四川省套作玉米茎腐病致病镰孢菌的分离与鉴定
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摘要
【目的】明确四川省套作玉米茎腐病镰孢菌的种类。【方法】2016—2017年采集仁寿、崇州、雅安、荣县4个县市套作玉米茎腐病样品进行分离与纯化,采用形态特征及基于r DNA-ITS和EF1-α序列分析鉴定镰孢菌种类,以幼苗刺伤接种法检测致病性。【结果】共获得103株纯培养物,其中71株为镰孢菌,包括层生镰孢菌、腐皮镰孢菌、轮枝镰孢菌、禾谷镰孢菌、尖孢镰孢菌、藤仓镰孢菌和木贼镰孢菌,分离频率依次为35.21%、16.9%、15.49%、11.27%、8.45%、7.04%和5.63%,且各地区镰孢菌种类及分离频率存在差异;致病性测定表明,禾谷镰孢菌致病性最强,层生镰孢菌和轮枝镰孢菌次之,尖孢镰孢菌和木贼镰孢菌致病性较弱,藤仓镰孢菌和腐皮镰孢菌致病性最弱。【结论】四川套作玉米茎腐病优势致病菌为层生镰孢菌、轮枝镰孢菌和禾谷镰孢菌。
【Objective】 This study aims to find out the population of Fusarium species causing maize stalk rot in maize soybean relay intercropping pattern in Sichuan Province.【Method】The samples were collected from the infected stem intercropping maizes in four cities including Renshou,Chongzhou,Ya'an,and Zigong in Sichuan province during 2016—2017. Fusarium species were obtained through tissue isolation and purification,and identified based on morphological characters and molecular identification based on PCR amplification of the gene fragements of rDNA ITS and translation elongation factor 1α. Pathogenicity test was performed on maize seedlings through wounding inoculation.【Result】We found that total 71 strains of Fusarium species were obtained from 103 isolates from diseased maize,and were identified into seven species including Fusarium proliferatum,F. solani,F. verticillioides,F.graminearum,F. oxysporum,F. fujikuroi and F. equiseti with the isolation frequencies for 35.21% 、16.9%、15.49%、11.27%、8.45%、7.04% and 5.63%. Pathogencity tests with representative strains of each Fusarium species showed that F. graminearum had the strongest pathogenicity on maize seedling,F.proliferatum,F. fujikuroi and F. verticillioides showed weaker pathogenicity,when F. oxysporum,F.solani and F. equiseti was the weakest pathogenicity.【Conclusion】This study indicates that maize stalk rot is predominantly caused by F. proliferatum,F. verticillioides,and F. graminearum in maize soybean relay intercropping in Sichuan province.
【Objective】 This study aims to find out the population of Fusarium species causing maize stalk rot in maize soybean relay intercropping pattern in Sichuan Province.【Method】The samples were collected from the infected stem intercropping maizes in four cities including Renshou,Chongzhou,Ya'an,and Zigong in Sichuan province during 2016—2017. Fusarium species were obtained through tissue isolation and purification,and identified based on morphological characters and molecular identification based on PCR amplification of the gene fragements of rDNA ITS and translation elongation factor 1α. Pathogenicity test was performed on maize seedlings through wounding inoculation.【Result】We found that total 71 strains of Fusarium species were obtained from 103 isolates from diseased maize,and were identified into seven species including Fusarium proliferatum,F. solani,F. verticillioides,F.graminearum,F. oxysporum,F. fujikuroi and F. equiseti with the isolation frequencies for 35.21% 、16.9%、15.49%、11.27%、8.45%、7.04% and 5.63%. Pathogencity tests with representative strains of each Fusarium species showed that F. graminearum had the strongest pathogenicity on maize seedling,F.proliferatum,F. fujikuroi and F. verticillioides showed weaker pathogenicity,when F. oxysporum,F.solani and F. equiseti was the weakest pathogenicity.【Conclusion】This study indicates that maize stalk rot is predominantly caused by F. proliferatum,F. verticillioides,and F. graminearum in maize soybean relay intercropping in Sichuan province.
引文
[1]YU C,SARAVANAKUMAR K,XIA H,et al.Occurence and virulence of Fusarium spp.associated with stalk rot of maize in North-East China[J].Physiological and Molecular Plant Pathology,2017,98:1-8.
[2]BORAH S N,GOSWAMIN D,SARMA H K,et al.Rhamnolipid Biosurfactant against Fusarium verticillioides to control stalk and ear rot disease of maiz[eJ].Frontiers in Microbiology,2016,7:1505.
[3]陈捷.我国玉米穗茎腐病病害研究现状与展望[J].沈阳农业大学报,2000,31(5):393-401.
[4]GAI X T,XUAN Y H,GAO Z G,et al.Diversity and pathogenicity of Fusarium graminearum species complex from maize stalk and ear rot strains in northeast China[J].Plant Pathology,2017,66(8):1267-1275.
