山东省多主棒孢对三种常用杀菌剂的敏感性监测及对氟吡菌酰胺的抗性

doi:10.3864/j.issn.0578-1752.2022.07.008

Abstract

Abstract:

【Objective】Corynespora target spot caused by Corynespora cassiicola is recognized as one of the three most serious worldwide diseases of cucumber, which seriously affects the yield and quality of cucumber. With the continuous use of commercial fungicides, resistance problem of C. cassiicola increased seriously. The objective of this study is to (i) clarify the resistance profile of C. cassiicola in cucumber from Shandong Province to three common fungicides; (ii) provide a theoretical basis for the fungicide control of Corynespora target spot of cucumber; (iii) provide data support for the screening of highly efficient mixed fungicides for the resistance management of C. cassiicola. 【Method】A total of 140 C. cassiicola isolates were isolated from Shandong Province. The sensitivity of C. cassiicola to difenoconazole, prochloraz and fluopyram was investigated using mycelium growth rate method. The relationship of SdhC-S73P and fluopyram-resistance was analyzed using Modeller v9.19 and AutoDock4.2.6 software. The optimal ratio of fluopyram and prochloraz was also screened using in vitro mycelial growth method and detached leaf assay. 【Result】After data outlier analysis and elimination, the EC₅₀ values of the 121 C. cassiicola isolates to difenoconazole ranged from 0.29 to 6.80 μg·mL ^-1, and produced an unimodal distribution with a mean of 2.44 μg·mL ^-1. The EC₅₀ values of the 129 C. cassiicola isolates to prochloraz ranged from 0.01 to 0.57 μg·mL^-1 with a mean of 0.16 μg·mL ^-1, and also produced an unimodal distribution. The EC₅₀ values of the 136 C. cassiicola isolates to fluopyram ranged from 0.56 to 47.54 μg·mL^-1, with a mean of 6.94 μg·mL ^-1. Many fluopyram-resistant isolates with different resistant factors were detected, and the mutation frequency of fluopyram-resistant isolates containing S73P in SdhC was 31.62%. Molecular docking results showed that after the S73P mutation occurred on the SdhC subunit, the rigidity of the amino acid increased, and the steric hindrance became larger, so that the hydrogen bond between fluopyram and Ser73 disappeared, so the affinity was reduced, leading to the occurrence of fluopyram resistance. No cross-resistance was detected between fluopyram and prochloraz. The highest synergistic effect of fluopyram and prochloraz was 1.84 at a ratio of 7﹕3. The control efficacy of the mixture (fluopyram﹕prochloraz=7﹕3) against Corynespora target spot was higher than that of single fungicide (fluopyram or prochloraz) at the same dose. 【Conclusion】The sensitivity of C. cassiicola from different regions to fungicide is different in Shandong Province. Most of C. cassiicola isolates showed fluopyram resistance and many point mutations in SdhB, SdhC and SdhD were detected. Among them, the frequency of S73P mutation in SdhC was the highest. The control efficacy of the mixture of fluopyram and prochloraz in 7﹕3 against Corynespora target spot in cucumber was more excellent compared with that of single fungicide. It is suggested that the fluopyram can be used alternately or in mixture with prochloraz to control the resistance of C. cassiicola to SDHI fungicides such as fluopyram.

Key words: Corynespora cassiicola, Corynespora target spot, fungicide sensitivity, S73P mutation, molecular docking, synergistic effect

LI GuiXiang,LI XiuHuan,HAO XinChang,LI ZhiWen,LIU Feng,LIU XiLi. Sensitivity of Corynespora cassiicola to Three Common Fungicides and Its Resistance to Fluopyram from Shandong Province[J].Scientia Agricultura Sinica, 2022, 55(7): 1359-1370.

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References 33

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基因 Gene	引物名称 Primer name	引物序列 Primer sequence (5′-3′)	引物长度 Length of primer (bp)	扩增片段大小 Length of product (bp)
SdhB	CcSdhB-F	ATGGCTTGCACACGCGCTTT	20	1106
SdhB	CcSdhB-R	CTACGTGAAAGCCATGCTC	19	1106
SdhC	CcSdhC-F	ATGGCTTCCCAGCGCGTCTT	20	629
SdhC	CcSdhC-R	TAAACAAACAGAGAATAGTA	20	629
SdhD	CcSdhD-F	ATGAAGCGCACCTCG CCAATC	21	938
SdhD	CcSdhD-R	AAAGATCCTAATAATCACATGTATCCAAAC	30	938

点突变类型 Point mutation type	菌株数量 Number of isolates	抑制中浓度 EC₅₀(μg·mL^-1)	比值 Ratio (%)
无突变No mutation	42	0.56-2.42	30.88
SdhB H278Y	9	0.56-1.19	6.62
SdhB I280V	19	1.58-24.63	13.97
SdhC S73P	43	1.88-47.54	31.62
SdhC N75S	5	1.42-11.31	3.68
SdhC H134R	8	0.61-18.84	5.88
SdhD G135V	10	2.66-18.65	7.35

Sensitivity of Corynespora cassiicola to Three Common Fungicides and Its Resistance to Fluopyram from Shandong Province

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