地黄内生菌的分离鉴定及其抑菌活性测定

Isolation, Identification and Antibacterial Activities of Endophytes in Fresh Rehmannia

对河南焦作产鲜地黄的内生菌进行分离、纯化,对其形态学、生化、16S r DNA序列进行分析及抑菌活性进行测定,初步了解该菌的相关特性并为其资源的开发利用提供参考。方法:对鲜地黄用营养琼脂培养基及玫瑰红钠琼脂培养基培养,用划线法进行分离与纯化,然后用革兰氏染色、生化管、16S r DNA序列分析鉴定、药敏试纸抑菌实验对内生菌及其抑菌性进行鉴定。结果:筛选得到的鉴定菌在革兰氏染色及镜检后鉴定显示1号、2号和3号菌均为梭状细菌,4号为杆状细菌;生化管实验的颜色变化显示1号、2号和3号菌为梭菌属细菌,4号菌为土壤农杆菌;16S r DNA的鉴定结果表明3号菌为苏云金芽孢杆菌,3号菌对三种病原微生物的抑菌作用不明显。结论:地黄中分离纯化得到的3号菌的抑菌活性不显著。

Objective： Endophytes of henan jiaozuo fresh Rehmannia get endophytes separation, purification, and then on the morphology, biochemistry, 16S rDNA identification and the determination of antibacterial activity, preliminary understanding the characteristics of the bacteria and provide data for the development and utilization of its resources. Methods： the fresh Rehmannia nourished with nutrient AGAR culture medium and rosy sodium, by exploring method for separation and purification, and then using gram staining, biochemical tube, PCR and drug susceptibility＇ test paper bacteriostatic experiment of endophytes and their antibacterial sex identifi- cation. Results： the identification of bacteria in screening for gram staining and microscopy after identification showed no. I, 2 and 3 were fusiform bacteria, 4 for rod-shaped bacteria; Biochemical tube experiments showed the color change of the no. 1, 2 and 3 bacte- ria for Clostridium bacteria, 4 bacteria for soil Agrobacterium; 16S rDNA identification results showed that the 3 bacteria for bacteria and fusobaeterium thuringiensis bacillus; And 3 isolates on the bacteriostatie action of three kinds of pathogenic microorganisms were not obvious. Conclusion：No.3 bacteria＇s antibacterial activity are not significant.