Two novel strains, SL014B61AT and SL014B11A, were isolated from an oil-polluted saline soil from Gudao in the coastal Shengli Oilfield, eastern China. Cells of strains SL014B61AT and SL014B11A were motile, Gram-negative and rod-shaped. Growth occurred at NaCl concentrations of between 0 and 15% and at temperatures of between 10 and 45 6C. Strain SL014B61AT had Q9 as the major respiratory quinone and C16: 0 (21.2%), C18: 1v9c (20.3%), C16: 1v7c (7.3%) and C16: 1v9c (6.4%) as predominant fatty acids. The G+C content of the DNA was 57.9 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain SL014B61AT belonged to the genus Marinobacter in the class Gammaproteobacteria. Strain SL014B61AT showed the highest 16S rRNA gene sequence similarity with Marinobacter bryozoorum (97.9%) and showed 97.8% sequence similarity to Marinobacter lipolyticus. DNA–DNA relatedness to the reference strains Marinobacter bryozoorum and Marinobacter lipolyticus was 35.5% and 33.8%, respectively. On the basis of these data, it is proposed that strains SL014B61AT and SL014B11A represent a novel species, Marinobacter gudaonensis sp. nov. The type strain is strain SL014B61AT (=DSM 18066T=LMG 23509T=CGMCC 1.6294T).

A thermophilic, methylotrophic methanogen, strain ZC-1T, was isolated from the Shengli oilfield, China. Cells of strain ZC-1T were motile cocci, 0.7-1.0 mm in diameter and always occurred in clusters of two to four cells. Lysis-susceptibility experiments and analysis of transmission electron micrographs of strain ZC-1T suggested the presence of a proteinaceous cell wall. Strain ZC-1T used methanol, methylamine and trimethylamine as substrates for methanogenesis. Optimal growth, with a doubling time of around 5 h, occurred at pH 6.0-6.5, 65 6C, 0.3-0.5 M NaCl and 0.05-0.20 M MgCl2. The DNA G+C content of this organism was 56 mol%. Analysis of 16S rRNA gene sequence and the inferred amino acid sequence of the mcrA gene of strain ZC-1T indicated that it is related specifically to members of the family Methanosaetaceae (90.6 and 76.6% sequence similarity, respectively). However, strain ZC-1T failed to grow with acetate as substrate for methanogenesis, which is a special characteristic of the family Methanosaetaceae. Based on these phenotypic and phylogenic characteristics, strain ZC-1T is proposed to represent a novel genus and species, for which the name Methermicoccus shengliensis gen. nov., sp. nov. is proposed. The type strain is ZC-1T (5CGMCC 1.5056T5DSM 18856T). Methermicoccaceae fam. nov. is also proposed.

末端限制性酶切片段长度多态性分析（terminal restriction hagment length polymorphism，T-RFLP）是近年来发展起来的、不依赖于培养的微生物群落分析方法之一。由于其在微生物群落结构分析方面的特点，包括分辨率高、易于实现自动化及互联网海量数据共享等优势，自1997年最先被报道以来得到了广泛的应用，成为环境微生物群落分析的最强有力的工具之一。本文详细介绍了T-RFLP技术的原理，并从环境样品群落DNA的提取、引物设计和PCR扩增、限制性酶切、电泳分离检测和T-RFLP图谱解析等5个方面讨论了用该技术解析环境微生物群落的方法和技巧，简述了近8a来国外T_RFLP技术在群落分析中的研究进展。类似于其他的分子微生物生态学技术，T-RFLP也有自身的缺陷，因此重点分析了该技术的局限性及相应的解决办法。

A Gram-negative, non-motile, rod-shaped bacterium, strain SS011B1-20T, was isolated from sediments of the South China Sea. Growth occurred at NaCl concentrations between 0 and 10% and at temperatures between 10 and 37 6C. Strain SS011B1-20T contained Q-10 as the major respiratory quinone and C18: 1v7c (81.2%), C16: 0 (7.0%) and C18: 1 methyl (4.3%) as the predominant fatty acids. The G+C content of the genomic DNA was 64.7 mol%. A phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain SS011B1-20T belonged to a clade within the genus Oceanicola in the Alphaproteobacteria, the highest sequence similarities being found with respect to Oceanicola batsensis (96.3%) and with Oceanicola granulosus (94.9%). Strain SS011B1-20T could be clearly distinguished from other Oceanicola species on the basis of the genotypic, phenotypic and phylogenetic data. Thus, it is proposed that strain SS011B1-20T represents a novel species of the genus Oceanicola, with the name Oceanicola nanhaiensis sp. nov. The type strain is SS011B1-20T (=LMG 23508T=CGMCC 1.6293T).

目前自然界中只有极少部分微生物能够得到培养，严重阻碍了对微生物生命活动规律的研究和微生物资源的开发。改进传统培养方法，采用新型培养技术，提高微生物可培养性，大量培养自然界中存在的微生物，从而更全面、准确地了解微生物细胞的生命规律、获悉微生物群落中各种微生物之间的动态相互作用和相互协调的规律，对环境微生物工艺进行准确地设计、精细地调控和高效地利用。简要介绍了微生物不可培养的原因，系统总结了有关提高微生物可培养性方法的最新研究进展，提出研究中存在的问题，并阐述了模拟自然环境条件、强调微生物相互关系是提高微生物可培养性的关键。