山东高密地区马铃薯疮痂病菌种类及致病性鉴定

doi:10.7668/hbnxb.20191183

摘要/Abstract

摘要： 为明确山东省高密地区马铃薯疮痂病的病原菌种类及其特征，2018-2019年从山东省高密市马铃薯主产区采集具有疮痂病病斑的马铃薯块茎，从病斑上共分离纯化到178个链霉菌菌株；采用薯片法、萝卜幼苗法及温室盆栽柯赫氏法则对马铃薯进行致病性试验，筛选到178个致病性较强的菌株。此外，将高密市5个地区所分离的马铃薯疮痂病菌株进行形态学鉴定、PCR分子鉴定和病原菌致病基因检测，之后根据16S rDNA基因的差异采用分子手段对所分离的致病菌株进行种类鉴定并构建系统发育树。结果显示：引起山东高密地区马铃薯疮痂病的病原菌主要包括4种链霉菌，分别为Streptomyces acidiscabies、Streptomyces scabies、Streptomyces luridiscabiei和Streptomyces griseoaurantiacus，与聚类分析结果相一致。其中Streptomyces acidiscabies为链霉菌优势种群且新菌株鉴定为Streptomyces griseoaurantiacus，为国内新报道病原菌，说明我国山东高密地区马铃薯疮痂病病原菌种类和分布较为复杂。

关键词: 马铃薯疮痂病, 链霉菌, 分类, 致病性分析, 多样性

Abstract: To identify and characterize the species of Streptomyces causing potato common scab in Shandong Gaomi. The typical scabby tubers were collected from potato growing areas in Shandong Gaomi from 2018 to 2019. After isolating and purifying,178 Streptomyces strains were obtained from scabby tubers. The pathogenicity of the isolates was determined on tuber slices,radish seedlings method and Koch's rule in greenhouse. A total of 178 strains were screened as pathogenic strains and could induce typical scab symptoms on immature potato. In addition,the potato scab strains isolated in 5 areas of Gaomi City were subjected to morphological identification,PCR molecular identification and pathogenic gene detection. These pathogenic strains were identified and phylogenetic tree was constructed on the basis of the differences in 16S rDNA sequences. Results revealed four kinds of Streptomyces species were found in Gaomi City,Shandong Province,including Streptomyces acidiscabies, Streptomyces scabies, Streptomyces luridiscabiei and Streptomyces griseoaurantiacus, which were consistent with the cluster analysis results. Among them, Streptomyces acidiscabies are dominant species and Streptomyces griseoaurantiacus are newly discovered common scab pathogens in China. The results suggestd that the Streptomyces species causing potato scab had obvious composition diversity and distribution complexity in Shandong Gaomi.

Key words: Potato common scab, Streptomyces, Classification, Pathogenic identification, Diversity

中图分类号:

S432.4

陈利达, 李磊, 谢学文, 石延霞, 柴阿丽, 李宝聚. 山东高密地区马铃薯疮痂病菌种类及致病性鉴定[J]. 华北农学报, 2020, 35(S1): 347-354. doi: 10.7668/hbnxb.20191183.

CHEN Lida, LI Lei, XIE Xuewen, SHI Yanxia, CHAI Ali, LI Baoju. Species and Pathogenicity Identification of Streptomyces Species Causing Potato Common Scab in Gaomi City, Shandong Province[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(S1): 347-354. doi: 10.7668/hbnxb.20191183.

