Pathogen Identification of Anthracnose Disease in Chili Pepper from Hebei Province and Their Sensitivity to Difenoconazole

doi:10.13304/j.nykjdb.2025.0201

Abstract

Abstract:

To identify the pathogen of anthracnose on Capsicum annuum L. in Jize and Guangzong county in Hebei province， diseased pepper fruits were collected. 71 strains were isolated， in which 32 strains from Jize county， and 39 from Guangzong county. By morphological and molecular identification， 24 strains were identified as Colletotrichum scovillei and 8 strains were as C. truncatum in 32 strains from Jize county， acounting for 75.00% and 25.00%， respectively， 32 strains were identified as Colletotrichum scovillei and 7 strains were as C. truncatum in 39 strains from Guangzong county， acounting for 82.05% and 17.95%， respectively. Fungicide difenoconazole was employed to assay the sensitivity of 46 tested strains， the median effect concentration （EC₅₀） were ranging from 0.143 0 to 0.434 8 μg·mL^-1， with average of 0.289 8 μg·mL^-1. Above results identified the pathogen of pepper anthracnose in south of Hebei province， and provided theoretical basis and fungicide recommendation for the prevention and control of pepper anthracnose caused by C. scovillei.

Key words: chili anthracnose disease, pathogen identification, difenoconazole, sensitivity to fungicide, Colletotrichum scovillei

摘要：

为明确河北省鸡泽县和广宗县辣椒炭疽病病原，以田间采集的辣椒炭疽病病果为材料，利用组织分离法获得分离物71株，其中鸡泽县32株，广宗县39株。通过形态学和分子生物学鉴定，确定鸡泽县的32株中有24株为斯高维尔炭疽菌（Colletotrichum scovillei），另8株为平头炭疽菌（C. truncatum），分离比例分别为75.00%和25.00%；广宗县的39株中有32株为斯高维尔炭疽菌，另7株为平头炭疽菌，分离比例分别为82.05%和17.95%。采用菌丝生长速率法测定了46株斯高维尔炭疽菌对苯醚甲环唑的敏感性，结果表明，苯醚甲环唑对这些菌株的半数效应浓度（median effect concentration，EC₅₀）为0.143 0~0.434 8 μg·mL^-1，平均0.289 8 μg·mL^-1。研究结果明确了河北省南部辣椒炭疽病的病原菌，为河北省辣椒生产中防治由斯高维尔炭疽菌引起的炭疽病提供理论依据和用药参考。

关键词: 辣椒炭疽病, 病原鉴定, 苯醚甲环唑, 药剂敏感性, 斯高威尔炭疽菌

CLC Number:

S432

Yanan ZHU, Bingxue WANG, Yijiao HOU, Yizhuo DAI, Zhangxin PEI, Likun DONG, Tongle HU, Yunqiang WANG, Wenxiang YANG, Na ZHANG. Pathogen Identification of Anthracnose Disease in Chili Pepper from Hebei Province and Their Sensitivity to Difenoconazole[J]. Journal of Agricultural Science and Technology, 2025, 27(10): 144-150.

朱亚楠, 王冰雪, 侯怡娇, 戴祎卓, 裴张新, 董立坤, 胡同乐, 王云强, 杨文香, 张娜. 河北省不同来源辣椒炭疽病病菌鉴定及其对苯醚甲环唑的敏感性[J]. 中国农业科技导报, 2025, 27(10): 144-150.

Figures/Tables 5

Table 1 Primers used in this research for identification of the Colletotrichum spp.

名称 Name	序列 Sequence （5’-3’）	参考文献 Reference
ITS1	TCCGTAGGTGAACCTGCGG	［18］
ITS4	TCCTCCGCTTATTATATGC	［18］
GDF1	GCCGTCAACGACCCCTTCATTGA	［19］
GDR1	GGGTGGAGTCGTACTTGAGCATGT	［19］
ACT-512F	ATGTGCAAGGCCGGTTTCGC	［20］
ACT-783R	TACGAGTCCTTCTGGCCCAT	［20］
CHS-79F	TGGGGCAAGGATGCTTGGAAGAAG	［20］
CHS-345R	TGGAAGAACCATCTGTGAGAGTTG	［20］

Fig. 1 Microscopic observation of GZ6 and JZ15-1 colonies， conidia and appressoriumA： Front and back morphology of GZ6 colon； B： Conidia of GZ6； C： Mycelial appressoria of GZ6； D： Front and back morphology of JZ15-1 colon； E： Conidia of JZ15-1； F： Mycelial appressoria of JZ15-1； Bar=10 μm

