葡萄炭疽病菌鉴定及同源性分析

doi:10.11924/j.issn.1000-6850.casb16110006

摘要/Abstract

摘要： 为揭示葡萄炭疽病菌种类和遗传特征，给深入研究该病提供科学理论依据。本研究对不同地区的26 株葡萄炭疽病菌通过形态学和rDNA-ITS 序列分析进行鉴定及同源性研究。利用真菌通用引物ITS4、ITS5 对病菌基因组进行PCR 扩增，将测序结果与NCBI 数据库上已知序列进行比对，同时利用Clustalx 1.83 和MEGA 5.0 软件进行聚类分析。结果表明：病菌菌落为白色或灰白色，菌丝呈绒状或絮状，分生孢子为圆柱形或圆筒状，单孢，无色；测得病菌ITS 序列为500 bp 左右，鉴定病原均为胶孢炭疽菌(Colletotrichum gloeosporioides)。不同地区葡萄炭疽菌的rDNA-ITS 序列同源性高，亲缘关系比较近，但是各菌株间存在着遗传差异，并且菌株之间差异与地理来源和品种无明显的相关性。

关键词: 肉鸡, 肉鸡, 肠毒综合症, 病因分析, 防治措施

Abstract: The objectives are to clarify fungal species and genetic characteristics of Colletotrichum gloeosporioides, and provide basis for further studying this disease. The identification and homology analysis of 26 strains of C. gloeosporioides from different districts were conducted by morphology and rDNA-ITS sequence analysis. PCR amplification was performed with fungal universal primers ITS4 and ITS5. We compared the sequences with the sequences available in NCBI, and cluster analysis was conducted with Clustalx 1.83 and MEGA 5.0 software. Results indicated that colony color was white or grey white, hypha exhibited velvet or flocculent, conidium showed cylindrical shape or cylindric shape, and was colorless; ITS sequence of pathogen was about 500 bp, all identified pathogen were C. gloeosporioides. Although grape anthrax from different districts exhibited high sequence homology and close genetic relationship, different strains showed genetic differences and this difference had no obvious correlation with geographical origin and varieties.

刘丽,刘长远,王辉,关天舒,于舒怡,李柏宏. 葡萄炭疽病菌鉴定及同源性分析[J]. 中国农学通报, 2017, 33(35): 132-136.