http://www.hbnxb.net/CN/Y2020/V35/IS1/347

参考文献

[1] Hadi M R,Balali G R. The effect of salicylic acid on the reduction of Rhizoctonia solani damage in the tubers of marfona potato cultivar[J]. American-Eurasian Journal of Agricultural and Environmental Sciences,2010,7(4):492-496.
[2] 农业部. 农业部关于推进马铃薯产业开发的指导意见[J]. 中华人民共和国农业部公报,2016(3):4-7.
Ministry of Agriculture. Guiding opinions of the ministry of agriculture on promoting the development of the potato industry[J]. Bulletin of The People's republic of China,2016(3):4-7.
[3] 韩鹏,付羚. 内蒙古马铃薯种植业发展现状、问题及对策研究[J]. 内蒙古财经大学学报,2017,15(6):8-11. doi:10.3969/j.issn.2095-5871.2017.06.003.
Han P,Fu L. Research on the present situation,problems and countermeasures of potato planting in Inner Mongolia[J]. Journal of Inner Mongolia University of Finance and Economics,2017,15(6):8-11.
[4] 秦军红,李文娟,卢肖平,谢开云. 世界马铃薯产业发展概况[C]//屈冬玉,陈伊里. 马铃薯产业与中国式主食. 哈尔滨:哈尔滨地图出版社,2016:7-14.
Qin J H,Li W J,Lu X P,Xie K Y. Overview of the development of the world potato industry[C]//Qu D Y,Chen Y L. Potato Industry and Chinese Style Food. Harbin:Harbin Map Publishing House,2016:7-14.
[5] Miyajima K, Tanaka F, Takeuchi T, Kuninaga S. Streptomyces turgidiscabies sp. nov[J]. International Journal of Systematic and Evolutionary Microbiology,1998,48(2):495-502. doi:10.1099/00207713-48-2-495.
[6] Liu D Q,Anderson N A,Kinkel L L. Biological control of potato scab in the field with antagonistic Streptomyces scabies[J]. Phytopathology,1995,85(7):827-831. doi:10.1094/Phyto-85-827.
[7] Kreuze J F,Suomalainen S,Paulin L,Valkonen J P T. Phylogenetic analysis of 16S rRNA genes and PCR analysis of the nec1 gene from Streptomyces spp. causing common scab,pitted scab,and netted scab in Finland[J]. Phytopathology,1999,89(6):462-469. doi:10.1094/PHYTO.1999.89.6.462.
[8] 张良,程林润,卞晓波,谢关林. 中国马铃薯疮痂病的研究与防控[J]. 浙江农业科学,2019,60(10):1778-1781.doi:10.16178/j.issn.0528-9017.20191024.
Zhang L,Cheng L R,Bian X B,Xie G L. Research and prevention of potato scab in China[J]. Zhejiang Agricultural Science s,2019,60(10):1778-1781.
[9] Locci R. Actinomycetes as plant pathogens[J]. European Journal of Plant Pathology,1994,100(3-4):179-200. doi:10.1007/BF01876235.
[10] Scholte K,Labruyere R E. Netted scab:a new name for an old disease in Europe[J]. Potato Research,1985,28(4):443-448. doi:10.1007/BF02357520.
[11] Bouchek-Mechiche K,Pasco C,Andrivon D,Jouan B. Differences in host range,pathogenicity to potato cultivars and response to soil temperature among Streptomyces species causing common and netted scab in France[J]. Plant Pathology,2000,49(1):3-10. doi:10.1046/j.1365-3059.2000.00419.x.
[12] Sagova-Mareckova M,Omelka M,Kopecky J. Sequential analysis of soil factors related to common scab of potatoes[J]. FEMS Microbiology Ecology,2016,93(1):201. doi:10.1093/femsec/fiw201.
[13] Thaxter R. Potato scab[R]. Connectict Agricultural Experiment Station Report,1892:153-160.
[14] Goyer C,Beaulieu C. Host range of streptomycete strains causing common scab[J]. Plant Disease,1997,81(8):901-904. doi:10.1094/pdis.1997.81.8.901.
