湘潭锰污染土壤中可培链霉菌多样性及两株新物种的多相分类
发布时间：2018-08-18 21:23
【摘要】：放线菌属于革兰氏阳性的一类原核微生物,它们分布于各种各样的环境。由于产生有益的代谢物,它们在医药、食品、工农业等多个领域上具有巨大的应用价值和潜力,因而其研究得到了研究人员的广泛关注,尤其是链霉菌。据报道,至2010年微生物产生的活性物质已有超过30000种被发现,其中由放线菌产生的具有生物活性的次级代谢产物达到13000多种,链霉菌产生的物质约7600种。本研究从湘潭锰矿污染区(N27°53'-28°03',E112°45'-112°55')及周边环境的土壤分离得到1200多株放线菌,通过结合菌落的培养特征和16S r RNA基因序列,最终得到51株菌落形态、特征不同的链霉菌。由此可知:湘潭锰污染土壤中存在丰富多样的链霉菌。初步的研究结果表明:菌株LUSFXJ,MKLv,MK 44,MK 45和MK47可能为潜在的链霉菌新物种。本研究选取菌株LUSFXJ和MK 44进行了多相分类鉴定,研究结果如下:(1)菌株LUSFXJ,分离自锰矿污染区或周边环境的土壤,16S r RNA基因序列以及孢子形态、结构特征和培养特征均表明:菌株LUSFXJ属于链霉菌。甲基萘醌包含MK-9(H_6),MK-9(H_8),MK-9(H_2)和MK-8(H_8)。磷脂类型包括双磷脂酰甘油(DPG)和一个未鉴定的极性脂。脂肪酸主要成分为iso-C_(1 5:0),anteiso-C_(1 5:0),iso-C_(16:0),C_(16:0)和summed Feature 3。菌株LUSFXJ的细胞壁含有meso-DAP,细胞水解产物含有核糖,甘露糖和葡萄糖。菌株LUSFXJ与进化关系最近的参照菌株的杂交率远远低于70%。根据形态特征、生理生化特征以及DNA的分子杂交结果,确定菌株LUSFXJ是链霉菌的一个新种,并将其命名为Streptomyces(2)菌株MK 44,分离自锰矿污染区的土壤。根据菌落的形态、培养特征、生理生化特征的比较和16S r RNA基因序列分析,我们可以得出:菌株MK 44属于链霉属,同时明显区别于同源性97%有效发表的种。其与选择的菌株S.speciali JCM 16611~T和S.mayteni JCM 16957~T的杂交值分别为30.90±0.28%和29.86±3.51%远远低于Wayne et al(1987年)提出的70%作为新种的界限。因此可以判断该菌为链霉菌的潜在新种,并将其命名为鹤岭链霉菌Streptomyces heling sp.nov.(=GDMCC 4.137~T=KCTC 39920~T)。
[Abstract]:Actinomycetes belong to a class of Gram-positive prokaryotes, which are distributed in various environments. Due to the production of beneficial metabolites, they have great application value and potential in many fields, such as medicine, food, industry and agriculture, so their research has been widely concerned by researchers, especially Streptomyces. It is reported that more than 30000 active substances have been found by microbes in 2010, of which more than 13000 are bioactive secondary metabolites produced by actinomycetes and 7600 by Streptomyces. In this study, more than 1200 strains of actinomycetes were isolated from the contaminated area of Xiangtan Manganese Mine (N27 掳53 ~ 28 掳03 掳E 112 掳45 掳-112 掳55') and the surrounding environment. By combining the culture characteristics of the colony and 16s r RNA gene sequence, 51 strains of Streptomyces with different characteristics were obtained. It can be seen that there are abundant and diverse streptomyces in Xiangtan Manganese contaminated soil. The preliminary results showed that the strains LUSFXJ MKLv4, MK44, MK45 and MK47 might be potential new species of Streptomyces. The results were as follows: (1) strain LUSFXJ, 16s r RNA gene sequence and spore morphology were isolated from manganese ore contaminated area or surrounding environment. The structure and culture characteristics showed that the strain LUSFXJ belonged to Streptomyces. Methyl naphthoquinone contains MK-9 (H6) MK-9 (Hs _ 8), MK-9 (H _ s _ 2) and MK-8 (H _ s _ 8). Phospholipid types include diphosphatidylglycerol (DPG) and an unidentified polar lipid. The main components of fatty acids are iso-C _ (1 5:0) anteiso-C _ (1 5:0), iso-C _ (16:0) C _ (16:0) and summed Feature _ 3. The cell wall of strain LUSFXJ contains meso-DAP.The cell hydrolysate contains ribose, mannose and glucose. The hybridization rate of LUSFXJ with the nearest reference strain was far lower than 70%. According to the morphological, physiological and biochemical characteristics and the results of molecular hybridization of DNA, strain LUSFXJ was identified as a new species of Streptomyces and named Streptomyces (2) strain MK44, which was isolated from the soil of manganese ore contaminated area. According to the comparison of colony morphology, culture characteristics, physiological and biochemical characteristics and the sequence analysis of 16s r RNA gene, we can conclude that strain MK44 belongs to Streptomyces, and is obviously different from the effective species with 97% homology. The crossbreeding values with the selected strains S.speciali JCM 16611T and S.mayteni JCM 16957T were 30.90 卤0.28% and 29.86 卤3.51% respectively, which were much lower than that of Wayne et al (1987 (70%). Therefore, the strain can be identified as a potential new strain of Streptomyces and named Streptomyces Streptomyces heling sp.nov. (GDMCC 4.137~T=KCTC 39920T).
