绣球叶斑病的病原菌分离鉴定与防治研究

doi:10.3969/j.issn.1004-1524.20241081

浙江农业学报 ›› 2026, Vol. 38 ›› Issue (1): 105-113.DOI: 10.3969/j.issn.1004-1524.20241081

绣球叶斑病的病原菌分离鉴定与防治研究

陈晶晶¹(), 张玉¹, 温联好¹, 刘光炎¹, 周梅¹, 李正敏¹, 熊海燕¹, 杨振翼², 杨曾华², 童译², 王超¹^,^*()

1.西南林业大学园林园艺学院,云南省功能性花卉资源及产业化技术工程研究中心,国家林业和草原局西南风景园林工程技术研究中心,云南省面向南亚东南亚经济林全产业链联合研发中心,云南省森林灾控预警与控制实验室,云南昆明 650224
2.昆明杨月季园艺有限责任公司,云南昆明 652501

收稿日期:2024-12-11 出版日期:2026-01-25 发布日期:2026-02-11
作者简介:*王超,E-mail:47188127@qq.com
陈晶晶,研究方向为园林植物与观赏园艺。E-mail:3327197360@qq.com
通讯作者: 王超
基金资助:
云南省重大科技专项(202202AE090028);云南省大学生创新训练计划(20200554052)

Isolation, identification and control of the pathogen causing leaf spot disease on hydrangea

CHEN Jingjing¹(), ZHANG Yu¹, WEN Lianhao¹, LIU Guangyan¹, ZHOU Mei¹, LI Zhengmin¹, XIONG Haiyan¹, YANG Zhenyi², YANG Zenghua², TONG Yi², WANG Chao¹^,^*()

1. Yunnan Province Engineering Research Center for Functional Flower Resources and Industrialization, Southwest Research Center for Landscape Architecture Engineering (State Forestry and Grassland Administration), Yunnan Province South and Southeast Asia Joint R&D Center of Economic Forest Full Industry Chain, Key Laboratory of Forest Disaster Warning and Control in Universities of Yunnan Province, College of Landscape and Horticulture, Southwest Forestry University, Kunming 650224, China
2. Kunming Yang Chinese Rose Gardening Co., Ltd., Kunming 652501, China

Received:2024-12-11 Online:2026-01-25 Published:2026-02-11
Contact: WANG Chao

摘要/Abstract

摘要：

为明确绣球叶斑病的致病菌及其生物学特性,探寻适宜的防治方法,本研究对绣球品种玛梅丽的叶斑病样本进行组织分离,纯化病原菌,基于柯赫氏法则验证,并利用基于ITS、LSU、TEF1等多基因序列构建的系统发育树分析,对病原菌进行鉴定。同时,采用菌丝生长速率法,研究温度、培养基、pH值、碳源、氮源、光周期对病原菌菌丝生长的影响,并测定其致死温度。此外,还测试了7种化学药剂对病原菌的抑制效果。结果显示,分离到的绣球叶斑病病原菌为神农架鹅膏菌(Amanita shennongjiana),其菌丝在25 ℃、马铃薯葡萄糖琼脂培养基,pH值为5,每日各12 h光、暗交替的条件下生长较好,其最适碳源为蔗糖,偏好不加氮,致死温度为65 ℃持续10 min。室内环境下,80%乙蒜素和80%波尔多液对该菌的抑制效果较好,其有效中浓度(EC₅₀)分别为1.948 9、1.591 8 mg·L^-1。本研究是对由神农架鹅膏菌引起的绣球叶斑病在中国的首次报道。

关键词: 绣球, 叶斑病, 神农架鹅膏菌(Amanita shennongjiana), 生物学特性, 药剂筛选

Abstract:

In order to identify the pathogenic fungus of hydrangea leaf spot disease and its biological characteristics, and explore suitable control methods, tissue isolation was conducted on leaf spot samples of hydrangea cultivar Mme E Mouilliere, and the pathogenic fungus was purified, and verified based on Koch’s rule. Phylogenetic tree analysis was constructed using multi-gene sequences (ITS, LSU, TEF1) to identify the pathogenic fungus. Meanwhile, the mycelial growth rate method was adopted to investigate the effects of temperature, culture media, pH value, carbon source, nitrogen source, and photoperiod on the mycelial growth of the pathogenic fungus, and its lethal temperature was determined. Additionally, the inhibitory effects of 7 chemical fungicides on the fungus were tested. The results showed that the pathogenic fungus causing hydrangea leaf spot disease was identified as Amanita shennongjiana. Its mycelia grew well under the following conditions: temperature of 25 ℃, potato dextrose agar (PDA) medium, pH value of 5, and 12 h light/12 h dark alternation daily. Its optimal carbon source was sucrose, with a preference for nitrogen-free conditions. The lethal temperature of the fungus was 65 ℃ for 10 minutes. Under laboratory conditions, 80% ethylicin and 80% Bordeaux mixture exhibited better inhibitory effects on the fungus, with their median effective concentrations (EC₅₀) being 1.948 9 mg·L^-1 and 1.591 8 mg·L^-1, respectively. This study is the first report of hydrangea leaf spot disease caused by Amanita shennongjiana in China.

