福建省玉米大斑病菌对吡唑醚菌酯不同敏感性菌株的遗传多样性与群体遗传结构分析  

Analysis of Genetic Diversity and Population Genetic Structure of Seto⁃sphaeria turcica Isolates with Different Sensitivities to Pyraclostrobin in Fujian Province

作　　者：代玉立[1] 甘林[1] 刘晓菲 兰成忠[1] 杨秀娟[1] DAI Yu-Li;GAN Lin;LIU Xiao-Fei;LAN Cheng-Zhong;YANG Xiu-Juan(Institute of Plant Protection,Fujian Key Laboratory for Monitoring and Integrated Management of Crop Pests,Fujian Engineering Research Center for Green Pest Management,Fujian Academy of Agricultural Sciences,Fuzhou 350013,China)

机构地区：[1]福建省农业科学院植物保护研究所/福建省作物有害生物监测与治理重点实验室/福建省作物有害生物绿色防控工程研究中心,福州350013

出　　处：《农业生物技术学报》2023年第3期603-616,共14页Journal of Agricultural Biotechnology

基　　金：福建省科技计划公益类专项(2020R1024003);国家重点研发计划(2021YFC2600402);福建省农业科学院“5511”协同创新工程(XTCXGC2021011,XTCXGC2021017);福建省农业科学院农作物重大有害生物灾变机制与绿色防控科技创新团队(CXTD2021002-1)。

摘　　要：玉米大斑病(northern corn leaf blight,NCLB)是严重影响玉米(Zea mays)产量和品质的叶部真菌病害。为明确福建省玉米大斑病菌(Setosphaeria turcica)对吡唑醚菌酯(pyraclostrobin)不同敏感性菌株的遗传多样性和遗传结构,本研究采用含药平板抑制菌落生长法评估福建省玉米大斑病菌对吡唑醚菌酯的敏感性,使用简单重复序列区间(inter-simP_(L)e sequence repeat,ISSR)标记技术分析吡唑醚菌酯不同敏感性菌株的遗传多样性和遗传结构。结果显示,吡唑醚菌酯对福建省62株玉米大斑病菌的菌落有效抑制中浓度(effective concentration for 50%colonial inhibition,EC50)值的范围为0.0021~1.2884μg/mL,平均值为(0.1274±0.2545)μg/mL,吡唑醚菌酯的频率分布曲线为连续单峰曲线且不符合正态分布(W=0.7422,P=0.0010<0.05)。吡唑醚菌酯敏感型、中间型和不敏感型菌株的EC50值的范围分别为0.0021~0.0112、0.0206~0.1000和0.1019~1.2884μg/mL,不敏感型菌株的抗性倍数介于3.01~38.01之间,方差分析表明不敏感型菌株的EC50平均值与敏感型和中间型菌株比较均存在显著的差异(P<0.05)。综上所述,福建省玉米大斑病菌已出现对吡唑醚菌酯敏感性下降的亚群体。遗传多样性分析表明,11条ISSR引物共扩增出66个位点,多态性位点百分比(percentage of polymorphic loci,P_(L))高达98.48%,敏感型群体的DNA多态性水平最低(P_(L)=60.61%),不敏感型群体的DNA多态性水平最高(P_(L)=68.18%),不敏感型群体的遗传多样性指数值均高于敏感型群体,表明不敏感型群体的遗传多样性更丰富。遗传分化(genetic differentiation,ΦPT)与基因流(gene flow,Nm)分析表明,吡唑醚菌酯敏感型、中间型和不敏感型两两群体间的遗传分化水平较低(ΦPT<0.054,P>0.37)且基因交流十分频繁(Nm>8.60)。聚类分析也表明吡唑醚菌酯不同敏感性的菌株随机聚于同一分支,说明病原菌群体的遗传分化与抗药性水平的相关性不明�Northern corn leaf blight(NCLB)is a crucial foliar fungal disease that seriously affects corn(Zea mays)yield and quality.To determine the genetic diversity and genetic structure of Setosphaeria turcica isolates with different sensitivities to pyraclostrobin in Fujian Province,the sensitivity of S.turcica isolates to pyraclostrobin in Fujian Province was evaluated using the method of measuring the colony growth inhibition on the fungicide-amended P_(L)ate,and genetic diversity and genetic structure of the S.turcica isolates with different sensitivities to pyraclostrobin was analyzed using inter-simP_(L)e sequence repeats(ISSR)markers in this study.The results showed that the range of effective concentration for 50%colonial inhibition(EC50)values of 62 S.turcica isolates to pyraclostrobin in Fujian Province was 0.0021~1.2884μg/mL,with the mean value of(0.1274±0.2545)μg/mL.The frequency distribution curve of pyraclostrobin was continuous and unimodal,and disobeyed the normal distribution(W=0.7422,P=0.0010<0.05).The ranges of EC50 value for S.turcica isolates with pyraclostrobin-sensitivity,-medium and-insensitivity were 0.0021~0.0112,0.0206~0.1000 and 0.1019~1.2884μg/mL,respectively.The resistant ratios of pyraclostrobin-insensitive isolates ranged from 3.01 to 38.01.Analysis of variance revealed that the mean EC50 value of pyraclostrobin-insensitive isolates was significant difference compared with those of isolates with pyraclostrobin-medium and-sensitivity,respectively(P<0.05).In conclusion,a subpopulation of S.turcica with decreased sensitivity to pyraclostrobin has been developed in Fujian Province.Genetic diversity analysis indicated that a total of 66 loci were amP_(L)ified using 11 ISSR primers,and the percentage of polymorphic loci(P_(L))reached as high as 98.48%.The lowest DNA polymorphism was detected from pyraclostrobin-sensitive population(P_(L)=60.61%),whereas the most abundant polymorphism was detected from those in pyraclostrobin-insensitive population(P_(L)=68.18%).The index values of genetic di

关 键 词：玉米大斑病菌 敏感性 吡唑醚菌酯 遗传多样性 群体遗传结构 

分 类 号：S432.1[农业科学—植物病理学]