黄瓜新炭疽病原的鉴定及生物学特性初探

doi:10.11924/j.issn.1000-6850.casb2020-0850

中国农学通报 ›› 2021, Vol. 37 ›› Issue (28): 96-108.doi: 10.11924/j.issn.1000-6850.casb2020-0850

黄瓜新炭疽病原的鉴定及生物学特性初探

韩帅¹(), 吴婕¹, 张河庆¹, 梁根云²(), 席亚东¹()

¹四川省农业科学院植物保护研究所/农业农村部西南作物有害生物综合治理重点实验室,成都 610066
²四川省农业科学院园艺研究所/蔬菜种质与品种创新四川省重点实验室,成都 610066

收稿日期:2020-12-29 修回日期:2021-07-27 出版日期:2021-10-05 发布日期:2021-10-28
通讯作者: 梁根云,席亚东
作者简介:韩帅,男,1988年出生,四川什邡人,助理研究员,硕士,研究方向为植物病理学。通信地址：610066 成都市静居寺路20号四川省农业科学院植物保护研究所,Tel：028-84590050,E-mail： shuaihan24@163.com。
基金资助:
四川省财政创新能力提升项目“黄瓜白粉病抗性基因研究与新材料创制”(2016GXTZ-005);四川省现代农业学科建设推进工程“蔬菜新病害的鉴定及重要病原菌致病机理研究”(2021XKJS087);四川省财政创新能力提升工程专项“无苦味华南型黄瓜及丛生南瓜优异种质创新及高效栽培技术研究”(2016SCHH-018)

Pathogen Identification and Biological Characteristics of a New Anthracnose on Cucumber

Han Shuai¹(), Wu Jie¹, Zhang Heqing¹, Liang Genyun²(), Xi Yadong¹()

¹Institute of Plant Protection, Sichuan Academy of Agriculture Sciences/ Key Laboratory of Integrated Pest Management on Crops in Southwest, Ministry of Agriculture and Rural Affairs, Chengdu 610066
²Horticulture Research Institute, Sichuan Academy of Agriculture Sciences/ Vegetable Germplasm Innovation and Variety Improvement Key Laboratory of Sichuan Province, Chengdu 610066

Received:2020-12-29 Revised:2021-07-27 Online:2021-10-05 Published:2021-10-28
Contact: Liang Genyun,Xi Yadong

摘要/Abstract

摘要：

明确引起黄瓜炭疽病的病原,旨在为该病的有效防治提供理论依据。通过形态学方法、柯赫氏法则验证,基于ITS、ACT和GDPH序列分析鉴定病原菌,并对该菌的生物学特性进行研究。结果表明,该菌回接黄瓜后的症状与田间症状一致。在PDA培养基上菌落为黑色,气生菌丝稀疏、灰白色;分生孢子长柱形,透明无隔;分生孢子盘着生黑色刚毛;孢子附着胞浅褐色,卵圆形,个别双生;菌丝附着胞深褐色,不规则形、浅裂或深裂。结合病原菌三基因联合建树的分析结果,可将其鉴定为Colletotrichum brevisporum。最适菌丝生长和产孢的温度25℃、pH 8,在PDA培养基上菌丝延伸最快,OA培养基上产孢最多,碳源中葡萄糖利于菌丝生长,可溶性淀粉和乳糖利于产孢,氮源中蛋白胨和酵母浸粉最适合菌丝生长,且后者产孢最多,致死温度为49℃。

关键词: 黄瓜炭疽病, Colletotrichum brevisporum, 病原鉴定, 多基因分子系统学, 生物学特性

Abstract:

Identifying the pathogen causing suspected anthracnose on cucumber can lay a foundation to control the disease effectively. Experiments were performed including Koch’s postulation test, biological characteristics identification using morphology and molecular methods based on ITS, ACT and GDPH sequences. The results showed that the symptom of inoculated cucumber leaves was the same as field symptom. The pathogen’s colony was black with sparse gray-white aerial mycelium. Conidia were long cylindrical, hyaline, aseptate; acervuli generated black seta; conidial appressoria were light brown and oval, several conidia produced twins; mycelial appressoria were dark brown and irregular, sometimes were deep or slightly lobed. Based on the above morphology description, the fungus was identified as Colletotrichum brevisporum according to the results of phylogenetic analyses of multi-locus sequences including ITS region, ACT and GDPH genes. The optimum temperature and pH for mycelial growth and spore reproduction were respectively 25℃ and 8, mycelium extended the fastest in PDA and the largest amount of conidium was produced in OA. Glucose was the most suitable carbon source for mycelia growth, soluble and lactose were most beneficial to reproduction spore. Peptone and yeast extract powder were conductive to mycelial growth and the latter was the best for making spores. The lethal temperature was 49℃.

Key words: cucumber anthracnose, Colletotrichum brevisporum, pathogenic identification, multi-gene phylogeny, biological characteristics

中图分类号:

S432.1

韩帅, 吴婕, 张河庆, 梁根云, 席亚东. 黄瓜新炭疽病原的鉴定及生物学特性初探[J]. 中国农学通报, 2021, 37(28): 96-108.

