基于高通量测序的乐安江冬季细菌群落特征分析

【目的】分析乐安江从上游至下游水体细菌群落结构组成变化,揭示细菌群落结构组成变化的影响因素。【方法】分析不同河段水体中C、N、P、Cu、Zn、As和Pb等化学指标。对水体DNA的16S rRNA基因进行高通量测序确定细菌群落特征。基于Bray-Curtis距离的采样点非度量多维尺度(NMDS)分析和聚类分析研究乐安江水体细菌群落结构差异,基于冗余分析(RDA)研究环境因子与细菌群落的关系。【结果】乐安江水体中C、N、P、Cu、Zn、As和Pb等化学指标含量中下游偏高。中游河水受德兴铜矿废水影响,细菌群落多样性降低,下游受农业、生活废水影响,细菌群落丰富度和多样性升高。水体中优势菌群为β-变形菌纲(Beta-proteobacteria,53.03%)、放线菌门(Actinobacteria,20.24%)和拟杆菌门(Bacteroidetes,14.75%)。中游受德兴铜矿废水影响,Beta-proteobacteria丰度增大,而Actinobacteria丰度减小;下游受微生物间捕食影响,Bacteroidetes丰度下降。在细菌群落与环境因子的关系中,DO是解释乐安江细菌群落结构变化的最佳环境因子。【结论】乐安江中游德兴铜矿废水和中下游农业、生活废水明显改变了水体细菌群落结构组成,使水体细菌群落特征从上游到下游发生显著变化。本研究为揭示人类活动对乐安江水生态环境的影响提供了参考性数据。     

海南海口温泉真菌、细菌多样性及其环境影响因素分析

【目的】海南海口含有丰富的温泉资源,对温泉微生物多样性进行研究,有助于进一步开发和利用海南温泉微生物资源。【方法】本文采用Illumina Hi Seq高通量测序技术对海口3个温泉[海甸岛荣域温泉(S1)、火山口开心农场温泉(S2)和西海岸海长流温泉(S3)]水样中微生物ITS序列和16Sr RNA基因V3-V4区进行测序及生物信息学分析,探究海口市3个不同区域的温泉真菌多样性与细菌多样性。【结果】(1)α多样性分析表明,真菌群落中,S3(29)S1(29)S2,而在细菌群落中,S2(29)S1(29)S3。β多样性分析表明,3个温泉真菌群落和细菌群落组成差异皆显著。(2)分类分析表明,温泉真菌群落优势菌门为子囊菌门(Ascomycota)和担子菌门(Basidiomycota),细菌群落优势菌门为变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)、Thermi、硝化螺旋菌门(Nitrospirae)、绿菌门(Chlorobi)、厚壁菌门(Firmicutes)、绿弯菌门(Chloroflexi)、放线菌门(Actinobacteria)。(3) CCA (Canonical correspondence analysis)分析表明,3个温泉的真菌群落主要影响因子是温度,细菌群落主要影响因子是总磷。【结论】海南省海口市温泉中含有丰富的微生物资源,其微生物群落组成受多种环境因子影响,且影响真菌和细菌的主要环境因子不同。     

DGGE分析东江流域农村饮用水源中微生物多样性及其与环境因子相关性

为评价东江流域农村饮用水源中微生物多样性及其与环境因子的相关性, 分别采集了集中式供水井、塘坝型水井、猪场附近水井、普通村落水井、水库水5种水样, 进行基因组总DNA的提取和主要理化指标的测定, 运用DGGE技术分析各水样总DNA的PCR产物。UPGMA聚类分析DGGE指纹图谱结果表明, 相同类型水样的微生物群落结构相似性较高, 聚集到一个分支上; 典型相关性分析(CCA)结果表明, 水体中总磷(TP)和总氮(TN)的浓度与微生物群落结构的关联度最高, 即磷和氮两种生命过程的基本元素对微生物群落影响最大; 序列分析表明农村饮用水源中微生物群落结构丰富, 包含了螺旋体门(Spirochaetes)、蓝藻门(Cyanobacteria)、变形菌门(Proteobacteria)、放线菌门(Actinobacteria)、酸杆菌门(Acidobacteria)5个门的细菌, 且每类水样拥有各自的优势菌。     

