菜用大豆炭疽病病原菌的分离鉴定与防治

doi:10.3969/j.issn.1004-1524.2022.12.11

浙江农业学报 ›› 2022, Vol. 34 ›› Issue (12): 2682-2688.DOI: 10.3969/j.issn.1004-1524.2022.12.11

菜用大豆炭疽病病原菌的分离鉴定与防治

刘娜(), 范翘楚, 周佳, 宋雅静, 张古文, 冯志娟, 卜远鹏, 王斌, 龚亚明()

农产品质量安全危害因子与风险防控国家重点实验室,浙江省农业科学院蔬菜研究所,浙江杭州 310021

收稿日期:2021-09-18 出版日期:2022-12-25 发布日期:2022-12-26
通讯作者: 龚亚明
作者简介:*龚亚明,E-mail:gongym07@163.com
刘娜(1985—),女,河北唐山人,博士,副研究员,从事豆类蔬菜种质创新与分子育种研究。E-mail:ln200811@163.com
基金资助:
农产品质量安全危害因子与风险防控国家重点实验室自主设计课题(2010DS700124-ZZ2005);浙江省重点研发课题(2021C02052);浙江省“十四五”蔬菜育种专项(2021C02065-6);浙江省基础公益研究计划(LGN20C150006);国家自然科学基金(31872114)

Identification and control of anthracnose in vegetable soybean

LIU Na(), FAN Qiaochu, ZHOU Jia, SONG Yajing, ZHANG Guwen, FENG Zhijuan, BO Yuanpeng, WANG Bin, GONG Yaming()

State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Institute of Vegetables, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China

Received:2021-09-18 Online:2022-12-25 Published:2022-12-26
Contact: GONG Yaming

摘要/Abstract

摘要：

炭疽病是菜用大豆生产上发生的常见真菌性病害,破坏力强,发病时在豆荚表面形成黑色斑点,严重影响鲜荚外观商品性和品质,已成为我国菜用大豆生产上面临的主要问题之一。为明确浙江省菜用大豆炭疽病病原菌的种类,本文对采集的菜用大豆炭疽病样本进行分离纯化培养,基于柯赫氏法则分离到引起菜用大豆炭疽病的病原菌,并通过病原菌的菌落形态特征结合其rDNA-ITS区域的序列分析对病原菌进行种类鉴定。结果表明,造成浙江省菜用大豆炭疽病的主要病原菌为平头炭疽菌(Colletotrichum truncatum)和胶孢炭疽菌(Colletotrichum gloeosporioides)。为筛选可以用于防控菜用大豆炭疽病的杀菌剂,测定了分离得到的病原菌平头炭疽菌Cts18和胶孢炭疽菌Cts22对生产中常用杀菌剂多菌灵和戊唑醇的敏感性,发现多菌灵对Cts18、Cts22的抑制中浓度(EC₅₀)分别为2.13 μg·mL^-1和96.12 μg·mL^-1,戊唑醇对Cts18、Cts22的EC₅₀分别为0.27 μg·mL^-1和0.63 μg·mL^-1。结果表明,平头炭疽菌Cts18相比胶孢炭疽菌Cts22对多菌灵和戊唑醇更加敏感,其中戊唑醇可作为生产中防控菜用大豆炭疽病的杀菌剂之一。

关键词: 菜用大豆, 炭疽病, 病原菌鉴定, 平头炭疽菌, 胶孢炭疽菌, 杀菌剂敏感性

Abstract:

Anthracnose disease is a common fungal disease of vegetable soybean, which forms black spots on the pod surface and affects the appearance and quality of fresh pod seriously. It has become one of economically important fungal diseases on vegetable soybean yield in China. To identify the pathogens causing vegetable soybean anthracnose in Zhejiang Province, we collected anthracnose disease samples and isolated the pathogens according to Koch's postulate. The pathogens were further identified based on morphological characteristics and rDNA-ITS sequence analysis in this study. Our results showed that two Colletotrichum species, Colletotrichum truncatum and Colletotrichum gloeosporioides, were the major pathogens causing vegetable soybean anthracnose in this region. To further guide the vegetable soybean anthracnose control using fungicides, we measured the sensitivity of representative strain C. truncatum Cts18 and C. Gloeosporioides Cts22 to commonly used fungicide carbendazim and tebuconazole. Results indicated that the concentration for 50% of maximal effect (EC₅₀) of carbendazim against C. truncatum Cts18 and C. gloeosporioides Cts22 were 2.13 μg·mL^-1 and 96.12 μg·mL^-1, respectively. The EC₅₀ of tebuconazole against Cts18 and Cts22 were 0.27 μg·mL^-1 and 0.63 μg·mL^-1, respectively. These results suggested that Cts18 was more sensitive than Cts22 towards the tested fungicides, and tebuconazole is a potential fungicide for managing vegetable soybean anthracnose in fields in this region.

