
出处
《沈阳农业大学学报》
CAS
CSCD
北大核心
2022年第6期641-649,共9页
Journal of Shenyang Agricultural University
基金
国家重点研发计划子课题项目(2019YFD1000200)
辽宁省重点研发项目(2020JH2/10200032)。
