Bacterial diversity in Arctic marine sediment determined by culture-dependent and-independent approaches

Bacterial diversity in surface sediment from the Arctic Ocean was investigated by culture-dependent and-independent approaches. Conventional culture-dependent techniques revealed 11 strains based on their distinct morphological characteristics on marine Zobell 2216E agar plates. Phylogenetic analysis showed that these isolates belonged to three major lineages of the Bacteria, γ-proteobacteria, Bacteroidetes and Actinobacteria, and that they included 10 genera. Most isolates were psychrotrophic, and NaCl was not necessary for their growth. Furthermore, they exhibited activity of at least one extracellular hydrolytic enzyme at 4°C and had various abilities to assimilate carbon sources. A total of 67 phylotypes were detected among 142 clones based on the 16S rRNA library of the total community DNA and grouped into nine major lineages of bacteria. Phylotypes affiliated with γ-, δ- and ε-proteobacteria accounted for 36.7%, 21.8% and 16.9% of the total clones, respectively. The rest of the clones belonged to Bacteroidetes, α-proteobacteria, Actinobacteria, Fusobacteria, Nitrospirae and an unclassified group.     

Response of Microbial Community to Petroleum Stress and Phosphate Dosage in Sediments of Jiaozhou Bay,China

The dynamic microcosms were used to evaluate the effect of oil spills on microbial ecological system in marine sedi- ment and the enhancement of nutrient on the oil removal. The function and structure of microbial community caused by the oil pollu-tion and phosphate dosage were simultaneously monitored by dehydrogenase activity assay and PCR-denaturing gradient gel elec- trophoresis （DGGE） techniques. The results indicated that the amount of total bacteria in all dynamic microcosms declined rapidly with incubation time. The number of petroleum-degrading bacteria and the activity of sediment dehydrogenase were gradually en-hanced by petroleum in the oil-treated microcosms, while they both showed no obvious response to phosphate dosage. In comparison, phosphate spiked heterotrophic bacteria and they showed a significant increase in amount. DGGE profiles indicated that petroleum dosage greatly changed community structure, and the bacteria belonged to class Deltaproteobacteria, and phyla Bacteroidetes and Chlorobi were enriched. This study demonstrated that petroleum input greatly impacted the microbial community structure and con- sequently the marine sediment petroleum-degrading activity was enhanced. Phosphate dosage would multiply heterotrophic bacteria but not significantly enhance the petroleum degradation.     

春季长江口及其邻近海域溶解有机物的分布、组成及降解行为研究

2021年3月对长江口及其邻近海域9个断面溶解氨基酸(THAA)、溶解有机碳(DOC)和溶解有机氮(DON)的分布和组成进行了调查研究。结果表明:THAA的浓度范围为1.24~4.71μmol/L,平均浓度为(2.61±0.85)μmol/L;DOC的浓度范围为118.17~450.58μmol/L,平均浓度为(149.73±33.34)μmol/L;DON的浓度范围为10.48~24.45μmol/L,平均浓度为(13.80±1.81)μmol/L。THAA、DOC和DON的水平分布表现出近岸高于远岸的特点,表明陆源输入对调查区域的THAA等分布有重要影响。而THAA与D-天门冬氨酸、D-谷氨酸、D-丝氨酸和D-丙氨酸的D/L比均呈显著负相关,与天门冬氨酸/β-丙氨酸(Asp/β-Ala)呈正相关,表明细菌活动是影响THAA浓度变化的重要因素。长江口及邻近海域的优势氨基酸为L型氨基酸,D型氨基酸和非蛋白型氨基酸的占比仅为7.34%。基于C/N比值分析,长江口及其邻近海域溶解有机物(DOM)的来源受到长江冲淡水和台湾暖流的共同影响。THAA-C%、THAA-N%、降解因子和反应活性指数表明:水平方向上盐度接近于0的淡水区域和盐度高于34的远海区域降解程度较高;垂直方向上随着深度的增加,有机物的降解程度逐渐升高。调查区域有机物的降解程度主要受到陆源输入和细菌活动的影响。     

