摘要
以48个鲜食葡萄品种为试材,通过室内离体叶盘接种霜霉病菌试验,对各品种的霜霉病抗性分级,结合对抗病相关基因PR1、PR5和NPR1的半定量RT-PCR试验,分析不同抗性级别品种在霜霉病菌侵染后表达模式的差异,以期为抗葡萄霜霉病的新品种选育提供参考依据。结果表明:48个品种中有30个品种对霜霉病高度敏感,7个品种对霜霉病敏感,4个品种中抗,6个品种高抗霜霉病,"金星无核"对霜霉病完全免疫。PR1和PR5基因在接菌前有本底表达,抗病和高抗品种接菌后均上调表达,感病品种接菌后无明显变化,仅"无核翠宝"和"红地球"有微量下调。NPR1基因接菌后在抗病品种中表达量上升,而在感病品种和高抗品种中均表现为半数品种上升,半数品种下降。
In order to provide reference for the breeding of downy mildew resistance varieties,the resistance to downy mildew of 48 table grape varieties were tested by leaf disc inoculation indoor.Meanwhile,the expression patterns of resistance-related genes PR1,PR5 and NPR1 to downy mildew were investigated through semi-quantitative RT-PCR.The results showed that 30 varieties were highly susceptible,7 susceptible,4 resistant,6 highly resistant,‘Venus Seedless’ immune to downy mildew.The difference of PR1 and PR5 expression were not significantly in susceptible varieties except ‘Wuhecuibao’ and ‘Red Globe’,which were reduced slightly after inoculation.The expression of PR1 and PR5 were higher in the resistant ones after downy mildew infection.In susceptible varieties and high-resistant varieties,the expression of gene NPR1 was up-regulated in 50% varieties and down-regulated in the other ones,while the expression was up-regulated in all the resistant varieties.
引文
[1] CHEN Z,SILVA H,KLESSIG D F.Active oxygen species in the induction of plant systemic acquired resistance by salicylic acid[J].Science,1993,262(17):1883-1886.
[2] FU Z Q,DONG X.Systemic acquired resistance:Turning local infection into global defense[J].Annual Review of Plant Biology,2013,64(1):839-863.
[3] PENNINCKX I A M A,EGGERMONT K,TERRAS F R G.Pathogen-induced systemic activation of a plant defensin gene in Arabidopsis follows a salicylic acid-independent pathway[J].The Plant Cell Online,1996,8(12):2309-2323.
[4] CASAGRANDE K,FALGINELLA L,CASTELLARIN S D,et al.Defence responses in Rpv3-dependent resistance to grapevine downy mildew[J].Planta,2011,234(6):1097-1109.
[5] HENANFF G L,HEITZ T,MESTRE P,et al.Characterization of Vitis vinifera NPR1 homologs involved in the regulation of pathogenesis-related gene expression[J].BMC Plant Biology,2009,9(1):54.
[6] 胡宗利,邓磊,姚楠,等.番茄PR-1和PR-5基因的表达特性分析[J].西南大学学报(自然科学版),2009,31(8):67-72.
[7] 张红志,蔡新忠.病程相关基因非表达子1(NPR1):植物抗病信号网络中的关键节点[J].生物工程学报,2005(4):511-515.
[8] SPOEL S H.NPR1 Modulates cross-talk between salicylate-and jasmonate-dependent defense pathways through a novel function in the cytosol[J].The Plant Cell Online,2003,15(3):760-770.
[9] 赵雪艳,张玮,严红,等.不同葡萄品种对霜霉病的田间苗期抗性评价[J].植物保护,2014,40(5):172-175.
[10] SU H,JIAO Y T,WANG F F,et al.Overexpression of VpPR10.1 by an efficient transformation method enhances downy mildew resistance in V.vinifera[J].Plant Cell Reports,2018,37(5):819-832.
[11] 肖婷,郭建,吴祥,等.不同品种葡萄对霜霉病的抗性研究[J].安徽农业科学,2014,42(33):11726-11727,11733.
[12] 栗小英.PR1、PR2和PR5基因参与小麦TcLr35成株抗叶锈病反应的表达分析[D].保定:河北农业大学,2015.
[13] 肖栋,韦艳萍,李英,等.不结球白菜病程相关蛋白基因BcPR5的克隆及表达分析[J].南京农业大学学报,2018,41(4):640-646.
[14] 张青,赵景梅,黄东益,等.大薯病程相关蛋白1(PR1)基因及其启动子序列的克隆与分析[J].分子植物育种,2018,16(7):2078-2084.
[15] 张玉,王杰,周世奇,等.烟草PR10蛋白生物活性及赤星病菌Alternaria alternata诱导下的表达分析[J].植物保护学报,2018,45(3):455-462.
[16] 何佳,葛露霞,金魏佳,等.青花菜病程相关蛋白基因BoPR1的克隆与表达分析[J].福建农业学报,2018,33(1):35-40.
[17] 吕梓剑,龙瑶,张效焓,等.水杨酸和乙烯信号转导在拟南芥相应灰霉菌侵染后生长发育及光合作用的影响[J].绿色科技,2018(10):14-15.
[18] 焦鹏.苹果NPR1基因家族的全基因组鉴定与表达分析[D].杨凌:西北农林科技大学,2016.
[19] 李霄,陈婷,鲍玲莉,等.甘蓝型油菜BnNPR1的序列及其环境信号响应分析[J].江苏农业科学,2018,46(14):19-24.
[20] DONG X N.NPR1,all things considered[J].Current Opinion in Plant Biology,2004,7(5):547-552.
[21] IAVICOLI A,BOUTET E,BUCHALA A,et al.Induced systemic resistance in Arabidopsis thaliana in response to root inoculation with Pseudomonas fluorescens CHA0[J].Mol Plant Microbe Interact,2003,16(10):851-858.
[22] PIETERSE C M,LOON L V.NPR1:The spider in the web of induced resistance signaling pathways[J].Current Opinion in Plant Biology,2004,7(4):456-464.