樱桃褐腐病菌对啶酰菌胺的敏感性及其对4种琥珀酸脱氢酶抑制剂的交互抗性

董怡; 李阿根; 毛程鑫; 张艳婷; 张传清; 刘亚慧

doi:10.16801/j.issn.1008-7303.2021.0180

樱桃褐腐病菌对啶酰菌胺的敏感性及其对4种琥珀酸脱氢酶抑制剂的交互抗性

doi: 10.16801/j.issn.1008-7303.2021.0180

董怡^1,,
李阿根²,
毛程鑫¹,
张艳婷¹,
张传清^1, ,,
刘亚慧^1, ,

1.
浙江农林大学农业与食品科学学院，杭州 311300
2.
杭州市余杭区农业生态与植物保护管理总站，杭州 311100

基金项目: 浙江省重点研发项目 (2020C02005).

详细信息

作者简介:
董怡, 1262406417@qq.com

通讯作者:
张传清, cqzhang9603@126.com

刘亚慧, liuyahui716@163.com.

中图分类号: S481.4；TQ450.13
计量
- 文章访问数: 85
- HTML全文浏览量: 30
- PDF下载量: 22
- 被引次数: 0
出版历程
- 收稿日期: 2020-12-29
- 录用日期: 2021-11-04
- 网络出版日期: 2021-12-08
- 刊出日期: 2022-04-10

Sensitivity of Monilinia fructicola to boscalid and cross-resistance to four succinate dehydrogenase inhibitors

1.
College of Agriculture and Food Science, Zhejiang Agriculture & Forestry University, Hangzhou 311300, China
2.
Yuhang’s Management Station for the Agricultural Ecology and Plant Protection of Hangzhou, Hangzhou 311100, China

Funds: Key R & D Program in Zhejiang Province (2020C02005).

摘要

摘要: 采用菌丝生长速率法测定了樱桃褐腐病菌Monilinia fructicola对啶酰菌胺的敏感性，同时研究了不同敏感性菌株的生物学性状，探究了琥珀酸脱氢酶B亚基的氨基酸突变与其对啶酰菌胺产生抗性的相关性，并分析了樱桃褐腐病菌对啶酰菌胺与其他3种琥珀酸脱氢酶抑制剂 (SDHIs)氯苯醚酰胺、氟唑菌苯胺和氟吡菌酰胺之间的交互抗性。结果表明：啶酰菌胺对樱桃褐腐病菌具有较好的抑制活性和治疗作用，但樱桃褐腐病菌已对其产生了一定的抗性，且抗性菌株具有较高的适合度。褐腐病菌SDHB亚基上的氨基酸点突变与其对啶酰菌胺的抗性之间无明显联系。交互抗药性分析表明，啶酰菌胺与氯苯醚酰胺、氟唑菌苯胺和氟吡菌酰胺之间均存在交互抗性。为了延缓抗药性的发生和发展，在樱桃褐腐病防治过程中啶酰菌胺应与SDHIs之外的其他类型杀菌剂进行合理的混用或轮用。
- 樱桃褐腐病 /
- 啶酰菌胺 /
- 敏感性 /
- 琥珀酸脱氢酶抑制剂 /
- 交互抗性
Abstract: The sensitivity of Monilinia fructicola to boscalid was determined by the mycelial growth rate method. Meanwhile, the study investigated the biological characteristics of isolates with different sensitivities, the correlation between the amino acid mutations of SDHB protein with resistance to boscalid, and the cross-resistance of M. fructicola between boscalid and other 3 tested succinate dehydrogenase inhibitors (SDHIs), Y12196, penflufen and fluopyram. The results showed that boscalid had a good inhibitory activity and therapeutic effect against M. fructicola. However, M. fructicola has developed a certain degree of resistance, and the resistant isolates have comparable fitness with the sensitive ones. The sequence analysis of SDHB gene showed that the amino acid point mutations of M. fructicola SDHB subunits were not significantly correlated with the resistance to boscalid. The cross-resistance studies indicated that there was cross-resistance between boscalid and the other three SDHIs, Y12196, penflufen, and fluopyram. In order to retard the occurrence and exacerbation of resistance, the rational mixes or rotations of boscalid with other kinds of fungicides rather than SDHIs are recommended during the prevention and control of cherry brown rot disease.
- cherry brown rot /
- boscalid /
- sensitivity /
- succinate dehydrogenase inhibitors /
- cross-resistance

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图 1 樱桃褐腐病菌 (n=28) 对啶酰菌胺的敏感性

Figure 1. The sensitivity of M. fructicola (n=28) to boscalid

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图 2 樱桃褐腐病菌 (n=28) 对啶酰菌胺的抗性倍数分布

Figure 2. The distribution of resistance ratio of M. fructicola (n=28) to boscalid

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图 3 樱桃褐腐病菌 (n=28) 对氯苯醚酰胺、氟唑菌苯胺和氟吡菌酰胺的敏感性(EC₅₀区间)分布

Figure 3. Distribution profile of M. fructicola sensitivity (EC₅₀) to Y12196, penflufen and fluopyram (n=28)

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图 4 樱桃褐腐病菌中啶酰菌胺与氯苯醚酰胺 (A) 、氟唑菌苯胺 (B) 和氟吡菌酰胺 (C) 的交互抗性

Figure 4. Cross-resistance between boscalid and Y12196 (A), penflufen (B) and fluopyram (C) in M. fructicola

