桃根癌病菌拮抗放线菌抑菌活性物质的分离及其初步鉴定

[目的]分离并初步鉴定从桃树高发根癌病土壤中获得的拮抗线菌G19菌株的抑菌活性物质。[方法]采用蛋白沉淀法对拮抗放线菌G19菌株的抑菌活性物质进行粗提,利用高效液相色谱仪、中压制备色谱仪对其进行分离、纯化,应用MALDI-TOFMS法进行分子量的测定,最后通过化学显色反应进行相关官能团的验证。[结果]经分离纯化后拮抗放线菌G19的抑菌物质被抨击出7个肽段,是分子量范围为900～1 300 Da,同时推断其含有一个Cys并携带H2O,Na+;官能团显色反应验证该物质为多肽且含有糖基。[结论]研究结果为拮抗放线菌G19菌株的抑菌物质结构的最终确定奠定了基础。

Isolation and Preliminary Identification of Antibacterial Active Substances of Antagonistic Actinomycetes against Peach Crown Gall(Agrobacterium tumefaciens)

[ Objective ] The paper was to isolate and preliminarily identify the antibacterial active substances of antagonistic actinomyeete strain G19 obtained from the soil highly affected by peach crown gall （Agrobacterium tumefaciens）. [ Method] The antibacterial substances of antagonistic actinomycete strain G19 were ex- tracted using protein precipitation method, then isolated and purified using high performance liquid chromatography and medium-pressure preparative chromatogra- phy. Its molecular weight was determined by MALDI-TOFMS method, and the related functional groups were verified through chemical color reaction. [ Result] Seven peptide portions were produced from the antibacterial substances of antagonistic actinomycete strain G19 with the molecular weights of 900 - 1 300 Da after isolation and purification. It could be also inferred that it contained Cys, and carried with H2O and Na＋. Color reaction of functional groups verified that the sub- stance was polypeptide containing glycosyl. [ Conclusion] The result provided basis for the final definition of the structure of antibacterial substances in antagonistic actinomycete strain G19.

抗桃根癌病菌的耐氰放线菌筛选及其抑菌作用研究

为了筛选耐氰并抗桃根癌病的微生物菌株,以不同浓度的KCN培养基作为选择培养基,以根癌农杆菌为靶标菌,通过平板对峙培养的方法,筛选出了耐氰放线菌G-19,其发酵液代谢产物对根癌农杆菌有明显的抑制作用,抑菌圈直径为21.66 mm。其抑菌作用主要表现在3个方面:使得病原菌的生长对数期缩短,菌体浓度处于低水平状态;使病原菌的细胞壁弯曲、皱缩,继之结构发生破裂,细胞质向边缘凝聚,中心变得稀薄,甚至形成空腔;使细胞膜的通透性增加,导致病原菌的细胞质液外泄;但未见到病原菌DNA的损伤。因此,认为G-19菌株抑菌的作用机理是使根癌病原菌的细胞壁结构破坏和膜透性增加。