半边红李炭疽病病害调查及病原菌鉴定

doi:10.11924/j.issn.1000-6850.casb2020-0328

中国农学通报 ›› 2021, Vol. 37 ›› Issue (15): 128-136.doi: 10.11924/j.issn.1000-6850.casb2020-0328

所属专题：植物保护

半边红李炭疽病病害调查及病原菌鉴定

王群¹(), 毕云青¹, 何成兴¹, 郭志祥¹, 钟德卫², 李君洪³, 陈乾昭², 游继华², 黄正会², 李进斌¹()

¹云南省农业科学院农业环境资源研究所,昆明 650205
²云南省昭通市绥江县农业农村局农技推广中心,云南绥江 657700
³绥江县渔业技术服务站,云南绥江 657700

收稿日期:2020-08-20 修回日期:2020-11-13 出版日期:2021-05-25 发布日期:2021-05-18
通讯作者: 李进斌
作者简介:王群,女,1970年出生,四川资阳人,副研究员,博士,研究方向：生物多样性及植物病害研究。通信地址：650205 云南省昆明市北京路2238号云南省农科院环资所,Tel：15288249540,E-mail： qunwang70@163.com。
基金资助:
云南省专家基层科研工作站—李进斌专家工作站(绥江县云人社通「2017」130号);国家自然科学基金“稻瘟病菌无毒基因Avr-Pi9在云南稻区的变异及微进化特点”(31860481);云南省人才培养项目“省学术技术带头人后备人才”(2015HB076)

Anthracnose Disease on Prunus salicina ‘Banbianhong’: Investigation and Pathogen Identification

Wang Qun¹(), Bi Yunqing¹, He Chengxing¹, Guo Zhixiang¹, Zhong Dewei², Li Junhong³, Chen Qianzhao², You Jihua², Huang Zhenghui², Li Jinbin¹()

¹Agricultural Environment and Resources Research Institute, Yunnan Academy of Agricultural Sciences, Kunming 650205
²Agricultural Technology Extension Center of Agricultural and Rural Bureau of Suijiang County, Suijiang Yunnan 657700
³Service Station of Fishery Technology of Suijiang County, Suijiang Yunnan 657700

Received:2020-08-20 Revised:2020-11-13 Online:2021-05-25 Published:2021-05-18
Contact: Li Jinbin

摘要/Abstract

摘要：

探明绥江半边红李炭疽病为害程度及病原菌的种类,可为李树炭疽病的防控提供依据。以绥江县2个主要种植区会仪镇和新滩镇的6个半边红李果园为研究对象,对炭疽病为害状况进行了调查,并对炭疽病原菌进行分离保存;利用病原菌分生孢子形态及核糖体ITS序列对绥江半边红李树炭疽病致病菌进行了种类鉴定和分析。病害调查表明,新滩镇比会仪镇的危害程度严重。单孢分离获得15株分离株,各分离株平板菌落形态略有差异,获得4类不同的分生孢子,经回接验证均致病。rDNA ITS序列系统进化分析初步推定半边红李炭疽病的病原菌为胶孢炭疽复合群(Colletotrichum gloeosporioides species complex)和暹罗炭疽复合群C. siamense species complex。该研究探查了半边红李炭疽病的为害状况,获得了李炭疽菌的单胞分离菌株,初步鉴定了半边红李炭疽菌归属的复合群。

关键词: 半边红李, 炭疽病, 炭疽菌, 形态鉴定, ITS

Abstract:

The aim is to investigate the anthracnose disease and the pathogen species in Prunus salicina ‘Banbianhong’ and provide a basis to prevent and control the disease. In this study, six P. salicina ‘Banbianhong’ orchards were taken as research objects in two major production areas, Huiyi and Xintan Town in Suijiang County, Yunnan Province, anthracnose disease was investigated and pathogenic fungi were isolated and stored. The pathogen species were identified and analyzed according to the spore morphology and ITS sequences of Colletotrichum. Disease investigation showed that the disease severity of anthracnose in Xintan was higher than that in Huiyi. A total of 15 single spore isolates were obtained, and the morphology of colonies on every strain plate had weak difference. Four kinds of conidia were obtained and all of them were pathogenic by Tie-back experiment. rDNA ITS sequence phylogenetic analysis showed that the pathogen of anthracnose disease in P. salicina ‘Banbianhong’ were Colletotrichum gloeosporioides species complex and C. siamense species complex. This study investigated the damage status of anthracnose disease in P. salicina ‘Banbianhong’, got the single spore isolates, and determined the complex groups of the pathogen of anthracnose disease in P. salicina ‘Banbianhong’.

