杨树叶枯病生物防治初步研究
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摘要
杨树是世界上分布广、适应性强的树种,主要分布在北半球温带、寒温带区域。杨树不仅能防风固沙,保持水土,改善环境,而且其木材是制造纤维板、刨花板、细木工板及造纸纸浆的优良材料,具有很高的社会、生态和经济效益,在我国深受群众喜爱。
杨树也是一种病虫害发生较多的树种,其中杨树叶枯病作为其中的一项主要病害,严重影响了杨树诸多方面效益。
本研究分离杨树叶枯病的病原菌,通过显微形态特征及分子鉴定将该菌鉴定为半知菌门、丝孢纲、丝孢目真菌,细链格孢菌(Alternaria alternata(Fr) Keissl),rDNA-ITS序列测定结果表明其同源性大于99%。
本文研究了杨树叶栖真菌的群落结构,应用形态学和分子生物学的方法对真菌进行了鉴定,并利用所分离到得真菌筛选出拮抗菌株,并进行了叶片回接等生防试验,筛选出具有防治价值较高的生防菌。
从杨树叶片上共分离得16属。子囊菌1属,有丝分裂菌15属,未具体鉴定到种。有些只加以编号。其中有丝分裂菌15属,包括：胶帚霉属(Gliocladium)、粘束孢属(Graphium)、枝孢属(Cladosporium)、盾壳霉属(Coniothyrium)、弯孢属(Curvularia)、拟青霉属(Paecilomyces)、附球孢属(Epicoccum)、被孢霉(Mortierella)、曲霉属(Aspergillus)青霉属(Penicillium)、镰刀菌属(Fusarium)、木霉属(Trichoderma)、孔球孢属(Gilmaniella)、色串孢属(Tolura)、轮枝孢属(Verticilium);子囊菌门(Ascomycota)一属：毛壳属(Chaetomium)。
根据拮抗生长半径、病原菌的被抑制率和相对抑制效果及菌后期的反寄生的综合评价可确定有效抑制杨树叶枯病病原菌的拮抗菌株,选用Trichoderma、Chaetomium、Tolura作为试验生防菌株。且Toluraa为腐生菌,拮抗实验表明,可以对杨树细链格孢菌抑制,后期具有反寄生作用。
室内接种试验结果表明,除接种病原Alternaria alternata的叶片分离出病原菌外,其余接种叶片并未分离到,说明T. atroviride、C. globosum和T. herbarum三种菌株对杨树叶枯病病原菌Alternaria alternata具有很好的抑制作用。浓度梯度生防试验表明,防治效果为T. herbarum>代森锰锌400倍液>T. atroviride>C. globosum,三种菌剂在50%喷洒浓度时,真菌多样性最大,且与CK对照组最为相似,说明在该浓度下叶栖真菌系统较其他浓度更为稳定。
Poplar is the species which are widely distributed in the world,adaptable and mainly in northern temperate,cold temperate regions. Poplar not only can check winds but also can fix drifting sand,conserve soil and water,improve the environment,and its wood is fine material in manufacturing fibreboard,particle board,block board and paper pulp with high social, ecological and economic benefits,it is very popular in China.
Poplar is also a species pest occurred more,poplar leaf blight disease is one of the main disease which has a serious impact on many aspects of poplar benefits.
This research is to separate the pathogen of poplar leaf blight,Strain was identified as fungi imperfecti,Trichosporon Gang,Trichosporon fungi present,Fine tenuis by microscopic morphology and molecular identification, rDNA-ITS sequencing results showed that the homology of GenBank is greater than 99%.
This paper studied the community structure of Poplar fungi of needles,morphological and molecular biology methods were used for identification of fungi,and use the isolated fungi selected antagonistic strains,take such leaves grouped joint test and select biocontrol bacteria with high value of prevention.
Isolated from the poplar leaves,we can get 1 ascomycetes genera,15 mitosis bacteria genera,these are not specifically identified to species. Some are only numbered. Among the 15 mitosis,bacteria,genera,including:Gliocladium,Graphium,Cladosporium,Coniothyrium,Curvula ria,Paecilomyces,Epicoccum,Mortierella,Aspergillus,Penicillium,Fusarium,Trichoderma,Gilm aniella,Tolura,Verticilium and the genera of Ascomycota:Chaetomium.
According to growth radius of antagonism,the inhibition rate of pathogenic bacteria,the relative inhibitory effect and the comprehensive evaluation of anti-parasitic in the late period of bacteria,It is determined that the antagonistic strains can inhibited the pathogen of poplar leaf blight disease effectively,and then selected Trichoderma,Chaetomium,Tolura as experimental biocontrol strains. Meawhile Tolura is saprophytic bacteria,the antagonistic experiments show that we can inhibit the alternaria tenuis of polar and prevent it from growing.
Inoculation tests showed that,in addition to the leaves of inoculated pathogen Alternaria alternata isolated from pathogens,the rest of inoculated leaves isolated nothing from it. It showed that Description T. atroviride,C. globosum and T. herbarum, this three strains has good inhibition to Alternaria alternata which is pathogenic bacteria of poplar leaf blight disease. The biocontrol experiments of concentration gradient show that control effect is the T. herbarum> mancozeb 400 dilution> T. atroviride> C. globosum, when the three kinds of microbial agents is in the spray concentration of 50%,the diversity of fungi is maximum,and it is most similar to the control group of CK. It indicates that the concentration of leaf fungi system is more stable than other concentrations on this concentration.