Key words: vegetable soybean, anthracnose, pathogen identification, Colletotrichum truncatum, Colletotrichum gloeosporioides, fungicide sensitivity

中图分类号:

S643.7

刘娜, 范翘楚, 周佳, 宋雅静, 张古文, 冯志娟, 卜远鹏, 王斌, 龚亚明. 菜用大豆炭疽病病原菌的分离鉴定与防治[J]. 浙江农业学报, 2022, 34(12): 2682-2688.

LIU Na, FAN Qiaochu, ZHOU Jia, SONG Yajing, ZHANG Guwen, FENG Zhijuan, BO Yuanpeng, WANG Bin, GONG Yaming. Identification and control of anthracnose in vegetable soybean[J]. Acta Agriculturae Zhejiangensis, 2022, 34(12): 2682-2688.

图/表 7

图1 菜用大豆炭疽病发病植株和不同发病程度豆荚

Fig.1 Anthracnose symptoms of vegetable soybean plants and pods

图2 病原菌Cts18 和Cts22的形态特征 A,Cts18的菌落形态;B,Cts22的菌落形态。

Fig.2 Colony morphology of strain Cts18 and Cts22 A, Morphological characteristics of Cts18; B, Morphological characteristics of Cts22.

图3 病原菌Cts18 和Cts22的致病性检测 A,对照;B,接种Cts18的叶片;C,接种Cts22的叶片。

Fig.3 Pathogenicity of strain Cts18 and Cts22 A, Control; B, Leaf inoculated with Cts18; C, Leaf inoculated with Cts22.

图4 菌株Cts18 和Cts22的ITS区域PCR扩增产物凝胶电泳

Fig.4 PCR product of ITS regions of Cts18 and Cts22 strain

图5 基于ITS构建的系统发育树

Fig.5 Phylogenetic tree of Collectotrichum species based on ITS sequences

表1 用于系统发育树构建的炭疽菌菌株的背景及其ITS序列长度

Table 1 The Colletotrichum species used in phylogenetic tree and their ITS sequence length

种 Species	菌株 Isolates	ITS序列长度 Length of ITS/bp	GenBank序列号 GenBank number
Colletotrichum truncatum	CF-2-1	558	MK611675.1
Colletotrichum capsici	GK17A	584	MK713419.1
Colletotrichum jasminigenum	Co-15-Coy	560	JQ658867.1
Colletotrichum dematium	IMI-080025	580	AB046608.1
Colletotrichum siamense	C-106	606	MG913611.1
Colletotrichum gloeosporioides	WZ-244	583	MN856366.1
Colletotrichum lindemuthianum	GY3	575	MF511774.1
Colletotrichum sichuanense	CMM2902	549	MF279904.1
Colletotrichum cliviae	AGMy0252	594	KX578803.1
Colletotrichum magnum	Cmj2	555	KT949410.1
Pseudevernia furfuracea	M1_H21	503	MN654498.1

表2 不同药剂对Cts18、Cts22的室内毒力测定

Table 2 Toxicity of carbendazim and tebutazolol against Cts18 and Cts22

化学药剂 Fungicides categories	菌株 Strain	毒力回归方程 Regression equation	EC₅₀/ (μg· mL^-1)	r
多菌灵 Carbendazim	Cts18	Y=0.25+0.69x²	2.13	0.90
	Cts22	Y=2.85+1.39x²	96.12	0.91
戊唑醇 Tebutazolol	Cts18	Y=0.46+0.81x²	0.27	0.98
	Cts22	Y=0.28+1.40x²	0.63	0.97

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菜用大豆炭疽病病原菌的分离鉴定与防治

Identification and control of anthracnose in vegetable soybean

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