非洲菊菌核病病原菌的分离鉴定及抑菌药剂筛选

曹奕鸯; 夏朝水; 陈昌铭; 陈玮婷; 甘玮欣; 林辉锋; 林发壮; 莫智龙

doi:10.19303/j.issn.1008-0384.2021.10.013

非洲菊菌核病病原菌的分离鉴定及抑菌药剂筛选

doi: 10.19303/j.issn.1008-0384.2021.10.013

三明市农业科学研究院花卉研究所，福建　三明　365051

基金项目: 中央引导地方科技发展专项（2019L3025）

详细信息

作者简介:
曹奕鸯（1984−），男，硕士，助理研究员，研究方向：花卉遗传育种（E-mail：caoyiyang@yeah.net）

通讯作者:
夏朝水（1982−），男，硕士，助理研究员，研究方向：花卉遗传育种（E-mail：86837432@qq.com）

中图分类号: S 436.8
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- 被引次数: 0
出版历程
- 收稿日期: 2021-07-09
- 修回日期: 2021-08-16
- 网络出版日期: 2021-10-23
- 刊出日期: 2021-10-28

Pathogen Identification and Fungicide Evaluation for Sclerotinia Stem Rot of Gerbera jamesonii

Flower Research Institute, Institute of Sanming Agricultural Sciences, Sanming, Fujian　365051, China

摘要

摘要: 目的分离鉴定福建省非洲菊种植过程中的主要病害菌核病的病原，筛选几种有效防治药剂进行轮换使用，避免农药的滥用及延缓病菌产生抗药性。方法采用组织分离法分离非洲菊菌核病病原菌，经病原菌株形态学、ITS分子鉴定及柯赫氏法则验证，确定致病的病原种类，并利用室内毒力测定法筛选对菌核病病原毒力高的药剂。结果分离获得1个非洲菊菌核病菌株C4-1；形态学、ITS及柯赫氏法则验证鉴定C4-1菌株为核盘菌Sclerotinia sclerotiorum；室内毒力测定结果显示，5种杀菌剂中25%咪鲜胺对核盘菌菌株C4-1的抑菌作用最强，EC₅₀值为0.0324 μg·mL⁻¹；其次为50%啶酰菌胺、40%菌核净、15%三唑醇，EC₅₀值分别为1.0337、1.8362、6.9408 μg·mL⁻¹；50%多菌灵的抑菌作用最弱，EC₅₀值为22.4349 μg·mL⁻¹。结论分离鉴定出福建省非洲菊菌核病病原为核盘菌Sclerotinia sclerotiorum，咪鲜胺、啶酰菌胺、菌核净对核盘菌毒力强，防治非洲菊菌核病过程中可轮换使用。
- 非洲菊 /
- 菌核病 /
- 核盘菌 /
- 鉴定 /
- 毒力测定
Abstract: Objective The pathogen of sclerotinia stem rot, a major disease of Gerbera jamesonii cultivated in Fujian, was identified. Effective fungicides that could be alternatively applied to curtail drug resistance were evaluated. Method The causative pathogen of sclerotinia stem rot was isolated from tissue of the diseased plant and verified by Koch’s rule as well as morphological observation and ITS sequence of the microbe. Toxicities of selected fungicides were determined in the laboratory. Result The isolated strain, C4-1, was identified as the culprit to be Sclerotinia sclerotiorum. Among the fungicides, hymexazol with an EC₅₀ of 0.032 4 μg·mL⁻¹ was most potent in inhibiting the pathogen growth. It was followed by boscalid with an EC₅₀ of 1.033 7 μg·mL⁻¹, dimetachlone with an EC₅₀ values of 1.8362 μg·mL⁻¹, triadimenol with an EC₅₀ of 6.9408 μg·mL⁻¹, and carbendazim the least effective agent with anEC₅₀ of 22.434 9 μg·mL⁻¹. Conclusion S. sclerotiorumwas isolated and identified as the pathogen that caused the sclerotinia stem rot disease on G. jamesonii in the province. Of the fungicides evaluated, hymexazol, boscalid, and dimetachlone, could be used alternatively for the disease control to minimize drug resistance by the pathogen.
- Gerbera jamesonii /
- sclerotinia stem rot /
- Sclerotinia sclerotiorum /
- identification /
- toxicity test