杨树也是一种病虫害发生较多的树种,其中杨树叶枯病作为其中的一项主要病害,严重影响了杨树诸多方面效益。
本研究分离杨树叶枯病的病原菌,通过显微形态特征及分子鉴定将该菌鉴定为半知菌门、丝孢纲、丝孢目真菌,细链格孢菌(Alternaria alternata(Fr) Keissl),rDNA-ITS序列测定结果表明其同源性大于99%。
本文研究了杨树叶栖真菌的群落结构,应用形态学和分子生物学的方法对真菌进行了鉴定,并利用所分离到得真菌筛选出拮抗菌株,并进行了叶片回接等生防试验,筛选出具有防治价值较高的生防菌。
从杨树叶片上共分离得16属。子囊菌1属,有丝分裂菌15属,未具体鉴定到种。有些只加以编号。其中有丝分裂菌15属,包括：胶帚霉属(Gliocladium)、粘束孢属(Graphium)、枝孢属(Cladosporium)、盾壳霉属(Coniothyrium)、弯孢属(Curvularia)、拟青霉属(Paecilomyces)、附球孢属(Epicoccum)、被孢霉(Mortierella)、曲霉属(Aspergillus)青霉属(Penicillium)、镰刀菌属(Fusarium)、木霉属(Trichoderma)、孔球孢属(Gilmaniella)、色串孢属(Tolura)、轮枝孢属(Verticilium);子囊菌门(Ascomycota)一属：毛壳属(Chaetomium)。
根据拮抗生长半径、病原菌的被抑制率和相对抑制效果及菌后期的反寄生的综合评价可确定有效抑制杨树叶枯病病原菌的拮抗菌株,选用Trichoderma、Chaetomium、Tolura作为试验生防菌株。且Toluraa为腐生菌,拮抗实验表明,可以对杨树细链格孢菌抑制,后期具有反寄生作用。
室内接种试验结果表明,除接种病原Alternaria alternata的叶片分离出病原菌外,其余接种叶片并未分离到,说明T. atroviride、C. globosum和T. herbarum三种菌株对杨树叶枯病病原菌Alternaria alternata具有很好的抑制作用。浓度梯度生防试验表明,防治效果为T. herbarum>代森锰锌400倍液>T. atroviride>C. globosum,三种菌剂在50%喷洒浓度时,真菌多样性最大,且与CK对照组最为相似,说明在该浓度下叶栖真菌系统较其他浓度更为稳定。
Poplar is the species which are widely distributed in the world,adaptable and mainly in northern temperate,cold temperate regions. Poplar not only can check winds but also can fix drifting sand,conserve soil and water,improve the environment,and its wood is fine material in manufacturing fibreboard,particle board,block board and paper pulp with high social, ecological and economic benefits,it is very popular in China.
Poplar is also a species pest occurred more,poplar leaf blight disease is one of the main disease which has a serious impact on many aspects of poplar benefits.
This research is to separate the pathogen of poplar leaf blight,Strain was identified as fungi imperfecti,Trichosporon Gang,Trichosporon fungi present,Fine tenuis by microscopic morphology and molecular identification, rDNA-ITS sequencing results showed that the homology of GenBank is greater than 99%.
This paper studied the community structure of Poplar fungi of needles,morphological and molecular biology methods were used for identification of fungi,and use the isolated fungi selected antagonistic strains,take such leaves grouped joint test and select biocontrol bacteria with high value of prevention.
Isolated from the poplar leaves,we can get 1 ascomycetes genera,15 mitosis bacteria genera,these are not specifically identified to species. Some are only numbered. Among the 15 mitosis,bacteria,genera,including:Gliocladium,Graphium,Cladosporium,Coniothyrium,Curvula ria,Paecilomyces,Epicoccum,Mortierella,Aspergillus,Penicillium,Fusarium,Trichoderma,Gilm aniella,Tolura,Verticilium and the genera of Ascomycota:Chaetomium.
According to growth radius of antagonism,the inhibition rate of pathogenic bacteria,the relative inhibitory effect and the comprehensive evaluation of anti-parasitic in the late period of bacteria,It is determined that the antagonistic strains can inhibited the pathogen of poplar leaf blight disease effectively,and then selected Trichoderma,Chaetomium,Tolura as experimental biocontrol strains. Meawhile Tolura is saprophytic bacteria,the antagonistic experiments show that we can inhibit the alternaria tenuis of polar and prevent it from growing.
Inoculation tests showed that,in addition to the leaves of inoculated pathogen Alternaria alternata isolated from pathogens,the rest of inoculated leaves isolated nothing from it. It showed that Description T. atroviride,C. globosum and T. herbarum, this three strains has good inhibition to Alternaria alternata which is pathogenic bacteria of poplar leaf blight disease. The biocontrol experiments of concentration gradient show that control effect is the T. herbarum> mancozeb 400 dilution> T. atroviride> C. globosum, when the three kinds of microbial agents is in the spray concentration of 50%,the diversity of fungi is maximum,and it is most similar to the control group of CK. It indicates that the concentration of leaf fungi system is more stable than other concentrations on this concentration.
引文
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