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

doi:10.3969/j.issn.1004-1524.20250254

Acta Agriculturae Zhejiangensis ›› 2026, Vol. 38 ›› Issue (4): 696-706.DOI: 10.3969/j.issn.1004-1524.20250254

• Plant Protection • Previous Articles Next Articles

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

Abstract

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)

CLC Number:

S432.44

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.

Figures/Tables 7

References 28

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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

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

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

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

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

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