Key words: Prunus salicina ‘Banbianhong’, anthracnose, Colletotrichum spp., morphological identification, ITS

中图分类号:

S463.65

王群, 毕云青, 何成兴, 郭志祥, 钟德卫, 李君洪, 陈乾昭, 游继华, 黄正会, 李进斌. 半边红李炭疽病病害调查及病原菌鉴定[J]. 中国农学通报, 2021, 37(15): 128-136.

Wang Qun, Bi Yunqing, He Chengxing, Guo Zhixiang, Zhong Dewei, Li Junhong, Chen Qianzhao, You Jihua, Huang Zhenghui, Li Jinbin. Anthracnose Disease on Prunus salicina ‘Banbianhong’: Investigation and Pathogen Identification[J]. Chinese Agricultural Science Bulletin, 2021, 37(15): 128-136.

图/表 6

果园	海拔/m	发病率/%	病情指数
会仪镇黄坪村1	380~500	0	0.0
会仪镇黄坪村2	500~700	30	31.1
会仪镇黄坪村3	700~900	30	30.0
合计		20	20.4
新滩镇鲢鱼10队	380~500	40	41.1
新滩镇鲢鱼12队	500~700	70	70.0
新滩镇鲢鱼17队	700~900	40	63.3
合计		50	78.5

表1. 基于0~9级分级标准的绥江6个半边红李果园炭疽病发病率及病情指数

图1. 半边红李炭疽病菌株18-5-B1、18-8-C1、18-8-D1和18-4-C2菌落及分生孢子形态 A和B列分别为菌落的正面和反面,C列为分生孢子形态图

菌株	长度/μm	宽度/μm	长宽比
18-4-C2	18.35±1.70(15.9~21.6) B	4.72±0.64(4.0~6.1) A	3.94±0.58(3.0~4.8) B
18-8-C1	13.35±1.14(11.9~15.1) C	4.00±0.55(3.3~4.8) B	3.40±0.62(2.9~4.6) B
18-8-D1	21.94±2.25(18.2~26.1) A	2.62±0.52(2.1~3.5) C	8.65±1.91(6.7~12.2) A
18-5-B1	14.65±1.94(12.8~19.1) C	4.35±0.53(3.6~5.2) AB	3.44±0.82(2.5~5.4) B

表2. 云南绥江半边红李炭疽病分离菌株分生孢子大小

图2. 半边红李炭疽病分离菌株18-8-C1、18-5-B1接种离体叶片9天后的发病症状

图3. 部分半边红李炭疽病菌分离株ITS引物PCR扩增电泳图 M：分子量标记,片段大小从上到下依次为2000 bp、1000 bp、750 bp、500 bp、250 bp和100 bp;A2、A5、A7、A8、A9、A12、B3、B5和B7分别代表菌株18-2-D2、18-7-D5、18-8-C1、18-8-D1、18-10-A2、18-020-6、18-8-1、18-9-2和18-10-5

