Sensitivity of Fusarium pseudograminearum isolates to fludioxonil in Henan Province, China
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摘要:
由假禾谷镰刀菌Fusarium pseudograminearum引起的小麦茎基腐病已成为重要的土传病害,并且影响小麦的品质和产量。为了明确中国河南省假禾谷镰刀菌对咯菌腈的敏感性，采用菌丝生长速率法测定了咯菌腈对2019年从河南省6个地市分离的105株假禾谷镰刀菌F. pseudograminearum 的敏感性，通过最小显著差异法(LSD)和SPSS聚类方法对测定结果进行了分析，并测定了假禾谷镰刀菌对多菌灵和戊唑醇的敏感性，分析了咯菌腈与这两种杀菌剂毒力的相关性。结果表明：咯菌腈对供试菌株的最低抑制浓度(MIC)为0.2400 μg/mL。敏感性频率分布图显示，EC50值范围在0.0027~0.0470 μg/mL，敏感性差异达17.41倍；敏感性频率分布为连续单峰曲线，平均EC50值为(0.0263 ± 0.0101) μg/mL，可作为假禾谷镰刀菌对咯菌腈的敏感性基线。方差分析结果显示，不同县市的小麦假禾谷镰刀菌对咯菌腈的敏感性差异较大，EC50值变化范围为0.0150~0.0335 μg/mL，其中咯菌腈对郑州中牟的敏感性最低和最高菌株的EC50值相差16.78倍。聚类分析结果显示，河南省小麦茎基腐病菌菌株对咯菌腈敏感性差异与菌株的地理来源无明显关联性。多菌灵和戊唑醇对病菌的平均EC50值分别为 (0.7881 ± 0.3153) μg/mL和(0.0886 ± 0.1453) μg/mL。病菌对咯菌腈与其对多菌灵和戊唑醇的敏感性之间无明显相关性。温室防效结果显示，用咯菌腈悬浮种衣剂对小麦进行拌种处理，2020年 (咯菌腈有效成分为75.0 μg/g)对小麦茎基腐病的防治效果可达58.00%，2021年 (咯菌腈有效成分为50.0 μg/g)的防治效果可达到63.69%。本研究结果可为咯菌腈在小麦茎基腐病防治中的合理使用提供依据，为病原菌对药剂的敏感性监测提供参考。
Abstract:Wheat crown rot caused by Fusarium pseudograminearum has become an important soil-borne disease and affects the quality and yield of wheat. In order to detect the sensitivity of F. pseudograminearum to fludioxonil in Henan Province, China, 105 isolates of F. pseudograminearum were collected from six cities in 2019. Sensitivity was determined by the mycelial growth rate method, then the methods of least-significant difference (LSD) and SPSS cluster were used for result analysis. The sensitivity of F. pseudograminearum to carbendazim and tebuconazole was determined and the correlation coefficient which existed between fludioxonil and the two fungicides, carbendazim and tebuconazole was analyzed. The results showed that the minimum inhibitory concentration (MIC) of fludioxonil was 0.2400 μg/mL. The sensitivity frequency distribution was a continuous single peak curve, and the EC50 value ranged from 0.0027 to 0.0470 μg/mL. The average EC50 value of (0.0263 ± 0.0101) μg/mL, could be used as the sensitivity baseline of the pathogen to fludioxonil. The variance analysis results showed that the sensitivity of the different cities to fludioxonil was different, and the EC50 value ranged from 0.0150 to 0.0335 μg/mL. The maximum EC50 value of the isolate from Zhongmu County Zhengzhou City was 16.78 times bigger than the minimum value. Cluster analysis showed that there was no significant correlation between the sensitivity of isolates to fludioxonil and their geographical origin. The mean EC50 values of carbendazimand tebuconazole against the pathogens were (0.7881 ± 0.3153) μg/mL and (0.0886 ± 0.1453) μg/mL, respectively. There was no significant correlation between the sensitivity of the isolates to fludioxonil, carbendazim and tebuconazole. The results of greenhouse trials showed that the control efficacy of fludioxonil suspension seed coat agent could reach 58.00% (75.0 μg a.i/g) in 2020 and 63.69% (50.0 μg a.i/g) in 2021 when used to treat wheat as a seed dressing agent. The results of this study provide the basis for the rational use of fludioxonil in the control of wheat crown rot and provide information for monitoring the sensitivity of pathogenic fungi to fungicides.
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Figure 2. Determination (A) and frequency (B) of sensitivity of F. pseudograminearum to fludioxonil
Figure 3. Hierarchical cluster analysis on EC50 values of fludioxonil to F. pseudograminearum from different areas in Henan Province
Figure 4. Correlation of the sensitivity of F. pseudograminearum to fludioxonil and carbendazim (A) or tebuconazole(B)
Table 1. Sensitivity of F. pseudograminearum from different areas in Henan Province to fludioxonil
Sampling site Strain name Tested isolate Frequency/% EC50 value/(μg/mL) Range EC50 (Max)/ EC50 (Min) Mean±SD Sensitive factor Luolong District, Luoyang City JFLF 13 12.38 0.0183-0.0445 2.43 0.0251 ± 0.0088 b 0.95 Wugang County, Luohe City JFLW 14 13.33 0.0084-0.0240 2.86 0.0150 ± 0.0037 c 0.57 Neixiang County, Nanyang City JFNN 5 4.76 0.0140-0.0459 3.28 0.0302 ± 0.0122 ab 1.15 Yucheng County, Shangqiu City JFSQ 6 5.71 0.0163-0.0405 2.48 0.0256 ± 0.0103 b 0.97 Hongqi District, Xinxiang City JFXH 21 20.00 0.0119-0.0453 3.81 0.0261 ± 0.0094 b 0.99 Yanjin District, Xinxiang City JFXY 20 19.05 0.0106-0.0470 4.43 0.0335 ± 0.0091 a 1.27 Zhongmu County, Zhengzhou City JFXS 26 24.76 0.0027-0.0453 16.78 0.0268 ± 0.0090 ab 1.02 Total 7 105 100 0.0027-0.0470 17.41 0.0263 ± 0.0101 ab 1.00 Note: Different letters in the same column indicate significant difference at P<0.05 by LSD test. 
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Table 2. Control efficacy of fludioxonil 25 g/L FSC on wheat crown rot in greenhouse
Year Treatments Dosage/(μg a.i/g) Disease index* Control efficacy/% 2020 T1
T2
T3
T4
CK12.5
25.0
50.0
75.0
030.00 ± 2.02 b
32.89 ± 3.42 b
31.43 ± 4.04 b
20.00 ± 4.04 c
47.62 ± 1.34 a37.00
30.93
34.00
58.00
—2021 T1
T2
T3
T4
CK12.5
25.0
50.0
75.0
012.14 ± 6.35 ab
13.33 ± 7.20 ab
9.52 ± 8.73 b
11.00 ± 3.91 ab
26.22 ± 14.24 a53.70
49.16
63.69
58.09
—Note: * Different letters in the same column indicate significant difference at P<0.05 by LSD test.
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