Han Shuai, Wu Jie, Zhang Heqing, Liang Genyun, Xi Yadong. Pathogen Identification and Biological Characteristics of a New Anthracnose on Cucumber[J]. Chinese Agricultural Science Bulletin, 2021, 37(28): 96-108.

图/表 8

图1. 田间及人工接种后发病症状 A.田间症状,B.菌饼接种后3天,C.菌饼接种后10天,D.分生孢子接种后10天

图2. 病原菌的形态学特征 A.菌落形态,B.分生孢子和孢子附着胞,C.分生孢子盘和刚毛,D.菌丝附着胞,标尺为20 μm

图3. 基于ITS、ACT和GDPH序列构建对C. brevisporum的系统发育树

图4. 不同培养基对病原菌菌丝生长和产孢量的影响不同小写字母表示在5%水平差异显著,不同大写字母表示在1%水平差异显著,下同

图5. 不同温度对病原菌菌丝生长和产孢量的影响

图6. 不同pH对病原菌菌丝生长和产孢量的影响

图7. 不同碳源对病原菌菌丝生长和产孢量的影响

图8. 不同氮源对病原菌菌丝生长和产孢量的影响

参考文献 25

[1]	郭予元. 中国农作物病虫害-中册(第三版)[M]. 北京: 中国农业出版社, 2014: 189.
[2]	肖敏, 曾向萍, 严婉荣, 等. 海南黄瓜新炭疽病病原的鉴定[J]. 分子植物育种, 2019, 16(22):7481-7486.
[3]	Noireung P, Phoulivong S, Liu F, et al. Novel Species of Colletotrichum Revealed by Morphology and Molecular Analysis[J]. Cryptogamie Mycologie, 2012, 33(3):347-362. doi: 10.7872/crym.v33.iss3.2012.347 URL
[4]	Liu F L, Tang G T, Zheng X J, et al. Molecular and phenotypic characterization of Colletotrichum species associated with anthracnose disease in peppers from Sichuan Province, China[J]. Sentific Reports, 2016, 6:32761.
[5]	Du Y X, Shi N N, Chen W L, et al. Identification of Colletotrichum brevisporum causing anthracnose on passion fruit[J]. Canadian Journal of Plant Pathology, 2017, 39:527-532. doi: 10.1080/07060661.2017.1367725 URL
[6]	Liu L P, Zhang L, Qiu P L, et al. First report of Colletotrichum brevisporum causing anthracnose on pumpkin in China[J]. Plant Disease, 2017, 102(5).
[7]	Cao X R, Xu X M, Che H Y, et al. Characteristics and distribution of Colletotrichum species in coffee plantations in Hainan, China[J]. Plant Pathology, 2019, 68(6).
[8]	Duan C H, Pan H R, Wang C C, et al. First Report of Colletotrichum brevisporum Causing Anthracnose on Papaya in Taiwan[J]. Plant Disease, 2017, 102(5).
[9]	Bezerra J P, Ferreira P V Barbosa L D F,, et al. First Report of Anthracnose on Chayote Fruits (Sechium edule) Caused by Colletotrichum brevisporum[J]. Plant Disease, 2015.
[10]	Vieira W A S, Nascimento R J, Michereff S J, et al. First Report of Papaya Fruit Anthracnose Caused by Colletotrichum brevisporum in Brazil[J]. Plant Disease, 2015, 97(12):1659-1659.
[11]	Narayan P, Hyang L, Ji L, et al. Endophytic Fungi from Lycium chinense Mill and Characterization of Two New Korean Records of Colletotrichum[J]. International Journal of Molecular Sciences, 2014, 15(9):15272-15286. doi: 10.3390/ijms150915272 pmid: 25170812
[12]	Chai A L, Zhao Q, Li B J, et al. First Report of Anthracnose Caused by Colletotrichum brevisporum on Gac (Momordica cochinchinensis) in Thailand[J]. Plant Disease, 2018.
[13]	方中达. 植病研究法(第3版)[M]. 北京: 中国农业出版社, 1998:122-124.
[14]	Yang Y L, Liu Z Y, Cai L, et al. Colletotrichum anthracnose of Amaryllidaceae[J]. Fungal Diversity, 2009, 39(2):123-146.
[15]	Sutton B C. The Coelomycetes; Fungi imperfecti with pycnidia, acervuli and stromata[J]. Commonwealth Mycological Institute, 1980.
[16]	Cai L, Hyde K D, Taylor P, et al. A polyphasic approach for studying Colletotrichum[J]. Fungal Diversity, 2009, 39(2):183-204.
[17]	White T J, Bruns T, Lee S, et al. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics[A].// Innis M A, Gelfand D, Sninsky J J, et al. PCR Protocols: a guide to methods and applications [M]. San Diego, California, USA: Academic Press, 1990:315-322.
[18]	Carbone I, Kohn L M. A method for designing primer sets for speciation studies in filamentous ascomycetes[J]. Mycologia, 1999, 91(3):553-556. doi: 10.2307/3761358 URL
[19]	Templeton M D, Rikkerink E H, Solon S L, et al. Cloning and molecular characterization of the glyceraldehyde-3-phosphate dehydrogenase-encoding gene and cDNA from the plant pathogenic fungus Glomerella cingulata[J]. Gene, 1992, 122:225-230. pmid: 1452034
[20]	杨友联, 刘永翔, 刘作易. 炭疽菌属真菌分类学研究进展[J]. 贵州农业科学, 2011, 39(1):152-157.
[21]	杨友联. 中国贵州、云南、广西炭疽菌属真菌多基因分子系统学研究[D]. 武汉:华中农业大学, 2010.
[22]	吴文平, 张志铭. 炭疽菌属(Colletotrichum Cda)发类学研究Ⅳ种的划分特征及评价[J]. 河北农业大学学报, 1995(2):93-99.
[23]	Du M, Schardl C L, Nuckles E M, et al. Using mating-type gene sequences for improved phylogenetic resolution of Collectotrichum species complexes[J]. Mycologia, 2005, 97(3):641-658. doi: 10.1080/15572536.2006.11832795 URL
[24]	Cannon P F, Damm U, Johnston P R, et al. Colletotrichum- current status and future directions[J]. Studies in Mycology, 2012, 73:181-213. doi: 10.3114/sim0014 pmid: 23136460
[25]	Weir B S, Johnston P R, Damn U. The Colletotrichum gloeosporioides species complex[J]. Studies Mycology, 2012, 73:115-180. doi: 10.3114/sim0011 URL

