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枣缩果病拮抗菌的鉴定及其拮抗效应

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[目的]发掘有效生防菌株以控制枣缩果病,并探究生防菌株的拮抗特性.[方法]采用梯度稀释法和平板对峙法筛选出2株对枣缩果病菌细极链格孢(Alternaria tenuissima)具有显著拮抗作用的菌株STO-12和STO-45,通过PCR扩增得到其16S rRNA基因序列,进行同源比对分析和分子系统树的构建,并结合形态学观察和生理生化实验对菌株进行鉴定.采用皿内对峙、显微观察及蛋白质抑菌试验等测定了菌株STO-45的拮抗效应.[结果]两拮抗菌株STO-12和STO-45均鉴定为枯草芽孢杆菌,但菌株STO-12能产生橘红色色素,且两菌株在大小、生理生化特征及系统发育关系上具有生理差异.这两株拮抗细菌的发酵液、发酵上清液及发酵滤液均对枣缩果病菌具有显著抑菌活性.STO-45发酵滤液在0.8%的低体积浓度下即达到抑制中浓度,可使病原菌A.tenuissima MY5的菌丝及芽管部分畸形膨大,发酵液中的蛋白类物质可能是其拮抗物质之一.[结论]拮抗菌株STO-12和STO-45对枣缩果病的生物防治均具有潜在的应用价值,其抑菌机制及生物制剂的开发利用值得进一步研究.
Identification of antagonistic bacteria against Alternaria tenuissima, and its effect on antagonism
[Objective] This study aimed to screen and characterize antagonistic bacteria against Alternaria tenuissima,the causal agent of jujube shrinking fruit disease,and to study their antagonistic effects.[Methods] Serial dilutions and a dual culture technique were used for screening bacteria.The bacteria were identified based on morphological,physiological and biochemical features,and a phylogenetic analysis was conducted on an alignment of 16S rRNA gene sequences.Evaluation of the control activity and efficacy of fermentation filtrates against A.tenuissima was carried out by a dual culture assay,microscopic inspection and a spore germination test.Crude active proteins were separated by an ammonium sulfate method.[Results] Two strains of bacteria,STO-12 and STO-45,that were isolated and screened from the soil exhibited excellent inhibition against the fungal pathogen.Based on molecular,morphological,physiological and biochemical features,the two strains were identified as Bacillus subtilis.However,the two strains differed in the production of a pigment that was shorter in length in STO-12 than in STO-45.The fermentation filtrate,the supernatant and the cell-free filtrate of both strains exhibited excellent inhibition against the strain MY5 with a dual culture assay.Further effects of the cell-free filtrate of STO-45 on the pathogen included inhibition of germ tube and hyphal growth,and abnormally swollen or degraded hyphae and germ tubes.Crude active extracellular proteins of STO-45 isolated by an ammonium sulfate method proved to restrain the growth of hyphae.[Conclusion] Both STO-12 and STO-45 strains of B.subtilis have potential for the biological control of jujube shrinking fruit disease.

Fruit shrink disease of Ziziphus jujubaAlternaria tenuissimaBacillus subtilisIdentificationInhibition rate

武自强、马荣、游崇娟、梁英梅

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北京林业大学林学院 北京100083

新疆农业大学林学与园艺学院林学院 新疆乌鲁木齐830052

北京林业大学标本馆 北京100083

枣缩果病 细极链格孢菌 枯草芽孢杆菌 鉴定 抑菌率

国家“十二五”科技支撑课题

2012BAD19B08

2015

微生物学通报
中科院微生物所 中国微生物学会

微生物学通报

CSTPCDCSCD北大核心
影响因子:1.052
ISSN:0253-2654
年,卷(期):2015.42(7)
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