深海独岛枝芽胞杆菌1A00493杀根结线虫活性物质研究  

Nematicidal Active Substance Produced by Deep-Sea Virgibacillus dokdonensis 1A00493

作　　者：郑娜[1] 黄典[1] 邵宗泽[2] 喻子牛[1] 张吉斌[1] 

机构地区：[1]农业微生物学国家重点实验室华中农业大学生命科学技术学院微生物农药国家工程研究中心,湖北武汉430070 [2]国家海洋局第三海洋研究所海洋生物遗传资源重点实验室,福建厦门361005

出　　处：《化学与生物工程》2017年第5期21-27,共7页Chemistry & Bioengineering

基　　金：国家重点基础研究发展计划项目(2013CB127504)

摘　　要：以一株具有杀南方根结线虫(Meloidogyne incongnita)活性的深海独岛枝芽胞杆菌(Virgibacillus dokdonensis)1A00493为研究对象,对其发酵上清液的杀线虫活性进行测定,并对杀线虫活性物质进行理化分析。利用正相硅胶柱、反相硅胶柱、LH-20葡聚糖凝胶柱分离以及示差折光高效液相色谱(RID-HPLC)对活性物质进行分离纯化,将纯化的活性物质进行液相色谱-质谱联用分析。结果表明,杀线虫活性物质耐高温、对蛋白酶和酸碱不敏感、硫酸铵沉淀物无杀线虫活性、3kDa超滤管滤过液有活性、易溶于甲醇和氯仿,推测活性物质是中强极性小分子物质。杀线虫活性物质主要有3种化合物,其中分子量为234Da的新型化合物很有可能是主要的活性物质。A deep-sea Virgibacillus dokdonensis 1A00493, which had nematicidal activity against Meloidogyne incongnita, was taken as the research object. The nematicidal activity of fermentation supernatant was de- termined,also the physical and chemical analyses on nematicidal active substance were carried out. The crude extracts of fermentation supernatant were isolated and purified by a positive silica gel column, followed by a re- verse silica gel column,a Sephadex LH-20 gel column and RID-HPLC. Results showed that, nematicidal active substance was insensitive to high temperature,protease and pH value, ammonium sulfate precipitation had no nematicidal activity,the supernatant filtered through a 3 kDa ultrafiltration tube remained active,and it was easily soluble in methanol and chloroform. It is speculated that the active substance is a moderate polarity com- pound with small molecules. The results of LC-MS show that,the nematicidal active substance most likely con- tain three compounds,and the active substance molecular weight may be 234 Da.

关 键 词：深海独岛枝芽胞杆菌 根结线虫 活性物质 分离纯化 

分 类 号：S482.3[农业科学—农药学]