米槠天然林土壤真菌对N2O产生的贡献

研究表明亚热带地区森林土壤硝化和反硝化能力都非常低,但却是全球N2O最大的自然源之一,因而推测极有可能存在N2O潜在源未被认识,有迹象表明真菌最有可能是这一潜在源的贡献者,但实际研究证据并不多．为此本研究以中亚热带地区典型常绿阔叶林——武夷山自然保护区米槠天然林土壤为对象,利用室内基质诱导呼吸选择抑制方法,研究在水分60％WHC和温度25℃条件下土壤真菌细菌活性比及其对N2O产生的相对贡献．结果显示真菌和细菌的活性比例分别是0．7±0．3和0．3±0．1,前者显著大于后者（P〈0．05）,真菌与细菌的活性比为2．00±1．00;与对照相比,添加放线菌酮（真菌抑制剂）处理N2O产生速率减少了50．8±11．3％,添加链霉素（细菌抑制剂）处理则减少了43．7±11．8％,前者减少是后者的1．15倍,但差异不显著．研究结果表明该米槠天然林土壤在给定实验条件下真菌活性比细菌的大,但二者对土壤N2O产生的贡献几乎相等．基质诱导呼吸选择抑制方法具有较多的限制条件,若得出可靠结论,还应该利用包括生物化学、分子生物学方法在内的多种方法进行相互验证．同时今后应加强真菌产生N2O途径及机理研究．     

Changes in groundwater bacterial community during cyclic groundwater-table variations

Column experiments containing an aquifer sand were subjected to static and oscillating water tables to investigate the impact of natural fluctuations and rainfall infiltration on the groundwater bacterial community just below the phreatic surface, and its association with the geochemistry. Once the columns were established, the continuously saturated zone was anoxic in all three columns. The rate of soil organic matter (SOM) mineralization was higher when the water table varied cyclically than when it was static due to the greater availability of NO₃⁻ and SO₄²⁻. Natural fluctuations in the water table resulted in a similar NO₃⁻ concentration to that observed with a static water table but the cyclic wetting of the intermittently saturated zone resulted in a higher SO₄²⁻ concentration. Rainfall infiltration induced cyclic water-table variations resulted in a higher NO₃⁻ concentration than those in the other two columns, and a SO₄²⁻ concentration intermediate between those columns. As rainwater infiltration resulted in slow downward displacement of the groundwater, it is inferred that NO₃⁻ and SO₄²⁻ were being mobilized from the vadose zone. NO₃⁻ was mainly released by SOM mineralization (which was enhanced by the infiltration of oxygenated rainwater), but the larger amount of SO₄²⁻ release required a second mechanism (possibly desorption). Different groundwater bacterial communities evolved from initially similar populations due to the different groundwater histories.     

枸杞岛海藻场冬季细菌群落结构分析

从枸杞岛海藻场和对照点贻贝场分别采取表层及底层水样,利用细菌通用引物,提取样品总DNA进行聚合酶链式反应(PCR)扩增,采用16SrDNA文库技术,对其细菌群落进行鉴定和分析。结果表明,海藻场表层和底层细菌群落分为5和6个门,贻贝场表层和底层细菌群落分为3和4个门,海藻场中细菌分类门数多于贻贝场内,优势菌均为变形细菌门,分别占克隆文库的比例为40.7%、50%、59.1%和48.1%。γ-变形菌纲为变形细菌门的优势亚群,各个文库中均含有许多未确定其分类的序列,即含有许多未被认知的微生物类群。变形菌门、厚壁菌门和放线菌门为所有水样共有,但在水层垂直分布及不同地理位置间存在较大差异。不同功能类群可能与其特殊生境密切相关。     

短期原油污染致使冻土活动层细菌群落显著改变——以加格达奇冻土活动层为例

生物降解与土著微生物群落结构、功能及其变化密切相关. 目前, 对于东北冻土土壤中的适冷降解菌了解不足. 新建成的中俄输油管道穿越中国东北的多年冻土区, 为相关研究提供了契机. 实验利用454高通量测序分析了加格达奇冻土活动层土壤在受控原油污染前后的微生物群落结构. 结果显示: 污染后的Proteobacteria和Firmicutes相对丰度显著升高, 优势类群包括Alicyclobacillus、Sphingomonas、Nevskia以及Bacillus. 群落以芳烃降解菌或者耐受油污环境的细菌为主. 这种变化与原油(尤其是芳烃)组分的生态毒害作用有关. 较高浓度的原油污染下, 群落中可耐受油污环境的细菌丰度相对更高.     

