吉林省和黑龙江省烟草赤星病病原鉴定
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摘要
为明确吉林省和黑龙江省烟草赤星病病原菌的种类,对吉林省15个烟区和黑龙江省7个烟区的烟草赤星病病原菌进行了分离鉴定,共分离获得312株致病菌株。通过形态学和分子生物学鉴定,共鉴定出4种链格孢属真菌,其中链格孢(Alternaria alternata)230株,细极链格孢(Alternaria tenuissima)78株,鸭梨链格孢(Alternaria yaliinficiens)2株,长柄链格孢(Alternaria longipes)2株。其中吉林省烟区鉴定出了链格孢、细极链格孢和鸭梨链格孢3种,黑龙江省烟区鉴定出了长柄链格孢、链格孢和细极链格孢3种。两个省区的优势种均为链格孢。此外,本试验利用5种基因序列片段(ITS、GDP、ACT、EF、RPB2)进行烟草赤星病菌的分子生物学鉴定,其中GDP基因仅能鉴定出链格孢和细极链格孢2种,RPB2基因能鉴定出4种链格孢,而ITS、EF、ACT基因序列比较保守,不适合区分这些链格孢种。本研究明确了吉林省和黑龙江省烟区引起烟草赤星病的病原菌种类,为下一步的定向防控奠定了基础。
In order to clarify the types of pathogens causing tobacco brown spot disease in Jilin and Heilongjiang Provinces, this study isolated and identified pathogens of tobacco brown spot disease from 15 major tobacco producing areas in Jilin Province and 7 major tobacco producing areas in Heilongjiang Province. A total of 312 pathogenic strains were isolated and identified. Four species of Alternaria were identified through morphological and molecular biological identification, including 230 strains of Alternaria alternata, 78 strains of Alternaria tenuissima, 2 strains of Alternaria yaliinficiens, and 2 strains of Alternaria longipes. A. alternata, A. tenuissima and A. yaliinficiens were identified in Jilin Province and A. longipes, A. alternata and A. tenuissima were identified in Heilongjiang Province. A. alternate and A. tenuissima were both identified in the two Province and A. alternata was the dominant species in both Province. In addition, five gene sequence fragments(ITS, GDP, ACT, EF, RPB2) were used for molecular biological identification in this study, in which the GDP gene can only identify A. alternata and A. tenuissima, RPB2 gene can identify all four species of Alternaria, while ITS, EF, ACT gene sequences are more conservative. So ITS, EF and ACT gene sequences are not suitable for distinguishing the various species of Alternaria. Through this study, the types of pathogens causing tobacco brown spot disease in Jilin Province and Heilongjiang Province were identified, which laid the foundation for the next stage of targeted prevention and control.
In order to clarify the types of pathogens causing tobacco brown spot disease in Jilin and Heilongjiang Provinces, this study isolated and identified pathogens of tobacco brown spot disease from 15 major tobacco producing areas in Jilin Province and 7 major tobacco producing areas in Heilongjiang Province. A total of 312 pathogenic strains were isolated and identified. Four species of Alternaria were identified through morphological and molecular biological identification, including 230 strains of Alternaria alternata, 78 strains of Alternaria tenuissima, 2 strains of Alternaria yaliinficiens, and 2 strains of Alternaria longipes. A. alternata, A. tenuissima and A. yaliinficiens were identified in Jilin Province and A. longipes, A. alternata and A. tenuissima were identified in Heilongjiang Province. A. alternate and A. tenuissima were both identified in the two Province and A. alternata was the dominant species in both Province. In addition, five gene sequence fragments(ITS, GDP, ACT, EF, RPB2) were used for molecular biological identification in this study, in which the GDP gene can only identify A. alternata and A. tenuissima, RPB2 gene can identify all four species of Alternaria, while ITS, EF, ACT gene sequences are more conservative. So ITS, EF and ACT gene sequences are not suitable for distinguishing the various species of Alternaria. Through this study, the types of pathogens causing tobacco brown spot disease in Jilin Province and Heilongjiang Province were identified, which laid the foundation for the next stage of targeted prevention and control.