图4. 基于4个半边红李炭疽病菌及Genbank中下载的139条炭疽菌ITS序列的ML系统进化分析分支上数值为大于50%的Bootstrap支持率

参考文献 45

[1]	成策, 陆俊, 韩文琪, 等. 李子果粒果汁饮料加工工艺研究[J]. 食品与机械, 2015,31(6):215-218.
[2]	Cannon P F, Buddie A G, Bridge P D. The typification of Colletotrichum gloeosporioides[J]. Mycotaxon, 2008,104(2):189-204.
[3]	Sutton B C. The genus Glomerella and its anamorph Colletotrichum[M]// Bailey J A and Jeger M J. Colletotrichum: biology, pathology control. Wallingford: CABI, 1992:1-26.
[4]	Corda A C I. Die Pilze Deutschlands[M] // Sturm J. Deutschlands flora in Abbildungen nach der Natur mit Beschreibungen. Nürnberg: Sturm, 1831:33-64.
[5]	Arx J V. Die Arten der Gattung Colletotrichum Cda[J]. Phytopathologische Zeitschrift, 1957,29:413-468.
[6]	Sutton B C. The Coelomycetes[M]// Fungi imperfecti with pycnidia acervuli and stromata. Kew, UK: CABI, 1980:523-537.
[7]	Cannon P F, Damm U, Johnston P R, et al. Colletotrichum - current status and future directions[J]. Studies in Mycology, 2012,73(1):181-213. doi: 10.3114/sim0014 pmid: 23136460
[8]	Damm U, Cannon P F, Liu F, et al. The Colletotrichum orbiculare species complex: important pathogens of field crops and weeds[J]. Fungal Diversity, 2013,61(1):29-59. doi: 10.1007/s13225-013-0255-4 URL
[9]	Damm U, Cannon P F, Woudenberg J H C, et al. The Colletotrichum acutatum species complex[J]. Studies in Mycology, 2012,73(73):37-113. doi: 10.3114/sim0010 URL
[10]	Damm U, Cannon P F, Woudenberg J H C, et al. The Colletotrichum boninense species complex[J]. Studies in Mycology, 2012,73(1):1-36. doi: 10.3114/sim0002 pmid: 23136457
[11]	Damm U, O'Connell R J, Groenewald J Z, et al. The Colletotrichum destructivum species complex-hemibiotrophic pathogens of forage and field crops[J]. Studies in Mycology, 2014,79(79):49-84. doi: 10.1016/j.simyco.2014.09.003 URL
[12]	Hyde K D, Cai L, Cannon P F, et al. Colletotrichum - names in current use[J]. Fungal Diversity, 2009,39:147-182.
[13]	Liu F, Cai L, Crous P W. The Colletotrichum gigasporum species complex[J]. Persoonia, 2014,33(1):83-97. doi: 10.3767/003158514X684447 URL
[14]	Sharma G, Kumar N, Weir B, et al. The ApMat marker can resolve Colletotrichum species: a case study with Mangifera indica[J]. Fungal Diversity, 2013,61(1):117-138. doi: 10.1007/s13225-013-0247-4 URL
[15]	Weir B S, Johnston P R, Damm U. The Colletotrichum gloeosporioides species complex[J]. Studies in Mycology, 2012,73(1):115-180. doi: 10.3114/sim0011 pmid: 23136459
[16]	Crouch J A. Colletotrichum caudatum s.l. is a species complex[J]. IMA Fungus, 2014,5(1):17-30. doi: 10.5598/imafungus.2014.05.01.03 URL
[17]	Lee H D, Kim D H, Jeon Y A, et al. Molecular and cultural characterization of Colletotrichum spp. causing bitter rot of apples in Korea[J]. The Plant Pathology Journal, 2007,23(2):37-44. doi: 10.5423/PPJ.2007.23.2.037 URL
[18]	侯丽冰, 贺伟, 刘小勇, 等. 我国几种松干锈菌亲缘关系的ITS 序列分析[J]. 北京林业大学学报, 2002,24(Z1):175-182.
[19]	Schoch C L, Seifert K A, Sabine H, et al. Nuclear ribosomal internal transcribed spacer (ITS) region as a universal DNA barcode marker for Fungi[J]. Proceedings of the National Academy of Sciences of the United States of America, 2012,109(16):6241-6246.
[20]	Dresler-Nurmi A, Kaijalainen S, LindstrÖM K, et al. Grouping of lignin degrading corticioid fungi based on RFLP analysis of 18S rDNA and ITS regions[J]. Mycological Research, 1999,103(8):990-996. doi: 10.1017/S0953756298008156 URL
[21]	Freeman S, Minz D, Jurkevitch E, et al. Molecular analyses of Colletotrichum species from almond and other fruits[J]. Phytopathology, 2000,90(6):608-614. doi: 10.1094/PHYTO.2000.90.6.608 pmid: 18944540
[22]	邓先琼, 郭立中, 张天晓. 布朗李炭疽病的病原鉴定及药效试验[J]. 华中农业大学学报, 2004,23(6):626-630.
[23]	Hassan O, Lee Y S, Chang T. Colletotrichum Species Associated with Japanese Plum (Prunus salicina) Anthracnose in South Korea[J]. Scientific Reports, 2019,9(1):1-12.
[24]	Montri P, Taylor P W J, Mongkolporn O. Pathotypes of Colletotrichum capsici, the causal agent of chili anthracnose, in Thailand[J]. Plant Disease, 2009,93:17-20. doi: 10.1094/PDIS-93-1-0017 URL