夏季达里诺尔湖浮游细菌群落表、底层结构特征及其关键驱动因子

浮游细菌是湖泊水体物质循环和能量流动过程中的关键要素, 影响着整个生态系统的迁移转化。为探索中国北方典型内陆封闭型湖泊浮游细菌群落结构对环境因子变动的响应, 以夏季内蒙古达里诺尔湖(以下简称“达里湖”)为例, 采集表、底层水共32个样品, 基于16S rRNA基因测序, 详细分析了浮游细菌群落结构的表、底层变化特征及其主要影响因素, 结果显示: (1)夏季达里湖水体中浮游细菌群落多样性和均匀度均呈现表层＞底层, 丰富度则呈底层＞表层。(2)浮游细菌群落以变形菌门(Proteobacteria)、放线菌门(Actinobacteria)、蓝细菌门(Cyanobacteria)和拟杆菌门(Bacteroidetes)四大优势菌门为主要组成部分, 且变形菌门含量在表、底层水体中均占绝对优势: 在底层水中占比46.46 %, 在表层水中占比36.21 %。(3)基于不同环境因子和浮游细菌优势菌门间的Pearson相关性分析, 表层水体中总溶解性固体(TDS)、溶解态无机磷(DIP)是多数优势菌门的主要驱动因子, 而底层水体中的总氮(TN)、叶绿素a(Chl a)和水温(WT)为主要驱动因子。总的来说, 达里诺尔湖表、底层浮游细菌群落结构与不同环境因子间存在显著相关, 而水体环境因子的表、底层差异是影响多数优势菌门演化的重要因素。     

珠江口水体浮游细菌种群多样性空间分布特征

为认识珠江口水体浮游细菌的多样性分布规律,运用聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)和多维尺度分析(MDS)的方法,研究了春季珠江口十个站位表底水层浮游细菌种群16SrRNA基因多样性特征。研究结果表明,珠江口浮游细菌种群具有丰富的基因多样性;不同站位细菌群落结构和优势种群变化显著:大多数站位表底层细菌群落结构比较相似,河口外站位(A12,A14和C5),表底层细菌群落结构差异性较大:多样性分析表明A14,B6和C5站位底层细菌多样性大于表层。遗传发育分析表明,序列归属于变形细菌(Proteobacteria),酸杆菌(Acidobacteria),蓝细菌(Cyanobacteria),厚壁细菌(Firmicutes)和放线菌(Actinobacteria)。变形细菌(Proteobacteria)种类最多,从河口内淡水区到河口外海水区都有大量分布,是珠江口水域占优势的菌群;同时也检测到种类丰富的放线菌(Actinobacteria)的存在,也是珠江口水域的优势菌群。     

汾河入黄口夏季微生物群落结构分析

【背景】河流交汇区日益成为流域生态治理的焦点和热点之一。【目的】探明汾河入黄口微生物群落结构及其主要环境影响因子。【方法】应用16S rRNA基因Illumina MiSeq高通量测序技术,分析了汾河入黄口夏季微生物群落结构,并利用典范对应分析(Canonical correspondence analysis,CCA)了解影响微生物群落的主要环境因子。【结果】多样性指数分析表明该区域微生物群落多样性较高。微生物多样性分析发现优势菌门为变形菌门(Proteobacteria)、厚壁菌门(Firmicutes)、拟杆菌门(Bacteroidetes)和放线菌门(Actinobacteria);在属分类水平上,相对丰度最高的菌属为芽孢杆菌属(Bacillus),其次为乳球菌属(Lactococcus)和hgcI_clade。Spearman相关性分析及典范对应分析表明环境因子对水体微生物群落结构具有显著影响。【结论】汾河与黄河微生物群落组成具有一定的差异,不同环境因子对不同微生物的影响程度不同,p H和溶解氧(Dissolved oxygen,DO)是汾河入黄口微生物群落结构的主要影响因子。     