基于浒苔暴发海水池塘的微生物生态特征研究

为研究浒苔对海水养殖环境微生物生态的影响,采用Illumia高通量测序技术测定冬季垦区进水区、浒苔暴发的池塘和排水区水体以及底泥中菌群结构,分析进水区、池塘养殖区和排水区的环境微生物结构演变过程。结果表明:（1）γ-变形细菌、α-变形细菌、放线菌、拟杆菌和δ-变形细菌为海水养殖区的优势细菌门类;（2）进水区水体和底泥微生物种类和多样性指数均高于浒苔池塘和排水区;浒苔暴发的池塘和排水区微生物群落结构较为相似;（3）浒苔池塘的水体微生物以异养细菌为主,底泥以脱硫杆菌等厌氧细菌为主,增加了底泥潜在产生硫化氢的危害;（4）进水区水体含有丰度较高的弧菌和鱼立克次氏体等条件致病性病原菌,而浒苔池塘中含量较低,说明浒苔对弧菌等有一定的抑制作用;但浒苔暴发的池塘和排水区中另一种潜在致病菌黄杆菌类的丰度较高,养殖品种务必防患此类病菌感染疾病的发生。本文从微生态学角度全面揭示浒苔对近岸海区、海水池塘和排水区微生态演变过程的影响,发现了浒苔具有抑制和增加不同致病菌的双重作用,这对水产养殖有一定指导,也为浒苔发生原因、预报、预防等研究提供一定理论参考。     

南海北部海水中氨基酸的分布及其对溶解有机物降解行为的指示研究

于2015年6月对南海北部海区5个断面共26个站位海水中溶解态氨基酸（THAA）、溶解有机碳（DOC）和叶绿素a（Chl a）的浓度进行了科学调查。结果表明:夏季南海北部海水中THAA的浓度范围为0.40~1.95 μmol/L,平均值为（0.80±0.40） μmol/L,THAA的水平分布总体上体现出近岸高、远海低的特点,表明陆源输入对南海北部海域表层THAA分布有重要影响。THAA在断面上的垂直分布呈现出由近岸至远岸、由表层至底层逐渐降低的趋势。THAA浓度与两种D型氨基酸（D-谷氨酸:D-Glu和D-丙氨酸:D-Ala）含量之间存在显著负相关性,与天门冬氨酸/β-丙氨酸（Asp/β-Ala）和谷氨酸/γ-氨基丁酸（Glu/γ-Aba）比值之间存在显著正相关性,表明细菌的消耗是影响南海海水中THAA浓度的重要因素。D-Ala作为细菌肽聚糖中相对稳定的氨基酸,根据其占DOC的含量估算南海海水中的细菌源有机碳对DOC的贡献率为（29.32±14.32）%,其水平分布显示出近岸低、远岸高的特点;而其垂直分布则呈现出从表层至底层逐渐增加的趋势。THAA占DOC百分比（THAA-C%）的变化范围为1.02%~5.49%,平均值为（2.97±1.38）%。THAA-C%、活性因子和降解因子的高值均出现在珠江口外围区域。随着海水深度增加3种降解因子的数值均显著降低,这表明底层海水中有机物比表层海水中的有机物降解程度更大。     

Pyrosequencing reveals specific associations of bacterial clades Roseobacter and Flavobacterium with the harmful dinoflagellate Cochlodinium polykrikoides growing in culture

Harmful algal blooms of the dinoflagellate Cochlodinium polykrikoides are of worldwide environmental concern. Previous field studies have demonstrated the association of such algal blooms with free‐living bacteria (FLB) and particle‐associated bacteria (PAB). In the present study, we report evidence for bacterial association with C. polykrikoides in laboratory culture. Sampling was performed at different growth stages (lag, exponential and stationary) and pyrosequencing was used to identify taxa. As a result, the community structures of FLB and PAB were elucidated and significant differences between FLB and PAB were identified by hierarchical cluster analysis and the similarity profiles test (p = .05). The relative abundances of bacterial operational taxonomic units distinctly changed in response to the different phases of C. polykrikoides growth. The Roseobacter genus Marivita and the Flavobacterium genus Winogradskyella were dominant in the FLB and PAB associated with C. polykrikoides, respectively. The Roseobacter clade, which has also exhibited associations with C. polykrikoides blooms in field samples, may influence host cell growth through the provision of vitamins.