Key words: hydrangea, leaf spot disease, Amanita shennongjiana, biological characteristic, screening of fungicide

中图分类号:

陈晶晶, 张玉, 温联好, 刘光炎, 周梅, 李正敏, 熊海燕, 杨振翼, 杨曾华, 童译, 王超. 绣球叶斑病的病原菌分离鉴定与防治研究[J]. 浙江农业学报, 2026, 38(1): 105-113.

CHEN Jingjing, ZHANG Yu, WEN Lianhao, LIU Guangyan, ZHOU Mei, LI Zhengmin, XIONG Haiyan, YANG Zhenyi, YANG Zenghua, TONG Yi, WANG Chao. Isolation, identification and control of the pathogen causing leaf spot disease on hydrangea[J]. Acta Agriculturae Zhejiangensis, 2026, 38(1): 105-113.

图/表 11

图1 绣球叶斑病病害早期(A、B)和后期(C)症状照片

Fig.1 Photos of early (A, B) and late (C) symptoms of hydrangea leaf spot disease

图2 病原菌的形态鉴定及其致病性 A、C,菌株活体回接前、后;B、D:不接菌的空白对照;E、H,菌丝;F,在PDA培养基上培养7 d的菌落特征;G,孢子。

Fig.2 Morphological identification and pathogenicity of pathogen A,C, Results before and after in vivo inoculation, respectively; B, D, Results without in vivo inoculation; E, H, Mycelium; F, Characteristics of colony cultured on PDA medium for 7 days; G, Basidiospore.

图3 基于分离菌株ITS序列、LSU基因和TEF1基因序列联合构建的系统发育树

Fig.3 The constructed phylogenetic tree of isolated strains based on sequences of ITS, LSU and TEF1 genes

图4 不同温度对病原菌菌丝生长的影响柱上无相同字母的表示差异显著(p<0.05)。下同。

Fig.4 Effect of different temperature on mycelial growth rate of pathogen Bars marked without the same letters indicate significant difference at p<0.05. The same as below.

图5 不同培养基对病原菌菌丝生长的影响 OMA,燕麦琼脂培养基;CMA,玉米琼脂培养基;LB,LB培养基;JZ,叶片煎汁琼脂培养基;PDA,马铃薯葡萄糖琼脂培养基。

Fig.5 Effect of different culture media on mycelial growth of pathogen OMA, Oatmeal agar medium; CMA, Cornmeal agar medium; LB, LB medium; JZ, Leaf extract agar medium; PDA, Potato dextrose agar medium.

图6 不同光周期对病原菌菌丝生长的影响 T1,每日24 h光照;T2,每日12 h光照与12 h黑暗交替;T3,每日24 h黑暗。

Fig.6 Effect of different photoperiods on mycelial growth rate of pathogen T1,24 h light per day; T2, 12 h light and 12 h dark per day; T3, 24 h dark per day.

图7 不同氮源对病原菌菌丝生长的影响

Fig.7 Effect of different nitrogen sources on mycelial growth of pathogen

图8 不同碳源对病原菌菌丝生长的影响

Fig.8 Effect of different carbon sources on mycelial growth of pathogen

图9 不同pH值对病原菌菌丝生长的影响

Fig.9 Effect of different pH values on mycelial growth of pathogen

图10 病原菌的致死温度

Fig.10 Lethal temperature of pathogen

表1 各杀菌剂对病原菌的室内毒力

Table 1 Toxicity of fungicides against pathogen

杀菌剂 Fungicide	回归方程 Regression equation	相关系数 Correlation coefficient(r)	EC₅₀/(mg·L^-1)
80%代森锰锌80% Mancozeb	y=3.26x-1.63	0.73	2.379 5
10%多抗霉素10% Polyoxin	y=0.30x+4.40	0.77	2.284 7
80%乙蒜素80% Ethylicin	y=4.09x-4.12	0.98	1.948 9
80%波尔多液80% Bordeaux mixture	y=0.27x+4.05	0.80	1.591 8
50%啶酰菌胺50% Boscalid	y=0.44x+4.18	0.93	2.717 2
80%嘧霉胺80% Pyrimethanil	y=0.38x+5.50	0.75	2.497 5
没食子酸丙酯Propyl gallate	y=1.04x+2.07	0.89	6.326 2

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绣球叶斑病的病原菌分离鉴定与防治研究

Isolation, identification and control of the pathogen causing leaf spot disease on hydrangea

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