放牧和刈割对大针茅草原土壤微生物群落结构及多样性的影响

放牧和刈割是内蒙古草原的两种主要利用方式,然而,长期放牧和刈割对大针茅草原土壤微生物群落的影响知之甚少,因此,以内蒙古大针茅草原为研究对象,设置放牧和刈割两种利用方式,以围封不利用为对照,基于高通量测序技术,研究大针茅草原在不同利用方式下土壤微生物组成及多样性的变化,并结合土壤理化因子进一步探究土壤微生物群落组成的主要影响因素。研究结果表明：不同利用方式下土壤细菌α多样性指数无显著差异,而刈割显著提高了土壤真菌Observed＿species、Chao1和ACE指数;土壤细菌群落的优势菌门是变形菌门(Proteobacteria)和放线菌门(Actinobacteria),土壤真菌群落的优势菌门是子囊菌门(Ascomycota)和被孢霉门(Mortierellomycota),不同利用方式下部分微生物类群的相对丰度差异显著,放牧显著提高了细菌群落的变形菌门、疣微菌门(Verrucomicrobia)和芽单胞菌门(Gemmatimonadetes)的相对丰度,刈割显著提高了真菌群落的担子菌门(Basidiomycota)相对丰度,此外,放牧和刈割均显著降低了厚壁菌门(Firmicutes...     

基于高通量测序的禁牧对土壤微生物群落结构的影响

麋鹿的采食、躺卧和践踏行为均会对栖息地土壤环境造成影响,进而影响土壤微生物群落结构。利用高通量测序技术,分析江苏大丰麋鹿国家级自然保护区禁牧点和补饲点土壤细菌和真菌群落结构差异,并结合土壤理化性质探究禁牧对土壤微生物群落结构的影响。结果表明细菌群落的优势菌门为变形菌门,真菌群落的优势菌门为子囊菌门。禁牧改变了土壤微生物群落结构,在门水平上提高了变形菌门、放线菌门和担子菌门的相对丰度,降低了绿弯菌门、厚壁菌门和子囊菌门的相对丰度,禁牧点与补饲点土壤微生物群落多样性的相似性较低。冗余分析中,细菌受土壤环境因子的影响大于真菌,其中土壤pH是影响细菌和真菌群落最大的土壤环境因子。研究揭示了禁牧对土壤微生物群落结构的影响,为保护区制定麋鹿生境恢复方案提供参考。     

鄱阳湖湖泊细菌群落组成及结构——以松门山为例

于2011年5月在鄱阳湖——松门山湖区采集底泥与表层水样,分别提取了表层水体浮游和底泥微生物基因组DNA,利用454高通量测序技术对细菌的16S rRNA基因进行了序列测定,分析了湖泊底泥细菌、水体浮游细菌群落结构特征。结果显示:底泥细菌OTUs(Operational Taxonomic Units)为1454,表层水体浮游细菌OTUs为269;底泥细菌群落比表层水体更加多样化,底泥细菌的物种数大大多于表层水体。同时,底泥细菌群落与表层浮游细菌群落结构存在显著差异。物种分类显示鄱阳湖底泥细菌种类隶属于20门,228属,其中优势种群为δ-变形菌纲(Deltaproteobacteria)、β-变形菌纲(Betaproteobacteria)和疣微菌门(Verrucomicrobia);表层水体浮游细菌隶属于13门116属,优势种群为β-变形菌纲、拟杆菌门(Bacteroidetes)和放线菌门(Actinobacteria)。结果进一步揭示,无论是浮游细菌群落还是沉积物细菌群落,优势细菌种群的基因型多样性更高。     

