毕氏海蓬子SbDREB基因的克隆与表达分析研究

以毕氏海蓬子的基因组为模板,通过PCR技术扩增到一个编码DREB蛋白AP2保守结构域的基因片段;根据该片段序列设计引物,以毕氏海蓬子经NaCl处理的植株肉质茎cDNA为模板,应用RACE技术获得该基因的cDNA全长,命名为SbDREB(GenBank登录号:JF894301)。SbDREB基因cDNA全长1206bp,包含一个编码284个氨基酸的完整开放阅读框。对氨基酸序列比对分析表明,该蛋白在靠近N端具有典型的AP2/EREBP保守结构域,且该结构域与一些高等植物DREB类转录因子的AP2区域具有高度同源性。进化树分析表明SbDREB属于DREB亚家族中的A-6亚族。实时荧光定量PCR结果显示:干旱、高盐和ABA能够诱导其表达,而低温则使其表达下调,表明该基因在毕氏海蓬子植株对干旱、盐和低温等非生物胁迫的应答中起作用。     

中国块菌要览及其保护策略

截至目前报道的中国块菌有28种,其中黑块菌4种,白块菌24种.具有重要经济价值和生态价值的黑块菌类有印度块菌复合种和中国夏块菌,其中印度块菌复合种与欧洲法国黑孢块菌属于姊妹类群,中国夏块菌与欧洲夏块菌也是姊妹类群.虽然欧洲意大利白块菌在中国尚未找到、波氏块菌在中国的存在尚待最终确认,但是,最近的更多白块菌新种的发现,说...     

Lipidomic changes during different growth stages of Nitzschia closterium f. minutissima

Ultra Performance Liquid Chromatography-Electrospray ionization-Quadrupole-Time of Flight Mass Spectrometry (UPLC-ESI-Q-TOF–MS) is a powerful lipidomic tool. In this study, we developed a UPLC/Q-TOF–MS based method to investigate the lipid metabolomic changes in different growth phases of Nitzschia closterium f. minutissima. The data classification and biomarker selection were carried out by using multivariate statistical analysis, including principal components analysis (PCA), projection to latent structures with discriminant analysis (PLS-DA), and orthogonal projection to latent structures with discriminant analysis (OPLS-DA). We discovered that the intercellular lipid metabolites were significantly different among exponential, early stationary and late stationary phases. Thirty-one lipid molecules were selected and identified as putative biomarkers, including free fatty acid, Harderoporphyrin, phosphatidylglycerol, 1,2-diacyglycerl-3-O-4′-(N,N-trimethy)-homoserine, triacylglycerol, cholesterol, sulfoquinovosyldiacylglycerol, lyso-sulfoquinovosyldiacylglycerol, monogalactosyldiacylglycerol, digalactosyldiacylglycerol and lyso-digalactosyldiacylglycerol. These lipids have been shown previously to function in energy storage, membrane stability and photosynthesis efficiency during the growth of diatoms. Further analysis on the putative biomarkers demonstrated that nitrate starvation played critical role in the transition from exponential phase to stationary phase in N. closterium. This study is the first one to explore the lipidomic changes of microalgae in different growth phases, which promotes better understanding of their physiology and ecology.     

互花米草入侵九段沙河口湿地对当地昆虫多样性的影响

为认识因互花米草(Spartinaalterniflora)入侵而给九段沙河口湿地昆虫多样性带来的影响,我们于2004年5月到2005年10月间用网捕和收割植株两种方法对3个典型植物群落中昆虫多样性作了连续调查。研究期间,共采集到昆虫11,300头。经鉴定为97个种,隶属于12目69科。互花米草群落中昆虫的物种数、个体数量以及Shannon-Wiener多样性指数均显著低于土著植物芦苇(Phragmitesaustralis)群落和海三棱藨草(Scirpusmariqueter)群落中的;而Simpson优势度指数较土著植物群落中高。聚类分析结果表明:芦苇与海三棱藨草群落中昆虫群落结构更为相似。互花米草的入侵将可能导致九段沙湿地昆虫多样性的降低和群落结构的改变。     

AiiA蛋白的可溶性表达及其抗菌活性研究

AHLs是革兰氏阴性细菌在增殖过程中产生的一类信号分子,与其致病性密切相关。AiiA蛋白作为一种胞内解酯酶．能水解致病菌产生的AHb分子,使内酯环开环后不能再激活某些胞外酶的表达．从而极大地减弱了细菌的致病性。本研究从苏云金芽孢杆菌LLB15中分离编码aiiA基因的质粒DNA。用PCR方法克隆面讲基因,并利用pET载体构建6-His融合表达质粒pET29a-aiiA,转化E.coli BL21（DE3）菌株,并筛选得到E.coli BL21（DE3）-pET29a-aiiA工程菌。在20℃的低温和0．8mmol／LIPTG条件下。经25h的诱导表达,获得了54．4ug／mL可溶性AiiA蛋白。通过镍柱亲和层析,在国内外首次纯化了带6-His标记的AiiA蛋白。水解活性和抗病性检测表明,该蛋白能水解AHLs分子．对胡萝卜欧文氏软腐病菌具有较强的抗病作用。     

