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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

赵兰凤, 张丽娟, 胡伟, 刘小锋, 李华兴   

  1. 华南农业大学资源环境学院, 广州 510642
  • 收稿日期:2012-09-12 修回日期:1900-01-01 出版日期:2013-08-08 发布日期:2013-08-08
  • 通讯作者: 李华兴,教授,E-mail:huaxli@scau.edu.cn。

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

ZHAO Lanfeng, ZHANG Lijuan, HU Wei, LIU Xiaofeng, LI Huaxing   

  1. College of Resources and Environment, South China Agricultural University, Guangzhou 510642
  • Received:2012-09-12 Revised:1900-01-01 Online:2013-08-08 Published:2013-08-08

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。

Abstract: 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 all treatments were as follow: no fertilizer and pathogen (CK1)delta proteobacterium, uncultured actinobacterium, Sphingomonas sp. and Streptomyces griseoruber strain. Bio-compound fertilizer could change the structure of soil bacterial community and these dominant species induced by bio-compound fertilizer might play an important role in control of banana wilt.

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