新疆艾比湖湿地博乐河入口处土壤细菌多样性分析

【目的】了解新疆艾比湖湿地国家级自然保护区非培养土壤细菌群落组成及多样性。【方法】采用非培养法直接从湿地土壤提取总DNA进行16S r RNA基因扩增,构建细菌16S r RNA基因克隆文库。使用MspⅠ和AfaⅠ限制性内切酶对阳性克隆进行16S r RNA基因扩增片段的限制性酶切分析(Amplified r DNA restriction analysis,ARDRA),挑取具有不同双酶切图谱的克隆进行测序,序列比对并构建16S r RNA基因系统发育树。【结果】从土壤细菌的16S r RNA基因文库中随机挑取75个不同谱型的克隆子,共得到58个OTUs,系统发育归类为8个细菌类群:绿弯菌门(Chloroflexi)、蓝藻门(Cyanobacteria)、变形菌门(Proteobacteria)、厚壁菌门(Firmicutes)、放线菌门(Actinobacteria)、拟杆菌门(Bacteroidetes)、疣微菌门(Verrucomicrob)和芽单胞菌门(Gemmatimonadetes)。其中,变形菌门为第一优势菌群,拟杆菌门为第二优势菌群,两者约占总克隆的65%。【结论】艾比湖湿地博乐河入口处土壤细菌多样性丰富,且存在一定数量的潜在微生物新种。     

大兴安岭典型永久冻土土壤细菌群落组成和多样性

【背景】土壤微生物是土壤生物中的重要成分,参与了土壤生态系统中关键的生物化学循环过程。但是关于寒温带多年冻土土壤微生物的研究还比较薄弱。【目的】探究大兴安岭多年冻土土壤中微生物的多样性和种群结构。【方法】利用MiSeq高通量测序技术对黑龙江大兴安岭地区呼中保护区落叶松冻土和樟子松林冻土土壤样品进行测序。【结果】在落叶松冻土和樟子松林冻土土壤中,相对丰度最高的优势菌群的组成基本一致,在门水平有疣微菌门(Verrucomicrobia)、变形菌门(Proteobacteria)、酸杆菌门(Acidobacteria)、浮霉菌门(Planctomycetes)、绿菌门(Chlorobi)、Parcubacteria、放线菌门(Actinobacteria)、拟杆菌门(Bacteroidetes)、厚壁菌门(Firmicutes)、芽单胞菌门(Gemmatimonadetes)10个细菌门类,其中,疣微菌门(Verrucomicrobia)在樟子松林土壤中的相对丰度较多,变形菌门(Proteobacteria)在落叶松林土壤中的相对丰度较多。通过α多样性分析可知,落叶松冻土土壤微生物的群落多样性高于樟子松林冻土,而且两者的细菌群落组成与结构差异性较大。【结论】为深入认识大兴安岭多年冻土区的土壤微生物群落结构组成以及影响因素提供数据支撑。     

溴代阻燃剂污染底泥与未污染底泥中细菌群落结构之比较分析

【目的】通过免培养的分子生物学方法,比较受溴代阻燃剂污染严重的河流底泥和与未污染水库底泥中细菌多样性差异,解析二者间细菌群落结构,为污染河流的治理与生物修复提供相关的理论依据。【方法】从中国贵屿溴代阻燃剂污染区练江底泥样品和未污染水库底泥样品中分别提取微生物总DNA,用细菌通用引物27F和1500R扩增16S rRNA基因,构建16S rRNA基因文库。用HhaⅠ和HinfⅠ2种限制性内切酶对克隆子进行扩增产物rDNA的限制性酶切分析(ARDRA),挑取不同的操作分类单元OUT中的克隆进行测序并构建系统发育树,比较代表克隆子的基本分类类群和生物多样性构成。【结果】未污染水库底泥中细菌群落组成主要包括变形细菌门(Proteobacteria)的α、β、γ和δ4个亚门、浮霉菌门(Planctomycetes)、酸杆菌门(Acidobacteria)、放线菌门(Actinobacteria)、绿弯菌门(Chloroflexi)、疣微菌门(Verrucomicrobia)和厚壁菌门(Firmicutes)。优势菌是酸杆菌,占文库克隆的30.2%。污染河流底泥中细菌群落组成包括变形细菌门(Proteobacteria)的α、β、δ和ε4个亚门、浮霉菌门(Planctomycetes)、绿菌门或拟杆菌门(Chlorobi orBacteroidetes)、酸杆菌门(Acidobacteria)、放线菌门(Actinobacteria)、绿弯菌门(Chloroflexi)、待定菌群candidatedivision OP01、candidate division OP08和candidate division WS3的相似菌。优势菌是ε-变形细菌和绿弯菌,占文库克隆的44.9%。【结论】受溴代阻燃剂污染河流底泥中的细菌群落具有较高水平的多样性,与未污染底泥有显著区别,主要体现在ε-变形细菌和绿弯菌在细菌群落中具有优势地位。这种优势种群的改变可能与污染物的过度富集具有一定的相关性,对于我们进一步探索和了解溴代阻燃剂的微生物修复具有一定的指导意义。     

