新疆尉犁县黑湖嗜盐嗜碱菌的分离及系统发育学分析

Isolation and phylogenetic analysis of haloalkalophilic bacteria in the black lake of Yuli County in Xinjiang

【目的】从新疆尉犁县黑湖中筛选分离获得嗜盐嗜碱微生物,并对筛选获得的微生物进行种属鉴定。【方法】采用传统分离鉴定技术,进行形态和生理生化特性研究和基于16S r RNA基因的序列分析。【结果】从样品中分离获得可培养嗜盐嗜碱菌25株,对其进行鉴定。根据生理生化特征、16S r RNA基因序列测定和系统发育分析表明,25株菌分布在古菌Halorubrum、Haloarcula、Natrialba、Halohasta和Halopiger等5个属。其中优势菌群为Halorubrum,次优势菌群为Natrialba。其中DH-66(KU663028)属于Halopiger属,16S r RNA基因序列同源性与该属的模式菌株Halopiger aswanensis 56T同源性最高,为95.75%,预示为潜在的新种(新种鉴定将另行报道)。25株嗜盐嗜碱菌生长条件实验表明,这些菌适应Na Cl的浓度范围为15%-30%、最适浓度为20%-25%,生长的p H范围为7.0-13.0、最适p H为9.0-10.0。各种水解酶类的分析表明,在分离的25株菌中产淀粉酶的菌有5株占20%、产蛋白酶的菌有4株占16%、产酯酶可水解吐温20的菌有15株占60%、可水解吐温40的有7株占28%、可水解吐温80的有4株占16%、产过氧化氢酶的菌有14株占56%。9株菌同时能产4种酶,2株菌同时能产3种酶。表明了嗜盐嗜碱菌产酶的多样性。19株菌硝酸盐还原为阳性。【结论】揭示了新疆尉犁县黑湖嗜盐嗜碱菌生理生化特性的多样性和系统发育多样性,而且蕴藏着较丰富的新的微生物类群,亟待系统研究和进一步开发利用。

[Objective] Screen and isolate Haloalkalophilic microorganisms from the black lake of Yuli County in Xinjiang and take the species identification analysis. [Methodsl Traditional isolation and identification techniques were used to study the morphological, physiological and biochemical characteristics and sequence analysis based on 16S rRNA gene were proceeded. [Resultsl 25 strains of culturable Haloalkalophilic bacteria were separated and identified from the sample. The physiological and biochemical characteristics, 16S rRNA gene sequence analysis and phylogenetic analysis showed that these microorganisms distributed in five genera which were Halorubrum, Haloarcula, Natrialba, Halohasta and Halopiger etc. Among them, the most dominant bacteria group is Halorubrum, next is Natrialba. Sequence homology of 16S rRNA gene were 95.75% respectively, herald some potential new species （Identification of new species not shown here）, the Growth condition experiment of 25 strains of culturable Haloalkalophilic bacteria show that concentration range of NaC1 was 15%-30%, with optimum concentration was 20%-25%, the pH range for growth was 7.0-13.0, optimum at 9.0-10.0. The results of physical and biochemical characteristics show that among 25 strains of culturable Haloalkalophilic bacteria, 5 strains belong to amylase producing bacteria, accounted for 20%, 4 strains belong to protease producing bacteria, accounted for 16%, 15 strains belong to esterase Twain hydrolysis 20 producing bacteria, accounted for 60%, 7 strains belong to Twain hydrolysis 40 producing bacteria, accounted for 28%, 4 straise belong to Twain hydrolysis 80 producing bacteria, accounted for 16%. 14 strains belong to hydrogen peroxide producing bacteria, accounted for 56%. 9 strains of these bacteria can produce 4 kinds of enzymes, 2 strains can produce 3 kinds of enzymes at the same time, showed the diversity of enzyme production of the alkaline bacteria. 19 strains of the bacteria were positive for nitrate reduction. [Conclusion] The study shows the physiological and biochemical characters diversity and phylogenetic diversity of Haloalkalophilic bacteria in the black lake of Yuli County in Xinjiang, and there are much more abundant new microbial groups in the lake. The study has the regional characteristics and potential application value, recommend that the microbia urgent need systematic research and further develope and utilize.