Fig. 2 Symptom before separation and inoculation of GZ6 and JZ15-1A： Symptom before separation of GZ6； B： Symptom inoculated GZ6； C： Symptom before separation of JZ15-1； D： Symptom inoculated JZ15-1

Fig. 3 Phylogenetic tree of Colletotrichum

Fig. 4 EC50 of 46 C. scovillei strains to difenoconazole

References 36

[1]	周黛媛, 张正海, 曹亚从, 等. 辣椒抗炭疽病遗传育种研究进展[J].中国蔬菜, 2022(2): 17-24.
	ZHOU D Y, ZHANG Z H, CAO Y C, et al.. Research progress on genetic breeding of papper resistant to anthracnose [J]. China Veget., 2022(2): 17-24.
[2]	乔立娟, 赵帮宏, 宗义湘,等.我国辣椒产业发展现状、趋势及对策[J].中国蔬菜, 2023(11): 9-15.
	QIAO L J, ZHAO B H, ZONG Y X, et al.. Development current situation, tendency, and countermeasure for China’s pepper industry [J]. China Veget., 2023(11): 9-15.
[3]	许艺, 李红, 方宏修, 等.四川辣椒产业现状及发展建议[J].辣椒杂志, 2020, 18(3): 1-5.
	XU Y, LI H, FANG H X, et al.. Development status and suggestions on pepper industry in Sichuan province [J]. J. China Capsicum, 2020, 18(3): 1-5.
[4]	侯思皓, 巩雪峰, 许艺, 等.辣椒炭疽病菌鉴定及炭疽菌胁迫下的生理变化[J].西南农业学报, 2024, 37(10): 2185-2193.
	HOU S H, GONG X F, XU Y, et al.. Pathogen identification of pepper anthracnose and physiological changes of pepper under anthracnose stress [J]. Southwest China J. Agric. Sci., 2024, 37(10): 2185-2193.
[5]	杨佳文, 赵尊练, 张管曲, 等.陕西线辣椒炭疽病原菌的鉴定及生物学特性研究[J].西北农业学报, 2017, 26(11): 1695-1705.
	YANG J W, ZHAO Z L, ZHANG G Q, et al.. Identification and biological characterization of Anthrax bacteria in Xianlajiao chili pepper in Shaanxi province [J]. Acta Agric. Bor-Occid. Sin., 2017, 26(11): 1695-1705.
[6]	贾令晨.鸡泽县辣椒产业特点及关键栽培技术应用研究[D].邯郸: 河北工程大学, 2020.
	JIA L C. The study of the industrial characteristics and application of key planting technology of pepperin Jize county [D]. Handan: Hebei University of Engineering, 2020.
[7]	朱晓琴, 方树贤, 刘冬梅, 等.辣椒炭疽病生防菌株的筛选、鉴定及其抑菌机理[J].植物保护学报, 2023, 50(4): 913-922.
	ZHU X Q, FANG S X, LIU D M, et al.. Screening, identification and antifungal mechanism of bacterial biocontrol strains against hot pepper anthracnose [J]. J. Plant Prot., 2023, 50(4): 913-922.
[8]	DE SILVA D D, GROENEWALD J Z, CROUS P W, et al.. Identification, prevalence and pathogenicity of Colletotrichum species causing anthracnose of Capsicum annuum in Asia [J/OL]. IMA Fungus, 2019, 10: 8 [2025-02-20]. .
[9]	DIAO Y Z, ZHANG C, LIU F, et al.. Colletotrichum species causing anthracnose disease of chili in China [J]. Persoonia, 2017, 38: 20-37.
[10]	夏花, 朱宏建, 周倩, 等.湖南芷江辣椒上一种新炭疽病的病原鉴定[J].植物病理学报, 2012, 42(2): 120-125.
	XIA H, ZHU H J, ZHOU Q, et al.. Pathogen identification of a new anthracnose of pepper in Zhijiang, Hunan [J]. Acta Phytopathol. Sin., 2012, 42(2): 120-125.
[11]	曾向萍, 赵志祥, 符美英, 等.海南辣椒炭疽病病原菌鉴定及其对嘧菌酯的敏感性分析[J].中国植保导刊, 2016, 36(5): 5-8, 81.
	ZENG X P, ZHAO Z X, FU M Y, et al.. Identification of pepper anthracnose pathogen in Hainan province and analysis on its sensitivity to azoxystrobin [J]. China Plant Prot., 2016, 36(5): 5-8, 81.