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图 1 非洲菊苗期菌核病感病过程及田间发病症状

Figure 1. Development of sclerotinia stem rot in gerbera seedlings and disease symptoms in the field

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图 2 菌株C4-1在PDA培养基中的生长过程

Figure 2. Growth of C4-1 in PDA medium

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图 3 菌核的萌发过程及子囊和子囊孢子的形态

注：A，菌核萌发初期；B，子囊盘开始形成； C，子囊盘形成；D，一个菌核萌发形成多个子囊盘； E，子囊盘形态；F，子囊孢子形态。

Figure 3. Germination process of sclerotia and the morphology of asci and ascospores

Note: A, primary stage of sclerotia germination; B, apothecia being formed; C, formed apothecia; D, geminated sclerotia with several apothecia; E, apothecia morphology; F, ascospore morphology.

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图 4 基于ITS核苷酸序列构建的核盘菌属系统发育树

Figure 4. Phylogenetic tree of Sclerotinia based on ITS nucleotide sequence

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表 1 供试杀菌剂母液及使用浓度

Table 1. Original and applied concentrations of fungicides under study

杀菌剂 Fungicide	剂型 Pesticide formulation	生产厂家 Manufacturer	母液质量浓度 Initial concentration/ （μg·mL⁻¹）	处理质量浓度 Treatment mass concentration/ （μg·mL⁻¹）
50% 多菌灵 50% Carbendazim	可湿性粉剂 Wettable Powder	江苏蓝丰生物化工股份有限公司 Jiangsu Lanfeng Biochemical Co., Ltd.	1000	3.125，6.25，12.5，25，50
25% 咪鲜胺 25% Prochloraz	水乳剂 Emulsion in Water	江苏辉丰生物农业股份有限公司 Jiangsu Huifeng Bio-agriculture Co., Ltd.	1000	0.0125，0.25，0.05，0.1，0.2
50% 啶酰菌胺 50% Boscalid	水分散粒剂 Water Dispersible Granule	巴斯夫（中国）有限公司 BASF China Co., Ltd.	1000	1.25，2.5，5，10，20
40% 菌核净 40% Dimethachlon	可湿性粉剂 Wettable Powder	江西禾益化工股份有限公司 Jiangxi Heyi Chemical Co., Ltd.	1000	0.5，1，2，4，8
15% 三唑醇 15% Triadimenol	可湿性粉剂 Wettable Powder	江苏剑牌农化股份有限公司 Jiangsu Sword Agrochemical Co., Ltd.	1000	1.25，2.5，5，10，20

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表 2 5种杀菌剂对非洲菊核盘菌菌株C4-1的毒力

Table 2. Toxicities of 5 fungicides on S. sclerotiorum C4-1

杀菌剂 Fungicide	剂型 Pesticide Formulation	毒力回归方程 Virulent regression equation	相关系数 Correlation coefficient, r	EC₅₀值 EC₅₀ value/（μg·mL⁻¹）
50% 多菌灵 50% Carbendazim	可湿性粉剂 Wettable powder	y=0.2272x−0.5196	0.9757	22.4349
25% 咪鲜胺 25% Prochloraz	水乳剂 Emulsion in water	y=0.1849x+1.4149	0.9698	0.0324
50% 啶酰菌胺 50% Boscalid	水分散粒剂 Water dispersible granule	y=0.1067x+0.4949	0.9756	1.0337
40% 菌核净 40% Dimethachlon	可湿性粉剂 Wettable powder	y=0.3821x+0.1650	0.9858	1.8362
15% 三唑醇 15% Triadimenol	可湿性粉剂 Wettable powder	y=0.2489x−0.1957	0.9947	6.9408

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参考文献(24)

