RFLP法研究生物复混肥及香蕉枯萎病对土壤细菌群落多样性的影响  

Effects of Bio-compound Fertilizer on Banana Wilt and Soil Bacterial Community Based on Analysis with RFLP

作　　者：赵兰凤[1] 张丽娟[1] 胡伟[1] 刘小锋[1] 李华兴[1] 

机构地区：[1]华南农业大学资源环境学院,广州510642

出　　处：《中国生物防治学报》2013年第3期406-416,共11页Chinese Journal of Biological Control

基　　金：国家自然科学青年基金项目(41001150)

摘　　要：本文采用PCR-RFLP法研究了施用生物复混肥和化肥对香蕉枯萎病以及土壤细菌群落多样性的影响。采用直接法提取土壤微生物总DNA,以细菌的16S rDNA通用引物27F/1492R扩增16S rDNA片段,建立16S rDNA克隆文库。从每个处理中筛选50个阳性克隆,阳性克隆子用限制性内切酶MspⅠ和RsaⅠ消化,获得酶切指纹图。结果表明,不同处理间病情指数为不接病原菌不施肥处理(CK1)<接病原菌施生物复混肥处理(BF)<接病原菌施化肥处理(CF)<接病原菌不施肥处理(CK2),表明香蕉枯萎病发病前期施用生物复混肥能降低病情指数。对不同处理的细菌16S rDNA阳性克隆子的酶切图谱进行聚类分析,供试50个克隆子取相似值为1.00时,CK1、CK2、CF和BF分别聚为29、21、11和25类,说明香蕉枯萎病的发生降低了土壤细菌群落多样性,施生物复混肥可以提高土壤细菌群落多样性。CF处理的Shannon-Wiener指数、Simpson指数和丰富度都低于其它处理,均匀度高于其它处理;而BF处理OTU种类、丰富度均低于CK1而高于CK2,Shannon-Wiener指数和Simpson指数均高于其它处理,表明香蕉枯萎病的发生和化肥的施用都降低了土壤细菌群落多样性,化肥的施用会引起某些土壤微生物的富集和一些微生物物种的丧失。生物复混肥一定程度上提高土壤细菌群落多样性,并且形成四种优势类型,这4种类型所占比例较高,分别为12%、12%、8%和10%。16S rDNA测序结果表明,生物复混肥处理的四种优势菌群分别与未培养的δ-变形菌纲uncultured delta proteobacterium、放线菌属uncultured actinobacterium、鞘氨醇单胞菌属Sphingomonas sp.和链霉菌Streptomyces griseoruber有很高的同源性。生物复混肥的施用引起了土壤细菌群落结构的变化,推测其特有的优势类型可能是生物复混肥在香蕉枯萎病防治中起作用的原因之一。In this study, PCR-RFLP was adopted to investigate the effect of bio-compound fertilizer and chemical fertilizer treatments on banana wilt and soil bacterial community. Total microbial genomic DNA was extracted by a direct method from four different soil treatments, respectively. The 16S rDNA of the extracted DNA were amplified using bacterial universal primers 27F and 1492R and a 16S rDNA clone library was constructed. A total of 50 positive clones were randomly chosen from the clone library to be examined. The PCR products were digested by Msp I and Rsa I, and their fingerprints were analyzed. Results indicated that disease indexes among the alltreatments were as follow： no fertilizer and pathogen （CK1）〈bio-compound fertilizer plus pathogen （BF）〈 chemical fertilizer plus pathogen （CF）〈pathogen only CK2. The fingerprints of 16S rDNA positive clones were analyzed using cluster analysis, CK1, CK2, CF and BF were clustered into 29, 21,11, and 25 classes when the similar value was 1.00. Indicating that banana wilt could reduce soil bacterial community, bio-compound fertilizer could increase bacterial community. The bacterial diversity indexes of Shannon-Wiener, Simpson and Richness were lower in CF treatment compared to other treatments, but the evenness was higher. The type of operational taxonomic units and richness were lower in BF treatment in comparison to CK1 and higher in comparison to CK2, and the bacterial diversity indexes of Shannon-Wiener and Simpson were higher than those of other treatments. This revealed that CF could induce specific bacteria enrichment and loss of some other bacteria. Most importantly, BF could increase soil bacterial community and induce 4 kinds of dominant species in soil. The four dominant species had a higher proportions which were 12%, 12%, 10% and 8% respectively. 16S rDNA analysis results showed that these 4 dominant species had separately high homology with uncultured delta proteobacterium, uncultured actinobacterium, Sphingomonas sp. and Streptomyces gris

关 键 词：香蕉枯萎病 生物复混肥 细菌群落多样性 RFLP 

分 类 号：S436.67[农业科学—农业昆虫与害虫防治]