PCR-DGGE技术解析东北稻田蓝藻群落结构

[目的]明确东北稻田生态系统蓝藻群落结构组成变化及主要驱动因子。[方法]采用蓝藻特异性引物GC-Cya-b-F371和Cya-R783对采自东北不同土壤类型稻田土壤及田面水提取的DNA进行PCR扩增,采用DGGE电泳和条带测序技术解析稻田蓝藻群落结构。[结果]多样性指数表明,土壤中蓝藻群落结构比田面水体丰富。图谱冗余分析(RDA)显示,稻田土壤中蓝藻群落结构具有明显的地域差异,土壤碳氮比值是驱动土壤蓝藻分布的主要环境因素;而田面水体蓝藻群落结构地域差异不明显,p H值是驱动水体蓝藻分布的主要环境因素。DGGE条带测序发现,稻田蓝藻群落结构组成以丝状颤藻亚纲蓝藻(Oscillatoriophycideae)为主,一些条带基因序列与已知蓝藻相似度低于97%。[结论]东北稻田生态系统蓝藻群落结构复杂多样,生存着一些未知的蓝藻种群。     

氮磷营养盐对四种淡水丝状蓝藻生长的影响

通过设定不同的氮、磷营养盐浓度,在实验室条件下研究了不同浓度的氮和磷对4种淡水丝状蓝藻:皮质颤藻、尖细颤藻、蛇形颤藻和坑形细鞘丝藻生长的影响.结果表明,皮质颤藻和坑形细鞘丝藻对高浓度的氮、磷有着较强的适应能力,在硝态氮浓度0.016mg·mL^-1～2.0mg·mL、磷浓度1.36mg·mL^-1～13.6μg·mL时生长较好.4株藻在只存在氨态氮的情况下生长都受到抑制.     

东南极格罗夫山土壤可培养细菌的分离鉴定及其产胞外酶和抗菌活性北大核心CSCD

【目的】获得东南极格罗夫山地区土壤中可培养细菌组成信息,分析菌株胞外水解酶产生及抗菌活性情况。【方法】稀释直接涂布平板法获得可培养细菌菌株并根据其16S rRNA基因序列对其进行系统发育分析。平板法初步鉴定菌株的胞外酶产生情况,琼脂块法分析菌株对5种供试菌的抗性。【结果】所有土壤样品共分离出39株可培养细菌,分属于厚壁菌门(Firmicutes,19株,48.7%)、变形菌门(Proteobacteria,分属于α-、β-、γ-变形菌纲,共计10株,25.6%)、放线菌门(Actinobacteria,8株,20.5%)、拟杆菌门(Bacteroidetes,1株,2.6%)和异常球菌-栖热菌门(Deinococcus-Thermus,1株,2.6%)等5门20个菌属,优势菌属为芽孢杆菌(Bacillus)。不同冻存温度的土壤样品分离菌株有所不同。33株细菌具有至少一种的胞外酶活力,其中产淀粉酶的菌株最多(25株,64.1%)。6株细菌可抑制至少一种供试菌的生长。【结论】格罗夫山土壤可培养细菌组成在大分类上与南极其他地区一致,但在属的水平上有所不同。分离出的具有胞外酶活性和抗菌活性的适冷菌株为进一步开发利用南极低温酶和抗菌活性物质提供了良好的菌种资源。     

