广西崇左市甘蔗梢腐病病原菌的分离鉴定及生物学特性测定
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摘要
【目的】明确广西崇左市甘蔗梢腐病病原菌种类及其生物学特性,为甘蔗梢腐病的综合防治提供理论依据。【方法】以采集自广西崇左市甘蔗种植区的甘蔗梢腐病病叶为材料,采用组织分离法进行病原菌单孢分离,结合科赫法则进行致病性测定,通过形态学特征观察及rDNA-ITS序列分析对病原菌进行种类鉴定;应用十字交叉法进行病原菌的生物学特性测定。【结果】分离获得的病原菌rDNA-ITS序列长度为496 bp,与轮枝镰刀菌(Fusarium verticillioides)的同源性达99%,与轮枝镰刀菌(登录号EU047)的遗传关系最近。病原菌的生物学特性测定结果表明,该菌菌丝最适生长温度为28℃,致死温度为70℃;菌丝生长最适pH为8,光暗交替条件下生长较快;菌丝生长最适氮源为胰蛋白胨,最适碳源为甘露醇。【结论】引起广西崇左市甘蔗梢腐病的病原菌是轮枝镰刀菌,该菌适宜在高温高湿、弱碱性、光暗交替及富含有机营养的环境条件下生长。
【Objective】The present experiment was conducted to investigate species and biological characteristics of the pathogen causing sugarcane pokkah boeng in Chongzuo,Guangxi in order to provide scientific basis for controlling sugarcane pokkah boeng.【Method】Sugarcane leaves with sugarcane pokkah boeng in Chongzuo sugarcane planting areas were collected. Conventional tissue separation was applied to isolate monospore from the pathogen,and the pathogenicity of the pathogen were investigated. Based on morphological characteristics observation and analysis of rDNA-ITS sequence,the pathogen of sugarcane pokkah boeng was identified. Biological characteristics of the pathogen were determined with crossing methods.【Result】The rDNA-ITS sequence of tested strains was 496 bp in length,which showed 99%homology with Fusarium verticillioides. It had the closest genetic relationship with Fusarium verticillioides(accession number EU047). According to the biological characteristics test,the pathogen could grow with an optimal condition at28 ℃,and the lethal temperature was 70 ℃. The best cultural condition for mycelial growth was pH 8,12 h dark+12 h light. The best nitrogen sources for the pathogen growth was tryptone,and the best carbon sources was mannito.【Conclusion】The pathogen causing sugarcane pokkah boeng in Chongzuo is identified as F. verticillioides. The optimal growth conditions for F. verticillioides is high temperature,high humidity,weak alkaline,alternating light and dark and full of organic nutrition.
【Objective】The present experiment was conducted to investigate species and biological characteristics of the pathogen causing sugarcane pokkah boeng in Chongzuo,Guangxi in order to provide scientific basis for controlling sugarcane pokkah boeng.【Method】Sugarcane leaves with sugarcane pokkah boeng in Chongzuo sugarcane planting areas were collected. Conventional tissue separation was applied to isolate monospore from the pathogen,and the pathogenicity of the pathogen were investigated. Based on morphological characteristics observation and analysis of rDNA-ITS sequence,the pathogen of sugarcane pokkah boeng was identified. Biological characteristics of the pathogen were determined with crossing methods.【Result】The rDNA-ITS sequence of tested strains was 496 bp in length,which showed 99%homology with Fusarium verticillioides. It had the closest genetic relationship with Fusarium verticillioides(accession number EU047). According to the biological characteristics test,the pathogen could grow with an optimal condition at28 ℃,and the lethal temperature was 70 ℃. The best cultural condition for mycelial growth was pH 8,12 h dark+12 h light. The best nitrogen sources for the pathogen growth was tryptone,and the best carbon sources was mannito.【Conclusion】The pathogen causing sugarcane pokkah boeng in Chongzuo is identified as F. verticillioides. The optimal growth conditions for F. verticillioides is high temperature,high humidity,weak alkaline,alternating light and dark and full of organic nutrition.
引文
陈宏州,杨红福,姚克兵,束兆林,周华飞,庄义庆.2018.水稻恶苗病病原菌鉴定及室内药剂毒力测定[J].植物保护学报,45(6):1356-1366.[Chen H Z,Yang H F,Yao K B,Shu Z L,Zhou H F,Zhuang Y Q.2018.Identification of rice bakanae pathogens and indoor toxicity test of fungicides[J].Journal of Plant Protection,45(6):1356-1366.]