山东地区盐碱土花生种子际土壤微生物群落结构的研究

【目的】以不同含盐量的滨海盐土、内陆盐碱土和中等肥力非盐碱土壤为实验对象,探讨花生种子在吸水膨胀与萌发过程中,不同类型盐碱土对种子际土壤微生物多样性变化的影响。【方法】采集不同含盐量的滨海盐土、内陆盐碱土和中等肥力非盐碱土壤,通过对各样品中细菌的16S r RNA基因的V3-V4区进行PCR扩增,利用Illumina Hiseq高通量测序技术对12份V3-V4高变区PCR产物进行测序,并对测序数据进行生物信息学分析。【结果】(1)盐碱土壤的种子际细菌群落多样性高于非盐碱土壤,且以东营青坨滨海盐土种子际土壤细菌群落多样性较高。(2)不同类型土壤样本微生物群落结构在纲水平存在明显差异。4种土壤类型种子际土壤细菌共分属于6个菌纲,分别为Proteobacteria、Actinobacteria、Actinobacteria、Bacteroidetes、Acidobacteria和Firmicutes菌纲,并均以Proteobacteria和Actinobacteria菌纲为主要菌纲。全样本菌落结构分析结果表明,4种类型土壤中不同吸胀时间内种子际微生物菌落在门、属水平上的类型和丰度差异最为显著(P0.05)。(3)beta多样性分析和各样本遗传距离(phylogenetic distances)聚类树图分析表明,4个土壤类型的12个土壤样本种子际土壤中微生物群落均可聚为2大类。【结论】土壤含盐量越高其种子际土壤细菌群落多样性较高。不同类型土壤样本微生物群落结构在纲水平存在明显差异,以Proteobacteria和Actinobacteria菌纲为主要菌纲。种子吸胀萌发时间影响种子际微生物菌落在门、属水平上的类型和丰度,但对相同土壤类型样本间遗传距离无影响。     

2017年中国植物科学若干领域重要研究进展

2017年中国植物科学继续保持高速发展态势, 重大成果频出, 具体表现在中国植物学家在国际顶级学术期刊发表的文章数量平稳上升。中国植物科学领域的研究工作者成果精彩纷呈, 如新型广谱抗病机制的发现、水稻广谱抗病遗传基础及机制和疫霉菌诱发病害成灾机制研究等。2017年中国生命科学领域十大进展评选中, 有两项植物科学领域的研究成果入选。水稻生物学、进化与基因组学和激素生物学等领域学科发展突出。另外, 值得一提的是, 长期从事高等植物与代谢途径调控分子网络研究和水稻品种设计育种的李家洋院士的研究成果“水稻高产优质性状形成的分子机理及品种设计”荣获2017年国家自然科学一等奖。这一具有重大国际影响的开创性贡献标志着中国植物科学在该领域的国际科学前沿居于引领和卓越地位。该文对2017年中国本土科学家在植物科学若干领域取得的重要研究成果进行了系统梳理, 旨在全面追踪和报道当前中国植物科学领域发展的最新前沿动态, 与广大读者共同分享我国科学家所取得的辉煌成就。     

2018年中国植物科学若干领域重要研究进展

2018年中国植物科学继续呈现快速发展的态势, 我国科学家在国际植物科学主流学术刊物发表论文数量大幅增加, 取得了多项具有重要影响的成果。调控植物生长-代谢平衡实现可持续农业发展入选2018年度中国科学十大进展; 中国被子植物区系进化历史研究入选2018年度中国生命科学十大进展。以水稻为代表的农作物和果蔬等经济作物研究在国际上已呈现出明显的优势, 若干领域已从“追赶”状态跨越到“领跑”地位。该文对2018年中国科学家在植物科学若干领域取得的重要研究成果进行了概括性评述, 旨在全面追踪和报道当前中国植物科学领域的发展前沿和热点, 展示中国科学家所取得的杰出成就。     

2018年中国植物科学若干领域重要研究进展

2018年中国植物科学继续呈现快速发展的态势, 我国科学家在国际植物科学主流学术刊物发表论文数量大幅增加, 取得了多项具有重要影响的成果。调控植物生长-代谢平衡实现可持续农业发展入选2018年度中国科学十大进展; 中国被子植物区系进化历史研究入选2018年度中国生命科学十大进展。以水稻为代表的农作物和果蔬等经济作物研究在国际上已呈现出明显的优势, 若干领域已从“追赶”状态跨越到“领跑”地位。该文对2018年中国科学家在植物科学若干领域取得的重要研究成果进行了概括性评述, 旨在全面追踪和报道当前中国植物科学领域的发展前沿和热点, 展示中国科学家所取得的杰出成就。     