[12]	GAO Y Y, LI X X, HE L F, et al.. Effect of pyrisoxazole on Colletotrichum scovillei infection and anthracnose on chili [J]. Plant Dis., 2020, 104(2): 551-559.
[13]	罗跃, 刘旭东, 韩磊, 等.贵州省辣椒炭疽病病原鉴定及协同增效药剂筛选[J].北方园艺, 2021(14): 21-30.
	LUO Y, LIU X D, HAN L, et al.. Identification of the pathogen of Capsicum anthracnose and screening of synergistic agents in Guizhou province [J]. North. Hortic., 2021(14): 21-30.
[14]	霍建飞, 姚玉荣, 任文来, 等.戊唑醇对2种辣椒炭疽菌的敏感性差异[J]. 安徽农业科学, 2020, 48(8): 151-152.
	HUO J F, YAO Y R, REN W L, et al.. The sensitivity difference of tebuconazole against Colletotrichum scovillei and Colletotrichum truncatum [J]. J. Anhui Agric. Sci., 2020, 48(8): 151-152.
[15]	姚福田.辣椒炭疽病菌对2种杀菌剂的敏感性及其对戊唑醇抗性机制研究[D].太谷: 山西农业大学,2023.
	YAO F T. Study on sensitivity to two fungicides and mechanism of resistance to tebuconazole in Colletotrichum scovillei [D]. Taigu: Shanxi Agricultural University, 2023.
[16]	王东平, 霍晓倩, 宗义湘, 等.“鸡泽辣椒”产业现状与高质量发展建议[J].北方园艺, 2024(20): 131-136.
	WANG D P, HUO X Q, ZONG Y X, et al.. Current status and high-quality development suggestions of the “Jize Pepper” industry [J]. North. Hortic., 2024(20): 131-136.
[17]	陈汝, 王金政, 薛晓敏, 等.苹果炭疽菌DNA改良提取法及PCR体系的优化[J].江西农业学报, 2017, 29(4): 69-71.
	CHEN R, WANG J Z, XUE X M, et al.. Comparison of several modified DNA extraction methods and optimization of PCR system for Colletotrichum gloeosporioides in apple [J]. Acta Agric. Jiangxi, 2017, 29(4): 69-71.
[18]	WHITE T J, BRUNS S, LEE S, et al.. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics [J]. PCR Protocols Guide Methods Appl., 1990(1): 315-322.
[19]	GUERBER J C, LIU B, CORRELL J C, et al.. Characterization of diversity in Colletotrichum acutatum sensu lato by sequence analysis of two gene introns, mtDNA and intron RFLPs, and mating compatibility [J]. Mycologia, 2003, 95(5): 872-895.
[20]	CARBONE I, KOHN L M. A method for designing primer sets for speciation studies in filamentous ascomycetes [J]. Mycologia, 1999, 91(3): 553-556.
[21]	LIU F, TANG G, ZHENG X, et al.. Molecular and phenotypic characterization of Colletotrichum species associated with anthracnose disease in peppers from Sichuan province, China [J/OL]. Sci. Rep., 2016, 6: 32761 [2025-02-20]. .
[22]	郑婕, 庄远航, 郭梓琪, 等.南雄辣椒炭疽病菌的分离鉴定与药剂筛选[J].中国蔬菜, 2022(10): 58-65.
	ZHENG J, ZHUANG Y H, GUO Z Q, et al.. Isolation, identification and fungicide screening of Capsicum anthracnose pathogen in Nanxiong county [J]. China Veget., 2022(10): 58-65.
[23]	王赛风.山西省辣椒炭疽病病原鉴定及其对常用杀菌剂的敏感性[D].太谷: 山西农业大学,2022.
	WANG S F. Identification of pathogen of chili anthracnose in Shanxi province and its sensitivity to common fungicides [D]. Taigu: Shanxi Agricultural University, 2022.
[24]	陈鹏宇.吉林省及内蒙古东部地区辣椒炭疽病的病原鉴定及Colletotrichum scovillei对3种杀菌剂的抗性风险评估[D].长春: 吉林农业大学, 2023.
	CHEN P Y. Pathogen identification of Capsicum anthracnose and risk assessment of resistance of Colletotrichum scovillei to three fungicides in Jilin province and eastern Inner Mongolia [D]. Changchun: Jilin Agricultural University, 2023.
