摘要本文报道了从东海海域分离出的以木质素为唯一碳源的菌株MY15的分离鉴定。通过革兰氏染色并用电镜对菌株形态进行观察；采用API20NE非肠道菌鉴定系统对菌株的血清学性状及生理生化特性进行测定；对菌株的16SrDNA基因序列进行PCR扩增和测序，将其与数据库中的菌株进行比对，选取同源性较高的构建了系统发育树。分析表明：菌株MY15为G-菌，直杆状，无芽孢，无鞭毛，营好氧生活；在海水固体营养培养基平板上形成的菌落为白色、圆形、隆起、表面光滑、易挑取；该菌株在35℃、250转/分钟的条件下摇床振荡培养24小时，测得最适pH为7.5，最适盐浓度为25克/升NaCl，在不同温度下培养24小时测得最适温度为35℃；由菌株MY15的系统发育分析可以看出，该菌株与海单胞菌属的亲缘关系最为接近。通过以上鉴定，证实菌株MY15属于海单胞菌属(Marinomonas)。(序列号：KT247807)。50714

关键词: 海单胞菌菌株  木质素  血清学特征  16SrDAN  系统发育树 

Isolation, Identification and Growth Characteristics of a Strain of MY15 with Degrading Lignin

Abstract

This thesis reported the isolation and characterization of a bacterial strain named MY15 that has the ability to use lignin as sole carbon source for growth from the East China Sea. By Gram Staining and Electron Microscope were used to observe the morphology of Strain; Non Enteric Bacteria Identification System to the Strains of Serological Properties and Physiological and Biochemical Characteristics were determined by API20NE; Through the Strains of 16S rDNA Gene Sequence was amplified by PCR and sequenced, Then compare the Strains with Known Strains in the Database,Selected homologous higher to constructed the Phylogenetic Tree.The analysis showed that the Strain MY15 was Gram negative, no bacillus, no flagella, Aerobic life ; In Seawater Solid Nutrient Plate formed colonies are white, round, uplift, smooth surface, easy picking; The Strain under the conditions of 35 ℃ and 250 r / min shaking cultured for 24 h, measured the Optimum pH was 7.5, the Optimum Salt Concentration Degree was 22.5g/L NaCl. Under the different temperature cultured 24h, measured the Optimal temperature is 35℃; Through The Phylogenetic analysis of Strain MY15 , the Strain most closely related to the Marinomonas . By the above identification, the Strain MY15 was identified as Marinomonas. (Serial Number: KT247807).

Key Words: Marinomonas  Lignin  Serological Characteristics  16SrDNA    Phylogenetic Trees

目  录

摘要 I

Abstract II

目  录 III

图清单 IV

表清单 IV

引言 1

1  材料与方法 2

1.1实验材料 2

1.2实验方法 3

2  结果与分析 5

2.1分离筛选结果 5

2.2菌株的形态与菌落特征 5

2.3生理生化特征 6

2.4血清学特征 8

2.5 DNA的(G+C) mol%含量 8

2.6菌株MY15的PCR扩增产物电泳分析及其基因序列 9

2.7系统发育分析 11

3  讨论 12

参考文献 13

致谢