湖北省蔬菜根际土壤中的镰孢菌分布特征
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摘要
为明确湖北省蔬菜产区镰孢菌(Fusarium spp.)的种类及分布特征,本研究从湖北省13个地级市蔬菜产区采集根际土壤样品55份,对土样中的镰孢菌进行分离、纯化和鉴定,获得271株镰孢菌;通过镰孢菌特异性引物扩增、测序等分子生物学技术,鉴定出尖孢镰孢菌(F. oxysporum)118株,占分离菌株43.6%;木贼孢菌(F. equiseti)46株,占17.0%;共享镰孢菌(F.commune)45株,占16.6%;茄病镰孢菌(F. solani)31株,占11.4%;藤仓镰孢菌(F. fujikuroi)11株,占4.1%;镰孢霉菌(F.delphinoides)6株,占2.2%;轮纹镰孢菌(F. concentricum)5株,占1.8%;三线镰孢菌(F. tricinctum)3株,占1.1%;镰孢菌属(Fusarium spp.)6株,占2.2%。通过与致病菌尖孢镰孢菌番茄专化型(F. oxysporum f. sp. lycopersici)菌株进行对峙培养,筛选出拮抗镰孢菌16株,包括尖孢镰孢菌(F. oxysporum)4株,茄病镰孢菌(F. solani)4株,轮纹镰孢菌(F. concentricum)3株,三线镰孢菌(F.tricinctum)3株,木贼孢菌(F. equiseti)和藤仓镰孢菌(F. fujikuroi)各1株。本研究明确了湖北省蔬菜产区的镰孢菌属真菌的种类和多样性特征,为蔬菜镰孢菌病害的防治提供科学依据。
To clarify the species and distribution of Fusarium species in vegetable growing areas in Hubei province,55 rhizosphere soil samples were collected from 13 cities of Hubei province,and Fusarium species strains from the samples were isolated,purified and identified.Total 271 Fusarium strains were obtained. Among these Fusarium strains,according to molecular biological techniques such as species-specific primers and sequencing,118 strains were identified as Fusarium oxysporum accounting for 43.6% of the total,46 strains as F. equiseti for17.0% of the total,45 strains as F. commune for 16.6% of the total,31 strains as F. solani for 11.4% of the total,11 strains as F. fujikuroi for 4.1% of the total,6 strains as F. delphinoides for 2.2% of the total,5 strains as F. concentricum for 1.8%,3 strains as F. tricinctum for1.1%,and 6 strains were Fusarium spp. for 2.2% of the total. Based on the confrontation culture with pathogenic F. oxysporum f. sp. lycopersici strain,16 strains of antagonistic Fusarium species were identified,including 4 strains of F. oxysporum,4 strains of F. solani,3 strains of F.concentricum,and 3 strains of F. tricinctum,while each of F. equiseti and F. fujikuroi was only one,respectively. In this study,the species and diversity of Fusarium species in vegetable producing areas of Hubei province were confirmed,which provides the theoretical basis for the accurate pathogen identification and scientific control of Fusarium disease.
To clarify the species and distribution of Fusarium species in vegetable growing areas in Hubei province,55 rhizosphere soil samples were collected from 13 cities of Hubei province,and Fusarium species strains from the samples were isolated,purified and identified.Total 271 Fusarium strains were obtained. Among these Fusarium strains,according to molecular biological techniques such as species-specific primers and sequencing,118 strains were identified as Fusarium oxysporum accounting for 43.6% of the total,46 strains as F. equiseti for17.0% of the total,45 strains as F. commune for 16.6% of the total,31 strains as F. solani for 11.4% of the total,11 strains as F. fujikuroi for 4.1% of the total,6 strains as F. delphinoides for 2.2% of the total,5 strains as F. concentricum for 1.8%,3 strains as F. tricinctum for1.1%,and 6 strains were Fusarium spp. for 2.2% of the total. Based on the confrontation culture with pathogenic F. oxysporum f. sp. lycopersici strain,16 strains of antagonistic Fusarium species were identified,including 4 strains of F. oxysporum,4 strains of F. solani,3 strains of F.concentricum,and 3 strains of F. tricinctum,while each of F. equiseti and F. fujikuroi was only one,respectively. In this study,the species and diversity of Fusarium species in vegetable producing areas of Hubei province were confirmed,which provides the theoretical basis for the accurate pathogen identification and scientific control of Fusarium disease.
