福建省大豆胶孢炭疽病菌对吡唑醚菌酯的抗性风险分析

阮宏椿; 黄艳琴; 陈巧红; 陈文乐; 兰成忠

福建省大豆胶孢炭疽病菌对吡唑醚菌酯的抗性风险分析

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/福建省农作物品种抗性工程技术研究中心，福建　福州　350013
2.
三明市种子站，福建　三明　365000
3.
三明市植保植检站，福建　三明　365000

基金项目: 福建省科技计划公益类专项（2021R1024002）；福建省农业科学院对外合作项目（DWHZ-2023-04）；福建省农业高质量发展超越“5511”协同创新工程项目（XTCXGC2021011、XTCXGC2021017）

详细信息

作者简介:
阮宏椿（1980 — ），女，硕士，副研究员，主要从事植物真菌病害防治研究，E-mail：hcruan@126.com

通讯作者:
兰成忠（1979 — ），男，博士，副研究员，主要从事植物病害防治技术研究，E-mail：lczhong7911@126.com

中图分类号: S435.29; S482.2
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出版历程
- 收稿日期: 2024-05-23
- 修回日期: 2024-08-20
- 网络出版日期: 2024-11-11

Risk Assessment on Resistance to Pyraclostrobin of Colletotrichum gloeosporioides

1.
Institute of Plant Protection, Fujian Academy of Agricultural Sciences/Fujian Key Laboratory for Monitoring and Integrated Management of Crop Pests/Fujian Crop Variety Resistance Engineering Technology Research Center, Fuzhou, Fujian　350013, China
2.
Sanming Seed Station, Sanming, Fujian　365000, China
3.
Sanming Plant Protection and Inspection Station, Sanming, Fujian　365000, China

摘要

摘要: 目的探明福建省大豆胶孢炭疽菌群体对吡唑醚菌酯的敏感程度及其与其他杀菌剂的交互抗性。方法采用菌丝生长速率法对2023年分离自福建省5个地区的112株大豆胶孢炭疽菌进行敏感性分析，以药剂驯化法获得抗性突变体，测定抗性突变体的遗传稳定性、适合度及对4种不同杀菌剂之间的交互抗性。结果吡唑醚菌酯对112个菌株的EC₅₀值介于0.0682～1.0309 µg·mL⁻¹，变异系数为15.12，敏感性频率分布为连续性单峰曲线，符合偏正态分布，EC₅₀平均值0.2036±0.1215 µg·mL⁻¹，可作为大豆胶孢炭疽菌对吡唑醚菌酯的敏感性基线；从2株大豆胶孢炭疽菌野生菌株中诱导获得4株中抗吡唑醚菌酯的突变体，1株低抗吡唑醚菌酯的突变体，突变频率为8.3×10⁻⁴，其抗药性性状能稳定遗传，抗性突变体与其亲本菌株相比，对温度的敏感性存在差异，抗性突变体菌丝的生长速率、产孢能力及致病力，通常略低于其亲本菌株的生存适应度，抗性突变体在田间可能存在一定适合度代价，但抗性突变体具有一定的自然竞争优势；吡唑醚菌酯与不同类杀菌剂多菌灵、苯醚甲环唑、咪鲜胺之间不存在交互抗性，与同属一类的杀菌剂啶氧菌酯存在明显的交互抗性。结论福建省大豆胶孢炭疽菌对吡唑醚菌酯具有较高的敏感性，但存在中等抗性风险，生产中可将吡唑醚菌酯与不同类杀菌剂多菌灵、苯醚甲环唑、咪鲜胺混合或交替使用，以延缓大豆胶孢炭疽菌抗药性的产生。
- 大豆胶孢炭疽菌 /
- 吡唑醚菌酯 /
- 敏感基线 /
- 抗性风险 /
- 交互抗性
Abstract: Objective Drug sensitivity to and cross-resistance of pyraclostrobin with other fungicides of Colletotrichum gloeosporioides that caused anthracnose in soybeans in Fujian Province were investigated. Methods Sensitivity to pyraclostrobin of 112 strains of C. gloeosporioides isolated from 5 regions in the province in 2023 was determined using the mycelial production rate method. Resistant mutants of the pathogen were obtained by fungicide domestication to determine their genetic stability, fitness, and possible cross-resistance to 4 fungicides. Results The EC₅₀ of pyraclostrobin for the 112 C. gloeosporioides strains ranged 0.0682 µg·mL⁻¹ to 1.0309 µg·mL⁻¹ with a coefficient of variation of 15.12 and averaged 0.203 6±0.121 5 µg·mL⁻¹ as the sensitivity baseline. The distribution of the sensitivity frequency was in a continuous unimodal function consistent with the continuous skewed normal distribution. Induced from two wild C. gloeosporioides, 4 moderate resistant strains and one low resistant strain were secured at a frequency of 8.3×10⁻⁴. The mutants stably inherited the drug resistance traits but differed from their parents in temperature sensitivity and were slightly lower on mycelial growth rate, sporulation ability, and pathogenicity. In the field, they might sacrifice other natural competitive advantages as a fitness cost in exchange for the drug resistance. Pyraclostrobin showed no cross-resistance with carbendazim, difenoconazole, and prochloraz but did with the same type of fungicide, picoxystrobin. Conclusion Although the C. gloeosporioides isolated in Fujian were relatively high in sensitivity to pyraclostrobin, they represented merely a moderate drug resistance risk. Nonetheless, in practice, it was plausible to blend pyraclostrobin or apply alternatively with fungicides such as carbendazim, difenoconazole, and prochloraz to delay such potential drawback.
- Colletotrichum gloeosporioides /
- pyraclostrobin /
- sensitivity baseline /
- resistance risk /
- cross-resistance