Role for dopamine in malonate-induced damage in vivo in striatum and in vitro in mesencephalic cultures

Defects in mitochondrial energy metabolism have been implicated in the pathology of several neurodegenerative disorders. In addition, the reactive metabolites generated from the metabolism and oxidation of the neurotransmitter dopamine (DA) are thought to contribute to the damage to neurons of the basal ganglia. We have previously demonstrated that infusions of the metabolic inhibitor malonate into the striata of mice or rats produce degeneration of DA nerve terminals. In the present studies, we demonstrate that an intrastriatal infusion of malonate induces a substantial increase in DA efflux in awake, behaving mice as measured by in vivo microdialysis. Furthermore, pretreatment of mice with tetrabenazine (TBZ) or the TBZ analogue Ro 4-1284 (Ro-4), compounds that reversibly inhibit the vesicular storage of DA, attenuates the malonate-induced DA efflux as well as the damage to DA nerve terminals. Consistent with these findings, the damage to both DA and GABA neurons in mesencephalic cultures by malonate exposure was attenuated by pretreatment with TBZ or Ro-4. Treatment with these compounds did not affect the formation of free radicals or the inhibition of oxidative phosphorylation resulting from malonate exposure alone. Our data suggest that DA plays an important role in the neurotoxicity produced by malonate. These findings provide direct evidence that inhibition of succinate dehydrogenase causes an increase in extracellular DA levels and indicate that bioenergetic defects may contribute to the pathogenesis of chronic neurodegenerative diseases through a mechanism involving DA.     

体内生物膜研究进展

生物膜是指由黏附于生命体或非生命体表面的微生物和其胞外聚合物所构成的复杂、多维的空间结构,在自然界中普遍存在。微生物在人类体内许多部位均可形成生物膜结构,如肺、心脏瓣膜、泌尿生殖道、肝胆系统、耳、鼻、皮肤等。越来越多的感染性疾病患者或动物模型体内均可检测出生物膜,如囊性纤维化肺炎、慢性中耳炎、细菌性心内膜炎、皮肤外伤后慢性感染、慢性鼻窦炎等。生物膜的形成与感染性疾病的关系及其在疾病的发生、发展、转归过程中的作用成为近年来研究的热点。目前体内生物膜检测技术主要有扫描电镜、激光共聚焦显微镜、荧光原位杂交技术等。就临床常见感染性疾病与生物膜的关系及体内生物膜检测方法展开综述。     

Changes in aquatic vegetation in Rhine floodplain streams in Alsace in relation to disturbance

Abstract. Changes are described in aquatic vegetation in oligotrophic, groundwater-fed Rhine floodplain streams in Alsace (eastern France), resulting from disturbance. Disturbance factors include changes in nutrients, either permanent ones - effluent from a waste water treatment plant or trout hatcheries - or periodic ones: flooding. Regular inputs of high levels of phosphate and ammonia modified the macrophyte vegetation in these streams. The floristic composition, which was characteristic of oligotrophic waters upstream of the eutrophicated sector, changed to that of a eutrophic situation as originally found downstream. Periodic disturbance by floods which normally occur once a year, irregularly eutrophicates the small streams, causing the development of a mixture of eutrophic and oligotrophic species. Six macrophyte communities are distinguished, indicating different trophic levels. The aquatic vegetation is adapted to the variations of phosphate and ammonia levels. Hence, aquatic macrophytes can be used as bio-indicators of fluctuations in water nutrient levels in relation to the type of disturbance.     

