强力霉素抗性基因目标供体菌、受体菌和接合子的筛选及鉴定

目的为进一步研究环境中病原微生物目标供体菌强力霉素抗性基因(Dox)水平传播机制奠定基础。方法以强力霉素抗性基因为筛选指标,通过药敏试验从7个菌株中筛选出遗传型分别为Dox~RX~S的候选供体菌和Dox~SX~R的候选受体菌(X代表不是强力霉素的抗生素),将供、受体菌共培养后筛选出遗传型为Dox~RX~R的接合子。分别测定X对目标供体菌、Dox对目标受体菌的最小抑菌浓度。对目标供、受体菌从形态学、生理生化、分子生物学和BIOLOG方面进行鉴定。结果研究的7个病原菌均为多重耐药,最多能耐受15种抗生素,对链霉素、萘啶酮酸和磺胺二甲嘧啶的耐药率最高(100%),对庆大霉素、诺氟沙星和环丙沙星的耐药率最低(0%)。目标供体菌、受体菌和接合子Con-Ⅱ基因型分别为Dox~RKan~S、Dox~SKan~R和Dox~RKan~R。Kan对目标供体菌的最小抑菌浓度为0.310 0μg/mL,Dox对目标受体菌的最小抑菌浓度为0.048 8μg/mL。目标供体菌(Dox~RKan~S)鉴定为Escherichia coli O157:H7(E.coli O157:H7),目标受体菌TR-M30-1(Dox~SKan~R)鉴定为产酸克雷伯菌。结论目标供体菌为E.coli O157:H7(Dox~RKan~S),目标受体菌为TR-M30-1(Dox~SKan~R),接合子为Con-Ⅱ(Dox~RKan~R)。     

基于BIOLOG技术分析氮沉降和降水对土壤微生物功能多样性的影响

土壤微生物是有机物分解和养分循环的主要介质,因此在维持土壤的功能多样性和持续性方面发挥着关键作用。气候变化驱动因素会影响土壤微生物的生理活动,引起其群落结构和功能多样性的改变,并对生物地球化学循环和气候―生态系统反馈产生连锁效应,其中氮沉降和降水是全球气候变化的研究热点。土壤氮(N)的有效性有可能通过改变微生物的群落组成以调节微生物对降水变化的响应,但目前关于N沉降和降水及其交互作用对土壤微生物群落功能多样性的影响机制仍不清楚。为了准确预测未来气候条件下生态系统的功能状况,需要更好地了解土壤微生物对环境变化的响应。基于BIOLOG技术综述了氮沉降和降水变化及其交互作用对土壤微生物功能多样性影响的相关研究进展,可以为进一步研究全球气候变化背景下地下生态学的发展提供参考。另外,分析阐述了当前工作中存在的一些主要瓶颈,并对未来的研究热点进行了探讨和展望。     

Genetic and physiological relatedness of antagonistic Trichoderma isolates against soil borne plant pathogenic fungi

In this study, the in vitro potential of 42 Trichoderma spp. were evaluated against four isolates of soil borne phytopathogenic fungi viz., Rhizoctonia solani, Macrophomina sp., Sclerotium rolfsii and Pythium aphanidermatum in dual culture techniques and through production of volatile and non-volatile inhibitors. In vitro screening results showed that the proportion of isolates with antagonistic activities was highest for the S. rolfsii followed by R. solani, Macrophomina sp. and P. aphanidermatum, respectively. The isolates TNT1, TNP2 and TWP1 showed consistent results in volatile and non-volatile activity in vitro against any of the two pathogens tested. Based on genomic finger prints, potential isolates showed no particular correlation between the origin of the isolates and the Random Amplified Polymorphic DNA (RAPD) groups could not be established. However, the polymorphism shown by the isolates did not correlate to their level of antagonism. Whereas, in physiology studies using BIOLOG (microbial identification system), three groups were formed, one group consists with 14 different Trichoderma species and two groups with two isolates each comprised of only T. koningii and T. viride.     