参考文献

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[3]沈淑琳.葡萄病害及其防治[M].中国林业出版社,1992.
[4]费关键,王西会,贾春雷,等.葡萄炭疽病的发生规律及综合防治技术[J]. 陕西农业科学, 2007, (03):176-177.
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[7]匡治州,许杨.核糖体rDNA-ITS序列在真菌学研究中的应用[J].生命的化学, 2004,24(2):120-122．
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[10]朱桂宁,蔡健和,胡春锦,等.广西山药炭疽病病原菌的鉴定与ITS序列分析[J].植物病理学报, 2007.37(6): 572-577.
[11]Abang, M. .M.,Winter,S.,Green, K. R.,et al. Molecular identification of Colletotrichum gloeosporioides causing yam anthracnose in Nigeria. Plant Pathology, 2002, 51(1): 63-71.
[12]Johnston, P. R., Jones, D.. Relationships among Colletotrichum isolates from fruit-rots assessed using rDNA sequences. Mycologia, 1997, 89:420-430.
[13]Sreenivasaprasad, S.. Phylogeny and systematics of 18 Colletotrichum species based on ribosomal DNA spacer sequences.Genome, 1996,39:499-512.
[14]方中达.植病研究法(第三版)[M].中国农业出版社.
[15]梁宏,张国珍,陈万权,等.小麦矮腥黑穗病菌与其近缘种的rDNA-ITS 序列分析[J].植物病理学报,2005,35(6)：181-183.
[16] WHITE T J, BRUNS T, LEE S．Analysis of phytogenetic relationships by amplification and direct sequencing of ribosomal RNA genes [M]//INNIS M A, GELFAND D H, SNINSKY J J, et al. PCR protocols：A guide to methods and applications.New York:Academic Press,1990.
[17]王辉,刘长远,赵奎华.辽宁地区辣椒疫病菌鉴定及同源性分析[J].沈阳农业大学学报,2011,42(4):433-437.
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[19]Sutton B C. The Coelomycetes Fungi Imperfecti with Pycnidia[J]. Acervuli and Stromata. CMI, Kew, Surrey,UK,1980,523-537.
[20]魏景超.真菌鉴定手册[M].上海科学技术出版社. 1978.
[21]雷百战,李国英.新疆葡萄炭疽病病原的鉴定及其生物学特性的研究[J].石河子大学学报(自然科学版), 2004,22(4):298-300.
[1]赵奎华.葡萄病虫害原色图鉴[M].北京:中国农业出版社, 2006.
[2]贺普超.葡萄学[M].中国农业出版社,1999.
[3]沈淑琳.葡萄病害及其防治[M].中国林业出版社,1992.
[4]费关键,王西会,贾春雷,等.葡萄炭疽病的发生规律及综合防治技术[J]. 陕西农业科学, 2007, (03):176-177.
[5]郑雪松,杨虹,李道棠,等.基因间隔序列(ITS)在细菌分类鉴定和种群分析中的应用[J].应用与环境生物学报, 2003, 9(6): 678-684.
[6]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.
[7]匡治州,许杨.核糖体rDNA-ITS序列在真菌学研究中的应用[J].生命的化学, 2004,24(2):120-122．
[8]马荣群,宋正旭,黄粤.葡萄炭疽菌和辣椒炭疽菌rDNA-ITS区序列比较[J].山东农业科学, 2009,3:41-42,46.
[9]杨腊英,黄华平,唐复润,等.香蕉炭疽菌rDNA-ITS区的分子鉴定与检测[J].植物病理学报, 2006,36(3):219-225.
[10]朱桂宁,蔡健和,胡春锦,等.广西山药炭疽病病原菌的鉴定与ITS序列分析[J].植物病理学报, 2007.37(6): 572-577.
[11]Abang, M. .M.,Winter,S.,Green, K. R.,et al. Molecular identification of Colletotrichum gloeosporioides causing yam anthracnose in Nigeria. Plant Pathology, 2002, 51(1): 63-71.
[12]Johnston, P. R., Jones, D.. Relationships among Colletotrichum isolates from fruit-rots assessed using rDNA sequences. Mycologia, 1997, 89:420-430.
[13]Sreenivasaprasad, S.. Phylogeny and systematics of 18 Colletotrichum species based on ribosomal DNA spacer sequences.Genome, 1996,39:499-512.
[14]方中达.植病研究法(第三版)[M].中国农业出版社.
[15]梁宏,张国珍,陈万权,等.小麦矮腥黑穗病菌与其近缘种的rDNA-ITS 序列分析[J].植物病理学报,2005,35（6）：181-183.
[16] WHITE T J, BRUNS T, LEE S．Analysis of phytogenetic relationships by amplification and direct sequencing of ribosomal RNA genes [M]//INNIS M A, GELFAND D H, SNINSKY J J, et al. PCR protocols：A guide to methods and applications.New York:Academic Press,1990.
[17]王辉,刘长远,赵奎华.辽宁地区辣椒疫病菌鉴定及同源性分析[J].沈阳农业大学学报,2011,42(4):433-437.
[18]陆家云.植物病原真菌学[M].北京:中国农业出版社,2001.
[19]刘锡进,郭英兰.中国真菌志[M].科学出版社. 1998.
[20]Sutton B C. The Coelomycetes Fungi Imperfecti with Pycnidia[J]. Acervuli and Stromata. CMI, Kew, Surrey,UK,1980,523-537.
[21]魏景超.真菌鉴定手册[M].上海科学技术出版社. 1978.
[22]雷百战,李国英.新疆葡萄炭疽病病原的鉴定及其生物学特性的研究[J].石河子大学学报(自然科学版), 2004,22(4):298-300.
[23]李利霞,赵奎华,刘长远,等.葡萄炭疽病菌SRAP遗传多样性分析[J].中国农学通报,2012,28(12):230-235.
[1]赵奎华.葡萄病虫害原色图鉴[M].北京:中国农业出版社, 2006.
[2]贺普超.葡萄学[M].中国农业出版社,1999.
[3]沈淑琳.葡萄病害及其防治[M].中国林业出版社,1992.
