凤凰单丛茶内生拮抗细菌的筛选与鉴定
Isolation and identification of endophytic bacteria from Fenghuang dancong tea
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摘要: 从潮州凤凰单丛茶树中分离筛选具有抑菌活性的内生细菌,为新型生物抗菌剂的研究开发提供理论基础。对凤凰单丛茶树的内生细菌进行分离纯化,采用双层平板法筛选对供试指示细菌具有抑制作用的菌株,平板对峙法筛选对供试植物病原菌具有抑制作用的菌株,并对拮抗细菌进行形态特征观察、生理生化试验及16S r DNA鉴定。结果表明:凤凰单丛茶树中共分离出内生细菌9株,其中FH01菌株抑菌效果最好,且对供试菌的抑菌谱广,对枯草芽孢杆菌、金黄色葡萄球菌的抑菌圈直径分别为14.92 mm和12.36 mm,对梨果黑斑病菌、核桃叶枯病菌、红枣黑斑病菌、立枯丝核菌的菌丝抑制率分别为82.10%、81.52%、75.06%、67.10%。经形态学、生理生化试验及16S r DNA鉴定,内生拮抗细菌FH01为甲基营养型芽孢杆菌(Bacillus methylotrophicus strain)。Abstract: The aim of this study was to isolate endophytic bacteria and screen endophytic antagonistic bacteria from Fenghuang dangcong tea tree.The study could provide theoretical basis for research and development of new biological antibacterial agent. The endophytic bacteria were isolated and purified from Fenghuang dangcong tea tree.The experiments of inhibition activity of endophytic bacteria against tested bacteria by double- plate method and against tested plant pathogenic fungi by flat- stand method were conducted. The endophytic bacteria were identified by morphology,physiological- biochemical experiments and the sequence of 16 S r DNA. The results showed that the total of 9 endophytic bacteria were isolated.The endophytic antagonistic bacterium FH01 showed obvious inhibition activity and had a wide antimicrobial spectrum to the tested germs,and the diameters of inhibition zones of FH01 strain against Bacillus subtilis and Staphylococcus aureus were 14.92 mm and 12.36 mm respectively.The hyphae inhibition rate of FH01 strain against Alternaria alternate,Sphaerulina juglandis,Alternaria sp and Rhizoctonia solani were 82.10%,81.52%,75.06%,67.10% respectively.The antagonistic bacterium FH01 was identified as Bacillus methylotrophicus strain by physiological- biochemical experiments and the sequence of16 S r DNA.
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