草莓根腐病病原菌的鉴定及其对杀菌剂的敏感性

doi:10.3969/j.issn.1004-1524.20240138

浙江农业学报 ›› 2025, Vol. 37 ›› Issue (2): 417-425.DOI: 10.3969/j.issn.1004-1524.20240138

草莓根腐病病原菌的鉴定及其对杀菌剂的敏感性

沈岚¹(), 杨肖芳², 张国芳¹^,^*()

1.宁波市农业科学研究院生物技术研究所,宁波市特色园艺作物品质调控与抗性育种重点实验室,浙江宁波 315040
2.浙江省农业科学院园艺研究所,浙江杭州 310021

收稿日期:2024-02-06 出版日期:2025-02-25 发布日期:2025-03-20
作者简介:*张国芳,E-mail: zhangguofang401@163.com
沈岚(1976—),男,浙江宁波人,学士,高级农艺师,从事草莓育种、栽培与病虫害防治研究。E-mail:shenlan5056@163.com
通讯作者: 张国芳
基金资助:
宁波市“科技创新2025”重大专项(2019B10021);宁波市重点研发计划(2023Z112);浙江省农业新品种选育重大科技专项(2021C02066-7)

Identification and fungicide sensitivity of the pathogen causing root rot on strawberry

SHEN Lan¹(), YANG Xiaofang², ZHANG Guofang¹^,^*()

1. Ningbo Key Laboratory of Characteristic Horticultural Crops in Quality Adjustment and Resistance Breeding, Institute of Biotechnology, Ningbo Academy of Agricultural Sciences, Ningbo 315040, Zhejiang, China
2. Institute of Horticulture, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China

Received:2024-02-06 Online:2025-02-25 Published:2025-03-20
Contact: ZHANG Guofang

摘要/Abstract

摘要： 根腐病是草莓上最严重的病害之一。为了明确近期发生在浙江省宁波市的草莓根腐病的病原菌,以受到病害的草莓根基部为材料,采用组织分离法和致病性测定获得病原菌,并基于形态学观察、ITS、TEF1-α序列,及系统发育分析进行病原菌鉴定。同时,采用生长速率法测定7种常规杀菌剂(咪鲜胺、吡唑醚菌酯、抑霉唑、溴菌腈、戊唑醇、嘧菌酯和氟啶胺)原药对该病原菌的室内毒力。结果表明,近期在宁波市引发草莓根腐病的病原菌为菜豆壳球孢(Macrophomina phaseolina)。该菌除可侵染草莓根基部外,还可侵染叶片和叶柄。室内毒力测定试验结果显示,对该菌毒力较强的是氟啶胺、戊唑醇、抑霉唑、咪鲜胺,有效中浓度(EC₅₀)分别为0.000 178、0.568、0.959、1.179 μg·mL^-1。这是浙江省首次报道M. phaseolina在草莓上引起病害。氟啶胺、戊唑醇、抑霉唑、咪鲜胺可用于防控M. phaseolina导致的草莓根腐病。

关键词: 草莓, 根腐病, 菜豆壳球孢(Macrophomina phaseolina), 室内毒力检测

Abstract:

Root rot is one of the most serious diseases on strawberry. To identify the pathogen causing root rot on strawberry in Ningbo, Zhejiang, China, diseased crown and rot were collected, and the pathogen was obtained by the tissue method and pathogenicity test. The pathogen was further identified by the morphological observation, ITS and TEF1-α sequence analysis and phylogenetic trees. Meanwhile, the toxicity of seven fungicides (prochloraz, pyraclostrobin, bromothalonil, prochloraz, azoxystrobin, tebuconazole, fluazinam) against this pathogen was determined in vitro. It was shown that the pathogen causing crown and root rot on strawberry in Ningbo was identified as Macrophomina phaseolina, which could also cause the leaf and petiole withered. Laboratory toxicity tests of 7 fungicides showed that fluazinam, tebuconazole, imazalil, prochloraz were the most virulent fungicides with median effective concentration (EC₅₀) of 0.000 178, 0.568, 0.959, 1.179 μg·mL^-1, respectively. To our knowledge, this is the first report of M. phaseolina causing root rot on strawberry in Zhejiang, China. Fluazinam, tebuconazole, imazalil, prochloraz are potential fungicides for the control of this disease caused by M. phaseolina on strawberry.

Key words: strawberry, root rot, Macrophomina phaseolina, laboratory toxicity determination

中图分类号:

S436.684

沈岚, 杨肖芳, 张国芳. 草莓根腐病病原菌的鉴定及其对杀菌剂的敏感性[J]. 浙江农业学报, 2025, 37(2): 417-425.