引文
[1]Ploetz RC. Management of Fusarium wilt of banana:A review with special reference to tropical race 4[J]. Crop Protection, 2015,73:7-15.
[2]符美英,肖彤斌,吴凤芝,等.南方根结线虫、香蕉穿孔线虫和尖孢镰孢菌4号生理小种对巴西蕉致病力的测定[J].广东农业科学, 2014, 41(1):56-60.
[3]李斌,郭红卫.镰刀菌毒素DON、NIV的细胞毒性和致突变、致畸、致癌研究进展[J].癌变.畸变.突变, 1999(4):48-49.
[4]Thangavelu R, Kumar KM, Devi PG, et al. Genetic diversity of Fusarium oxysporum f. sp. cubense isolates(Foc)of India by Inter Simple Sequence Repeats(ISSR)analysis[J]. Molecular Biotechnology, 2012, 8:32-38.
[5]Ogawa K. Studies on Fusarium wilt of sweet potato(Ipomoea baralas L.)[J]. Bullelin of the National Agricultural Research Cenfer, 1988, 10:123-127.
[6]Larkin RP, Fravel DR. Mechanisms of action and dose response relationships governing biological control of Fusarium wilt of tomato by nonpathogenic Fusarium spp.[J]. Phytopathology, 1999, 89:1152-1161.
[7]张向民.镰孢菌属分类学研究历史与现状[J].菌物研究,2005, 3(2):59-62.
[8]洪坚平.山西镰刀菌[M].北京:中国林业出版社, 2015.
[9]贺字典,高增贵,张志雯,等.河北蔬菜保护地土壤镰孢菌种群分布研究[J].北方园艺, 2009(10):21-25.
[10]方中达.植病研究方法[M].北京:中国农业出版社, 1998.
[11]Murry M, Thompson W. Rapid isolation of high molecular weight plant DNA[J]. Nucleic Acids Research, 1980, 8(19):4321-4326.
[12]Donnell OK, Kistler HC, Cigelnik E, et al. Panama disease of banana:Concordant multiple evoltionary origins of the fungus causing evidence from nuclear and mitochondrial gene genealogies[J]. Proceedings of the National Academy of Science of the United States of America, 1998, 95(5):2044-2049.
[13]俞大绂,方中达.云南省蚕豆枯萎病和茎腐病的研究[J].科学通报, 1950, 2(1):73-77.
[14]李明远,张涛涛,李兴红,等.十字花科蔬菜枯萎病及其病原鉴定[J].植物保护, 2003, 29(3):44-45.
[15]张汝祥.安徽省大豆镰孢根腐病菌鉴定及生物学特性研究[D].合肥:安徽农业大学, 2012.
[16]蓝江林,苏明星,葛慈斌,等.尖孢镰刀菌的分布与西瓜枯萎病病程的相关性[J].中国农学通报, 2007, 23(12):302-305.
[17]费诺亚.蓝莓根腐病病原菌鉴定及生物学特性研究[D].沈阳:沈阳农业大学, 2018.
[18]吴安忠,程崖芝,巫升鑫,等.烟草镰刀菌根腐病的病原鉴定[J].中国烟草学报, 2018, 24(2):135-140.
[19]曾莉莎,赵志慧,吕顺,等.香蕉上的镰孢菌种类及其系统发育关系[J].菌物学报, 2013, 32(4):617-632.
[20]曾莉莎,郑芝波,吕顺,等.荷花腐败病病原菌的形态学与多基因分子系统学鉴定[J].植物病理学报, 2017, 47(2):162-173.
[21]姜伊.大豆根腐病主要致病菌种类及防控技术[D].哈尔滨:黑龙江农业大学, 2014.
[22]李丹.甘肃省中西部地区镰孢菌的分类研究[D].兰州:甘肃农业大学, 2008.
[23]吴品珊,严进, Rupe JC,等.大豆猝死综合症病菌检测技术研究[J].植物病理学报, 2005, 35(6):28-34.
[24]张丽.黑龙江省大豆尖镰孢抗源筛选、致病机理及遗传多样性分析[D].哈尔滨:东北农业大学, 2014.
[25]王健生.大豆尖镰孢菌的快速分子检测与遗传多样性分析及品种抗性研究[D].南京:南京农业大学, 2012.
