绣球博大蓝叶斑病病原菌的分离鉴定与防治

doi:10.3969/j.issn.1004-1524.20250254

浙江农业学报 ›› 2026, Vol. 38 ›› Issue (4): 696-706.DOI: 10.3969/j.issn.1004-1524.20250254

绣球博大蓝叶斑病病原菌的分离鉴定与防治

张玉¹(), 陈晶晶¹, 温联好¹, 杨振翼², 杨曾华², 童译², 王超¹^,^*()

¹ 西南林业大学国家林业和草原局西南风景园林工程技术研究中心, 云南省功能性花卉资源及产业化技术工程研究中心,云南省森林灾害预警与控制实验室,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:2025-04-02 出版日期:2026-04-25 发布日期:2026-05-08
作者简介:*王超,E-mail:47188127@qq.com
张玉,研究方向为园艺学。E-mail:2316923973@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项目(20200554052)

Isolation, identification, and control of the pathogen causing leaf spot disease on Hydrangea macrophylla Bo Dalan

ZHANG Yu¹(), CHEN Jingjing¹, WEN Lianhao¹, YANG Zhenyi², YANG Cenghua², TONG Yi², WANG Chao¹^,^*()

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

Received:2025-04-02 Published:2026-04-25 Online:2026-05-08
Contact: WANG Chao

摘要/Abstract

摘要：

为明确绣球博大蓝叶斑病的病原菌,并探究其生物学特性和有效防治药剂,为病害的科学防控提供理论依据,本研究采用组织分离法对采集自云南省玉溪市南国山花绣球种质资源基地的博大蓝叶斑病病样进行病原菌分离,结合柯赫氏法则验证其致病性;通过形态学观察和多基因序列(ITS、LSU、TUB)联合分析对病原菌进行鉴定,采用菌丝生长速率法,研究不同培养基、碳源、氮源、温度、光照和pH值条件对病原菌菌丝生长的影响;并选取9种常用杀菌剂测定其对病原菌的室内毒力。结果显示,从感病叶片上分离得到的主要病原菌为高粱附球菌(Epicoccum sorghinum)。生物学特性测定结果表明,该菌菌丝生长的最适条件为以麦芽糖为碳源、蛋白胨和牛肉浸膏为氮源的PDA培养基,在温度25 ℃、pH值7.0和全光照环境下菌丝生长最佳。室内毒力测定结果显示,10%苯醚甲环唑水分散性粒剂(WG)的抑菌效果最好,其EC₅₀值为1.252 2 mg·L^-1。综上可知,引起绣球博大蓝叶斑病的病原菌为高粱附球菌,10%苯醚甲环唑WG对该病原菌具有较好的抑制作用,可作为田间防治的候选药剂。

关键词: 绣球博大蓝, 叶斑病, 病原菌, 高粱附球菌, 生物学特性, 10%苯醚甲环唑水分散性粒剂

Abstract:

To identify the pathogen causing leaf spot disease on Hydrangea macrophylla Bo Dalan and investigate its biological characteristics and effective control agents, thereby providing a theoretical basis for the scientific management of this disease, this study isolated the pathogen from diseased samples collected at the Nanguo Shanhua Hydrangea Germplasm Resource Base in Yuxi City, Yunnan Province, using the tissue isolation method. The pathogenicity was verified according to Koch’s postulates. The pathogen was identified based on morphological observations and multi-gene sequence (ITS, LSU, TUB) analysis. The effects of different culture media, carbon sources, nitrogen sources, temperature, light conditions, and pH value on the mycelial growth of the pathogen were studied using the mycelial growth rate method. Additionally, nine commonly used fungicides were selected to determine their in vitro toxicity against the pathogen. The results showed that the primary pathogen isolated from the infected leaves was Epicoccum sorghinum. The biological characteristics tests indicated that the optimal conditions for mycelial growth were PDA medium with maltose as the carbon source and peptone and beef extract as nitrogen sources, at a temperature of 25 ℃, pH value 7.0, and under continuous light. The in vitro toxicity tests revealed that 10% difenoconazole water dispersible granules (WG) had the best inhibitory effect, with an EC₅₀ value of 1.252 2 mg·L^-1. In conclusion, the pathogen causing leaf spot disease on H. macrophylla Bo Dalan was E. sorghinum, and 10% difenoconazole WG exhibited a strong inhibitory effect against this pathogen, making it a potential candidate for field control.

Key words: Hydrangea Bo Dalan, leaf spot disease, pathogen, Epicoccum sorghinum, biological characteristic, 10% difenoconazole water dispersible granules (WG)

中图分类号:

S432.44

张玉, 陈晶晶, 温联好, 杨振翼, 杨曾华, 童译, 王超. 绣球博大蓝叶斑病病原菌的分离鉴定与防治[J]. 浙江农业学报, 2026, 38(4): 696-706.

