解淀粉芽孢杆菌SQR9根际互作微生物的初步鉴定
毕业论文关键词：解淀粉芽孢杆菌SQR9；生物膜；微生物互作
Preliminary identification of cucumber rhizospheric microorganisms that interact with Bacillus amyloliquefaciens SQR9
Abstract：Bacillus amyloliquefaciens SQR9, isolated and identified by our laboratory, was proved to be an outstanding PGPR (plant growth-promoting rhizobacteria) strain by its abilities to inhibit the growth of FOC (Fusarium oxysporum f. sp. cucumerinum J. H. Owen) and decrease the incidence of Fusarium wilt of cucumber. Although in most environments, especially in the rhizophere, multi-species biofilms are dominate, single-species biofilms are the focus of most current researches under laboratary condition. Researches on the interaction between B. amyloliquefaciens SQR9 and other rhizospheric microorganisms are limited. To explore such interaction, the number of B. amyloliquefaciens SQR9 in mixed-species biofilms were monitored by real-time PCR. Results showed that the biofilm formation abilities of SQR9 was improved by incubating with microorganisms from black soil, indicate that some unkown microorganisms may be involved in this process. The microbial communities in biofilm and liquid media were analyzed by high-throughput sequencing technology. Results indicate that the abundance of Stenotrophomonas in bioflim is significantly higher than that in liquid media. Other microflora also existed in biofilm, but played a dominant role in liquid media. We speculate that the interaction of Stenotrophomonas and SQR9 was involved in the biofilm formation and root colonization.
Key words: Bacillus amyloliquefaciens SQR9；biofilm；microbial interactions
目 录
摘要1
关键词1
Abstract1
Key words1
引言1
1材料与方法2
1.1材料 2
1.2方法 2
1.2.1黄瓜根际土的分离2
1.2.2黄瓜根际土菌体细胞的提取3
1.2.3生物膜共培养3
1.2.4提取生物膜和培养液中的DNA3
1.2.5定量PCR分析生物膜及游离细胞中菌株SQR9的数量3
1.2.6高通量测序5
2结果与分析5
2.1绝对定量PCR5
2.1.1引物特异性验证5
2.1.2转化子鉴定5
2.1.3标准曲线制作6
2.1.4生物膜及培养液中菌株SQR9的绝对定量6
2.2生物膜基因组高通量测序7
3讨论 8
致谢9
参考文献9
解淀粉芽孢杆菌SQR9根际互作微生物的初步鉴定\引言:
黄瓜土传枯萎病是黄瓜种植区主要的连作障害之一，由真菌尖孢镰刀菌黄瓜专化型FOC引起，严重影响黄瓜的产量和品质，研究以微生物防控为核心的综合防控具有重大意义。解淀粉芽孢杆菌SQR9是从黄瓜土传枯萎病发病区的健康植株根际分离的一株根际促生菌（PGPR），能强烈抑制尖孢镰刀菌，在盆栽试验和大田试验中表现出良好的防控黄瓜土传枯萎病和促进黄瓜生长的效果[1-2]。同时该菌株在实验室条件下表现了优异的生物膜形成、根际定殖、趋化性、可产生多种次生代谢产物（主要为抗生素类物质）等性状，是研究PGPR菌株在根际生态行为及植物-微生物互作的优良菌株[3-5]。
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