[15] Wanner L A,Kirk W W. Streptomyces -from basic microbiology to role as a plant pathogen[J]. American Journal of Potato Research,2015,92(2):236-242. doi:10.1007/s12230-015-9449-5.
[16] 杨亚东,胡韵菲,栗欣如,马力阳,王道龙,罗其友. 中国马铃薯种植空间格局演变及其驱动因素分析[J].农业技术经济,2017(8):39-47. doi:10.13246/j.cnki.jae.2017.08.004.
Yang Y D,Hu Y F,Li X R,Ma L Y,Wang D,Luo Q L. Analysis of the evolution of potato planting spatial pattern in China and its driving factors[J]. Agricultural Technology and Economy,2017(8):39-47.
[17] 邢莹莹,吕典秋,魏琪,万书明,董学志,邱彩玲,金光辉. 黑龙江省部分地区马铃薯疮痂病菌种类及致病性鉴定[J]. 植物保护,2016,42(1):26-32,50. doi:10.3969/j.issn.0529-1542.2016.01.005.
Xing Y Y,Lü D Q,Wei Q,Wan S M,Dong X Z,Qiu C L,Jin G H. Species and pathogenicity identification of Streptomyces species causing potato coommon scab in part of Heilongjiang Province[J]. Plant Protection,2016,42(1):26-32,50.
[18] Lambert D H,Loria R. Streptomyces acidiscabies sp.nov[J]. International Journal of Systematic and Evolutionary Microbiology,1989,39(4):393-396. doi:10.1099/00207713-39-4-393.
[19] 赵伟全,杨文香,李亚宁,刘大群,孟庆芳,张汀. 中国马铃薯疮痂病菌的鉴定[J]. 中国农业科学,2006,39(2):313-318. doi:10.3321/j.issn:0578-1752.2006.02.014.
Zhao W Q,Yang W X,Li Y N,Liu D Q,Meng Q F,Zhang T. Characterization and identification on the pathogen of potato scab in China[J]. Scientia Agricultura Sinica.2006,39(2):313-318.
[20] 杨梦平,王瑞仙,杜魏甫,巩晨,于德才,赵伟全,宋双伟,张红骥. 云南省马铃薯疮痂病致病链霉菌种类组成研究[J]. 植物病理学报,2018,48(4):445-454. doi:10.13926/j.cnki.apps.000134.
Yang M P,Wang R X,Du W F,Gong C,Yu D C,Zhao W Q,Song S W,Zhang H Y. The pathogenic Streptomyces species causing potato scab in Yunnan Province[J]. Acta Phytopathologica Sinic, 2018,48(4):445-454.
[21] 张萌,赵伟全,于秀梅,杨文香,张汀,刘大群. 中国马铃薯疮痂病病原菌16S rDNA的遗传多样性分析[J]. 中国农业科学,2009,42(2):499-504. doi:10.3864/j.issn.0578-1752.2009.02.015.
Zhang M,Zhao W Q,Yu X M,Yang W X,Zhang T,Liu D Q. Genetic diversity of potato scab pathogens in China based on 16S rDNA sequences[J]. Scientia Agricultura Sinica, 2009,42(2):499-504.
[22] 陈君. 山东地区马铃薯疮痂病病原菌的多样性及主要致病类型研究[D]. 泰安:山东农业大学,2017.
Chen J. Study on the diversity and main pathogenic types of potato scab in Shandong area[D]. Taian:Shandong Agricultural University,2017.
[23] Hiltunen L H,Weckman A,Ylhäinen A,Rita H,Richter E,Valkonen J P T. Responses of potato cultivars to the common scab pathogens, Streptomyces scabies and S. turgidiscabies[J]. Annals of Applied Biology,2005,146(3):395-403. doi:10.1111/j.1744-7348.2005.040083.x.
[24] Faucher E,Savard T,Beaulieu C. Characterization of actinomycetes isolated from common scab lesions on potato tubers[J]. Canadian Journal of Plant Pathology,1992,14(3):197-202. doi:10.1080/07060669209500874.
[25] 刘炳辉,曹远银,闫建芳,齐小辉,程浩,刘秋. 6种链霉菌基因组DNA提取方法比较[J].河南农业科学,2008,37(10):86-89. doi:10.3969/j.issn.1004-3268.2008.10.024.
Liu B H,Cao Y Y,Yan J F,Qi X H,Cheng H,Liu Q. Comparison among six methods of genome DNA extraction from Streptomyces[J]. Journal of Henan Agricultural Science,2008,37(10):86-89.