SHEN Lan, YANG Xiaofang, ZHANG Guofang. Identification and fungicide sensitivity of the pathogen causing root rot on strawberry[J]. Acta Agriculturae Zhejiangensis, 2025, 37(2): 417-425.

图/表 8

表1 各杀菌剂的供试浓度

Table 1 Effective concentration of fungicides

杀菌剂 Fungicides	供试浓度 Effective concentration/(μg·mL^-1)
咪鲜胺Prochloraz	150.0、30.0、4.0、1.0、0.2
吡唑醚菌酯	50、20、10、5、1
Pyraclostrobin
溴菌腈Bromothalonil	30、20、10、8、5
抑霉唑Imazalil	5.00、2.00、1.00、0.10、0.01
嘧菌酯Azoxystrobin	200、150、50、10、5
戊唑醇Tebuconazole	15.0、10.0、5.0、1.0、0.5
氟啶胺Fluazinam	0.050 0、0.001 0、0.000 8、0.000 5、0.000 1

图1 草莓根腐病田间症状 A,枯萎植株;B、C,发病早期植株。

Fig.1 Symptoms of root rot on strawberry in the field A, Wilted plants; B, C, Diseased plants.

图2 分离物菌株的菌落与显微形态特征 A、B,培养5 d后的菌落;C,菌核;D,菌丝;E,正在释放孢子的分生孢子器;F,分生孢子(椭圆形至倒卵形)。

Fig.2 Colony and microscopic characteristics of the isolate A, B, Colony cultured for 5 days; C, Sclerotium; D, Mycelium; E, Pycnidia releasing conidia; F, Conidia (elliptic to obova).

图3 草莓人工接种XXmy-1菌株10 d后的症状 A、C,对照;B,接种XXmy-1 10 d后的症状;D,接种XXmy-1 10 d后的草莓根、基部;E,接种XXmy-1 10 d后的草莓叶柄。

Fig.3 Symptoms of strawberry after artificial inoculation with XXmy-1 for 10 d A, C, Control; B, 10 d after inoculation with XXmy-1; D, Root and crown of strawberry inoculated with XXmy-1 for 10 d; E, Petiole of strawberry inoculated with XXmy-1 for 10 d.

图4 草莓不同部位人工接种XXmy-1菌株后的症状 A、C、E,对照;B、D,接种XXmy-1 48 h后的症状;F,接种XXmy-1 72 h后的症状;G,分生孢子器;H,分生孢子。

Fig.4 Symptoms of strawberry after artificial inoculation with XXmy-1 at different parts A, C, E, Control; B, D, 48 h after inoculation with XXmy-1; F, 72 h after inoculation with XXmy-1; G, Pycnidia; H, Conidia.

图5 分离物XXmy-1、XXmy-2、XXmy-3、XXmy-4的ITS和TEF1-α序列PCR电泳图 A,ITS序列的PCR扩增产物电泳图;B,TEF1-α序列的PCR扩增产物电泳图;M,2 kb DNA ladder;1~4,XXmy-1、XXmy-2、XXmy-3、XXmy-4;5~8,XXmy-1、XXmy-2、XXmy-3、XXmy-4。

Fig.5 PCR products of partial ITS and TEF1-α amplification of the isolates A, PCR products of partial ITS sequences amplification; B, PCR products of partial TEF1-α sequences amplification; M, 2 kb DNA ladder; 1-4, ITS fragment of XXmy-1, XXmy-2, XXmy-3, XXmy-4; 5-8, TEF1-α fragment of XXmy-1, XXmy-2, XXmy-3, XXmy-4.

图6 基于ITS和TEF1-α序列以最大似然法联合构建的系统发育树

Fig.6 Maximum-likelihood tree based on ITS and TEF1-α sequences

表2 7种杀菌剂对M. phaseolina的毒力

Table 2 Toxicity of seven fungicides to M. phaseolina

药剂 Fungicide	回归方程 Regression equation	EC₅₀/(μg·mL^-1)	相关系数 Correlation coefficient
咪鲜胺Prochloraz	y=4.893+1.083x	1.179	0.988
吡唑醚菌酯Pyraclostrobin	y=4.859+0.258x	3.549	0.957
溴菌腈Bromothalonil	y=2.566+2.669x	8.078	0.946
抑霉唑Imazalil	y=5.011+0.613x	0.959	0.944
嘧菌酯Azoxystrobin	y=3.095+0.953x	98.793	0.966
戊唑醇Tebuconazole	y=5.407+1.331x	0.568	0.963
氟啶胺Fluazinam	y=10.967+1.592x	0.000 178	0.927

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草莓根腐病病原菌的鉴定及其对杀菌剂的敏感性

Identification and fungicide sensitivity of the pathogen causing root rot on strawberry

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