引文
[1]张艳令.烟草病害自动识别诊断系统的研究[D].泰安：山东农业大学，2015.ZHANG Y L. Research on automatic identification and diagnosis system of tobacco diseases[D]. Tai'an:Shandong Agricultural University, 2015.
[2]任民，王志德，牟建民，等.我国烟草种质资源的种类与分布概况[J].中国烟草科学，2009,30（增刊）：8-14.REN M, WANG Z D, MOU J M, et al. The types and distribution of tobacco germplasm resources in China[J].Chinese Tobacco Science, 2009, 30(Supplement):8-14.
[3]成巨龙，安德荣.陕西烟草赤星病的病原鉴定[J].陕西农业科学，1997(3):18.CHENG J L, AN D R. Identification of the pathogen of tobacco brown spot disease in Shaanxi[J]. Shaanxi Journal of Agricultural Sciences, 1997(3):18.
[4]祖艳青，蒋士君，王海涛，等.河南省烟草赤星病病原鉴定[J].中国烟草学报，2013,19(4):73-77.ZU Y Q, JIANG S J, WANG H T, et al. Identification of tobacco brown spot pathogen in Henan[J]. Acta Tabacaria Sinica, 2013, 19(4):73-77.
[5]关博元，肖崇刚，陈国康，等.重庆市烟草赤星病菌致病力测定[J].植物保护，2007,33(3):62-64.GUAN B Y, XIAO C G, CHEN G K, et al. Studies on the pathogenicity differentiation of Alternaria alternata in Chongqing[J]. Plant Protection, 2007, 33(3):62-64.
[6]胡中会，赵立华，严恩平.云南烟草赤星病菌及近似种rDNA-ITS序列分析[J].云南农业大学学报，2012,27(5):670-676.HU Z H, ZHAO L H, YAN E P. rDNA-ITS sequence analysis of Alternaria alternata in Yunnan and their similar species[J]. Journal of Yunnan Agricultural University, 2012, 27(5):670-676.
[7]耿莉娜，帅红，赵敏，等.重庆巫溪烟区烟草赤星病病原鉴定[J].江苏农业科学，2016,44(8):181-183.GENG L N, SHUAI H, ZHAO M, et al. Identification of tobacco brown spot pathogen in Wuxi county of Chongqing[J]. Jiangsu Agricultural Science, 2016, 44(8):181-183.
[8]唐承成，曾琛，姜于兰，等.贵州省烟草赤星病病原菌的鉴定[J].湖北农业科学，2015,54(15):3656-3658.TANG C C, ZENG C, JIANG Y L, et al. Identification of pathogenic fungi of tobacco brown spot in Guizhou Province[J]. Hubei Agricultural Science, 2015, 54(15):3656-3658.
[9]杨涛，黎妍妍，郑露，等.湖北烟区烤烟赤星病病原鉴定[J].中国烟草科学，2017,38(5):32-38.YANG T, LI Y Y, ZHENG L, et al. Identification of pathogens causing flue-cured tobacco brown spot in Hubei Province[J]. Chinese Tobacco Science, 2017, 38(5):32-38.
[10]胡晓莉，蒋彩虹，耿锐梅，等.山东部分地区烟草赤星病病原菌的形态学与分子生物学鉴定[J].江苏农业科学，2018,46(11):82-86.HU X L, JIANG C H, GENG R M, et al. Morphological and molecular biological identification of pathogens causing tobacco brown spot disease in some areas of Shandong Province[J]. Jiangsu Agricultural Science,2018, 46(11):82-86.