Fibromodulin gene is expressed in human epidermal keratinocytes in culture and in human epidermis in vivo

Fibromodulin is a small leucine-rich proteoglycan that has a central role in the maintenance of collagen fibrils structure, and in regulation of TGF-β biological activity. Although, it is mainly found in cartilage and tendon, little is known regarding the expression of the fibromodulin gene in other cell types. By RT-PCR, real time PCR and immunohistochemistry, we describe the expression of the fibromodulin gene and the presence of the protein in human epidermal keratinocytes (HEK), both in culture and in normal human epidermis. Our results show, for the first time, that fibromodulin gene is constantly expressed in HEK during culture time. Immunostaining showed that fibromodulin is located intracytoplasmically in basal and stratified keratinocytes of the growing colonies, confluent cultures, and epidermis in vivo. The expression and intracellular localization of fibromodulin in HEK is a new finding and opens new possible biological roles for the SLRP family.     

Morphine-potentiated platelet aggregation in in vitro and platelet plug formation in in vivo experiments

The detailed mechanisms underlying morphine-signaling pathways in platelets remain obscure. Therefore, we systematically examined the influence of morphine on washed human platelets. In this study, washed human platelet suspensions were used for in vitro studies. Furthermore, platelet thrombus formation induced by irradiation of mesenteric venules with filtered light in mice pretreated with fluorescein sodium was used for an in vivo thrombotic study. Morphine concentration dependently (0.6, 1, and 5 microM) potentiated platelet aggregation and the ATP release reaction stimulated by agonists (i.e., collagen and U46619) in washed human platelets. Yohimbine (0.1 microM), a specific alpha(2)-adrenoceptor antagonist, markedly abolished the potentiation of morphine in platelet aggregation stimulated by agonists. Morphine also potentiated phosphoinositide breakdown and intracellular Ca(2+) mobilization in human platelets stimulated by collagen (1 microg/ml). Moreover, morphine (0.6-5 microM) markedly inhibited prostaglandin E(1) (10 microM)-induced cyclic AMP formation in human platelets, while yohimbine (0.1 microM) significantly reversed the inhibition of cyclic AMP by morphine (0.6 and 1 microM) in this study. The thrombin-evoked increase in pH(i) was markedly potentiated in the presence of morphine (1 and 5 microM). Morphine (2 and 5 mg/g) significantly shortened the time require to induce platelet plug formation in mesenteric venules. We concluded that morphine may exert its potentiation in platelet aggregation by binding to alpha(2)-adrenoceptors in human platelets, with a resulting inhibition of adenylate cyclase, thereby reducing intracellular cyclic AMP formation followed by increased activation of phospholipase C and the Na(+)/H(+) exchanger. This leads to increased intracellular Ca(2+) mobilization, and finally potentiation of platelet aggregation and of the ATP release reaction.     

SSTR5 ablation in islet results in alterations in glucose homeostasis in mice

Somatostatin (SST) peptide is a potent inhibitor of insulin secretion and its effect is mediated via somatostatin receptor 5 (SSTR5) in the endocrine pancreas. To investigate the consequences of gene ablation of SSTR5 in the mouse pancreas, we have generated a mouse model in which the SSTR5 gene was specifically knocked down in the pancreatic beta cells (betaSSTR5Kd) using the Cre-lox system. Immunohistochemistry analysis showed that SSTR5 gene expression was absent in beta cells at three months of age. At the time of gene ablation, betaSSTR5Kd mice demonstrated glucose intolerance with lack of insulin response and significantly reduced serum insulin levels. Insulin tolerance test demonstrated a significant increase of insulin clearance in vivo at the same age. In vitro studies demonstrated an absence of response to SST-28 stimulation in the betaSSTR5Kd mouse islet, which was associated with a significantly reduced SST expression level in betaSSTR5Kd mice pancreata. In addition, betaSSTR5Kd mice had significantly reduced serum glucose levels and increased serum insulin levels at 12 months of age. Glucose tolerance test at an older age also indicated a persistently higher insulin level in betaSSTR5Kd mice. Further studies of betaSSTR5Kd mice had revealed elevated serum C-peptide levels at both 3 and 12 months of age, suggesting that these mice are capable of producing and releasing insulin to the periphery. These results support the hypothesis that SSTR5 plays a pivotal role in the regulation of insulin secretion in the mouse pancreas.