[26] Osborne C A,Galic M,Sangwan P,Janssen P H. PCR-generated artefact from 16S rRNA gene-specific primers[J]. FEMS Microbiology Letters,2005,248(2):183-187. doi:10.1016/j.femsle.2005.05.043.
[27] Edwards U,Rogall T,Blöcker H,Emde M,Böttger E C. Isolation and direct complete nucleotide determination of entire genes. Characterization of a gene coding for 16S ribosomal RNA[J]. Nucleic Acids Research,1989,17(19):7843-7853. doi:10.1093/nar/17.19.7843.
[28] 杨德洁,关欢欢,于秀梅,李寿如,赵伟全,刘大群. 2013-2017年我国北方马铃薯产区疮痂病病原菌组成分析[J]. 河南农业科学,2018,47(8):72-77. doi:10.15933/j.cnki.1004-3268.2018.08.012.
Yang D J,Guan H H,Yu X M,Li S R,Zhao W Q,Liu D Q. Analysis of potato scab pathogens in Northern China from 2013 to 2017[J]. Journal of Henan Agricultural Science,2018,47(8):72-77.
[29] 魏琪,闵凡祥,王晓丹,高云飞,杨帅,董学志,王文重,张抒,吕典秋,宿飞飞,张静华,李辉. 一种马铃薯疮痂病病原菌种类鉴定通用引物及检测方法:黑龙江,CN106957923A[P]. 2017-07-18.
Wei Q,Min F X,Wang X D,Gao Y F,Yang S,Dong X Z,Wang W Z,Zhang S,Lü D Q,Su F F,Zhang J H,Li H. Universal primer for detecting species of potato scab pathogen and its detection method:Heilongjiang,CN106957923A[P].2017-07-18.
[30] 王甄,沈艳芬,肖春芳,高剑华,陈巧玲. 马铃薯疮痂病病原菌的分离纯化方法及分子鉴定方法:湖北,CN107586749A[P].2018-01-16.
Wang Z,Shen Y F,Xiao C F,Gao J H,Chen Q L. Isolation and purification method and molecular identification method of potato scab pathogen:Hubei,CN107586749A[P]. 2018-01-16.
[31] 杜娟,任娟,赵思峰,任毓忠. 新疆马铃薯疮痂病病原的鉴定[J]. 石河子大学学报(自然科学版),2010,28(4):414-417. doi:10.3969/j.issn.1007-7383.2010.04.004.
Du J,Ren J,Zhao S F,Ren Y Z. Identification of pathogen on potato scab in Xinjang[J]. Journal of Shihezi University (Natural science),2010,28(4):414-417.
[32] Song J,Lee S C,Kang J W,Baek H J,Suh J W,Affiliations V. Phylogenetic analysis of Streptomyces spp. isolated from potato scab lesions in Korea on the basis of 16S rRNA gene and 16S 23S rDNA internally transcribed spacer sequences[J]. International Journal of Systematic and Evolutionary Microbiology,2004,54(1):203-209. doi:10.1099/ijs.0.02624-0.
[33] Lehtonen M J,Rantala H,Kreuze J F,Bång H,Kuisma L,Koski P,Virtanen E,Vihlman K,Valkonen J P T. Occurrence and survival of potato scab pathogens (Streptomyces species) on tuber lesions:quick diagnosis based on a PCR-based assay[J]. Plant Pathology,2004,53(3):280-287. doi:10.1111/j.0032-0862.2004.01009.x.
[34] Wanner L A. A survey of genetic variation in Streptomyces isolates causing potato common scab in the United States[J]. Phytopathology,2006,96(12):1363-1371. doi:10.1094/PHYTO-96-1363.
[35] Bukhalid R A,Chung S Y,Loria R. nec1,a gene conferring a necrogenic phenotype,is conserved in plant-pathogenic Streptomyces spp. and linked to a transposase pseudogene[J]. Molecular Plant-Microbe Interactions,1998,11(10):960-967. doi:10.1094/MPMI.1998.11.10.960.
[36] Loria R,Bukhalid R A,Fry B A,King R. Plant pathogenicity in the genus Streptomyces[J]. Plant Disease,1997,81(8):836-846. doi:10.1094/PDIS.1997.81.8.836.