ZHANG Yu, CHEN Jingjing, WEN Lianhao, YANG Zhenyi, YANG Cenghua, TONG Yi, WANG Chao. Isolation, identification, and control of the pathogen causing leaf spot disease on Hydrangea macrophylla Bo Dalan[J]. Acta Agriculturae Zhejiangensis, 2026, 38(4): 696-706.

图/表 7

表1 PCR扩增所用引物

Table 1 Primers used for PCR amplification

基因名称 Gene name	正向引物序列(5'→3') Forward primer sequence(5'→3')	反向引物序列(5'→3') Reverse primer sequence(5'→3')	扩增片段长度/bp Amplicon length/bp
ITS	TCCGTAGGTGAACCTGCGG	TCCTCCGCTTATTGATATGC	560
LSU	ACCCGCTGAACTTAAGC	TCCTGAGGGAAACTTCG	870
TUB	GGTAACCAAATCGGTGCTGCTTTC	ACCCTCAGTGTAGTGACCCTTGGC	460

图1 博大蓝田间叶斑病症状 A,田间健康叶片;B、C,田间感病叶片与发病症状。

Fig.1 Field symptoms of leaf spot disease on Bo Dalan A, Healthy leaves in the field; B and C, Diseased leaves in the field and the associated symptom manifestations.

图2 病原菌在PDA培养基上的形态特征 A、B和C,菌落形态;D,菌丝形态;E,厚垣孢子;F,分生孢子。

Fig.2 Morphological characteristics of the fungal pathogen on PDA medium A, B and C, Colony morphology; D, Mycelial morphology; E, Chlamydospores; F, Conidia.

图3 绣球叶斑病病原菌接种后症状 A、B和C,室内离体接种后叶片发病症状;D、E和F,棚内活体接种后叶片发病症状。

Fig.3 Symptoms of hydrangea leaves after pathogen inoculation A, B and C, Leaf disease symptoms after in vitro inoculation indoors; D, E and F, Leaf disease symptoms after in vivo inoculation in the greenhouse.

图4 基于ITS序列、LSU与TUB基因的多基因系统发育树括号内为该菌株所对应引物在NCBI数据库中的登录号。

Fig.4 Multigene phylogenetic tree based on ITS sequence, LSU and TUB genes The accession number of the primer corresponding to this strain in the NCBI database is indicated in parentheses.

图5 温度、光照条件、培养基、氮源、碳源、致死温度、pH值对Epicoccum sorghinum菌丝生长的影响 PDA,马铃薯葡萄糖琼脂培养基;LB,溶菌肉汤培养基;OMA,燕麦片琼脂培养基;CMA,玉米粉琼脂培养基;Czapek,察氏培养基。

Fig.5 Effects of temperature, light conditions, culture medium, nitrogen source, carbon source, lethal temperature, and pH value on mycelial growth of Epicoccum sorghinum PDA, Potato dextrose agar medium; LB, Luria-Bertani broth medium; OMA, Oatmeal agar medium; CMA, Corn meal agar medium; Czapek, Czapek medium.

表2 9种供试杀菌剂的毒力回归方程

Table 2 Toxicity regression equations of the nine tested fungicides

药剂 Fungicide	毒力回归方程 Toxicity regression equation	抑制中浓度/ (mg·L^-1) EC₅₀/(mg·L^-1)	相关系数 Correlation coefficient
50%腐霉利·福美双WP 50% Thiram + procymidone WP	y=0.764 5x+3.794 2	1.577 3	0.969 3
72%霜脲氰·代森锰锌WP 72% Mancozeb + gymoxanil WP	y=0.979 4x+3.378 7	1.655 5	0.972 4
70%烯酰吗啉·霜脲氰WG 70% Dimethomorph + cymoxanil WG	y=0.435 4x+3.925 9	2.466 6	0.932 6
80%烯酰吗啉WG 80% Dimethomorph WG	y=2.477 6x-1.655 0	2.686 1	0.951 5
69%烯酰吗啉·代森锰锌WP 69% Dimethomorph + mancozeb WP	y=1.119 3x+3.117 4	1.681 9	0.992 0
10%多抗霉素WP 10% Polyoxin WP	y=2.136 4x+0.230 8	2.232 4	0.950 6
50%多菌灵WP 50% Carbendazim WP	y=5.854 0x-3.121 9	1.387 4	0.776 9
70%嘧霉胺WG 70% Pyrimethanil WG	y=6.639 6x-7.112 9	1.824 3	0.785 7
10%苯醚甲环唑WG 10% Difenoconazole WG	y=0.886 3x+3.890 1	1.252 2	0.910 0

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绣球博大蓝叶斑病病原菌的分离鉴定与防治

Isolation, identification, and control of the pathogen causing leaf spot disease on Hydrangea macrophylla Bo Dalan

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