基于esp基因和JCV标记的MST方法在五大流域典型水源地中的应用

非点源污染目前已经成为影响水体环境的重要污染。微生物源示踪技术(microbial source tracking,MST)是解决非点源污染的一项新技术,它可以确定污染的宿主来源。肠球菌esp基因和多瘤病毒JCV可以作为检测水体中人源粪便污染的分子标记,其灵敏度和特异性都很高。为分析五大流域水源地是否受到人源粪便的污染,对辽河、海河、淮河、长江和黄河五大流域典型水源地水样进行采集和检测,选取人源粪便特异病原微生物肠球菌的esp基因和多瘤病毒JCV建立了相应的MST分子检测方法。结果表明,五大流域的典型水源地采样点均有可能受到了人源粪便的污染,可为当地相关部门提供技术支持和数据参考。     

紫色土丘陵坡地土壤微生物群落的季节变化

采用稀释平板法和Biolog-ECO微平板技术,以剌槐（Robinia pseudoacacia）天然次生林为研究对象,研究湖南省衡阳市紫色土丘陵坡地表层（0~10 cm）土壤微生物群落的季节变化特征。结果表明：1）夏季,土壤微生物总数,细菌数量、真菌数量和放线菌数量最高,分别为16.34×106、16.09×106、10.43×104和14.64×104 cfu·g-1干土,春季次之：11.61×106、11.45×106、5.00×104和10.65×104 cfu·g-1干土,秋季最低：5.87×106、5.78×106、4.67×104和4.08×104 cfu·g-1干土,春、夏和秋3季的差异达显著水平（P<0.05）;2）在培养168 h时,土壤微生物C源平均颜色变化率（Average well color development, AWCD）以夏季最高（1.20）,春季次之（0.88）,秋季最低（0.83）;3）土壤微生物功能多样性表现为夏季明显高于春、秋2季（P<0.05）,夏季的Patrick丰富度指数（R）（28）、Shannon-Wiener指数（H）（3.22）、Simpson指数（D）（0.96）和McIntosh指数（U）（8.20）显著高于春、秋2季（P<0.05）,而春、秋2季的R（23,24）、H（3.06,3.08）、D（0.95,0.95）和U（5.90,5.91）无显著差异（P>0.05）;4）氨基酸类、聚合物类和羧酸C源类是衡阳紫色土丘陵坡地土壤微生物偏好且利用率较高的C源类型;5）主成分分析表明,土壤微生物群落的C源利用可分为2类,一类在夏季,另一类在春、秋2季,其得分系数的分布范围分别为（2.59~6.00,2.43~5.09）和（-7.65~-1.90,-6.38~-3.43）。研究结果为科学评价湖南省衡阳市紫色土丘陵坡地土壤生境质量退化和恢复过程中微生物特征的变化提供了本底值参考。     

湖南石门雄黄矿区矿样中的砷氧化细菌的分离及鉴定

绿色的生物学方法处理含砷废水,对于缓解工业发展所带来砷污染问题,提高居民健康生活质量具有重要意义。以湖南石门雄黄矿区的尾矿池积水（SY）、地下400 m处的矿坑水（NY）和尾矿池底泥表层（XY）3个样本为研究对象,利用细菌富集和多次传代的方法,从这3个环境样本中获得可氧化三价砷的3个菌群。在砷质量浓度为l~5 g·L-1的培养基中分别检测上述3个菌群的耐受能力,发现在砷质量浓度高达5 g·L-1的条件下,上述菌群均能在2~3 d内达到109 cells·mL-1。利用上述菌群处理l g·L-1亚砷酸钠溶液,SY、XY和NY菌群可分别在25、20和35 h内将三价砷完全氧化,并且使溶液中的总砷质量浓度降低66.7%。进一步对3个菌群进行多次平板分离,利用五价砷和硝酸银的颜色反应,获得11株可氧化三价砷的目的菌株。对这些菌株16S rDNA进行NCBI的BLASTN序列比对发现,菌株SMY24、SMY33、SMY22、SMY32、SMY21和SMY31均属于假单胞菌属（Pseudomonas）,菌株SMY104属于不动杆菌属（Acinetobacter）,菌株SMY17、SMY25、SMY9和SMY1在所有的已知序列中,最大的相似率只有91%,最小的相似率为76%,且都是未曾报道纯培养的细菌。对这些菌株三价砷氧化能力进行测定发现,其中菌株SMY104和SMY21在20 h内能将三价砷几乎完全氧化。通过对这些菌株三价砷氧化酶基因进行初步检测,发现了多个菌株具有三价砷氧化酶基因（AoxB）。     

Molecular diversity of arbuscular mycorrhizal fungi at a large-scale antimony mining area in southern China