乳扇制品中乳酸杆菌的分离鉴定及16S rRNA序列同源性分析

目的乳扇是云南大理白族的一种传统乳制品,明确大理乳扇制品中乳杆菌的多样性及优势种群分布,为科学利用奠定基础。方法采用表型鉴定及16S r RNA鉴定方法,对10个家庭作坊的大理乳扇制品中的乳杆菌进行了分离鉴定。结果共分离到50株乳杆菌,通过表型鉴定为8个种,包括植物乳杆菌10株、德氏乳杆菌7株、发酵乳杆菌6株、干酪乳杆菌6株、棒状乳杆菌4株、鼠乳杆菌2株、弯曲乳杆菌3株和食果糖乳杆菌2株;06422和06430两株表型鉴定未能定种,进一步通过16S r RNA鉴定为植物乳杆菌和马酒乳杆菌,06422株与植物乳杆菌L.arizonensin、L.pentosus和L.plantarum P158的同源性分别是100%、100%和99.9%,与乳杆菌属其它种的同源性为83.4%(L.gallinarum)至93.5%(L.brevis),06430株与L.kefiranofaciens.subsp.Kefirgranum的16S r RNA同源性是99.9%,与乳杆菌属其它种的同源性为83.7%(L.plantarum P158)至96.3%(L.acidophilus)。结论大理乳扇制品中有9种乳杆菌,其优势种群为植物乳杆菌、德氏乳杆菌、发酵乳杆菌和干酪乳杆菌等四种。     

一株芽孢杆菌的分离和鉴定

从中国农业科学院北京畜牧兽医研究所鸡舍附近土壤中分离到一株芽孢杆菌P-25,并进行了分子鉴定。通过形态鉴定、革兰氏染色、生理生化测定、16SrRNA序列分析和系统发育树构建,确定该菌株为蜡状芽孢杆菌(Bacillus cereus),其16SrRNAGenBank登录号为GU271135。     

中华草龟荧光假单胞菌的分子鉴定及分析

从患病草龟病灶分离到几株优势菌,对该菌进行革兰氏染色、镜检,在普通肉汤培养基上进行观察。对分离菌株进行生化鉴定及16S rRNA基因序列的克隆、测定,并对其DNA进行测序,在NCBI上进行比对,构建系统进化树,对该菌的基因序列进行分析比对,基于形态学、生理学、生物化学和基因序列分析,将这些菌株鉴定为荧光假单胞菌。药敏实验结果表明该菌对复方新诺明、氧氟沙星、恩诺沙星、诺氟沙星高度敏感,对卡那霉素、四环素、庆大霉素、新霉素、氟苯尼考、氯霉素中度敏感,对头孢噻吩、甲氧苄啶有耐药性。     

系统发育树的构建方法北大核心CSCD

构建系统树是为了鉴定菌株的分类学地位,应该使用正确的方法构建。具体要求如下：1.将鉴定菌的16S rRNA序列递交GenBank,用Blast软件搜索相似的16S rRNA,然后一起构树。2.采用能反应分支长度的软件（如NJ法）,并用Boostrap值分析分支聚类的稳定性。     

系统发育树的构建方法北大核心CSCD

构建系统树是为了鉴定菌株的分类学地位,应该使用正确的方法构建。具体要求如下：1.将鉴定菌的16S rRNA序列递交GenBank,用Blast软件搜索相似的16S rRNA,然后一起构树。     

利用mtDNA 16S rRNA 序列差异鉴定江西青岚湖的河蚌物种

对从青岚湖采集的10属14种河蚌的19个样本进行了16S rRNA的序列测定,并同CenBank中鄱阳湖流域相同物种河蚌的序列比较,分析了基于Kimura 2-parameter模型参数得到的遗传距离,并构建了它们的UPGMA树。结果显示,所有用于比较的河蚌种间遗传距离变化范围在0．0274—0．2290,平均为0．1325,种内遗传距离在0．0034—0．0068之间,平均为0．0045,种间遗传距离远大于种内距离。表明以16S rRNA作为遗传标记,可以在物种水平上对青岚湖的河蚌进行准确的鉴定。     

一株纤维素降解细菌的筛选、鉴定及产酶条件分析

目的筛选高活性的纤维素降解细菌,并进行初步鉴定和产纤维素酶条件分析。方法采集吉首旗帜山松树林的土壤样品,通过富集培养和刚果红平板染色法筛选分离纤维素降解细菌;通过形态观察、生理生化特性检测和基于16S rRNA基因序列的系统发育分析对分离的菌株进行初步鉴定。利用单因素实验对产纤维素酶条件进行优化。结果分离获得1株高活性纤维素降解细菌JDM11,初步鉴定其为Bacillus velezensis;菌株JMD11产纤维素酶最佳培养温度、最适初始pH和培养时间分别为28℃、7.0～7.5和32h,在该条件下其滤纸酶(FPase)和羧甲基纤维素酶(CMCase)活力分别为260.32U/ml和651.75U/ml。结论菌株JDM11是1株高活性纤维素降解的Bacillus velezensis。     