[26]张连洪,燕继晔,赵文生,等.稻瘟菌无毒基因AVR-Pikm的定位[J].植物病理学报, 2006, 36(2):116-122.
[2]符美英,肖彤斌,吴凤芝,等.南方根结线虫、香蕉穿孔线虫和尖孢镰孢菌4号生理小种对巴西蕉致病力的测定[J].广东农业科学, 2014, 41(1):56-60.
[3]李斌,郭红卫.镰刀菌毒素DON、NIV的细胞毒性和致突变、致畸、致癌研究进展[J].癌变.畸变.突变, 1999(4):48-49.
[4]Thangavelu R, Kumar KM, Devi PG, et al. Genetic diversity of Fusarium oxysporum f. sp. cubense isolates(Foc)of India by Inter Simple Sequence Repeats(ISSR)analysis[J]. Molecular Biotechnology, 2012, 8:32-38.
[5]Ogawa K. Studies on Fusarium wilt of sweet potato(Ipomoea baralas L.)[J]. Bullelin of the National Agricultural Research Cenfer, 1988, 10:123-127.
[6]Larkin RP, Fravel DR. Mechanisms of action and dose response relationships governing biological control of Fusarium wilt of tomato by nonpathogenic Fusarium spp.[J]. Phytopathology, 1999, 89:1152-1161.
[7]张向民.镰孢菌属分类学研究历史与现状[J].菌物研究,2005, 3(2):59-62.
[8]洪坚平.山西镰刀菌[M].北京:中国林业出版社, 2015.
[9]贺字典,高增贵,张志雯,等.河北蔬菜保护地土壤镰孢菌种群分布研究[J].北方园艺, 2009(10):21-25.
[10]方中达.植病研究方法[M].北京:中国农业出版社, 1998.
[11]Murry M, Thompson W. Rapid isolation of high molecular weight plant DNA[J]. Nucleic Acids Research, 1980, 8(19):4321-4326.
[12]Donnell OK, Kistler HC, Cigelnik E, et al. Panama disease of banana:Concordant multiple evoltionary origins of the fungus causing evidence from nuclear and mitochondrial gene genealogies[J]. Proceedings of the National Academy of Science of the United States of America, 1998, 95(5):2044-2049.
[13]俞大绂,方中达.云南省蚕豆枯萎病和茎腐病的研究[J].科学通报, 1950, 2(1):73-77.
[14]李明远,张涛涛,李兴红,等.十字花科蔬菜枯萎病及其病原鉴定[J].植物保护, 2003, 29(3):44-45.
[15]张汝祥.安徽省大豆镰孢根腐病菌鉴定及生物学特性研究[D].合肥:安徽农业大学, 2012.
[16]蓝江林,苏明星,葛慈斌,等.尖孢镰刀菌的分布与西瓜枯萎病病程的相关性[J].中国农学通报, 2007, 23(12):302-305.
[17]费诺亚.蓝莓根腐病病原菌鉴定及生物学特性研究[D].沈阳:沈阳农业大学, 2018.
[18]吴安忠,程崖芝,巫升鑫,等.烟草镰刀菌根腐病的病原鉴定[J].中国烟草学报, 2018, 24(2):135-140.
[19]曾莉莎,赵志慧,吕顺,等.香蕉上的镰孢菌种类及其系统发育关系[J].菌物学报, 2013, 32(4):617-632.
[20]曾莉莎,郑芝波,吕顺,等.荷花腐败病病原菌的形态学与多基因分子系统学鉴定[J].植物病理学报, 2017, 47(2):162-173.
[21]姜伊.大豆根腐病主要致病菌种类及防控技术[D].哈尔滨:黑龙江农业大学, 2014.
[22]李丹.甘肃省中西部地区镰孢菌的分类研究[D].兰州:甘肃农业大学, 2008.
[23]吴品珊,严进, Rupe JC,等.大豆猝死综合症病菌检测技术研究[J].植物病理学报, 2005, 35(6):28-34.
[24]张丽.黑龙江省大豆尖镰孢抗源筛选、致病机理及遗传多样性分析[D].哈尔滨:东北农业大学, 2014.
[25]王健生.大豆尖镰孢菌的快速分子检测与遗传多样性分析及品种抗性研究[D].南京:南京农业大学, 2012.
[26]张连洪,燕继晔,赵文生,等.稻瘟菌无毒基因AVR-Pikm的定位[J].植物病理学报, 2006, 36(2):116-122.