Arbuscular mycorrhizal fungi (AMF) have great potential for assisting heavy metal hyperaccumulators in the remediation of contaminated soils. However, little information is available about the community composition of AMF under natural conditions in soils contaminated by antimony (Sb). The objective of this study was to investigate the characteristics of AMF molecular diversity, and to explore the effects of Sb content and soil properties on the AMF community structure in an Sb mining area. Four Sb mine spoils and one adjacent reference area were selected from around the Xikuangshan mine in southern China. The association of AMF molecular diversity and community composition with the rhizosphere soils of the dominant plant species was studied by Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis (PCR-DGGE). Results from all five studied sites showed that the diversity of AMF decreased with increasing Sb concentration. Principal component analysis (PCA) indicated that the AMF community structure was markedly different among these groups. Further redundancy analysis (RDA) showed that Sb contaminationwas the dominating factor influencing the AMF community structure in the Sb mine area. However, the multivariate analysis showed that, apart from the soil Sb content, extractable nitrogen content and organic matter content also attributed to AMF sequence distribution type. Some AMF sequences were only found in the highly contaminated area and these might be ideal candidates for improving phytoremediation efficiency in Sb mining regions. Gene sequencing analysis revealed that most species were affiliated with Glomus, suggesting that Glomus was the dominant AMF genus in the studied Sb mining area.     

A screening assay for thyroid hormone signaling disruption based on thyroid hormone-response gene expression analysis in the frog Pelophylax nigromaculatus

Amphibian metamorphosis provides a wonderful model to study the thyroid hormone (TH) signaling disrupting activity of environmental chemicals, with Xenopus laevis as the most commonly used species. This study aimed to establish a rapid and sensitive screening assay based on TH-response gene expression analysis using Pelophylax nigromaculatus, a native frog species distributed widely in East Asia, especially in China. To achieve this, five candidate TH-response genes that were sensitive to T3 induction were chosen as molecular markers, and T3 induction was determined as 0.2 nmol/L T3 exposure for 48 hr. The developed assay can detect the agonistic activity of T3 with a lowest observed effective concentration of 0.001 nmol/L and EC50 at around 0.118–1.229 nmol/L, exhibiting comparable or higher sensitivity than previously reported assays. We further validated the efficiency of the developed assay by detecting the TH signaling disrupting activity of tetrabromobisphenol A (TBBPA), a known TH signaling disruptor. In accordance with previous reports, we found a weak TH agonistic activity for TBBPA in the absence of T3, whereas a TH antagonistic activity was found for TBBPA at higher concentrations in the presence of T3, showing that the P. nigromaculatus assay is effective for detecting TH signaling disrupting activity. Importantly, we observed non-monotonic dose-dependent disrupting activity of TBBPA in the presence of T3, which is difficult to detect with in vitro reporter gene assays. Overall, the developed P. nigromaculatus assay can be used to screen TH signaling disrupting activity of environmental chemicals with high sensitivity.     

Cloning and expression analysis of geosmin synthase gene from Aphanizomenon gracile北大核心CSCD

Geosmin is a secondary metabolite responsible for earthy odors. The occurrence of geosmin has great impact on the quality of water environment. The gene essential for geosmin biosynthesis have been identified in several species. But little is known about the mechanism of geosmin synthesis in Aphanizomenon gracile. This study attempted to clone the gene involved in geosmin biosynthesis of Aphanizomenon gracile a nd a nalyze t he geosmin production u nder d ifferent e nvironments. T he high-efficiency thermal asymmetric interlaced PCR (hiTAIL-PCR) was used to amplify the full-length of geosmin synthase gene from Aphanizomenon gracile (WH-1). Real time PCR (RT-PCR) was applied to quantify the geosmin production in different light and temperature. As a result, geo, a geosmin synthase gene from Aphanizomenon gracile (WH-1) was cloned by hiTAIL-PCR. The full-length open reading frame (ORF) of geo was 2 262 bp, coding for a protein of 753 amino acids. Meanwhile, WH-1 was treated with different environment conditions and mRNA expression levels of geo were quantified by RT-PCR. It was found that low temperature (15 °C), high light intensity (35 μmol m-2 s-1) and continuous light illumination were beneficial to the expression of geo. The successful amplification of geosmin synthase gene verified that hiTAIL-PCR is an effective and simple procedure of low cost. The result provides fundamental knowledge on the monitoring and prevention for odorants.     