【学位授予单位】：湖南科技大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：X172;Q939.132
本文编号：2190723
[Abstract]:Actinomycetes belong to a class of Gram-positive prokaryotes, which are distributed in various environments. Due to the production of beneficial metabolites, they have great application value and potential in many fields, such as medicine, food, industry and agriculture, so their research has been widely concerned by researchers, especially Streptomyces. It is reported that more than 30000 active substances have been found by microbes in 2010, of which more than 13000 are bioactive secondary metabolites produced by actinomycetes and 7600 by Streptomyces. In this study, more than 1200 strains of actinomycetes were isolated from the contaminated area of Xiangtan Manganese Mine (N27 掳53 ~ 28 掳03 掳E 112 掳45 掳-112 掳55') and the surrounding environment. By combining the culture characteristics of the colony and 16s r RNA gene sequence, 51 strains of Streptomyces with different characteristics were obtained. It can be seen that there are abundant and diverse streptomyces in Xiangtan Manganese contaminated soil. The preliminary results showed that the strains LUSFXJ MKLv4, MK44, MK45 and MK47 might be potential new species of Streptomyces. The results were as follows: (1) strain LUSFXJ, 16s r RNA gene sequence and spore morphology were isolated from manganese ore contaminated area or surrounding environment. The structure and culture characteristics showed that the strain LUSFXJ belonged to Streptomyces. Methyl naphthoquinone contains MK-9 (H6) MK-9 (Hs _ 8), MK-9 (H _ s _ 2) and MK-8 (H _ s _ 8). Phospholipid types include diphosphatidylglycerol (DPG) and an unidentified polar lipid. The main components of fatty acids are iso-C _ (1 5:0) anteiso-C _ (1 5:0), iso-C _ (16:0) C _ (16:0) and summed Feature _ 3. The cell wall of strain LUSFXJ contains meso-DAP.The cell hydrolysate contains ribose, mannose and glucose. The hybridization rate of LUSFXJ with the nearest reference strain was far lower than 70%. According to the morphological, physiological and biochemical characteristics and the results of molecular hybridization of DNA, strain LUSFXJ was identified as a new species of Streptomyces and named Streptomyces (2) strain MK44, which was isolated from the soil of manganese ore contaminated area. According to the comparison of colony morphology, culture characteristics, physiological and biochemical characteristics and the sequence analysis of 16s r RNA gene, we can conclude that strain MK44 belongs to Streptomyces, and is obviously different from the effective species with 97% homology. The crossbreeding values with the selected strains S.speciali JCM 16611T and S.mayteni JCM 16957T were 30.90 卤0.28% and 29.86 卤3.51% respectively, which were much lower than that of Wayne et al (1987 (70%). Therefore, the strain can be identified as a potential new strain of Streptomyces and named Streptomyces Streptomyces heling sp.nov. (GDMCC 4.137~T=KCTC 39920T).
【学位授予单位】：湖南科技大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：X172;Q939.132
【参考文献】
相关期刊论文 前10条
1 林杨;王德明;文仕知;;湘潭锰矿重金属污染湿地修复研究[J];环境科学与技术;2014年05期
2 全哲学;;追寻被“遗漏”的微生物[J];微生物学通报;2013年01期
3 王保军;刘双江;;环境微生物培养新技术的研究进展[J];微生物学通报;2013年01期
4 云锟;吴德超;翁建兵;朱佳文;;湖南湘潭锰矿区蔬菜及菜园土重金属污染研究[J];四川环境;2012年06期
5 唐静;林雁冰;段晓丹;梁建强;韦革宏;;西安市北郊污灌区土壤中抗铅链霉菌的多样性分析[J];农业环境科学学报;2009年06期
6 李军;刘云国;彭晖冰;;锰矿废弃地重金属污染土壤的评价及修复措施探讨[J];环境保护科学;2009年02期
7 刘云国;周海舟;冯宝莹;樊霆;潘翠;;锰污染稻田土壤微生物群落结构和遗传多样性研究[J];湖南文理学院学报(自然科学版);2007年04期
8 闫文德;向建林;田大伦;;湖南湘潭矿业废弃地土壤特性研究[J];林业科学;2006年04期
9 姜怡;段淑蓉;唐蜀昆;陈华红;李文均;徐丽华;;稀有放线菌分离方法[J];微生物学通报;2006年01期
10 傅丰海;;湘潭锰矿的酸性水污染及其防治对策[J];湖南地质;1993年04期
相关博士学位论文 前1条
1 林雁冰;金属矿区及污水灌溉区抗重金属放线菌的筛选及吸附机理研究[D];西北农林科技大学;2010年
相关硕士学位论文 前6条
1 胡火;不同地点红树林土壤链霉菌的地理分布及新种鉴定[D];华中农业大学;2011年
2 纪翠平;非培养方法研究邯邢铁矿矿区放线菌多样性[D];河北大学;2010年
3 唐清畅;锰矿尾渣污染土壤的植物修复以及土壤改良研究[D];湖南大学;2009年
4 武丽花;植被修复对湘潭锰矿矿区废弃地土壤环境效应的影响[D];中南林业科技大学;2008年
5 周海舟;应用PCR-DGGE技术研究锰污染农田土壤微生物群落遗传多样性[D];湖南大学;2008年
6 谢荣秀;湘潭锰矿废弃地土壤退化及其植被恢复的研究[D];中南林学院;2005年
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