[5]FUMERO M V,REYNOSO M M,CHULZE S.Fusarium temperatum and Fusarium subglutinans isolated from maize in Argent-ina[J].International Journal of Food Microbiology,2015,199:86.
[6]VARELA C P,CASAL O A,PADIN M C,et al.First report of Fusarium temperatum causing seedling blight and stalk rot on maize in Spain[J].Plant Disease,2017,97(9):1252.
[7]WANG J H,ZHANG J B,LI H P,et al.Molecular identification,mycotoxin production and comparative pathogenicity of Fusarium temperatum isolated from maize in China[J].Journal of Phytopathology,2014,162(3):147-157.
[8]BRUNS T,PROCTOR R,MUNKVOLD G.The role of mycotoxins produced by Fusarium verticillioides and Fusarium graminearum in maize seedling infection[J].Phytopathology,2011,101(6):22.
[9]ZHANG L,WANG J,ZHANG C,et al.Analysis of potential fumonisin-producing Fusarium species in corn products from three main maize-producing areas in eastern China[J].Journal of the Science of Food and Agriculture,2013,93(3):693-701.
[10]KUHNEM P R,WARD T J,SILVA C N,et al.Composition and toxigenic potential of the Fusarium graminearum species complex from maize ears,stalks and stubble in Brazi[lJ].Plant Pathology,2016,65(7):1185-1191.
[11]ZHANG H,BRANKOVICS B,LUO W,et al.Crops are a main driver for species diversity and the toxigenic potential of Fusarium isolates in maize ears in China[J].World Mycotoxin Journal,2016,9(5):1-16.
[12]杨文钰,雍太文,任万军,等.发展套作大豆,振兴大豆产业[J].大豆科学,2008,27(1):1-7.
[13]高仁才,杨文钰,杨峰,等.行距配置对套作大豆冠层光环境及其形态特征和产量的影响[J].大豆科学,2015,34(4):611-615.
[14]YANG F,WANG X C,LIAO D P,et al.Yield response to different planting geometries in maize soybean relay strip intercropping systems[J].Agronomy Journal,2015,107(1):296-304.
[15]李春霞,苏俊,龚士堔,等.玉米茎腐病接种方法的研究[J].玉米科学,2001,9(2):72-74.
[16]LESLIE J F,SUMMERELL B A.The Fusarium laboratory ma-nua[lM].Oxford:Blackwell Publish Ltd,2006:22-30.
[17]何宛芹,付瑶,鲁雯璐,等.大豆根腐病致病镰孢菌的多重PCR检测技术[J].植物保护学报,2017,44(4):609-616.
[18]WHITE T J,BRUNS T D,LEE S B,et al.Analysis of phylogenetic relationships by amplification and direct seaquencing of ribosomal RNA genes[J].Pcr Protocols A Guide to Methods&Applications,1990(2):15-22.
[19]CARBONE I,KOHN L M.A method for designing primer sets for speciation studies in filamentous ascomycetes[J].Mycologia,1999,91,3:553-556.
[20]戴浩.四川省套作大豆根腐病镰孢菌分离鉴定及其侵染引起的大豆响应机制研究[D].成都:四川农业大学,2017.
[21]晋齐鸣,宋淑云,张伟,等.不同耕作方式玉米田病害发生情况调查[J].东北农业科学,2006,31(3):55-56.
[22]魏荐郦.四川省小麦赤霉病与玉米茎腐病相互关系的研究[D].成都:四川农业大学,2015.
[23]袁虹霞,闵营辉,张丹丹,等.河南省玉米茎腐病病原菌分离与致病性测定[J].玉米科学,2011,19(6):122-124.
[24]何婧.玉米茎腐病病原菌分离鉴定与镰孢菌群体遗传多样性研究[D].乌鲁木齐:新疆农业大学,2010.
[25]ZHANG H,LEE T V D,WAALWIJK C,et al.Population analysis of the Fusarium graminearum species complex from wheat in china show a shift to more aggressive isolates[J].PLoS One,2012,7(2):e31722.
[26]LEE J,KIM H,JEON J J,et al.Population structure of and mycotoxin production by Fusarium graminearum from maize in South Korea[J].Applied and Environmental Microbiology,2012,78(7):2161-2167.
[27]MALIHIPOUR A,GILBERT J,PIERCEY-NORMORE M,et al.Molecular phylogenetic analysis,trichothecene chemotype patterns,and variation in aggressiveness of Fusarium isolates causing head blight in wheat[J].Plant Disease,2012,96(7):1016-1025.
[28]KENDRA D.Asymptomatic colonization of soybean(Glycine Max)by Fusarium verticillioides[J].Optics&Precision Engineering,2013,21(8):2008-2015.
[2]BORAH S N,GOSWAMIN D,SARMA H K,et al.Rhamnolipid Biosurfactant against Fusarium verticillioides to control stalk and ear rot disease of maiz[eJ].Frontiers in Microbiology,2016,7:1505.
[3]陈捷.我国玉米穗茎腐病病害研究现状与展望[J].沈阳农业大学报,2000,31(5):393-401.
