Analysis of Genetic Diversity and Population Genetic Structure of Setosphaeria turcica Isolates with Different Sensitivities to Pyraclostrobin in Fujian Province
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
Abstract：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 plate, and genetic diversity and genetic structure of the S. turcica isolates with different sensitivities to pyraclostrobin was analyzed using inter-simple 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 0.002 1~1.288 4 μg/mL, with the mean value of (0.127 4±0.254 5) μg/mL. The frequency distribution curve of pyraclostrobin was continuous and unimodal, and disobeyed the normal distribution (W=0.742 2, P=0.001 0<0.05). The ranges of EC50 value for S. turcica isolates with pyraclostrobin-sensitivity, -medium and -insensitivity were 0.002 1~0.011 2, 0.020 6~0.100 0 and 0.101 9~1.288 4 μg/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 amplified using 11 ISSR primers, and the percentage of polymorphic loci (PL) reached as high as 98.48%. The lowest DNA polymorphism was detected from pyraclostrobin-sensitive population (PL=60.61%), whereas the most abundant polymorphism was detected from those in pyraclostrobin-insensitive population (PL=68.18%). The index values of genetic diversity for the insensitive population were higher than those of the sensitive population, suggesting that genetic diversity in the insensitive population of S. turcica was more diverse than those of sensitive and mediate populations. The results of genetic differentiation (ΦPT) and gene flow (Nm) analysis indicated that low genetic differentiation (ΦPT<0.054, P>0.37) with frequent gene exchange (Nm>8.60) were detected among pyraclostrobin-sensitive, -mediate and -insensitive populations. Clustering analysis also indicated that S. turcica isolates with different sensitivities to pyraclostrobin were randomly clustered in the same branch, suggesting no obvious correlation between genetic differentiation and the level of fungicide resistance. Analysis of molecular variance (AMOVA) revealed that the all source of genetic variation in Fujian S. turcica populations with different sensitivities to pyraclostrobin was derived from within populations. Principal coordinates (PCoA) and population genetic structure analysis indicated that the 3 S. turcica populations with different sensitivities to pyraclostrobin could be divided into 2 genetic groups, with a high similar genetic structure being observed among the 3 different populations. The results from this study provide a reference for monitoring of fungicide resistance of S. turcica in the field and for ecological regulation of northern corn leaf blight by using resistant cultivars.
代玉立, 甘林, 刘晓菲, 兰成忠, 杨秀娟. 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[J]. 农业生物技术学报, 2023, 31(3): 603-616.
DAI Yu-Li, GAN Lin, LIU Xiao-Fei, LAN Cheng-Zhong, YANG Xiu-Juan. Analysis of Genetic Diversity and Population Genetic Structure of Setosphaeria turcica Isolates with Different Sensitivities to Pyraclostrobin in Fujian Province. 农业生物技术学报, 2023, 31(3): 603-616.
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