小单孢菌属和拟诺卡氏菌属.ppt

稀有放线菌的分离与培养 报告人：张晓敏 报 告 内 容 分离培养基的选择 小单孢菌形态与生理生化特征 拟诺卡氏菌形态与生理生化特征 小单孢菌、拟诺卡氏菌与链霉菌区别 分离培养基的选择 淀粉-酪蛋白硝酸盐琼脂培养基 溶胶几丁质琼脂培养基 甘露醇-利福平琼脂培养基 淀粉-酪蛋白硝酸盐琼脂培养基 starch-casein nitrate agar ： starch 10g ，sodium caseinate 0.3g，KNO3 2g，agar 15g， PH 7.0-7.4. 37 μg novobiocin，223μg mycostatin 30 ℃ 培养 21天 Micromonospora chaiyaphumensis sp. nov Micromonospora eburnea sp. nov 溶胶几丁质琼脂培养基 基础培养基：K2HPO4 0.7g，KH2PO4 0.3 g，MgSO4·7H2O 0.5g，FeSO4·7H2O 0.01g，ZnSO4 0.001g，琼脂10 克，蒸馏水1.0L.PH7.0 121 ℃ 蒸汽灭菌20 min,每100 mL加20 mL胶体几丁质悬浮液和5 mg放线菌酮 甘露醇-利福平琼脂培养基 mannitol-rifampicin agar ： 甘露醇 10g，酵母提取物 7g，酪蛋白水解物 2g，蛋白胨 1g， NaCl 1 g， CaCO3 0.2 g，制霉菌素 100mg,利福平 5 mg 27℃ 培养6周 Nocardia acidivoran Micromonospora gyrB基因序列分析 和 DNA–DNA 杂交 15 species: Micromonospora aurantiaca, Micromonospora carbonacea, Micromonospora chalcea, M. chersina, Micromonospora coerulea, Micromonospora echinospora, Micromonospora gallica, Micromonospora halophytica, Micromonospora inositola, M. matsumotoense, Micromonospora nigra, M. olivasterospora, Micromonospora pallida, Micromonospora purpureochromogenes and M. rosaria. 小单孢菌形态与生理生化特征 Aerobic, Gram-positive, mesophilic, non-motile actinomycete well-developed and branched substrate hyphae. Aerial mycelium was absent. Spore chains were absent. Mycolic acids were absent. Acyl type of cell wall muramic acid is glycolyl. Fatty-acid type:3b. The predominant menaquinone: MK-9（H4）、MK-9（H4H6H8） Characteristic whole-cell sugars are xylose and arabinose. Phospholipids :P-II型 . Cell-wall peptidoglycan :CW-II (glutamic acid,alanine and meso-diaminopimelic acid). DNA G+C(mol%): 71%-73% . well-developed and branched substrate hyphae. Single, non-motile spores are formed on substrate hyphae ，0.45 mm in diameter. Aerial mycelium is absent. The spore- surface appears rough and nodular. Colonies are yellowish white and turn greyish black after sporula