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表 1 用于扩增美澳型核果链核盘菌SDHB基因的引物

Table 1. Primers used for the amplification of SDHB in M. fructicola

编号 Serial number	序列 Sequence (5′-3′)	长度 Length/bp	目的基因 Target gene
CQ110	ATCTCTCCGCACCACCAG	884	SDHB
CQ111	CTTGATCTCCGCAATCGCC	884	SDHB

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表 2 啶酰菌胺对樱桃褐腐病的防治效果

Table 2. The control effects of boscalid on cherry brown rot

菌株 Isolate	啶酰菌胺质量浓度 Mass concentration of boscalid/(µg/mL)	防治效果 Control efficacy/%
Mf19-3-8^S	10	0.01±7.17 d
	50	76.75±8.70 b
	100	100.00±0.00 a
Mf19-3-1^LR	10	−1.09±4.61 d
	50	100.00±0.00 a
	100	100.00±0.00 a
Mf19-1-7^LR	10	10.64±5.21 d
	50	88.30±10.53 ab
	100	100.00±0.00 a
Mf19-5-10^MR	10	6.74±1.59 d
	50	33.71±5.73 c
	100	88.76±9.67 ab
注：^S代表啶酰菌胺敏感菌株，^LR代表啶酰菌胺低抗菌株，^MR代表啶酰菌胺中抗菌株。^同列数据后不同字母表示该列所有处理的防治效果在P=0.05水平显著性差异。“±” 代表3次重复的防治效果之间的标准误差。Note: The superscript letter^"S"adjacent to the isolate number represents boscalid-sensitivity.^"LR" represents boscalid low-resistance. ^"MR" represents boscalid medium-resistance. ^The values in a column followed by different lowercase letters indicate the control efficacy of all treatments is significantly different at the P =0.05 level. "±" represents the standard error between three repetitions of control efficacy.

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表 3 对啶酰菌胺不同敏感性的樱桃褐腐病菌的生物学特性

Table 3. Biological characteristics of M. fructicola with different sensitivities to boscalid

菌株 Isolate	平均生长速率 Average growth rate/(mm/d)	产孢量 Sporulation quantity/ ( × 10⁶个/mL)	孢子萌发率 Spore germination rate/%
菌株 Isolate	平均生长速率 Average growth rate/(mm/d)	产孢量 Sporulation quantity/ ( × 10⁶个/mL)	0.5 h	1.5 h	2.5 h
Mf19-4-7^S	10.27±0.17 a	1.05±0.86 cde	23.44±4.94 b	52.28±1.46 bc	86.81±2.93 ab
Mf19-3-7^S	9.71±0.67 c	0.75±0.55 de	11.81±0.95 b	32.90±5.00 c	62.91±8.28 c
Mf19-3-8^S	10.25±0.29 ab	0.40±0.51 e	74.55±12.77 a	96.67±5.60 a	100.00±3.12 a
Mf19-1-7^LR	9.96±0.88 bc	2.50±1.26 a	17.83±5.05 b	60.43±4.60 b	90.28±1.12 ab
Mf19-3-1^LR	10.27±0.17 a	2.05±1.03 abc	6.86±4.73 b	42.18±3.33 bc	80.46±0.00 bc
Mf19-4-6^MR	9.97±0.53 bc	0.07±0.17 f	—	—	—
Mf19-5-9^MR	10.21±0.33 ab	2.05±0.37 ab	13.85±3.03 b	62.25±2.64 b	83.80±1.96 ab
Mf19-5-10^MR	10.21±0.44 ab	1.55±0.80 bcd	0.95±1.17 b	36.22±4.90 c	80.66±3.65 bc
注：^S代表啶酰菌胺敏感菌株，^LR代表啶酰菌胺低抗菌株，^MR代表啶酰菌胺中抗菌株。^同列数据后不同字母表示在P=0.05水平显著性差异。“±” 代表3次重复的平均生长速率、产孢量及孢子萌发率的标准误。Note: The superscript letter^"S"adjacent to the isolate number represents boscalid-sensitivity. ^"LR" represents boscalid low-resistance.^"MR"represents boscalid medium-resistance. Values in a column followed by different lowercase letters are significantly different at P = 0.05 level. "±" represents the standard error among three repetitions of average growth rates, sporulation quantities, and spore germination rates.

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表 4 樱桃褐腐病菌对啶酰菌胺的EC₅₀及氨基酸突变位点

Table 4. EC₅₀ of boscalid and amino acid mutation sites in M. fructicola

菌株 Isolate	EC₅₀/(µg/mL)	氨基酸突变 Mutation of amino acid
Mf19-3-7^S	1.05	—
Mf19-3-8^S	1.30	I117V，R237P
Mf19-3-10^S	1.34	—
Mf19-4-7^S	2.65	A133V，R237P
Mf19-4-10^S	1.02	—
Mf19-1-7^LR	10.17	—
Mf19-3-1^LR	5.68	A133V，R237P
Mf19-4-6^MR	25.14	—
Mf19-5-9^MR	17.761	A133V，R237P
Mf19-5-10^MR	30.59	A133V，R237P
注：^S代表啶酰菌胺敏感菌株，^LR代表啶酰菌胺低抗菌株，^MR代表啶酰菌胺中抗菌株。“—” 表示该菌株中未发生氨基酸突变。Note: The superscript letter^"S" adjacent to the isolate number represents boscalid-sensitivity. ^"LR"represents boscalid low-resistance. ^"MR" represents boscalid medium-resistance. "—" means that no amino acid point mutations had occurred in this strain.

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