[37] Bouchek-Mechiche K,Gardan L,Normand P,Jouan B. DNA relatedness among strains of Streptomyces pathogenic to potato in France:description of three new species, S. europaeiscabiei sp. nov. and S. stelliscabiei sp. nov. associated with common scab,and S. reticuliscabiei sp. nov. associated with netted scab[J]. International Journal of Systematic and Evolutionary Microbiology,2000,50(1):91-99. doi:10.1099/00207713-50-1-91.
[38] Natsume M,Komiya M,Koyanagi F,Tashiro N,Kawaide H,Abe H. Phytotoxin produced by Streptomyces sp. causing potato russet scab in Japan[J]. Journal of General Plant Pathology,2005,71(5):364-369. doi:10.1007/s10327-005-0211-6.
[39] Park D H,Yu Y M,Kim J S,Cho J M,Hur J H,Lim C K. Characterization of Streptomycetes causing potato common scab in Korea[J]. Plant Disease,2003,87(11):1290-1296. doi:10.1094/pdis.2003.87.11.1290.
[40] St-Onge R,Goyer C,Coffin R,Filion M. Genetic diversity of Streptomyces spp. causing common scab of potato in eastern Canada[J]. Systematic and Applied Microbiology,2008,31(6-8):474-484. doi:10.1016/j.syapm.2008.09.002.
[41] 李驰,刘艳,梁燕,邢星,张建平,李得宙,张笑宇. 马铃薯疮痂病病原菌鉴定及其生物学特性[J]. 农业生物技术学报,2019,27(5):897-907. doi:10.3969/j.issn.1674-7968.2019.05.013.
Li C,Liu Y,Liang Y,Xing X,Zhang J P,Li D Z,Zhang X Y. Identification and biological characteristics of pathogen in potato common scab[J]. Journal of Agricultural Biotechnology,2019,27(5):897-907.
[42] 杜魏甫.云南省马铃薯疮痂病菌鉴定及品种资源抗性评价[D]. 昆明:云南农业大学,2016.
Du W F. The identification of potato scab pathogen and evaluation of variety resource resistance in Yunnan Province[D]. Kunming:Yunna Agricultural University,2016.
[43] 王甄,沈艳芬,肖春芳,高剑华,张远学,叶兴枝,李大春. 湖北恩施马铃薯疮痂病的致病菌鉴定[C]//中国植物病理学会.中国植物病理学会2017年学术年会论文集,2017.
Wang Z,Shen Y F,Xiao C F,Gao J H,Zhang Y X,Ye X Z,Li D C. Pathogen identification of potato scab in Enshi,Hubei[C]//Chinese Society of Plant Pathology. Proceedings of the 2017 Annual Conference of Chinese Society of Plant Pathology,2017.
[44] 殷修鲁. 我国马铃薯疮痂病病原菌多样性及主要致病类型分析[D].泰安:山东农业大学,2018.
Yin X L. Diversity and main pathogenic types of pathogen isolates from potato scab in China[D]. Taian:Shandong Agricultural University,2018.
[45] 王丹,姚晓东,李新凤,王美琴,郝晓娟,王建明. 山西晋城马铃薯疮痂病病原菌鉴定[J]. 山西农业大学学报(自然科学版),2015,35(5):495-498. doi:10.3969/j.issn.1671-8151.2015.05.009.
Wang D,Yao X D,Li X F,Wang M Q,Hao X J,Wang J M. Identification of the pathogen of potato scab in Jincheng,Shanxi Province[J]. Journal of Shanxi Agricultural University(Natural Science Edition),2015,35(5):495-498.
[46] Zhou B,Zhang M S,Ma X K. First report of Streptomyces bottropensis causing potato common scab in Hebei Province,China[J]. Plant Disease,2017,101(3):1-2.doi:10.1094/PDIS-05-16v0671-PDN.
[47] 崔凌霄,杨成德,魏立娟,薛莉,张俊莲.甘肃省定西市马铃薯疮痂病新病原 Streptomyces galilaeus 的分离、鉴定及生物学特性研究[J]. 植物保护,2018,44(5):206-211. doi:10.16688/j.zwbh.2018109.
Cui L X,Yang C D,Wei L J,Xue L,Zhang J L. Isolation,identification and biological characteristics of Streptomyces galilaeus,a new pathogen of potato scab in Dingxi City,Gansu Province[J]. Plant Protection,2018,44(5):206-211.

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