Impact of allelopathic rice seedlings on rhizospheric microbial populations and their functional diversity

Field performance of rice allelopathic potential is indirectly regulated by the microflora in the rhizosphere. The present study aimed to investigate the dynamics of microbial populations and their functional diversities in the seedling rhizospheres of rice cultivars with varied allelopathic activities by employing agar plate bioassay, fumigation and BIOLOG analysis. Rice cultivars significantly affected the microbial carbon content in their associated rhizospheric soil. The microbial carbon contents were ranked in a decreasing order as Iguape Cateto (441.0 mg·kg^–1) > IAC47 (389.7 mg·kg^–1) > PI312777 (333.2 mg·kg^–1) > Lemont (283.8 mg·kg^–1) with the nil-rice control soil of 129.3 mg·kg^–1. Similarly, the respiration rate of the soils was 1.404, 1.019, 0.671 and 0.488 μgC·g^–1· h^–1 for PI312777, Iguape Cateto, IAC47 and Lemont, respectively. The respiration rate was only 0.304 μ gC·g^–1·h^–1 for the control soil. The microbial flora in the rhizospheric soil of different rice cultivars was dominated by bacteria (58.4%–65.6%), followed by actinomycete (32.2%–39.4%) and fungi (2.2%–2.8%). BIOLOG analysis showed that the value of Average Well Color Development (AWCD) differed significantly among rice cultivars. It was always the highest in the rhizospheric soil of the strongly allelopathic rice cv. PI312777, and the lowest in the rhizospheric soil of the poorly allelopathic rice cv. Lemont. The AWCD value reached the maximum in all the sampled soils after 144 hours of incubation. The AWCD values from the rhizospheric soils of PI312777, IAC47, Iguape Cateto and Lemont were 1.89, 1.79, 1.60 and 1.43 times higher than that of the control soil. Principal Component Analysis (PCA) identified 3 principal component factors (PCF) in relation to carbon sources, accounting for 70.1%, 11.3% and 7.0% of the variation, respectively. 19 categories of carbon sources were significantly positively correlated to the 3 principal components. Phenolic acids, carbohydrates, amino acids and amides were significantly correlated to the principal component 1, phenolic acids, carbohydrates and fatty acids to the principal component 2, and carbohydrates and hydroxylic acids to the principal component 3. Amino acids and amides were the two main carbon sources separating the 3 principal component factors. In addition, the total microbial population in the rhizospheric soil was significantly positively correlated with AWCD, microbial biomass carbon, microbial respiration and Shannon index. There was a significantly positive correlation between the total microbial population and the inhibition rate (IR) on the root length of lettuce owing to the different allelopathic activities of the rice cultivars. These results suggest that changes in microbial population, activity and functional diversity in the rhizospheres are highly cultivar-dependent. These changes might play an important role in governing the rice allelopathic activity in the field.     

新疆地震断裂带次生植物根际土壤微生物碳源利用

利用BIOLOG技术研究了新疆富蕴地震断裂带6种次生植物对根际土壤微生物碳源利用的影响.结果表明:多数植物根际土壤养分显著高于对照.6种根际土壤的平均颜色变化率差异显著,且均高于对照.次生植物不影响根际土壤微生物碳源利用的丰富度,但改变了其优势度和均匀度.不同处理根际土壤微生物碳源利用存在差异,主要体现在对糖类、氨基酸类、羧酸类的利用上.断裂带上次生植物的出现使土壤微生物利用碳源的类型由酚类向糖类、羧酸类转变.土壤速效钾含量与土壤微生物对聚合物(r=-0.84)、胺类(r=-0.83)的利用呈负相关.新疆地震断裂带的次生植物能显著增强土壤微生物的碳源利用能力,改变碳源利用类型.密刺蔷薇和蓍在提高土壤微生物碳源利用能力和改善土壤养分方面效果最佳.     

Phylogenetic and functional diversity of bacterioplankton during Alexandrium spp. blooms

The phylogenetic and functional diversity of the bacterioplankton assemblage associated with blooms of toxic Alexandrium spp. was studied in three harbours of the NW Mediterranean. Denaturing gradient gel electrophoresis and DNA sequence analysis revealed the presence of a bacterium within the Roseobacter clade related to the presence of Alexandrium cells. Phylogenetic diversity was affected by the presence of Alexandrium spp., geographic situation and seasonality. In contrast, functional diversity, assessed with Biolog plates, was clearly affected by seasonality, but not by the presence of Alexandrium, indicating that the presence of the bacterium associated with the blooms was not enough to modify the metabolic pattern of the bacterioplankton assemblage.     

Cultivable bacterial composition and BIOLOG catabolic diversity of biofilm communities developed on Phragmites australis