[25]	霍建飞, 姚玉荣, 郝永娟, 等.天津市宁河区辣椒炭疽病病原鉴定及防治药剂筛选[J].北方园艺, 2020(3): 1-7.
	HUO J F, YAO Y R, HAO Y J, et al.. Identification of pepper anthracnose and screening of fungicides against it in Ninghe of Tianjin [J]. North. Hortic., 2020(3): 1-7.
[26]	满益龙.湖南省辣椒炭疽病菌遗传多样性研究[D].长沙:湖南农业大学, 2016.
	MAN Y L. Genetic diversity of Colletotrichum spp. strains causing anthracnose of Capsicum annuum L. plants in Hunan [D]. Changsha: Hunan Agricultural University, 2016.
[27]	姚道忠, 李运超, 姚洁, 等.啶氧菌酯对辣椒炭疽病菌室内抑制与田间防效[J].安徽农业大学学报, 2019, 46(6):1028-1033.
	YAO D Z, LI Y C, YAO J, et al.. Control effect of picoxystrobin on Collerotrichum gloeosporioides indoor and field [J]. J. Anhui Agric. Univ., 2019, 46(6): 1028-1033.
[28]	高杨杨, 禾丽菲, 李北兴, 等.山东省辣椒炭疽病病原菌的鉴定及高效防治药剂的筛选[J].中国农业科学, 2017, 50(8): 1452-1464.
	GAO Y Y, HE L F, LI B X, et al.. Identification of the pathogen causing pepper anthracnose in Shandong province and screening of highly effective fungicides [J]. Sci. Agric. Sin., 2017, 50(8): 1452-1464.
[29]	王妮, 尹显慧, 彭丽娟, 等.辣椒炭疽病病原鉴定及其杀菌剂毒力测定[J].植物保护, 2019, 45(4): 216-223.
	WANG N, YIN X H, PENG L J, et al.. Identification of the pathogen and toxicity test of fungicides to Capsicum anthracnose [J]. Plant Prot., 2019, 45(4): 216-223.
[30]	杨力, 林锦辉, 赖添财.不同种类杀菌剂对辣椒炭疽病的田间防效[J].农药科学与管理, 2023, 44(4): 60-64.
	YANG L, LIN J H, LAI T C. Field control effects of different fungicides on pepper anthracnose [J]. Pestic. Sci. Admin., 2023, 44(4): 60-64.
[31]	王爱萍, 席淑文, 张国福, 等.苯并烯氟菌唑对尖孢炭疽菌的毒力及对其所致辣椒炭疽病的防效[J].植物保护学报, 2023, 50(3): 830-838.
	WANG A P, XI S W, ZHANG G F, et al.. Inhibitory activity of benzovindiflupyr against fruit rot pathogen Colletotrichum acutatum and its control efficacy to pepper anthracnose [J]. Acta Phytophy. Sin., 2023, 50(3): 830-838.
[32]	张文静, 姚福田, 吕红, 等.山西省辣椒炭疽病菌对4种杀菌剂的敏感性及对戊唑醇的抗性风险评估[J/OL].植物病理学报,2024:1-11[2025-02-20]..
	ZHANG W J, YAO F T, LYU H, et al.. Sensitivity of Colletotrichum scovillei causing pepper anthracnose in Shanxi province tothe 4 fungicides and resistance risk assessment to tebuconazole [J/OL]. Acta Phytopathol. Sin. 2024:1-11 [2025-02-20]. .
[33]	ZHANG C, DIAO Y Z, WANG W Z, et al.. Assessing the risk for resistance and elucidating the genetics of Colletotrichum truncatum that is only sensitive to some DMI fungicides [J/OL]. Front. Microbiol., 2017, 8: 1779 [2025-2-20]. .
[34]	刘倩, 马连杰, 张慧, 等.辣椒炭疽病生防菌株TN2的筛选鉴定与抑菌效果[J]. 生物技术通报, 2025, 41(1): 287-297.
	LIU Q, MA L J, ZHANG H, et al.. Screening, identification and control effects of biocontrol strain TN2 against pepper anthracnose [J]. Biotechnol. Bull., 2025, 41(1): 287-297.
[35]	李叶彤,马玉坤,杨仕佳,等.抗辣椒炭疽病菌盘长孢状刺盘孢的木霉菌的筛选[J].菌物学报, 2023, 42(12): 2374-2387.
	LI Y T, MA Y K, YANG S J, et al.. Screening of antagonistic Trichoderma strains against Colletotrichum gloeosporioides in pepper [J]. Mycosystema, 2023, 42(12): 2374-2387.
[36]	LE T T, CONG V K, SANGPUEA K, et al.. Efficacy of Bacillus subtilis for controlling anthracnose in chilli [J]. Agric. Nat. Res., 2023, 2: 223-232.