Structure and dynamics of the periphytic algae of Jinyang Lake in Shanxi Province, North China

The structure and dynamics of periphytic algae in Jinyang Lake, the largest man-made lake in Taiyuan, Shanxi, have been studied by examining the periphytic algae samples collected from four sampling stations, during March 2006 to February 2007. In total, 227 species of periphytic algae (including varieties), belonging to three divisions, 27 families, 62 genera, have been identified. Diatoms were the dominant periphytic algae group. Taken the Shannon–Weaver, Margalef and Simpson diversity index together, it showed obvious temporal and spatial dynamics among different seasons and different sampling stations. The highest periphytic community structure evenness appeared at sampling station I in spring, summer and autumn, while at sampling station III in winter. The average biomass of periphytic algae varied from seasons. Our data showed that major factors affecting the periphytic algae in Jinyang Lake was water temperature, nutrient levels as well as human activities. In addition, the main effect caused by thermal power plant has been discussed.     

Structure and dynamics of the periphytic algae of Jinyang Lake in Shanxi Province, North China

The structure and dynamics of periphytic algae in Jinyang Lake, the largest man-made lake in Taiyuan, Shanxi, have been studied by examining the periphytic algae samples collected from four sampling stations, during March 2006 to February 2007. In total, 227 species of periphytic algae (including varieties), belonging to three divisions, 27 families, 62 genera, have been identified. Diatoms were the dominant periphytic algae group. Taken the Shannon–Weaver, Margalef and Simpson diversity index together, it showed obvious temporal and spatial dynamics among different seasons and different sampling stations. The highest periphytic community structure evenness appeared at sampling station I in spring, summer and autumn, while at sampling station III in winter. The average biomass of periphytic algae varied from seasons. Our data showed that major factors affecting the periphytic algae in Jinyang Lake was water temperature, nutrient levels as well as human activities. In addition, the main effect caused by thermal power plant has been discussed.     

Description of two new species of Nostoc (Nostocales,Cyanobacteria) from central Mexico,using morphological,ecological, and molecular attributes

The present study describes two new Nostoc species, N. montejanii and N. tlalocii, based on a polyphasic approach that combines morphological, ecological, and genetic characteristics. The five investigated populations, including those from newly collected material from central Mexico, were observed to possess morphological features characteristic of the Nostoc genus. Results showed that both new species are strictly associated with running water, and they show clear differences in their habitat preferences. The 16S rRNA gene sequences of the five strains displayed between 98% and 99% similarity to the genus Nostoc sensu stricto. The 16S rRNA gene phylogenetic analyses inferred using Bayesian inference, maximum likelihood, and parsimony methods, placed these five strains in two separate clades distinct from other Nostoc species. The secondary structures of the 16S–23S internal transcribed spacer rRNA region in the two new species showed >10.5% dissimilarities in the operons when compared with other Nostoc species. In addition, clear morphological differences were observed between the two Mexican species, including the color of the colonies (black in N. montejanii and green in N. tlalocii), the size of the cells (greater in N. montejanii), and the number of polyphosphate granules present in the cells (one in N. montejanii and up to four in N. tlalocii).     

Nucleotide Sequences of Fragments of 16S rRNA of the Baikal Natural Populations and Laboratory Cultures of Cyanobacteria

Nucleotide sequences were determined for the 16S rRNA gene 5"-terminal regions of 29 strains of picoplankton cyanobacteria from the lake Baikal in Russia and four strains from the lake Constance (Bodensee) in Germany. Sequences of a 387-bp region of the gene of cultivated strains designated A, B, C, D, and E had only a few (1–3) substitutions as compared with the 3-27 sequence of the clones obtained earlier by cloning from a pooled sample of Baikalian winter picoplankton. The specific 4-33 sequence obtained earlier from a dominant species of Baikalian picoplankton was not found in cultivated cyanobacteria.     

