生防枯草芽孢杆菌的分离与鉴定

摘 要：本研究对分离得到的生防BCM菌株经PCR扩增后，测定其16SrRNA基因序列，与基因库中基因序列进行同源性比较，BCM菌株与枯草芽孢杆菌Bacillus subtilis的相似性达99%。结合其形态观察及生理生化实验综合分析，得出BCM菌株为枯草芽孢杆菌。此外，BCM菌株对辣椒疫病病菌的拮抗作用和防治效果也进行了研究。结果表明BCM对辣椒疫病病菌有拮抗作用；温室防病试验中，BCM喷雾处理对辣椒疫病具有明显的防治效果，防治效果达到 52.32 ％。该菌株对作物疫病有一定的生防潜力。

关键词：枯草芽孢杆菌；16SrRNA；鉴定；生防

 

Isolation and Identification of Bio-control Bacillus subtilis

Abstract: The sequence 16SrRNA gene of isolated BCM strains was determined after PCR amplification. Similarity of BCM isolate with Bacillus subtillis through homology gene sequence contrast in gene bank was 99%. And the isolate in morphological and physiological and biochemical characters was also identified. The result showed that BCM attributed to Bacillus subtillis. Besides, the antagonism and effect of BCM on Phytoophthora capsici were studied．Results revealed that BCM had some antagonism to P.capsici, and impact of BCM on chili blight in greenhouse trial was 52.32 %.. So, BCM will have some utilization value for controlling crop disease, caused by P.capsici.

Key words: Bacillus subtilis;16SrRNA; Dentification; Bio-control

目  录

摘 要 1

关键词 1

1  前言 2

2  材料及方法 3

2.1  材料 3

2.1.1  实验菌种 3

2.1.2  实验试剂及仪器 3

2.1.3  培养基 3

2.2  方法 4

2.2.1  细菌的初步分离及纯化 4

2.2.2  细菌的形态鉴定 4

2.2.3  细菌的生化鉴定 4

2.2.4  细菌的16SrRNA检测法 4

2.2.4.1  模板DNA的提取 5

2.2.4.2  琼脂糖凝胶电泳 5

2.2.4.3  PCR产物的回收及DNA测序 5

2.2.5  菌种鉴定 6

2.2.6  枯草芽孢杆菌BCM的生防效果测定 6

2.2.6.1  生防菌抑菌作用的检测 6

2.2.6.2  生防菌温室防治辣椒疫病试验 7

3 结果与分析 7

3.1  枯草芽孢杆菌BCM的形态鉴定 8

3.2  枯草芽孢杆菌BCM的生化鉴定 8

3.3  枯草芽孢杆菌BCM的16SrRNA检测 9

3.4  枯草芽孢杆菌的生防效果测定 9

3.4.1  生防菌抑菌作用的检测 10

3.4.2  生防菌温室防治辣椒疫病试验 10

4 讨论...……………………………………………………………………………………..11

参考文献 12

致谢 14