黄萎病对海/陆嫁接棉苗叶片中H2O2、抗病相关酶活性和GST、PAL基因表达的影响  

Effects of Verticillium dahliae on H2O2 content,activities of disease resisitance-related enzymes,gene expression of GST and PAL in leaves of upland/sea island cotton grafted seedlings

作　　者：张明菊 张浩 朱华国 夏启中 ZHANG Mingju;ZHANG Hao;ZHU Huaguo;XIA Qizhong(College of Biology and Agricultural Resources/Hubei Province Key Laboratory of Germplasm Improvement and Comprehensive Utilization of Economic Forest Tree Resources,Huanggang Normal University,Huanggang 438000,China)

机构地区：[1]黄冈师范学院生物与农业资源学院/经济林木种质改良与资源综合利用湖北省重点实验室,黄冈438000

出　　处：《华中农业大学学报》2020年第3期28-37,共10页Journal of Huazhong Agricultural University

基　　金：国家科技支撑计划项目(2011BAD35B05);湖北省自然科学基金重点项目(2012FFA066);湖北省自然科学基金一般项目(2008CDB091);湖北省教育厅重点项目(D20082701,D20102903)。

摘　　要：为探讨棉花黄萎病抗性机制,以抗黄萎病的海岛棉材料Pima90(R)为砧木,以在湖北省感病的陆地棉品种鄂棉22(S)为接穗,获得海/陆嫁接棉苗(S/R),研究海岛棉Pima90(R)、陆地棉鄂棉22(S)和海/陆嫁接棉苗(S/R)接种后的抗病反应,测定叶片中的H2O2含量,比较三者POD、PAL酶活性水平和PAL、GST基因表达模式的差异。结果显示:以黄萎病菌大丽轮枝菌菌株V991接种棉苗根部,海/陆嫁接棉苗(S/R)的萎蔫程度介于砧木Pima90(R)和接穗鄂棉22(S)之间,表明利用抗病海岛棉砧木(R)对感病陆地棉(S)嫁接可提高棉苗对黄萎病的抗性;接种后,鄂棉22(S)、Pima90(R)和海/陆嫁接棉苗(S/R)三者叶片中的H2O2含量、POD和PAL活性均有明显升高;其中感病品种鄂棉22(S)中H2O2含量和POD、PAL活性上升相对较缓慢,抗病品种海岛棉Pima90(R)上升较快,而海/陆嫁接棉苗(S/R)上升幅度介于R和S二者之间,且与抗病砧木Pima90(R)表现出相似的趋势;接种后不同时段对GST和PAL基因的real-time PCR分析结果显示,GST基因表达在Pima90(R)、鄂棉22(S)和海/陆嫁接棉苗(S/R)均表现出先缓慢上升后下降的趋势;在感病品种鄂棉22(S)中高水平表达持续时间短(8~16 h),而在抗病海岛棉品种Pima90(R)和海/陆嫁接棉苗(S/R)中,高水平表达持续时间较长(8~48 h);PAL基因在Pima90(R)、鄂棉22(S)和嫁接棉苗(S/R)中具有相似的表达模式,即均表现为上升-下降-上升(-下降)的表达趋势,在8 h和48 h出现2个表达峰值;在感病品种鄂棉22(S)中PAL表达量较低,2个峰值较小;而在抗病海岛棉品种Pima90(R)中,PAL表达水平高,2个峰值较大;海/陆嫁接棉苗(S/R)中PAL表达量介于R和S二者之间。Gossypium barbadense/G.hirsutum grafted cotton seedlings(S/R)were obtained by using G.hirsutum cotton Pima90(R)resistant to Verticillium wilt as the rootstock and G.barbadense cotton variety Emian 22(S)susceptible to Verticillium wilt in Hubei Province as scion.The disease resistance after incubation of Pima90(R),Emian 22(S)and grafted cotton seedlings(S/R)was studied.H2O2 content in leaves was determined.The differences in the levels of POD/PAL enzyme activity and the expression patterns of PAL/GST genes were compared.Results showed that wilting degree of the G.barbadense/G.hirsutum grafted cotton(S/R)after the incubation of damaged cotton root with pathogen suspensions was between that of the rootstock Pima90(R)and scion Emian 22(S),indicating that grafting improved the resistance of cotton seedlings to Verticillium wilt.H2O2 contents in leaves and the enzymatic activities of POD and PAL in Emian 22(S),Pima90(R)and the grafted cotton(S/R)were obviously increased after incubation with Verticillium dahliae Kleb.However,H2O2 content in leaves,POD and PAL activities increased slowly in Emian 22(S),rapidly in Pima90(R),and moderately in the S/R grafted cotton.Pima90(R)and the grafted cotton(S/R)showed similar tendency of changing.Results of real-time PCR showed that the mRNA expression level of GST gene in three cottons increased firstly and then decreased.The duration of GST gene expression for Emian 22(S),Pima90(R)and grafted cotton(S/R)was 8-16 h and 8-48 h,respectively.The patterns of PAL gene expression in the three cottons were similar.Expression level of PAL gene was increased firstly,then decreased,and increased subsequently,with twice peak at 8 h and 48 h.The peak value in Emian 22(S),Pima90(R)and grafted cotton(S/R)was low,high,and moderate.

关 键 词：棉花 海/陆嫁接棉苗 黄萎病 H2O2 抗病相关酶 GST、PAL基因 

分 类 号：S562.503.4[农业科学—作物学]