| 摘要: |
| 【目的】从源自秦岭的土壤微生物中筛选具有抗病毒活性的生防菌并进行分类鉴定，对其抗烟草花叶病毒（TMV）的作用方式进行研究并对发酵条件进行优化，为防治烟草花叶病毒病提供新资源。【方法】采用土壤分离法分离生防菌，采用形态学和分子生物学方法对其进行分类鉴定。采用无菌过滤法制备F01菌株发酵液，以心叶烟为材料，采用半叶枯斑法测定生防菌F01菌株抗TMV的作用方式，并采用单因素法优化F01菌株的发酵体系。【结果】从秦岭土壤中分离到1株生防放线菌株F01，经形态学和分子生物学鉴定为链霉菌。F01菌株发酵液对烟草花叶病毒具有良好的防治效果,其抗病作用方式包括体外病毒钝化和体内诱导抗病性，对TMV的抑制率为74.2%。优化后的发酵体系为：以高氏1号为培养基，以小米粉和酪蛋白胨为碳、氮源，在28 ℃、pH 8.0条件下培养5 d，发酵液对TMV有显著的抑制效果，抑制率为81.7%。【结论】分离得到的链霉菌F01菌株具有良好的抗TMV活性，优化后的发酵条件简单，发酵产物对TMV有显著抑制效果，具有进一步开发为抗TMV工程菌株的潜力。 |
| 关键词: 烟草花叶病毒 链霉菌 发酵条件优化 生物防治 |
| DOI： |
| 分类号: |
| 基金项目:陕西省自然科学基础研究计划项目（2019JM-060）；陕西省科技重大专项（2020zdzx03-01-04） |
|
| Anti TMV activity of F01 isolate of Streptomyces and the optimization of fermentation system |
|
HAO Xing’an,YANG Han
|
| Abstract: |
| 【Objective】For the purpose of exploring potential antiviral resource of microorganism,an actinomycete strain was screened and isolated from the soil of Qinling Mountains.The isolated strain and its anti TMV activity were identified, the antiviral effect was analyzed,and the fermentation system was optimized.【Method】The soil microorganism was isolated using the gradient dilute method,and the 16S rRNA gene from isolated colony was amplified and sequenced for homologous analysis.The bacterial free broth of the isolated strain fermentation was prepared using sterilizing filter,while the method of half leaf was adopted for screening antiviral activity using Nicotiana glutinosa.The fermentation system was optimized using single factor method.【Result】 The isolated F01 strain was identified as a Streptomyces spp.by morphology and molecular biology.The broth of F01 fermentation showed strong antiviral active to TMV with both inactive effect to virus and induced system resistance in host.The antiviral rate of F01 strain was 74.2% by half-leaf method in lab.The optimal conditions for F01 fermentation included the Gauze’s synthetic medium No.1 with rice flour and casein peptone as carbon and nitrogen sources,28 ℃ and pH 8.0 for 5 days.The TMV inhibition rate was 81.7%.【Conclusion】The isolated Streptomyces F01 strain showed strong anti-TMV activity and could be cultured in a simple optimum fermentation system.The F01 strain had potential to be explored as an engineering strain for anti-TMV. |
| Key words: tobacco mosaic virus Streptomyces fermentation system optimization biological control |