小溪自然保护区非盐环境土壤中嗜盐和耐盐菌多样性

【目的】研究湖南小溪国家级自然保护区普通非盐环境(ordinary non-saline environment)土壤样品中可培养嗜盐及耐盐细菌(含放线菌)多样性。【方法】采用纯培养法和基于16S rRNA基因序列的系统发育分析对样品中嗜盐及耐盐细菌多样性进行研究。【结果】用补充5%-20%(w/v)NaCl的MA、ISP2、ISP5、NA和HAA培养基从土壤样品中分离到114株细菌,其中8株为中度嗜盐菌,19株为轻度嗜盐菌,87株为耐盐菌。根据形态观察和部分生理生化实验结果去冗余,选取61个代表性菌株进行基于16S rRNA基因序列的系统发育多样性分析。结果表明,这些菌株属于细菌域(Bacteria)的3个大的系统发育类群(门;phylum)(Actinobacteria,Firmicutes,Proteobacteria)的16个科、18个属,代表了41个物种。多数菌株属于Firmicutes门(38株,62.3%)和Actinobacteria门(18株,29.5%)。大多数菌株与其系统发育关系最密切的已知物种的典型菌株之间存在一定的遗传差异(16S rRNA基因序列相似性为96.9%-99.8%),其中有7个菌株(JSM070026,JSM081004,JSM081006,JSM081008,JSM083058,JSM083085,JSM084035)代表7个潜在新种(potential novel species)。【结论】研究结果表明,湖南小溪国家级自然保护区普通非盐环境土壤中存在较为丰富的可培养嗜盐及耐盐细菌多样性,并且潜藏着较多新的微生物类群(物种)。     

Aptamer selection for the detection of Escherichia coli K88

In this study, the first group of single-stranded DNA aptamers that are highly specific to enterotoxigenic Escherichia coli (ETEC) K88 was obtained from an enriched oligonucleotide pool by the SELEX (Systematic Evolution of Ligands by Exponential Enrichment) procedure, during which the K88 fimbriae protein was used as the target and bovine serum albumin as counter targets. These aptamers were applied successfully in the detection of ETEC K88. They were then grouped under different families based on the similarity of their secondary structure and the homology of their primary sequence. Four sequences from different families were deliberately chosen for further characterization by fluorescence analysis. Having the advantage of high sensitivity, fluorescence photometry was selected as single-stranded DNA quantification method during the SELEX process. Aptamers with the highest specificity and affinity were analyzed to evaluate binding ability with E. coli. Since ETEC K88 is the only type of bacterium that expressed abundant K88 fimbriae, the selected aptamers against the K88 fimbriae protein were able to specifically identify ETEC K88 among other bacteria. This method of detecting ETEC K88 by aptamers can also be applied to bacteria other than ETEC K88.     

福建南亚热带雨林封禁30年的群落特征

福建南靖县和溪南亚热带雨林,是我国南亚热带东段境内目前保存较完整的地带性植被类型之一,是福建南亚热带的代表植被［１,２］,于１９６２年起由国家对其作永久性封禁,今为福建省自然保护区。现将该群落特征作定量分析,并与１９６５年的群落特征［３］进行全面比较...     

藏猪小肠形态、消化酶及微生物多样性研究

[目的]肠道是动物的主要消化器官,同时也是机体抵抗外源病原菌的重要屏障,已有研究表明,动物的品种、饲养方式、生长阶段均会影响动物的肠道菌群结构,但对舍饲和放牧饲养条件下藏猪的肠道菌群结构,以及藏猪和长白、约克与杜洛克三元杂交猪(DLY猪)的肠道菌群结构是否有差异,尚未见报道.[方法]本研究选取6-7月龄的放牧藏猪、舍饲...     

棕黑腐质霉(Humicola fuscoatra)导致真菌性腹膜炎1例

报道1例由棕黑腐质霉属(Humicola fuscoatra)导致的真菌性腹膜炎。此菌分离自1名长期腹膜透析患者的腹水。腐质霉属在自然界广泛存在,棕黑腐质霉导致的人类感染罕见。现对棕黑腐质霉的真菌学特点进行研究,并进行分子测序。体外药物敏感性试验结果对伊曲康唑的MIC为0.008μg/mL,伏立康唑的MIC为0.016μg/mL,两性霉素B的MIC为1.5μg/mL。患者拔除腹透管,改行血液透析。口服伊曲康唑0.1 g/12 h,28 d后病情明显改善,出院。