黄瓜新炭疽病原的鉴定及生物学特性初探

Pathogen Identification and Biological Characteristics of a New Anthracnose on Cucumber

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

图/表 8

参考文献 25

相关文章 15

编辑推荐

Metrics

本文评价

关于本刊

作者中心

审者中心

在线期刊

联系我们

[1]	魏日凤, 彭成彬, 陈美霞, 张政雄, 阮俊峰, 刘伟. 柳叶蜡梅真菌病害分离鉴定及生物学特性[J]. 中国农学通报, 2022, 38(3): 127-133.
[2]	贺泽霖, 陈晶, 倪洪涛, 张树权, 胡莹莹, 李岑. 药用植物水飞蓟的研究进展[J]. 中国农学通报, 2022, 38(18): 128-132.
[3]	任春燕, 刘杰, 罗明华, 聂忠扬, 黄宁, 赵海燕, 唐良德. 天敌昆虫—蠋蝽的研究进展[J]. 中国农学通报, 2022, 38(12): 100-109.
[4]	黄蔚, 崔丽红, 谢王超, 肖国强. 西瓜新炭疽病病原菌分子鉴定及其生物学特性初探[J]. 中国农学通报, 2022, 38(1): 131-136.
[5]	李燕, 李洪涛, 叶良超, 周俊祥, 罗明. 苹果细菌性枯枝病菌的生物学特性及抑菌药剂筛选[J]. 中国农学通报, 2021, 37(4): 112-119.
[6]	张国辉, 李向阳, 顾焕先, 张文华, 佀胜利, 李荣玉, 李明. 黔东南州红心猕猴桃叶尖干枯病病原鉴定[J]. 中国农学通报, 2021, 37(3): 145-149.
[7]	张卢慧, 赵志强, 赵全新, 郭庆元. 辣椒细菌性果实条斑病菌生物学特性及抑菌药剂筛选[J]. 中国农学通报, 2021, 37(25): 125-131.
[8]	李亚, 蔡永军, 余沁涵, 孔少连, 周晓晴, 张颖荷. 柠檬叶片溃疡病菌的分离鉴定及室内防治药剂筛选[J]. 中国农学通报, 2021, 37(24): 146-153.
[9]	钟全福, 陈斌, 樊海平, 林煜. 似鮈研究进展及前景展望[J]. 中国农学通报, 2021, 37(23): 133-137.
[10]	沈艳, 何鹏搏, 何鹏飞, 吴毅歆, 孔宝华, 李兴玉, Shahzad Munir, 何月秋. 番茄产后灰霉病的病原鉴定及生物防治[J]. 中国农学通报, 2021, 37(13): 102-107.
[11]	乔志文, 王积琛, 李彦丽. 甘蓝夜蛾研究进展[J]. 中国农学通报, 2020, 36(18): 147-153.
[12]	丁鑫, 郭巍, 沈希辉, 程建明, 孙萌. 4个国外接骨木属种质资源引种与生物学特性研究[J]. 中国农学通报, 2020, 36(16): 73-78.
[13]	徐太东,梁巧兰,吴琼,李嘉明,张强艳,康甲余,连芸芸,安贤. 甜瓜叶斑病病原鉴定及室内药剂筛选[J]. 中国农学通报, 2019, 35(20): 104-111.
[14]	黄立飞,刘伟明,刘也楠,何贤彪. 甘薯茎基部腐烂病调查及病原鉴定[J]. 中国农学通报, 2019, 35(18): 135-141.
[15]	刘满光,张娣,路文雅. 松枯梢病病原菌的研究进展[J]. 中国农学通报, 2018, 34(27): 97-101.