黄河三角洲自然保护区秋冬季水鸟群落组成与生境关系分析

于2014年10月至2015年1月采用样线调查法对黄河三角洲自然保护区秋季迁徙期和越冬期水鸟群落结构进行调查研究,调查共包括8条样线40个观测点.结果表明:(1)秋季迁徙期共记录水鸟6目14科54种,国家Ⅰ级保护鸟类5种,国家Ⅱ级保护鸟类8种;越冬期共记录水鸟5目7科31种,国家Ⅰ级保护鸟类1种,国家Ⅱ级保护鸟类4种.(2)2个时期雁形目水鸟种类和数量均占优势,豆雁(Anser fabalis)、斑嘴鸭(Anas poecilorhycha)、赤膀鸭(Anas strepera)和绿头鸭(Anas platyrhynchos)为优势物种,丹顶鹤(Grus japonensis)、东方白鹳(Ciconia boyciana)和大大鹅(Cygnus cygnus)等珍稀水鸟也有一定的种群数量.(3)秋季迁徙期水鸟种数、数量和ShannonWiener多样性指数多于或高于越冬期.(4)5种典型生境之间水鸟种类和数量存在差异,天然水域是水鸟群落的主要分布区.(5)自然生境内的水鸟种数、数量和Shannon-Wiener多样性指数一般多于或高于人工生境.相似性分析结果表明,自然生境之间水鸟群落结构相似程度高于人工生境.     

Influencing diversity beliefs through a personal testimonial,promotion-focused approach

Although researchers have highlighted the importance of diversity beliefs (i.e., team members' perceived value of diversity) for the elaboration of information in teams, little attention has been paid to whether and how diversity beliefs can be shaped. Drawing on theory and research on team diversity beliefs, we propose that diversity beliefs are more effectively influenced by interventions using a promotion (compared with a prevention) focus toward diversity and personal testimonial (compared with factual) knowledge. Results from an experiment conducted with 175 teams revealed that both a promotion focus and personal testimonial knowledge independently contributed to more positive diversity beliefs and consequently increased team elaboration of task-relevant information as well as integration of different perspectives. Our results reveal key factors that can influence diversity beliefs and underscore the pivotal role of diversity beliefs in improving the extent to which team members elaborate information and integrate diverse perspectives.     

闽江河口互花米草海向入侵对湿地土壤nirS型反硝化微生物群落结构及多样性的影响

2020年秋季,在闽江河口鳝鱼滩东部的互花米草分布区,由陆向海方向选择互花米草海向入侵前的光滩(MF)、入侵1～2年的互花米草湿地(SAN)和入侵6～7年的互花米草湿地(SA)为研究对象.基于高通量测序技术,探讨了互花米草海向入侵对土壤nir S型反硝化微生物群落结构及多样性的影响.结果表明,变形菌门(Proteobacteria)均是互花米草不同入侵阶段土壤中nir S型反硝化微生物的优势门(90.41%～97.36%),其次是厚壁菌门(Firmicutes)和放线菌门(Actinobacteria),三者在SAN和MF土壤中的丰度均与SA存在显著差异(p<0.05).不同入侵阶段土壤中共有的nir S型反硝化细菌属为假单胞菌属(Pseudomonas)、芽孢杆菌属(Bacillus)、偶氮弧菌属(Azoarcus)、陶厄氏菌属(Thauera)、Sulfurifustis属、副球菌属(Paracoccus)、鲁杰氏菌属(Ruegeria)、红肠命菌属(Rubrivivax)、趋磁螺菌属(Magnetospirillum)和福格斯氏菌属(Vogesella),其中,假单胞菌属是...     

典型氢氧化细菌固碳特性种间差异及胞外有机物对固碳过程影响

化能自养细菌对全球CO₂固定具有重要的意义.研究了4种典型氢氧化细菌（HOB,分别是Alcaligenes hydrogenophilus DSM 2625、Pelomonas saccharophila DSM 654、Variovorax paradoxus DSM 30034和Acidovorax facilis DSM 649）的固碳特性及其种间差异性,并探究了胞外游离有机碳（EFOC）对不同种类HOB固碳效率差异性的影响.结果表明：①不同HOB自养培养过程中表观固碳量随时间呈显著差异性,其中,DSM 2625生长速度较快,其平均固碳量分别是DSM 654、DSM 649和DSM 30034的6.30、8.76和7.02倍.②不同HOB之间cbbL基因转录量与 表观固碳量的相关系数为0.980 （p<0.05）,cbbL基因转录量是造成不同菌种之间表观固碳效率差异的关键因素.③HOB细胞蛋白质含量与cbbL基因丰度和转录量之间也存在显著正相关,相关系数分别为0.999（p<0.01）和0.976（p<0.05）,即化能自养细菌CO₂同化途径所固定的有机碳进一步参与细胞蛋白质的合成,进而影响其生长速度和表观固碳效率.④HOB在自养培养过程中产生的EFOC对cbbL基因转录效率产生反馈抑制作用,且不同HOB之间,EFOC/TOC比例越低,表观固碳效率越高.