Biofilm samples formed on submerged young and old stems of reed, Phragmites australis (Cav.) Trin ex Steudel were taken during summer at different sites of Lake Velencei, Hungary. BIOLOG GN microplates were used to analyze the patterns of sole carbon source utilizations by microbial communities. From the carbon sources, carbohydrates and amino acids were preferred by all microbial communities. In the case of the old reed stem samples, higher number of carbohydrates, carboxylic acids and polymers were used than in young samples. Biofilm bacterial communities from the old reed samples of the nature conservation area of the lake used the highest number of (≥50% of the available) substrates. In principal component analysis (PCA), the metabolic potential of the microbial communities from the middle open water region of the lake showed the smallest variability. The variability within metabolic potential of the reed stem microbial communities from a given sampling site was the largest in the case of samples originating from the western, reed-covered nature conservation area. A total of 251 bacterial isolates obtained after serial dilutions and plating onto different media were characterized by traditional phenotypic tests. The strains showed high activities mainly in the hydrolysis of certain biopolymers (gelatine and casein). PCA was used to evaluate the phenotypic variability of strain groups of different sampling sites. The two open water regions were similar to each other, and separated from the western reed covered part of the lake. Similarly to the BIOLOG community-level physiological profiles, strain groups of the young and old reed stem samples originating from the nature conservation area had the largest metabolic potential. On the basis of 16S rDNA sequence analysis, 23 representative strains with different ARDRA patterns were identified. The cultivation-based investigations of bacterial diversity showed characteristic differences in the number of identified taxa in connection with the sampling sites. No characteristic differences could be observed according to medium or sample type (young, first year and more than 1-year old stems) among the identified species. 16S rDNA sequence comparisons resulted in the identification of the genera Aureobacterium, Arthrobacter, Kocuria, Microbacterium, Micrococcus, Rhodococcus, Bacillus, Marinibacillus, Rhodobacter, Defluvibacter, Pseudomonas, Klebsiella, Serratia and Aeromonas. The results of the cultivation-based and BIOLOG investigations revealed characteristic differences in the bacterial community composition and activities of the open water region and the reed covered nature conservation part of the lake.     

不同森林恢复类型对土壤微生物群落的影响

为了评价不同森林恢复类型与方式对南方红壤丘陵区退化生态系统土壤微生物群落的影响,借助氯仿熏蒸法、平板涂抹法和BIOLOG检测法,比较研究了4种森林恢复类型土壤微生物的群落特征．结果表明,4种森林恢复类型土壤微生物生物量碳、细菌数量差异显著,2项指标均以天然次生林土壤最高,人工林次之,荒地最差;碳源平均颜色变化率(AWCD法)和微生物代谢多样性指数(丰富度和多样性)在5种植被类型的土壤中也有明显差异,其趋势与微生物量碳、细菌数量基本相同;天然次生林土壤微生物群落利用碳源的整体能力和功能多样性比人工林和荒地强．相关分析表明,0～20和20～40cm土壤微生物的代谢多样性与根系生物量紧密相关(r=0．933,P<0．05;r=0．925,P<0．05)．自然恢复更有利于改善土壤微生物的结构和功能．     

Functional succession of bacterioplankton on the basis of carbon source utilization ability by BIOLOG plates

Changes of bacterioplankton diversity in lake water were followed in triplicate, continuous-flow experimental tanks. Most probable numbers (MPN) were obtained for 95 different carbon sources using BIOLOG plates and were used to characterize bacterioplankton diversity. During 70 days of incubation, MPN declined for 15 of the 95 substrates while three of 95 appeared to be newly used, indicating functional succession in the bacterioplankton. Total bacterial cell abundance was constant from day 7 to day 70 of the incubation period. The succession of species composition of phyto- and zooplankton was also observed and suggested some involvement by phyto- and zooplankton species in the changes of bacterioplankton diversity. Thus, BIOLOG-based MPN assays is a simple but sensitive method for characterizing the changes in the bacterioplankton carbon utilization profile and is also useful for tracing the functional succession of bacterioplankton diversity within a community.     

Characterization of halophiles isolated from solar salterns in Baja California, Mexico

Solar salterns are extreme hypersaline environments that are five to ten times saltier than seawater (150–300 g L⁻¹ salt concentration) and typically contain high numbers of halophiles adapted to tolerate such extreme hypersalinity. Thirty-five halophile cultures of both Bacteria and Archaea were isolated from the Exportadora de Sal saltworks in Guerrero Negro, Baja California, Mexico. 16S rRNA sequence analysis showed that these cultured isolates included members belonging to the Halorubrum, Haloarcula, Halomonas, Halovibrio, Salicola, and Salinibacter genera and what may represent a new archaeal genus. For the first time, metabolic substrate usage of halophile isolates was evaluated using the non-colorimetric BIOLOG Phenotype MicroArray™ plates. Unique carbon substrate usage profiles were observed, even for closely related Halorubrum species, with bacterial isolates using more substrates than archaeal cultures. Characterization of these isolates also included morphology and pigmentation analyses, as well as salinity tolerance over a range of 50–300 g L⁻¹ salt concentration. Salinity optima varied between 50 and 250 g L⁻¹ and doubling times varied between 1 and 12 h. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.