[4]费关键,王西会,贾春雷,等.葡萄炭疽病的发生规律及综合防治技术[J]. 陕西农业科学, 2007, (03):176-177.
[5]郑雪松,杨虹,李道棠,等.基因间隔序列(ITS)在细菌分类鉴定和种群分析中的应用[J].应用与环境生物学报, 2003, 9(6): 678-684.
[6]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.
[7]匡治州,许杨.核糖体rDNA-ITS序列在真菌学研究中的应用[J].生命的化学, 2004,24(2):120-122．
[8]马荣群,宋正旭,黄粤.葡萄炭疽菌和辣椒炭疽菌rDNA-ITS区序列比较[J].山东农业科学, 2009,3:41-42,46.
[9]杨腊英,黄华平,唐复润,等.香蕉炭疽菌rDNA-ITS区的分子鉴定与检测[J].植物病理学报, 2006,36(3):219-225.
[10]朱桂宁,蔡健和,胡春锦,等.广西山药炭疽病病原菌的鉴定与ITS序列分析[J].植物病理学报, 2007.37(6): 572-577.
[11]Abang, M. .M.,Winter,S.,Green, K. R.,et al. Molecular identification of Colletotrichum gloeosporioides causing yam anthracnose in Nigeria. Plant Pathology, 2002, 51(1): 63-71.
[12]Johnston, P. R., Jones, D.. Relationships among Colletotrichum isolates from fruit-rots assessed using rDNA sequences. Mycologia, 1997, 89:420-430.
[13]Sreenivasaprasad, S.. Phylogeny and systematics of 18 Colletotrichum species based on ribosomal DNA spacer sequences.Genome, 1996,39:499-512.
[14]方中达.植病研究法(第三版)[M].中国农业出版社.
[15]梁宏,张国珍,陈万权,等.小麦矮腥黑穗病菌与其近缘种的rDNA-ITS 序列分析[J].植物病理学报,2005,35（6）：181-183.
[16] WHITE T J, BRUNS T, LEE S．Analysis of phytogenetic relationships by amplification and direct sequencing of ribosomal RNA genes [M]//INNIS M A, GELFAND D H, SNINSKY J J, et al. PCR protocols：A guide to methods and applications.New York:Academic Press,1990.
[17]王辉,刘长远,赵奎华.辽宁地区辣椒疫病菌鉴定及同源性分析[J].沈阳农业大学学报,2011,42(4):433-437.
[18]陆家云.植物病原真菌学[M].北京:中国农业出版社,2001.
[19]刘锡进,郭英兰.中国真菌志[M].科学出版社. 1998.
[20]Sutton B C. The Coelomycetes Fungi Imperfecti with Pycnidia[J]. Acervuli and Stromata. CMI, Kew, Surrey,UK,1980,523-537.
[21]魏景超.真菌鉴定手册[M].上海科学技术出版社. 1978.
[22]雷百战,李国英.新疆葡萄炭疽病病原的鉴定及其生物学特性的研究[J].石河子大学学报(自然科学版), 2004,22(4):298-300.
[23]李利霞,赵奎华,刘长远,等.葡萄炭疽病菌SRAP遗传多样性分析[J].中国农学通报,2012,28(12):230-235.
[1]赵奎华.葡萄病虫害原色图鉴[M].北京:中国农业出版社, 2006.
[2]贺普超.葡萄学[M].中国农业出版社,1999.
[3]沈淑琳.葡萄病害及其防治[M].中国林业出版社,1992.
[4]费关键,王西会,贾春雷,等.葡萄炭疽病的发生规律及综合防治技术[J]. 陕西农业科学, 2007, (03):176-177.
[5]郑雪松,杨虹,李道棠,等.基因间隔序列(ITS)在细菌分类鉴定和种群分析中的应用[J].应用与环境生物学报, 2003, 9(6): 678-684.
[6]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.
[7]匡治州,许杨.核糖体rDNA-ITS序列在真菌学研究中的应用[J].生命的化学, 2004,24(2):120-122．
[8]马荣群,宋正旭,黄粤.葡萄炭疽菌和辣椒炭疽菌rDNA-ITS区序列比较[J].山东农业科学, 2009,3:41-42,46.
[9]杨腊英,黄华平,唐复润,等.香蕉炭疽菌rDNA-ITS区的分子鉴定与检测[J].植物病理学报, 2006,36(3):219-225.
[10]朱桂宁,蔡健和,胡春锦,等.广西山药炭疽病病原菌的鉴定与ITS序列分析[J].植物病理学报, 2007.37(6): 572-577.
[11]Abang, M. .M.,Winter,S.,Green, K. R.,et al. Molecular identification of Colletotrichum gloeosporioides causing yam anthracnose in Nigeria. Plant Pathology, 2002, 51(1): 63-71.
[12]Johnston, P. R., Jones, D.. Relationships among Colletotrichum isolates from fruit-rots assessed using rDNA sequences. Mycologia, 1997, 89:420-430.
[13]Sreenivasaprasad, S.. Phylogeny and systematics of 18 Colletotrichum species based on ribosomal DNA spacer sequences.Genome, 1996,39:499-512.
[14]方中达.植病研究法(第三版)[M].中国农业出版社.
[15]梁宏,张国珍,陈万权,等.小麦矮腥黑穗病菌与其近缘种的rDNA-ITS 序列分析[J].植物病理学报,2005,35（6）：181-183.
[16] WHITE T J, BRUNS T, LEE S．Analysis of phytogenetic relationships by amplification and direct sequencing of ribosomal RNA genes [M]//INNIS M A, GELFAND D H, SNINSKY J J, et al. PCR protocols：A guide to methods and applications.New York:Academic Press,1990.
[17]王辉,刘长远,赵奎华.辽宁地区辣椒疫病菌鉴定及同源性分析[J].沈阳农业大学学报,2011,42(4):433-437.
[18]陆家云.植物病原真菌学[M].北京:中国农业出版社,2001.
[19]刘锡进,郭英兰.中国真菌志[M].科学出版社. 1998.
[20]Sutton B C. The Coelomycetes Fungi Imperfecti with Pycnidia[J]. Acervuli and Stromata. CMI, Kew, Surrey,UK,1980,523-537.
[21]魏景超.真菌鉴定手册[M].上海科学技术出版社. 1978.
[22]雷百战,李国英.新疆葡萄炭疽病病原的鉴定及其生物学特性的研究[J].石河子大学学报(自然科学版), 2004,22(4):298-300.
[23]李利霞,赵奎华,刘长远,等.葡萄炭疽病菌SRAP遗传多样性分析[J].中国农学通报,2012,28(12):230-235.