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高婧.2016.向日葵枯萎病菌的分离鉴定、致病力分化以及侵染过程的研究[D].呼和浩特:内蒙古农业大学.[Gao J.2016.The study on the pathogen isolation and identification,pathogenicity differentiation and infection process of sunflower wilt[D].Hohhot:Inner Mongolia Agricultural University.]
何姗珊,方位宽,经艳,谭芳,韦永定,韦士评,梁朝旭,李鸣.2016.甘蔗梢腐病原菌分离纯化与ITS序列鉴定[J].广西糖业,(4):10-14.[He S S,Fang W K,Jing Y,Tan F,Wei Y D,Wei S P,Liang C X,Li M.2016.Isolation of sugarcane pokkah boeng pathogen and ITS sequence identification[J].Guangxi Sugar Industry,(4):10-14.]
胡美姣,杨波,李敏,杨冬平,蒲金基,张正科,高兆银.2013.海南芒果果腐病病原菌鉴定及其生物学特性研究[J].热带作物学报,34(8):1564-1569.[Hu M J,Yang B,Li M,Yang D P,Pu J J,Zhang Z K,Gao Z Y.2013.Identification and biological characteristics of mango fruit rot pathogen[J].Chinese Journal of Tropical Crops,34(8):1564-1569.]
李杨瑞,杨丽涛.2009.20世纪90年代以来我国甘蔗产业和科技的新发展[J].西南农业学报,22(5):1469-1476.[Li Y R,Yang L T.2009.New developments in sugarcane industry and technologies in China since 1990s[J].Southwest China Journal of Agricultural Sciences,22(5):1469-1476.]
刘丹.2016.甘肃省设施辣椒镰刀菌根腐病病原鉴定及抗病种质资源筛选[D].兰州:甘肃农业大学.[Liu D.2016.Studies on pathogens and resistance to fusarium root rot on greenhouse pepper in Gansu,China[D].Lanzhou:Gansu Agricultural University.]
刘丽萍,方鹏鹏.2017.马铃薯干腐病镰刀菌的分离及生物学特性研究[J].陇东学院学报,28(5):62-66.[Liu L P,Fang P P.2017.The isolation and biological characteristic of Fusarium associated with potato dry rot[J].Journal of Longdong University,28(5):62-66.]
卢文浩.2007.甘蔗梢腐病的发生及综合防治措施[J].作物杂志,(2):92.[Lu W H.2007.The damage of sugarcanes pokkah boeng and the main control strategies[J].Crops,(2):92.]
曲田丽,辛惠普.2003.绿豆根腐病菌生物学特性研究[J].黑龙江八一农垦大学学报,15(4):109-114.[Qu T L,Xin H P.2003.The biological characteristics of the root rot pathogens in mung[J].Journal of Heilongjiang Bayi Agricultural University,15(4):109-114.]
单红丽,李文凤,张荣跃,王晓燕,李婕,仓晓燕,尹炯,罗志明,黄应昆.2018.甘蔗梢腐病暴发流行原因及产量糖分损失测定[J].中国糖料,40(3):40-42.[Shan H L,Li WF,Zhang R Y,Wang X Y,Li J,Cang X Y,Yin J,Luo ZM,Huang Y K.2018.Analysis on epidemic reason of sugarcane pokahh boeng and its losses on yield and sucrose content[J].Sugar Crops of China,40(3):40-42.]
孙嘉曼,张进忠,韦弟,蓝霞,郭文峰,卢江.2016.一种新的香蕉黑斑病病原菌鉴定及生物学特性研究[J].植物病理学报,46(1):119-123.[Sun J M,Zhang J Z,Wei D,Lan X,Guo W F,Lu J.2016.Identification and biological characterization of the pathogen causing a new black spot disease on Musa spp.[J].Acta Phytophologica Sinica,46(1):119-123.]
王芝涵,王春伟,高海馨,荆琦,余廷濠,王燕.2019.引起玉米穗腐病的禾谷镰刀菌LAMP快速检测方法的建立[J].江苏农业学报,35(3):581-585.[Wang Z H,Wang C W,Gao H X,Jing Q,Yu T H,Wang Y.2019.Development of loop-mediated isothermal amplification(LAMP)assay for the detection of the Fusarium graminearum causing ear rot of maize[J].Jiangsu Journal of Agricultural Sciences,35(3):581-585.]
张斌,杨晓云,陈志谊.2016.番茄枯萎病致病镰刀菌种类鉴定及优势种群的研究[J].植物病理学报,46(4):561-565.[Zhang B,Yang X Y,Chen Z Y.2016.Identification of the pathogenic and dominant Fusarium species causing tomato Fusarium wilt[J].Acta Phytophologica Sinica,46(4):561-565.]
