玫瑰黄链霉菌活性代谢产物诱导烟草灰霉病抗性机制研究  

Mechanism of resistance to tobacco gray mold induced by the active metabolite of Streptomyces roseoflavus

作　　者：刘海涛 王娇 刘大群[1,2] 李亚宁[1] LIU Hai-tao;WANG Jiao;LIU Da-qun;LI Ya-ning(College of Plant Protection,Hebei Agricultural University/Innovation Center for Biological Control of Crop Diseases and Insect Pests of Hebei Province/National Engineering Research Center for Agriculture in Northern Mountainous Areas,Baoding,Hebei 071001,China;Graduate School,Chinese Academy of Agricultural Sciences,Beijing 100081,China)

机构地区：[1]河北农业大学植物保护学院/河北省农作物病虫害生物防治技术创新中心/国家北方山区农业工程技术研究中心,河北保定071001 [2]中国农业科学院研究生院,北京100081

出　　处：《南方农业学报》2022年第6期1560-1567,共8页Journal of Southern Agriculture

基　　金：国家自然科学基金面上项目(31972316)。

摘　　要：【目的】解析温室条件下玫瑰黄链霉菌Men-myco-93-63活性代谢产物Roflamycoin和Men-myco-A(简称R&M)诱导烟草灰霉病抗性的作用机制,为利用R&M绿色防控烟草灰霉病提供理论依据。【方法】通过80 mg/L R&M注射4~5叶期本生烟草叶片后过敏反应(HR)染色验证及100 mg/L R&M注射4~5叶期本生烟草叶片后抗病标志基因hsr203J的表达验证R&M的诱抗作用;采用相关试剂盒检测烟草处理(分别为80 mg/L R&M喷雾处理和清水处理)后非药剂接触烟叶内过氧化氢酶(CAT)和脂氧合酶(LOX)的活性变化,以及防御相关物质过氧化氢(H_(2)O_(2))的含量变化。采用实时荧光定量PCR(qRT-PCR)检测烟草处理(处理1,用80 mg/L R&M溶液灌根处理4~5叶期本生烟草2 d后,于上部第3片叶采用灰霉菌孢子悬浮液摩擦接种;处理2,清水灌根处理4~5叶期本生烟草2 d后,于上部第3片叶采用清水摩擦接种)后烟草叶片木质素合成基因PAL及病程相关蛋白(PR)基因PR1、PR5相对表达量的变化。【结果】80 mg/L R&M诱导处理后,烟叶中CAT活性2 d后逐步提高;LOX活性第3 d后显著提高(P<0.05,下同),第7 d达最高值;H_(2)O_(2)含量在第1 d迅速增加后降低;PAL基因相对表达量在24 h后显著上调,为清水对照的25.37倍;病程相关蛋白基因PR1、PR5相对表达量分别在24和72 h达最高值,分别为清水对照的11.23和11.17倍。【结论】R&M通过提高烟叶中相关防御酶活性、协调活性氧水平及增强抗病相关基因的表达等提高烟草植株的抗病性。玫瑰黄链霉菌Men-myco 93-63产生的活性代谢产物R&M具有作为植物病害诱抗剂的潜力,有望应用于植物灰霉病等重要病害的绿色防控。【Objective】To study the mechanism of resistance to tobacco gray mold induced by the active metabolite Roflamycoin and Men-myco-A(R&M)of Streptomyces roseoflavus Men-myco-93-63 under greenhouse conditions,so as to provide theoretical basis for the ecofriendly use of R&M to prevent and control tobacco gray mold.【Method】Eighty mg/L R&M was injected to 4-5 leaf stage of Nicotiana benthamiana to conduct hypersensitive response(HR)staining verification and 100 mg/L R&M was injected to 4-5 leaf stage of N.benthamiana to get expression of the disease resis-tance marker gene hsr203J respectively to verify the induced disease resistance of R&M.Changes of catalase(CAT)activity,lipoxygenase(LOX)activity and content of hydrogen peroxide(H_(2)O_(2)),a defense-related substance,in non-phar-maceutically treated tobacco leaves of N.benthamiana were detected by kits(treatments of 80 mg/L R&M spray and wa-ter treatment).Quantitative real-time PCR(qRT-PCR)technology was used to detect changes of relative expression of lig-nin synthesis-related gene PAL and pathogenesis-related protein(PR)genes PR1 and PR5 after tobacco treatments(treat-ment 1,80 mg/L R&M solution for root treatment for 2 d,friction inoculation with Botrytis cinerea spore suspension on the upper third leaves;treatment 2,root irrigation with water for 2 d,friction inoculation with water on the upper third leaves).【Result】Catalase(CAT)activity in tobacco leaves gradually increased after induction treatment with 80 mg/L R&M;while lipoxygenase(LOX)activity increased significantly after 3 d(P<0.05,the same below),and both reached the highest on 7 d.H_(2)O_(2) content increased rapidly on the 1 d and then decreased.Relative expression of PAL gene signifi-cantly was up-regulated after 24 h,which was 25.37 folds of that of CK.Relative expression of PR genes,PR1 and PR5,reached the highest at 24 h and 72 h respectively,which were 11.23 times and 11.17 times of those of the CK.【Conclu-sion】R&M enhances disease resistance of tobacco plants through increasing the

关 键 词：玫瑰黄链霉菌MEN-MYCO-93-63 Roflamycoin Men-myco-A 烟草灰霉病 诱导抗病性 PR基因 

分 类 号：S435.72[农业科学—农业昆虫与害虫防治]