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图 1 福建省大豆胶孢炭疽菌对吡唑醚菌酯的敏感性频率分布

Figure 1. Frequency distribution of EC₅₀ of pyraclostrobin for C. gloeosporioides from Fujian

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图 2 吡唑醚菌酯与多菌灵、苯醚甲环唑、咪鲜胺和啶氧菌酯的交互抗性

Figure 2. Cross-resistance between pyraclostrobin and carbendazim, phenylimidazole, prochloraz, and propiconazole

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表 1 福建不同地区大豆胶孢炭疽菌对吡唑醚菌酯的敏感性

Table 1. Sensitivity to pyraclostrobin of C. gloeosporioides isolated from regions in Fujian

采集地区 Sampling region	菌株数 Number of isolates	吡唑醚菌酯EC₅₀值 EC₅₀ of Pyraclostrobin/(µg·mL⁻¹)		菌株间变异系数 Variation factor
采集地区 Sampling region	菌株数 Number of isolates	范围 Range	平均值 Mean	菌株间变异系数 Variation factor
闽东地区 Mindong region	24	0.0682～0.3777	0.1662±0.0633 a	5.54
闽南地区 Minnan region	31	0.0763～0.3506	0.1845±0.0751 a	4.60
闽西地区 Minxi region	27	0.0801～0.6428	0.2451±0.1359 a	8.02
闽北地区 Minbei region	15	0.1284～1.0309	0.2387±0.2269 a	8.03
闽中地区 Minzhong region	15	0.0725～0.3007	0.1933±0.0670 a	4.15
总计 Total	112	0.0682～1.0309	0.2036±0.1215 a	15.12
表中数据为平均数±标准差。同列数据后不同小写字母分别表示经Duncan氏新复极差法检验差异显著（P＜0.05）。下同。 Data are mean±SD; those with different lowercase letters on same column indicate significant difference at P＜0.05 in Duncan's new multiple range test.

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表 2 吡唑醚菌酯抗性突变体连续转代培养后EC₅₀和抗性倍数的变化

Table 2. Changes on EC₅₀ and resistance multiple of pyraclostrobin-resistant C. gloeosporioides mutants after continuous generational transference

菌株 Strain	第一代 1^st generation		第三代 3^rd generation		第五代 5^th generation		第七代 7^th generation
菌株 Strain	EC₅₀/(µg·mL⁻¹)	RM	EC₅₀/(µg·mL⁻¹)	RM	EC₅₀/(µg·mL⁻¹)	RM	EC₅₀/(µg·mL⁻¹)	RM
CG23017	0.1349	/	0.1396	/	0.1369	/	0.1344	/
CG23017-1	1.7673	13.10	1.7823	12.77	1.7121	12.5	1.6661	12.39
CG23017-2	0.6674	4.95	0.5275	3.78	0.4891	3.57	0.4376	3.26
CG23017-3	1.9268	14.29	1.9084	13.67	1.8966	13.85	1.8612	13.85
CG23052	0.1611	/	0.1647	/	0.1623	/	0.1580	/
CG23052-1	3.0151	18.71	3.0198	18.34	2.9742	18.33	2.9574	18.72
CG23052-2	2.7140	16.84	2.7191	16.51	2.7200	16.76	2.6425	16.73
RM为抗性倍数。 RM: Resistance multiple.

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表 3 抗性突变体及其亲本菌株对温度的敏感性测定

Table 3. Mycelial growth of resistant mutants and parents under different temperatures

菌株 Strain	菌落直径 Colony diameter/mm
菌株 Strain	16 ℃	20 ℃	25 ℃	28 ℃	35 ℃
CG23017	31.33±0.58 cd	43.33±1.53 c	59.33±2.31 c	63.67±1.53 b	20.67±1.53 a
CG23017-1	28.67±0.58 e	41.33±1.15 cd	58.33±1.15 c	60.67±1.53 b	18.67±3.06 ab
CG23017-2	25.33±1.15 f	37.67±1.15 e	56.67±1.53 c	60.33±2.31 b	19.33±1.53 ab
CG23017-3	29.67±1.53 de	39.33±1.53 de	57.33±1.53 c	60.67±2.31 b	19.67±1.15 ab
CG23052	35.67±2.08 a	48.33±1.15 a	72.67±0.58 a	74.67±1.53 a	17.67±1.53 ab
CG23052-1	34.33±1.53 ab	45.67±1.15 b	69.67±1.15 b	71.67±1.53 a	18.33±0.58 ab
CG23052-2	32.33±1.53 bc	46.33±0.58 ab	68.33±1.53 b	71.33±2.52 a	16.67±2.52 b

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表 4 抗性突变体及其亲本菌株菌丝生长速率、产孢能力、致病力测定

Table 4. Mycelial growth rate, sporulation per area, and disease index of resistant mutants and parents

菌株 Strain	菌丝生长速率 Mycelium growth rate/(mm·d⁻¹)	单位面积产孢量 Sporulation per area/(×10³个·mm⁻²)	病情指数 Disease index
CG23017	9.1±0.22 b	4.58±0.27 ab	47.52±2.34 a
CG23017-1	8.67±0.22 b	4.55±0.19 ab	40.37±4.56 a
CG23017-2	8.62±0.33 b	4.32±0.08 b	27.08±4.83 b
CG23017-3	8.67±0.33 b	4.50±0.26 ab	42.48±10.06 a
CG23052	10.67±0.22 a	4.77±0.25 a	41.48±8.82 a
CG23052-1	10.24±0.22 a	4.74±0.23 a	26.63±6.32 b
CG23052-2	10.19±0.36 a	4.61±0.2 ab	37.43±0.89 ab

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