津巴布韦烟叶中淀粉酶和蛋白酶产生菌的分离及鉴定

目的:从津巴布韦烟叶中分离产蛋白酶菌和产淀粉酶能力最高的菌株,并对其进行鉴定。方法:采用淀粉富集培养基和酪蛋白富集培养基分别分离津巴布韦烟叶中的产淀粉酶和产蛋白酶菌株,通过生理生化实验和16SrRNA序列分析鉴定分离的菌株。结果:产蛋白酶菌株菌体不透明、表面有褶皱,蛋白酶酶活为52.10±0.13 U/mL;产淀粉酶菌株菌体表面呈黏状,淀粉酶酶活为3.69±0.07 U/mL;产蛋白酶与产淀粉酶的2株菌均与枯草芽孢杆菌的16S rRNA序列有100%的相似性,结合生理生化指标初步鉴定为枯草芽孢杆菌。结论:获得的2株菌在降解烟叶的蛋白质和淀粉过程中可能起重要作用。     

Viili乳制品中干酪乳杆菌的分离鉴定

从引进Viili乳制品中筛选、鉴定出3种优良乳酸菌。采用平板分离法从Viili乳制品中分离3株乳酸菌,通过表型1、6S rRNA的PCR扩增、克隆、测序鉴定。3株乳酸菌的表型鉴定结果符合伯杰细菌鉴定手册中乳杆菌属的干酪乳杆菌,16S rRNA序列同源性分析结果表明3株分离菌与干酪乳杆菌的同源性分别为99.93%、100.00%9、9.78%,从Viili乳制品分离到3株干酪乳杆菌。     

Culturable bacteriorhodopsin-producing haloarchaea of Tuz Lake (Turkey)

Abstract

Lake Tuz, the largest hypersaline lake in Turkey, has a great variety of microbial communities that adapted to its extreme environment and produce many industrially important compounds such as photosensitive bacteriorhodopsins. So far, the information about the bacteriorhodopsin-producing haloarchaea species of the lake is still limited. In the present study, archaeal bacteriorhodopsin producers were isolated from three locations of the lake. Their bacteriorhodopsin-producing capability was validated by the purification and SDS-PAGE analysis of the delipidated bacteriorhodopsin molecule. The active isolates were identified by the sequencing of partial 16S rDNA gene regions. According to the results, 11 bacteriorhodopsin-producing isolates grouped in Halobacterium salinarum (4), Halobacterium sp. (3), Haloarcula salaria (2), Haloarcula sp. (1) and Halorubrum sp. (1). Our research demonstrated that Lake Tuz is an important natural source of bacteriorhodopsin-producing haloarchaea and the isolates can be valuable for the related technological applications.     

1株罗汉杉内生放线菌的鉴定、活性分析及抗生素生物合成基因的筛查

对1株从罗汉杉分离的内生放线菌LCB-0297进行系统学鉴定,在ISP3培养基上其气生菌丝灰烬色,有粉褐色可溶性色素,电镜观察孢子丝呈螺旋状,孢子椭球形表面多刺,通过其形态、生理生化特征初步确定为链霉菌。16S rRNA基因序列分析显示,放线菌LCB-0297与Streptomyces lanatus NBRC 12787（T）最为相似,相似性为98.531%,但在N-J树上仍在较远的2个分支上,确定为Streptomyces sp.。通过滤纸片法、SRB法进行抑菌、细胞毒活性检测,显示出较强的抑菌及抗癌活性。用PCR的方法对其多种抗生素生物合成基因进行筛查,该菌株含有Ⅰ型聚酮合成酶（PKS-Ⅰ）、非核糖体多肽合成酶（NRPS）、Ⅱ型聚酮合成酶（PKS-Ⅱ）的基因,含有多种潜在的抗生素合成基因。