[11]邱小燕，汤智鹏，张敏，等.一种适用于多数植物病原真菌的单孢分离方法[J].安徽农业科学，2011,39(9):5263-5264.QIU X Y, TANG Z P, ZHANG M, et al. Research on the isolation method of single spore of most plant pathogenic fungi[J]. Anhui Agricultural Sciences, 2011, 39(9):5263-5264.
[12]高崇，吴国贺，李佰霖，等.延边烟区烟草菌核病病原菌鉴定及抗性种质筛选[J].中国烟草科学，2018,39(2):69-74.GAO C, WU G H, LI B L, et al. Identification of tobacco sclerotinia rot in Yanbian and screening for resistant germplasms[J]. Chinese Tobacco Science, 2018, 39(2):69-74.
[13]张恒，朱迪，何现梅，等.烤烟苗期枯萎病原菌鉴定及室内药剂毒力测定[J].中国烟草科学，2018,39(6):36-42.ZHANG H, ZHU D, HE X M, et al. Identification pathogen of tobacco seedling wilt and toxicity test in laboratory[J]. Chinese Tobacco Science, 2018, 39(6):36-42.
[14] ROBERTS R G. Alternaria yaliinficiens sp. nov. on Ya Li pear fruit:from interception to identification[J]. Plant Disease, 2005, 89:134-145.
[15]魏景超.真菌鉴定手册[M].上海：上海科学技术出版社，1979.WANG J C. Fungal identification manual[M]. Shanghai:Shanghai Scientific&Technical Publishers, 1979.
[16]方中达.植病研究方法[M].北京：中国农业出版社，1998.FANG Z D. Research methods on plant diseases[M].Beijing:China Agriculture Press, 1998.
[17]李朋华.部分链格孢属真菌的形态学及多基因鉴定[D].郑州：河南农业大学，2016.LI P H. Identifiation of partial Alternaria by morphology and multiple-gene analysis[D]. Zhengzhou:Henan Agricultural University, 2016.
[18] SIMMONS E G. Alternaria:an identification manual[M].Utrecht:CBS Fungal Biodiversity Centre, 2007:1-585.
[19]张天宇.中国真菌志：链格孢属[M].北京：科学出版社，2003:1-220.ZHANG T Y. Flora fungorum sinicorum:Alternaria[M].Beijing:Science Press, 2003:1-220.
[2]任民，王志德，牟建民，等.我国烟草种质资源的种类与分布概况[J].中国烟草科学，2009,30（增刊）：8-14.REN M, WANG Z D, MOU J M, et al. The types and distribution of tobacco germplasm resources in China[J].Chinese Tobacco Science, 2009, 30(Supplement):8-14.
[3]成巨龙，安德荣.陕西烟草赤星病的病原鉴定[J].陕西农业科学，1997(3):18.CHENG J L, AN D R. Identification of the pathogen of tobacco brown spot disease in Shaanxi[J]. Shaanxi Journal of Agricultural Sciences, 1997(3):18.
[4]祖艳青，蒋士君，王海涛，等.河南省烟草赤星病病原鉴定[J].中国烟草学报，2013,19(4):73-77.ZU Y Q, JIANG S J, WANG H T, et al. Identification of tobacco brown spot pathogen in Henan[J]. Acta Tabacaria Sinica, 2013, 19(4):73-77.
[5]关博元，肖崇刚，陈国康，等.重庆市烟草赤星病菌致病力测定[J].植物保护，2007,33(3):62-64.GUAN B Y, XIAO C G, CHEN G K, et al. Studies on the pathogenicity differentiation of Alternaria alternata in Chongqing[J]. Plant Protection, 2007, 33(3):62-64.
[6]胡中会，赵立华，严恩平.云南烟草赤星病菌及近似种rDNA-ITS序列分析[J].云南农业大学学报，2012,27(5):670-676.HU Z H, ZHAO L H, YAN E P. rDNA-ITS sequence analysis of Alternaria alternata in Yunnan and their similar species[J]. Journal of Yunnan Agricultural University, 2012, 27(5):670-676.