[1]	肖建梅. 非洲菊组培技术 [J]. 中国花卉园艺, 2005(8):14−15. XIAO J M. Tissue Culture of Gerbera Jamesonii [J]. China Flowers & Horticulture, 2005(8): 14−15.(in Chinese)
[2]	郭方其, 陈文海, 叶琪明, 等. 不同非洲菊品种在浙北地区的适应性评价 [J]. 浙江农业科学, 2020, 61(10):2056−2059. GUO F Q, CHEN W H, YE Q M, et al. Evaluation of Adaptability of Different Gerbera jamesonii Cultivars in Northern Zhejiang Province [J]. Journal of Zhejiang Agricultural Sciences, 2020, 61(10): 2056−2059.(in Chinese)
[3]	MATHERON M E. First Report of Sclerotinia sclerotiorum on African Daisy in Arizona [J]. Plant Disease, 1994(78): 924D.
[4]	WOLCAN S M. Occurrence of Crown Rot of Gazania and Gerbera Caused by Scleroticnia sclerotiorum in Argentina [J]. Journal of Plant Pathology, 2004, 86(3): 263−264.
[5]	FOLK G, TUSNÁDI CS K. A New Disease of Gerbera in Hungary, Sclerotinia Wilt [J]. Növényvédelem, 1985, 21(12): 557−561.
[6]	VIGODSKY H. Methods for Controlling Sclerotinia Rot of Gerbera and Studies on the Disease Cycle in Israel [J]. lsrael J agric Res, 1969, 19: 185−194.
[7]	周韦成, 赵建军. 上海地区非洲菊真菌病害鉴定 [J]. 生物灾害科学, 2015, 38(4):339−344. doi: 10.3969/j.issn.2095-3704.2015.04.014 ZHOU W C, ZHAO J J. Identification of Pathogenic Fungi Gerbera jamesonii in Shanghai [J]. Biological Disaster Science, 2015, 38(4): 339−344.(in Chinese) doi: 10.3969/j.issn.2095-3704.2015.04.014
[8]	王涛. 非洲菊病虫害综合防治技术 [J]. 中国植保导刊, 2004, 24(5):25−26. doi: 10.3969/j.issn.1672-6820.2004.05.010 WANG T. Integrated Pest Control Techniques of Gerbera jamesonii [J]. China Plant Protechion, 2004, 24(5): 25−26.(in Chinese) doi: 10.3969/j.issn.1672-6820.2004.05.010
[9]	刘峰. 非洲菊主要病害及其防治 [J]. 特种经济动植物, 2004, 7(9):40. doi: 10.3969/j.issn.1001-4713.2004.09.036 LIU F. The Main Diseases and Control of Gerbera jamesonii [J]. Special Economic Animal and Plant, 2004, 7(9): 40.(in Chinese) doi: 10.3969/j.issn.1001-4713.2004.09.036
[10]	张志轩, 王洪习, 汪妮. 豫北地区非洲菊鲜切花常见病虫害及综合防治技术 [J]. 现代园艺, 2007(6):25−27. doi: 10.3969/j.issn.1006-4958.2007.06.016 ZHANG Z X, WANG H X, WANG N. Common Diseases and Pests of Gerbera Cut Flower in Northern Henan and Integrated Control Techniques [J]. Modern Horticulture, 2007(6): 25−27.(in Chinese) doi: 10.3969/j.issn.1006-4958.2007.06.016
[11]	夏朝水. 福建主栽非洲菊品种资源的综合评价与应用[D]. 福州: 福建农林大学, 2014. XIA C S. Comprehensive Evaluation and Application of Fujian Main Cultivated Varieties of Gerbera jamesonii Resources [D]. Fuzhou: Fujian Agriculture and Forestry University, 2014. (in Chinese)
[12]	方中达. 植病研究方法[M]. 3版. 北京: 中国农业出版社, 1998.
[13]	孙敬贤, 张鲁刚. 大白菜菌核病病原菌鉴定及其主要生物学特性 [J]. 华北农学报, 2015, 30(S):321−328. SUN J X, ZHANG L G. Identification and Main Biological Characteristics of Sclerotinia Rot Pathogen in Chinese Cabbage [J]. Acta Agriculturae Boreali-Sinica, 2015, 30(S): 321−328.(in Chinese)
[14]	张薇, 魏海雷, 张力群, 等. 紫花苜蓿菌核病病原鉴定及其主要生物学特性 [J]. 草业学报, 2005, 14(2):69−75. doi: 10.3321/j.issn:1004-5759.2005.02.013 ZHANG W, WEI H L, ZHANG L Q, et al. Identification and Control of Medicago sativa Sclerotinia Stem Rot Occurred in South Jiangsu Hilly Area [J]. Acta Prataculturae Sinica, 2005, 14(2): 69−75.(in Chinese) doi: 10.3321/j.issn:1004-5759.2005.02.013
