土木香内生细菌的分离鉴定及抑菌活性检测
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摘要
利用组织分离法从用药植物土木香根、茎和叶中分离得到内生细菌32株.采用纸片法对菌株进行了抑菌活性研究,并运用生理生化与分子生物学方法进行种类鉴定.抑菌试验结果表明,10株内生菌发酵液对大肠杆菌、枯草芽孢杆菌以及禽多杀性巴氏杆菌均有不同程度的抑菌活性.鉴定结果表明,菌株YIR-5,YIR-7,YIR-8,YIR-9为假单胞菌属菌种,菌株YIR-3,YIR-6为沙雷氏菌,菌株YIR-4,YIS-1,YIS-2分别为水生拉恩氏菌、薄壁芽孢杆菌和蜡样芽孢杆菌,而菌株YIL-3归属于芽孢杆菌属.土木香内生细菌组成丰富,莫海威芽孢杆菌(Bacillus mojavensis)YIS-1对大肠杆菌、枯草芽孢杆菌和禽多杀性巴氏杆菌的抑菌活性最强.
Using the tissue separation method, 32 endophytic bacteria were isolated from the roots, stems and leaves of Inula helenium L., and antibacterial activiy of these strains were determined by using the agar disc diffusion test. Identification of these strains was determined by their morphological, physiological-biochemical properties and phylogenetic analysis based on 16 S rRNA sequences. The results showed that the 10 endophytic bacteria had remarkable antibacterial activities to Escherichia coli, Bacillus subtilis and Pasteurella multocida. The endophytic strains YIR-5, YIR-7, YIR-8 and YIR-9 were classified as genus Pseudomonas, strains YIR-3 and YIR-6 as Serratia plymuthica, strains YIR-4, YIS-1 and YIS-2 as Rahnella aquatilis, Bacillus halotolerans and Bacillus cereus respectively, and the strain YIL-3 as the genus Bacillus. There are a variety of endophytic bacteria from Inula helenium L., and Bacillus mojavensis YIS-1 has the greatest antibacterial activity for escherichia coli, bacillus subtilis, and avian pasteurella multocida.
Using the tissue separation method, 32 endophytic bacteria were isolated from the roots, stems and leaves of Inula helenium L., and antibacterial activiy of these strains were determined by using the agar disc diffusion test. Identification of these strains was determined by their morphological, physiological-biochemical properties and phylogenetic analysis based on 16 S rRNA sequences. The results showed that the 10 endophytic bacteria had remarkable antibacterial activities to Escherichia coli, Bacillus subtilis and Pasteurella multocida. The endophytic strains YIR-5, YIR-7, YIR-8 and YIR-9 were classified as genus Pseudomonas, strains YIR-3 and YIR-6 as Serratia plymuthica, strains YIR-4, YIS-1 and YIS-2 as Rahnella aquatilis, Bacillus halotolerans and Bacillus cereus respectively, and the strain YIL-3 as the genus Bacillus. There are a variety of endophytic bacteria from Inula helenium L., and Bacillus mojavensis YIS-1 has the greatest antibacterial activity for escherichia coli, bacillus subtilis, and avian pasteurella multocida.
引文
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[3] BALLIO A,BOSSA F,DIGIOGIO P,et al.Sructure of the Pseudomycins,New Lipodepsipeptides Produced by Pseudomonas Syringae MSU16H[J].FEBS Letter,1994,355:96-100.
[4] MILLER C M,MILLER R V,GARTON-KENNY D,et al.Ecomycins,Unique Antimycotics from Pseudomonas Viridiflava[J].Journal of Applied Microbiology,1998,84(6):937-944.
[5] 国家药典委员会.中华人民共和国药典:一部[M].北京:中国医药科技出版社,2015:253.
[6] HUO Y,SHI H,LI W,et al.HPLC Determination and NMR Structural Elucidation of Sesquiterpene Lactones in Lnula Helenium[J].Journal of Pharmaceutical and Biomedical Analysis,2010,51(4):942-946.
[7] 张嫚丽,霍长虹,刘丽,等.土木香药材HPLC指纹图谱研究及土木香内脂和异土木香内酯测定[J].中草药,2010,41(9):15-39.
[8] DORN D C,ALEXENIZER M,HENGSTLER J G,et al.Tumor Cell Specific Toxicity of Lnula Helenium Extracts[J].Phytotherapy Research,2006,20(11):970-980.
[9] LI Y,NI ZY,ZHU MC,et al.Antitumour Activities of Sesquiterpene Lactones from Lnula Helenium and Lnula Japonica[J].Zeitschrift Für Naturforschung C,2012,67(7/8):375-380.
[10] HUO Y,SHI H M,WANG M Y,et al.Chemical Constituents and Pharmacological Properties of Radix Inulae[J].Die Pharmazie,2008,63(10):699-703.
[11] RHOADES K R,RIMLER R B.Capsular Groups of Pasteurella Multocida Isolated from Avian Hosts[J].Avian Diseases,1987,31(4):895-898.
[12] MYINT A,CARTER G R,JONES T O.Prevention of Experimental Haemorrhagic Septicaemia with a Live Vaccine[J].The Veterinary Record,1987,120(2):500-501.
[13] AVAKIAN A P,DICK J W,DERIEUX W T.Fowl Cholera Immunity Induced by Various Vaccines in Broiler Minibreeder Chickens Determined by Enzyme-Linked Immunosorbent Assay[J].Avian Diseases,1994,33(1):97-102.
[14] 东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001.