张玉娟.2009.甘蔗梢腐病病原分子检测及甘蔗组合、品种的抗病性评价[D].福州:福建农林大学.[Zhang Y J.2009.Molecular detection and resistance evaluation for pokkah boeng disease of sugarcane[D].Fuzhou:Fujian Agriculture and Forestry University.]
朱迎迎,高兆银,李敏,陈亮,胡美姣.2016.火龙果镰刀菌果腐病病原菌鉴定及生物学特性研究[J].热带作物学报,37(1):164-171.[Zhu Y Y,Gao Z Y,Li M,Chen L,Hu M J.2016.Identification and biological characteristics of dragon fruit(Hylocereus undatus Britt)fusarium rot pathogen[J].Chinese Journal of Tropical Crops,37(1):164-171.]
Bao Y X,Sun H J,Li Y F,Duan Z Z.2016.First report of Fusarium oxysporum isolate gx3 causing sugarcane pokkah boeng in Guangxi of China[J].Plant Disease,100(8):1785.
Khani K T,Alizadeh A,Nejad R F 2013.Pathogenicity of Fusarium proliferatum,a new causal agent of pokkah boeng in sugarcane[J].Proceedings of the International Society Sugracane Technology,28:1-5.
Lin Z,Wei J J,Xu S Q,Guo Q,Wang X,Qiu Y X,Deng ZH,Huang J H,Charles A P,Chen B S,Chen R K,Zhang M Q.2014.Species-specific detection and identification of Fusarium species complex,the causal agent of sugarcane pokkah boeng in China[J].PLoS One,9(8):e104195.
Mohammadi A,Nejad R F,Mofrad N N.2012.Fusarium verticillioides from sugarcane,vegetative compatibility groups and pathogenicity[J].Plant Protection Science,48(3):80-84.
Siti N.2007.Pathogenicity and aethiology of Fusarium species associated with pokkah boeng disease on sugarcane[D].Penang:University Sains Malaysia.
番华彩,郭志祥,白亭亭,徐胜涛,杨佩文,尹可锁,郑泗军,潘艳华,李迅东,曾莉.2018.香蕉棒孢霉叶斑病病原菌生物学特性测定[J].南方农业学报,49(3):481-487.[Fan H C,Guo Z X,Bai T T,Xu S T,Yang P W,Yin K S,Zheng S J,Pan Y H,Li X D,Zeng L.2018.Biological characteristics of pathogenic bacteria causing banana leaf spot disease[J].Journal of Southern Agriculture,49(3):481-487.]
高婧.2016.向日葵枯萎病菌的分离鉴定、致病力分化以及侵染过程的研究[D].呼和浩特:内蒙古农业大学.[Gao J.2016.The study on the pathogen isolation and identification,pathogenicity differentiation and infection process of sunflower wilt[D].Hohhot:Inner Mongolia Agricultural University.]
何姗珊,方位宽,经艳,谭芳,韦永定,韦士评,梁朝旭,李鸣.2016.甘蔗梢腐病原菌分离纯化与ITS序列鉴定[J].广西糖业,(4):10-14.[He S S,Fang W K,Jing Y,Tan F,Wei Y D,Wei S P,Liang C X,Li M.2016.Isolation of sugarcane pokkah boeng pathogen and ITS sequence identification[J].Guangxi Sugar Industry,(4):10-14.]
胡美姣,杨波,李敏,杨冬平,蒲金基,张正科,高兆银.2013.海南芒果果腐病病原菌鉴定及其生物学特性研究[J].热带作物学报,34(8):1564-1569.[Hu M J,Yang B,Li M,Yang D P,Pu J J,Zhang Z K,Gao Z Y.2013.Identification and biological characteristics of mango fruit rot pathogen[J].Chinese Journal of Tropical Crops,34(8):1564-1569.]
李杨瑞,杨丽涛.2009.20世纪90年代以来我国甘蔗产业和科技的新发展[J].西南农业学报,22(5):1469-1476.[Li Y R,Yang L T.2009.New developments in sugarcane industry and technologies in China since 1990s[J].Southwest China Journal of Agricultural Sciences,22(5):1469-1476.]
刘丹.2016.甘肃省设施辣椒镰刀菌根腐病病原鉴定及抗病种质资源筛选[D].兰州:甘肃农业大学.[Liu D.2016.Studies on pathogens and resistance to fusarium root rot on greenhouse pepper in Gansu,China[D].Lanzhou:Gansu Agricultural University.]
