本研究采用平板对峙法、菌丝生长速率法和菌核萌发抑制法筛选出一株细菌LB6-2,对立枯丝核菌的菌丝生长抑制率为 71.76%,菌核萌发抑制率为 100.00%.通过观察形态学特征、分析理化性质结合 16S rDNA扩增序列分析对该菌株鉴定为萎缩芽胞杆菌Bacillus atrophaeus(OK639005.1).采用大孔树脂法提取发酵产物并进行HPLC制备,对获得的 14 个组分进行活性测定确定其活性组分,从活性组分中制备获得 3 个单品化合物 LBN4-2、LBN4-4、LBN4-5,经 HPLC-MS、NMR 分析和活性测定结果表明,化合物LBN4-2 为Cyclo(Pro-Val)、LBN4-4为Cyclo(D-Leu-L-Pro)和LBN4-5为Cyclo(L-prolyl-D-phenylalanyl),3个化合物的浓度在 4 mmol/L时对立枯丝核菌菌丝生长的抑制率分别为 86.16%、84.87%和 83.10%,浓度在 2 mmol/L以上时,对立枯丝核菌菌核萌发的抑制率为 100%.由此可见,萎缩芽胞杆菌LB6-2具有作为防治立枯丝核菌生防菌剂研发的潜力.
Bacteria LB6-2 was selected against Rhizoctonia solani by plate face-off method,mycelium growth rate method and sclerotium germination inhibition method.It showed the inhibition rate of mycelium growth and sclerotium germination of strain LB6-2 was 71.76%and 100.00%.The strain was identified Bacillus atrophaeus(OK639005.1)by observing morphological characteristics,analyzing physicochemical properties and 16S rDNA amplification.The fermentation products were extracted by macroporous resin method and prepared by HPLC.Fourteen active components were determined by activity determination,and the single compounds were prepared and analyzed by HPLC-MS,NMR and activity determination.The compounds LBN4-2,LBN4-4 and LBN4-5 were identified as Cyclo(Pro-Val),Cyclo(D-Leu-L-Pro)and Cyclo(L-prolyl-D-phenylalanyl),respectively.The activity assay of the three compounds showed that the inhibition rates of hyphal growth were 86.16%,84.87%and 83.10%at concentration was 4 mmol/L,respectively.While the concentration of the three compounds were above 2 mmol/L,the inhibition rates of hyphal germination were 100%.In summary,Bacillus atrophaeus LB6-2 has the potential to be used as a biocontrol agent against Rhizoctonia solani.
罗文芳;周军辉;何伟;许建军
新疆农业科学院植物保护研究所/农业农村部西北荒漠作物有害生物综合治理重点实验室/新疆农业生物安全重点实验室,乌鲁木齐 830091
植物保护学
立枯丝核菌;萎缩芽胞杆菌;活性物质;结构鉴定
Rhizoctonia solani;Bacillus atrophaeus;active substance;structure identification
《中国生物防治学报》 2024 (002)
378-387 / 10
新疆农业科学院青年科技骨干创新能力培养项目(xjnkq-2021016);新疆维吾尔自治区重大科技专项(2022A02005-3);新疆现代农业产业技术体系(XJARS-07);新疆农业科学院稳定支持专项(xjnkywdzc-2022004)
10.16409/j.cnki.2095-039x.2024.02.013
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