[7]耿莉娜，帅红，赵敏，等.重庆巫溪烟区烟草赤星病病原鉴定[J].江苏农业科学，2016,44(8):181-183.GENG L N, SHUAI H, ZHAO M, et al. Identification of tobacco brown spot pathogen in Wuxi county of Chongqing[J]. Jiangsu Agricultural Science, 2016, 44(8):181-183.
[8]唐承成，曾琛，姜于兰，等.贵州省烟草赤星病病原菌的鉴定[J].湖北农业科学，2015,54(15):3656-3658.TANG C C, ZENG C, JIANG Y L, et al. Identification of pathogenic fungi of tobacco brown spot in Guizhou Province[J]. Hubei Agricultural Science, 2015, 54(15):3656-3658.
[9]杨涛，黎妍妍，郑露，等.湖北烟区烤烟赤星病病原鉴定[J].中国烟草科学，2017,38(5):32-38.YANG T, LI Y Y, ZHENG L, et al. Identification of pathogens causing flue-cured tobacco brown spot in Hubei Province[J]. Chinese Tobacco Science, 2017, 38(5):32-38.
[10]胡晓莉，蒋彩虹，耿锐梅，等.山东部分地区烟草赤星病病原菌的形态学与分子生物学鉴定[J].江苏农业科学，2018,46(11):82-86.HU X L, JIANG C H, GENG R M, et al. Morphological and molecular biological identification of pathogens causing tobacco brown spot disease in some areas of Shandong Province[J]. Jiangsu Agricultural Science,2018, 46(11):82-86.
[11]邱小燕，汤智鹏，张敏，等.一种适用于多数植物病原真菌的单孢分离方法[J].安徽农业科学，2011,39(9):5263-5264.QIU X Y, TANG Z P, ZHANG M, et al. Research on the isolation method of single spore of most plant pathogenic fungi[J]. Anhui Agricultural Sciences, 2011, 39(9):5263-5264.
[12]高崇，吴国贺，李佰霖，等.延边烟区烟草菌核病病原菌鉴定及抗性种质筛选[J].中国烟草科学，2018,39(2):69-74.GAO C, WU G H, LI B L, et al. Identification of tobacco sclerotinia rot in Yanbian and screening for resistant germplasms[J]. Chinese Tobacco Science, 2018, 39(2):69-74.
[13]张恒，朱迪，何现梅，等.烤烟苗期枯萎病原菌鉴定及室内药剂毒力测定[J].中国烟草科学，2018,39(6):36-42.ZHANG H, ZHU D, HE X M, et al. Identification pathogen of tobacco seedling wilt and toxicity test in laboratory[J]. Chinese Tobacco Science, 2018, 39(6):36-42.
[14] ROBERTS R G. Alternaria yaliinficiens sp. nov. on Ya Li pear fruit:from interception to identification[J]. Plant Disease, 2005, 89:134-145.
[15]魏景超.真菌鉴定手册[M].上海：上海科学技术出版社，1979.WANG J C. Fungal identification manual[M]. Shanghai:Shanghai Scientific&Technical Publishers, 1979.
[16]方中达.植病研究方法[M].北京：中国农业出版社，1998.FANG Z D. Research methods on plant diseases[M].Beijing:China Agriculture Press, 1998.
[17]李朋华.部分链格孢属真菌的形态学及多基因鉴定[D].郑州：河南农业大学，2016.LI P H. Identifiation of partial Alternaria by morphology and multiple-gene analysis[D]. Zhengzhou:Henan Agricultural University, 2016.
[18] SIMMONS E G. Alternaria:an identification manual[M].Utrecht:CBS Fungal Biodiversity Centre, 2007:1-585.
[19]张天宇.中国真菌志：链格孢属[M].北京：科学出版社，2003:1-220.ZHANG T Y. Flora fungorum sinicorum:Alternaria[M].Beijing:Science Press, 2003:1-220.
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