刘丽萍,方鹏鹏.2017.马铃薯干腐病镰刀菌的分离及生物学特性研究[J].陇东学院学报,28(5):62-66.[Liu L P,Fang P P.2017.The isolation and biological characteristic of Fusarium associated with potato dry rot[J].Journal of Longdong University,28(5):62-66.]
卢文浩.2007.甘蔗梢腐病的发生及综合防治措施[J].作物杂志,(2):92.[Lu W H.2007.The damage of sugarcanes pokkah boeng and the main control strategies[J].Crops,(2):92.]
曲田丽,辛惠普.2003.绿豆根腐病菌生物学特性研究[J].黑龙江八一农垦大学学报,15(4):109-114.[Qu T L,Xin H P.2003.The biological characteristics of the root rot pathogens in mung[J].Journal of Heilongjiang Bayi Agricultural University,15(4):109-114.]
单红丽,李文凤,张荣跃,王晓燕,李婕,仓晓燕,尹炯,罗志明,黄应昆.2018.甘蔗梢腐病暴发流行原因及产量糖分损失测定[J].中国糖料,40(3):40-42.[Shan H L,Li WF,Zhang R Y,Wang X Y,Li J,Cang X Y,Yin J,Luo ZM,Huang Y K.2018.Analysis on epidemic reason of sugarcane pokahh boeng and its losses on yield and sucrose content[J].Sugar Crops of China,40(3):40-42.]
孙嘉曼,张进忠,韦弟,蓝霞,郭文峰,卢江.2016.一种新的香蕉黑斑病病原菌鉴定及生物学特性研究[J].植物病理学报,46(1):119-123.[Sun J M,Zhang J Z,Wei D,Lan X,Guo W F,Lu J.2016.Identification and biological characterization of the pathogen causing a new black spot disease on Musa spp.[J].Acta Phytophologica Sinica,46(1):119-123.]
王芝涵,王春伟,高海馨,荆琦,余廷濠,王燕.2019.引起玉米穗腐病的禾谷镰刀菌LAMP快速检测方法的建立[J].江苏农业学报,35(3):581-585.[Wang Z H,Wang C W,Gao H X,Jing Q,Yu T H,Wang Y.2019.Development of loop-mediated isothermal amplification(LAMP)assay for the detection of the Fusarium graminearum causing ear rot of maize[J].Jiangsu Journal of Agricultural Sciences,35(3):581-585.]
张斌,杨晓云,陈志谊.2016.番茄枯萎病致病镰刀菌种类鉴定及优势种群的研究[J].植物病理学报,46(4):561-565.[Zhang B,Yang X Y,Chen Z Y.2016.Identification of the pathogenic and dominant Fusarium species causing tomato Fusarium wilt[J].Acta Phytophologica Sinica,46(4):561-565.]
张玉娟.2009.甘蔗梢腐病病原分子检测及甘蔗组合、品种的抗病性评价[D].福州:福建农林大学.[Zhang Y J.2009.Molecular detection and resistance evaluation for pokkah boeng disease of sugarcane[D].Fuzhou:Fujian Agriculture and Forestry University.]
朱迎迎,高兆银,李敏,陈亮,胡美姣.2016.火龙果镰刀菌果腐病病原菌鉴定及生物学特性研究[J].热带作物学报,37(1):164-171.[Zhu Y Y,Gao Z Y,Li M,Chen L,Hu M J.2016.Identification and biological characteristics of dragon fruit(Hylocereus undatus Britt)fusarium rot pathogen[J].Chinese Journal of Tropical Crops,37(1):164-171.]
Bao Y X,Sun H J,Li Y F,Duan Z Z.2016.First report of Fusarium oxysporum isolate gx3 causing sugarcane pokkah boeng in Guangxi of China[J].Plant Disease,100(8):1785.
Khani K T,Alizadeh A,Nejad R F 2013.Pathogenicity of Fusarium proliferatum,a new causal agent of pokkah boeng in sugarcane[J].Proceedings of the International Society Sugracane Technology,28:1-5.
Lin Z,Wei J J,Xu S Q,Guo Q,Wang X,Qiu Y X,Deng ZH,Huang J H,Charles A P,Chen B S,Chen R K,Zhang M Q.2014.Species-specific detection and identification of Fusarium species complex,the causal agent of sugarcane pokkah boeng in China[J].PLoS One,9(8):e104195.
Mohammadi A,Nejad R F,Mofrad N N.2012.Fusarium verticillioides from sugarcane,vegetative compatibility groups and pathogenicity[J].Plant Protection Science,48(3):80-84.
Siti N.2007.Pathogenicity and aethiology of Fusarium species associated with pokkah boeng disease on sugarcane[D].Penang:University Sains Malaysia.
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