[25]	Awa O C, Samuel O, Oworu O O, et al. First report of fruit anthracnose in Mango caused by Colletotrichum gloeosporioides in Southwestern Nigeria[J]. International Journal of Scientific & Technology Research, 2012,1(4):30-34.
[26]	曹若彬. 果树病理学[M]. 北京: 中国农业出版社, 1992.
[27]	Li Z, Liang Y M, Tian C M. Characterization of the causal agent of poplar anthracnose occurring in the Beijing region[J]. Mycotaxon, 2012,120(1):277-286. doi: 10.5248/120.277 URL
[28]	方中达. 植物病理研究方法[M]. 北京: 中国农业出版社, 1979.
[29]	Tai T H, Tanksley S D. A rapid and inexpensive method for isolation of total DNA from dehydrated plant tissue[J]. Plant Molecular Biology Reporter, 1990,8(4):297-303. doi: 10.1007/BF02668766 URL
[30]	White T J, Bruns T, Lee S, et al. Analysis of phylogenetic relationships by amplification and direct sequencing of ribosomal RNA genes[M] // Innis MA, Gelfand D H, Sninsky J J, et al. PCR protocols: a guide to methods and applications. San Diego, USA: Academic Press, 1990:315-322.
[31]	Thompson J D, Gibson T J, Plewniak F, et al. The CLUSTALX windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools[J]. Nucleic Acids Research, 1997,25(24):4876-4882. doi: 10.1093/nar/25.24.4876 URL
[32]	Kumar S, Stecher G, Tamura K. MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets[J]. Molecular Biology and Evolutionary 2016,33(7):1870-1874. doi: 10.1093/molbev/msw054 URL
[33]	Tamura K, Stecher G, Peterson D, et al. MEGA6: Molecular Evolutionary Genetics Analysis Version 6.0[J]. Molecular Biology & Evolution, 2013,30(4):2725-2729.
[34]	Prihastuti H, Cai L, Chen H, et al. Characterization of Colletotrichum species associated with coffee berries in northern Thailand[J]. Fungal Diversity, 2009,39:89-109.
[35]	Yang Y L, Liu Z Y, Cai L, et al. Colletotrichum anthracnose of Amaryllidaceae[J]. Fungal Diversity, 2009,39(2):123-146.
[36]	Chen Y, Qiao W, Zeng L, et al. Characterization, pathogenicity, and phylogenetic analyses of Colletotrichum species associated with brown blight disease on Camellia sinensis in China[J]. Plant Disease, 2017,101(6):1022-1028. doi: 10.1094/PDIS-12-16-1824-RE URL
[37]	Grammen A, Wenneker M, Van Campenhout J, et al. Identification and pathogenicity assessment of Colletotrichum isolates causing bitter rot of apple fruit in Belgium[J]. European Journal of Plant Pathology, 2019,153(1):47-63. doi: 10.1007/s10658-018-1539-z pmid: WOS:000484796700003
[38]	Diao Y Z, Zhang C, Liu F, et al. Colletotrichum species causing anthracnose disease of chili in China[J]. Persoonia, 2017,38:20-37. doi: 10.3767/003158517X692788 URL
[39]	Diao Y Z, Zhang C, Xu J P, et al. Genetic differentiation and recombination among geographic populations of the fungal pathogen Colletotrichum truncatum from chili peppers in China[J]. Evolutionary Applications, 2015,8:108-118. doi: 10.1111/eva.2015.8.issue-1 URL
[40]	Kanto T, Uematsu S, Tsukamoto T, et al. Anthracnose of sweet pepper caused by Colletotrichum scovillei in Japan[J]. Journal of General Plant Pathology, 2014,80:73-78. doi: 10.1007/s10327-013-0496-9 URL
[41]	Liao C Y, Chen M Y, Chen Y K, et al. Characterization of three Colletotrichum acutatum isolates from Capsicum spp.[J]. European Journal of Plant Pathology, 2012,133:599-608. doi: 10.1007/s10658-011-9935-7 URL
[42]	Sharma G, Shenoy B D. Colletotrichum fructicola and C. siamense are involved in chilli anthracnose in India[J]. Archives of Phytopathology and Plant Protection, 2014,47(10):1179-1194. doi: 10.1080/03235408.2013.833749 URL
[43]	Than P P, Prihastuti H, Phoulivong S, et al. Chilli anthracnose disease caused by Colletotrichum species[J]. Journal of Zhejiang University Science B, 2008,9(10):764-778. doi: 10.1631/jzus.B0860007 URL
[44]	向梅梅, 张云霞, 刘霄. 炭疽菌属真菌分类的研究进展[J]. 仲恺农业工程学院学报, 2017,30(1):59-66.
[45]	Sharma G, Pinnaka A K, Shenoy B D. Resolving the Colletotrichum siamense species complex using ApMat marker[J]. Fungal Diversity, 2015,71(1):247-264. doi: 10.1007/s13225-014-0312-7 URL

半边红李炭疽病病害调查及病原菌鉴定

Anthracnose Disease on Prunus salicina ‘Banbianhong’: Investigation and Pathogen Identification

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