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猕猴桃细菌性溃疡病生防放线菌的筛选与鉴定
关鹏1,贺志有1,代小娟1,秦培钢2
1.渭南职业技术学院,陕西 渭南 714026;2.新疆医科大学 基础医学院,新疆 乌鲁木齐 830011
摘要:
对分离自新疆巴里坤盐湖和达坂城盐湖的179株放线菌进行拮抗丁香假单胞杆菌猕猴桃致病变种筛选,从中筛选到8株具有拮抗作用的放线菌,其中ZBW8-1、E14-3和ZB5-6抗菌活性最显著,其抑菌圈直径分别为27.5 mm、23.6 mm和26.2 mm。针对这3株放线菌进行菌落形态观察,16s rRNA扩增、测序及系统进化分析,并对其进行抗生素合成相关基因的PCR鉴定。结果表明,放线菌ZBW8-1、E14-3和ZB5-6分别与糖霉菌属(Glycomyces)的Glycomyces fuscus TRM 49117、原小单孢菌属(Promicromonospora)的维也纳原小单孢菌(Promicromonospora vindobonensis)V45T以及拟诺卡氏菌属(Nocardiopsis)的Nocardiopsis terrae YIM 90022T 16s rRNA序列同源性最高,分别为99.86%、99.10%和99.86%。在系统进化关系上,这3株菌也分别与上述菌株聚类在同一个分支上。ZBW8-1、E14-3和ZB5-6菌株中均携带有NRPS基因,其中ZBW8-1和ZB5-6菌株中还含有PKS-II基因,且ZBW8-1菌株中还含有PKS-I基因。对这3株抗菌活性放线菌进行进化地位分析及抗生素合成相关功能基因的鉴定,将为研究这些菌株中的活性次生代谢产物奠定理论基础。
关键词:  放线菌  丁香假单胞杆菌猕猴桃致病变种  抗菌活性  系统进化树  功能基因
DOI:10.14188/j.ajsh.2024.03.008
分类号:Q939.96
基金项目:渭南市科技计划项目(2021ZDYF-JCYJ-299);陕西省教育厅2021年度一般专项科研计划项目(21JK0639)
Screening and identification of actinomycetes for biocontrol of kiwifruit bacterial canker
GUAN Peng1, HE Zhiyou1, DAI Xiaojuan1, Qin Peigang2
1.Weinan Vocational and Technical College, Weinan 714026, Shaanxi, China;2.Department of Basic Medicine, Xinjiang Medical University, Urumchi 830011, Xinjiang, China
Abstract:
179 actinomycete strains isolated from Barkol and Dabancheng salt lakes in Xinjiang were screened for antagonistic effects against Pseudomonas syringae pv. actinidiae. Eight actinomycete strains with antagonistic effects were identified, among which ZBW8-1, E14-3, and ZB5-6 showed the most significant antimicrobial activity, with the diameters of inhibition zones of 27.5 mm, 23.6 mm, and 26.2 mm, respectively. Colony morphology observation, 16s rRNA amplification and sequencing, and phylogenetic analysis based on 16s rRNA sequences were conducted on these three strains of actinomycetes. PCR identification of antibiotic synthesis-related genes was performed on these strains. The results show that the actinomycetes ZBW8-1, E14-3, and ZB5-6 have the highest 16s rRNA sequence homology with Glycomyces fuscus TRM 49117(99.86%), Promicromonospora vindobonensis V45T(99.1%), and Nocardiopsis terrae YIM 90022T(99.86%), respectively. In terms of evolutionary relationships, these three strains are also clustered on the same branch as the aforementioned strains. ZBW8-1, E14-3, and ZB5-6 strains all carry the NRPS gene, among which ZBW8-1 and ZB5-6 strains also contain the PKS-Ⅱ gene, and ZBW8-1 strain also contains the PKS-I gene. Analyzing the evolutionary status of these active actinomycetes and identifying their functional genes related to antibiotic synthesis will lay a theoretical foundation for studying the bioactive secondary metabolites in these strains.
Key words:  actinomycete  Pseudomonas syringae pv. actinidiae  antimicrobial activity  phylogenetic tree  functional gene