[15]	刘春来, 文景芝, 杨明秀, 等. rDNA-ITS在植物病原真菌分子检测中的应用 [J]. 东北农业大学学报, 2007, 38(1):101−106. doi: 10.3969/j.issn.1005-9369.2007.01.022 LIU C L, WEN J Z, YANG M X, et al. Application of rDNA-ITS in Molecular Test of Phytopathogenic fungi [J]. Journal of Northeast Agricultural University, 2007, 38(1): 101−106.(in Chinese) doi: 10.3969/j.issn.1005-9369.2007.01.022
[16]	李克宁, 龚贤弟. 荷兰进口的扶郎花上发生严重菌核病 [J]. 植物检疫, 1987, 1(4):316. LI K N, GONG X D. Severe Sclerotinia on Gerbera Imported from the Netherlands [J]. Plant Quarantine, 1987, 1(4): 316.(in Chinese)
[17]	卢灿华, 蔺忠龙, 甄安忠, 等. 香料烟灰霉病和菌核病病原鉴定及室内药剂筛选 [J]. 中国烟草科学, 2020, 41(6):68−75. LU C H, LIN Z L, ZHEN A Z, et al. Causal Pathogen Identification of Gray Mold Rot and Sclerotinia Rot on Oriental Tobacco and Indoor Screening of Fungicides [J]. Chinese Tobacco Science, 2020, 41(6): 68−75.(in Chinese)
[18]	孙雅楠, 哀嘉彬, 唐爱妙, 等. 浙江桑葚菌核病的病原菌鉴定及其对4种杀菌剂的抗性检测 [J]. 果树学报, 2020, 37(12):1934−1940. SUN Y N, AI J B, TANG A M, et al. Identification of Pathogenic Fungi Causing Mulberry Fruit Sclerotiniose and Their Resistance to Four Fungicides in Zhejiang [J]. Journal of Fruit Science, 2020, 37(12): 1934−1940.(in Chinese)
[19]	顾玉阳, 王黎娜, 袁娟, 等. 扁豆菌核病病原菌鉴定及其生物学特性 [J]. 中国蔬菜, 2020(10):68−76. GU Y Y, WANG L N, YUAN J, et al. Identification of Hyacinth Bean Sclerotinia Pathogen and Its Biological Characteristics [J]. China Vegetables, 2020(10): 68−76.(in Chinese)
[20]	BOLAND G J, HALL R. Index of plant hosts of Sclerotinia sclerotiorum [J]. Canadian Journal of Plant Pathology, 1994, 16(2): 93−108. doi: 10.1080/07060669409500766
[21]	桂勇武, 高年春, 魏启舜. 非洲菊常见病虫害综合防治技术 [J]. 现代农业科技, 2006(20):52−53. doi: 10.3969/j.issn.1007-5739.2006.20.043 GUI Y W, GAO N C, WEI Q S. Integrated Control Techniques of Common Diseases and Pests of Gerbera [J]. Modern Agricultural Science and Technology, 2006(20): 52−53.(in Chinese) doi: 10.3969/j.issn.1007-5739.2006.20.043
[22]	吴仁锋, 杨绍丽. 蔬菜菌核病的识别及防治 [J]. 长江蔬菜, 2016(5):42−43. doi: 10.3865/j.issn.1001-3547.2016.05.020 WU R F, YANG S L. Identification and Control of Sclerotinia sclerotiorum of Vegetables [J]. Journal of Changjiang Vegetables, 2016(5): 42−43.(in Chinese) doi: 10.3865/j.issn.1001-3547.2016.05.020
[23]	乔昕, 余洋, 陈杰, 等. 川渝地区油菜菌核病菌对多菌灵的抗性监测及抗性机理研究 [J]. 西南大学学报(自然科学版), 2016, 38(5):26−30. QIAO X, YU Y, CHEN J, et al. Molecular Mechanism and Monitoring on Carbendazim Resistance of Sclerotinia sclerotiorum Obtained from the Blight Stems of Rape in Chongqing and Sichuan [J]. Journal of Southwest University (Natural Science), 2016, 38(5): 26−30.(in Chinese)
[24]	乔广行, 李兴红, 周莹, 等. 北京地区不同寄主蔬菜菌核病菌抗药性比较 [J]. 江苏农业科学, 2014, 42(12):155−157. QIAO G X, LI X H, ZHOU Y, et al. Comparative Study on Resistance to Drugs of Sclerotinia sclerotiorum with Different Host Vegetables [J]. Jiangsu Agricultural Sciences, 2014, 42(12): 155−157.(in Chinese)