[4]GAI X T,XUAN Y H,GAO Z G,et al.Diversity and pathogenicity of Fusarium graminearum species complex from maize stalk and ear rot strains in northeast China[J].Plant Pathology,2017,66(8):1267-1275.
[5]FUMERO M V,REYNOSO M M,CHULZE S.Fusarium temperatum and Fusarium subglutinans isolated from maize in Argent-ina[J].International Journal of Food Microbiology,2015,199:86.
[6]VARELA C P,CASAL O A,PADIN M C,et al.First report of Fusarium temperatum causing seedling blight and stalk rot on maize in Spain[J].Plant Disease,2017,97(9):1252.
[7]WANG J H,ZHANG J B,LI H P,et al.Molecular identification,mycotoxin production and comparative pathogenicity of Fusarium temperatum isolated from maize in China[J].Journal of Phytopathology,2014,162(3):147-157.
[8]BRUNS T,PROCTOR R,MUNKVOLD G.The role of mycotoxins produced by Fusarium verticillioides and Fusarium graminearum in maize seedling infection[J].Phytopathology,2011,101(6):22.
[9]ZHANG L,WANG J,ZHANG C,et al.Analysis of potential fumonisin-producing Fusarium species in corn products from three main maize-producing areas in eastern China[J].Journal of the Science of Food and Agriculture,2013,93(3):693-701.
[10]KUHNEM P R,WARD T J,SILVA C N,et al.Composition and toxigenic potential of the Fusarium graminearum species complex from maize ears,stalks and stubble in Brazi[lJ].Plant Pathology,2016,65(7):1185-1191.
[11]ZHANG H,BRANKOVICS B,LUO W,et al.Crops are a main driver for species diversity and the toxigenic potential of Fusarium isolates in maize ears in China[J].World Mycotoxin Journal,2016,9(5):1-16.
[12]杨文钰,雍太文,任万军,等.发展套作大豆,振兴大豆产业[J].大豆科学,2008,27(1):1-7.
[13]高仁才,杨文钰,杨峰,等.行距配置对套作大豆冠层光环境及其形态特征和产量的影响[J].大豆科学,2015,34(4):611-615.
[14]YANG F,WANG X C,LIAO D P,et al.Yield response to different planting geometries in maize soybean relay strip intercropping systems[J].Agronomy Journal,2015,107(1):296-304.
[15]李春霞,苏俊,龚士堔,等.玉米茎腐病接种方法的研究[J].玉米科学,2001,9(2):72-74.
[16]LESLIE J F,SUMMERELL B A.The Fusarium laboratory ma-nua[lM].Oxford:Blackwell Publish Ltd,2006:22-30.
[17]何宛芹,付瑶,鲁雯璐,等.大豆根腐病致病镰孢菌的多重PCR检测技术[J].植物保护学报,2017,44(4):609-616.
[18]WHITE T J,BRUNS T D,LEE S B,et al.Analysis of phylogenetic relationships by amplification and direct seaquencing of ribosomal RNA genes[J].Pcr Protocols A Guide to Methods&Applications,1990(2):15-22.
[19]CARBONE I,KOHN L M.A method for designing primer sets for speciation studies in filamentous ascomycetes[J].Mycologia,1999,91,3:553-556.
[20]戴浩.四川省套作大豆根腐病镰孢菌分离鉴定及其侵染引起的大豆响应机制研究[D].成都:四川农业大学,2017.
[21]晋齐鸣,宋淑云,张伟,等.不同耕作方式玉米田病害发生情况调查[J].东北农业科学,2006,31(3):55-56.
[22]魏荐郦.四川省小麦赤霉病与玉米茎腐病相互关系的研究[D].成都:四川农业大学,2015.
[23]袁虹霞,闵营辉,张丹丹,等.河南省玉米茎腐病病原菌分离与致病性测定[J].玉米科学,2011,19(6):122-124.
[24]何婧.玉米茎腐病病原菌分离鉴定与镰孢菌群体遗传多样性研究[D].乌鲁木齐:新疆农业大学,2010.
[25]ZHANG H,LEE T V D,WAALWIJK C,et al.Population analysis of the Fusarium graminearum species complex from wheat in china show a shift to more aggressive isolates[J].PLoS One,2012,7(2):e31722.
[26]LEE J,KIM H,JEON J J,et al.Population structure of and mycotoxin production by Fusarium graminearum from maize in South Korea[J].Applied and Environmental Microbiology,2012,78(7):2161-2167.
[27]MALIHIPOUR A,GILBERT J,PIERCEY-NORMORE M,et al.Molecular phylogenetic analysis,trichothecene chemotype patterns,and variation in aggressiveness of Fusarium isolates causing head blight in wheat[J].Plant Disease,2012,96(7):1016-1025.
[28]KENDRA D.Asymptomatic colonization of soybean(Glycine Max)by Fusarium verticillioides[J].Optics&Precision Engineering,2013,21(8):2008-2015.