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水霉拮抗放线菌的分离、筛选与鉴定
引用本文:陈飘,罗玉双,张永丽,杨敏,张紫根.水霉拮抗放线菌的分离、筛选与鉴定[J].激光生物学报,2016(4):379-385.
作者姓名:陈飘  罗玉双  张永丽  杨敏  张紫根
作者单位:1. 湖南文理学院,水产高效健康生产湖南省协同创新中心,环洞庭湖水产健康养殖及加工湖南省重点实验室,动物学湖南省高校重点实验室,湖南 常德 415000;2. 湖南文理学院,水产高效健康生产湖南省协同创新中心,环洞庭湖水产健康养殖及加工湖南省重点实验室,动物学湖南省高校重点实验室,湖南 常德 415000; 湖南应用技术学院,湖南 常德 415000
基金项目:湖南省高校创新平台开放基金项目(12K121);微生物分子生物学湖南省重点实验室开放基金项目(2013-02);湖南省大学生研究性学习和创新性实验计划项目(湘教通【2014】248号);湖南文理学院芙蓉学院研究性学习和创新性实验计划项目(2014)
摘    要:目的:以珍珠健康养殖水体底泥为材料分离放线菌,筛选对水产动物水霉病病原菌有抗菌活性的放线菌。方法:采用稀释涂布法,选用萘啶酮酸和放线菌酮双抗平板分离获得放线菌;以黄颡鱼和湘云鲫鱼卵水霉病原菌为靶标菌,采用琼脂块法测试所分离菌株抗水霉菌活性及其稳定性;对拮抗活性强的放线菌采用形态观察和16S r DNA序列分析进行分类鉴定。结果:从分离获得的27株放线菌中筛选出3株对水霉病原菌有拮抗活性的菌株QF1、DNC17和QHV2,其中QHV2抗菌活性与稳定性最好;形态学观察与16S r DNA序列分析结果表明QF1、DNC17和QHV2均属于链霉菌属(Streptomycete sp.),分别鉴定为Streptomyces diastatochromogenes、Streptomyces variabilis和Streptomyces collinus。结论:3株放线菌对水霉病原菌具有较好的拮抗活性,具有开发成抗水霉药物的潜在价值。

关 键 词:水霉  拮抗菌  放线菌  分离与筛选  分子鉴定

Isolation,Screening and Identification of Antagonistic Actinomycetes against Saprolegniasis
Abstract:Objective:Isolation,screening and identification of antagonistic actinomycetes against Saprolegniasis from the sediment in healthy aquaculture water of pearl.Methods:Using double resistant screen plate containing nalidixic acid and cycloheximide,dilution and spread plate method was applied to isolate actinomycetes.Taking pathogenic sapro-legnia isolated from the fish eggs of Pelteobagrus fulvidraco and Triploid crucian carp as test bacteria,Antimicrobial ac-tivity against saprolegnia and its stability were assessed using agar diffusion method.The isolates with high antimicrobial activity were identified by morphology and 16S rDNA gene sequence analysis.Results:3 strains of antagonistic actino-mycetes,named as QF1,DNC17,QHV2,were screened from the 27 actinomycetes which were isolated from the sedi-ment samples.Among the 3 isolates,QHV2 strain had the best antibacterial activity and stability.Based on morphology and 16S rDNA gene sequencing,QF1,DNC17 and QHV2 strains all belonged to the genus Streptomyces,and were iden-tified as Streptomyces diastatochromogenes,Streptomyces variabilis and Streptomyces collinus respectively.Conclusion:Taken together,the 3 isolated actinomycetes have higher antagonistic activity against saprolegnia,suggesting that they have potential value of the medicine development for the preventing and controlling of Saprolegniasis.
Keywords:Saprolegnia  antagonistic bacteria  actinomycetes  isolation and screening  molecular identification
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