南极三洋藓(Sanionia uncinata)细菌群落组成分析

【背景】南极苔藓中蕴藏着丰富的极地微生物资源,然而目前对南极苔藓细菌群落组成的研究较少,限制了对这类极端生境微生物资源的开发和应用。【目的】在南极长城站不同地点随机取样,揭示南极三洋藓(Sanioniauncinata)的细菌群落结构组成。【方法】采用IlluminaHiSeq高通量测序技术对3个苔藓样品的16S rRNA基因V4区进行测序及生物信息学分析。【结果】3个苔藓样品中共获得细菌总优化序列273 367条。基于97%序列相似度,细菌优化序列聚类为9 579个OTU。分类地位明确的细菌可归为14门27纲50属。优势类群是变形菌门(Proteobacteria)(30.70%)、拟杆菌门(Bacteroidetes)(19.67%)、疣微菌门(Verrucomicrobia)(12.43%)、蓝细菌门(Cyanobacteria)(10.55%)及放线菌门(Actinobacteria)(9.36%)。在属水平上,56.73%属于未知细菌。【结论】南极三洋藓(Sanionia uncinata)中具有丰富多样的细菌,该研究为今后极地微生物研究奠定了基础。     

干旱区高寒湿地逆行演替下土壤微生物群落结构的研究

【目的】探究高寒湿地逆行演替对土壤性质与微生物群落结构的影响。【方法】以新疆巴音布鲁克天鹅湖高寒湿地为研究对象,依托逆行演替典型样带(沼泽-沼泽化草甸-草甸),利用高通量测序技术分析各演替区土壤微生物群落结构。【结果】高寒湿地逆行演替改变了土壤微生物在分类操作单元(operational taxonomic unit,OTU)水平上的物种组成,致使草甸区的微生物ACE、Chao1、Simpson、Shannon多样性指数显著低于沼泽区和沼泽化草甸区(P<0.05);随着演替发生,变形菌门(Proteobacteria)、酸杆菌门(Acidobacteria)、拟杆菌门(Bacteroidetes)、子囊菌门(Ascomycota)的相对丰度均减少,放线菌门(Actinobacteria)、芽单胞菌门(Gemmatimonadetes)、担子菌门(Basidiomycota)、被孢霉门(Mortierellomycota)的相对丰度增加;主坐标法分析(principal coordinates analysis,PCoA)排序分析显示,土壤微生物群落在各逆行演替都出现不同程度的离散...     

贫营养湖泊花神湖和紫霞湖浮游细菌群落结构分析

以南京市花神湖和紫霞湖两个贫营养型湖泊为研究对象,通过构建花神湖和紫霞湖16S rRNA基因克隆文库探讨了浮游细菌群落结构组成的变化。结果表明,花神湖和紫霞湖两湖泊水体中浮游细菌群落结构相似,主要隶属于放线菌门(Actinobacteria)、蓝藻门(Cyanobacteria)、α-变形菌门(Alphaproteobacteria)、β-变形菌门(Betaproteobacteria)、杆菌门(Bacteroidetes)、浮霉菌门(Planctomycetes)、疣微菌门(Verrucomicrobia)和芽单胞菌门(Gemmatimonadetes),其中放线菌门(Actinobacteria)、蓝藻门(Cyanobacteria)、β-变形菌门(Betaproteobacteria)是优势细菌类群。两个湖泊水体中75%的细菌与GenBank中已有的未培养细菌同源性高于97%,同时在两个克隆文库中还发现了6个淡水细菌新类群。通过对低纬度区域贫营养型湖泊浮游细菌群落结构的分析,加深了我们对浮游细菌多样性的了解,表明湖泊浮游细菌多样性有待进一步认识。     

五大连池不同火山喷发沉积物的细菌群落多样性研究

【目的】揭示五大连池火山区的细菌多样性。【方法】运用Illumina Miseq高通量测序技术解析五大连池不同火山喷发沉积物中细菌的群落组成和分布规律。【结果】五大连池火山区沉积物中的细菌主要包含厚壁菌门(Firmicutes)、变形菌门(Proteobacteria)、疣微菌门(Verrucomicrobia)、酸杆菌门(Acidobacteria)、绿弯菌门(Chloroflexi)、放线菌门(Actinobacteria)等23个细菌门类,其中,厚壁菌门、变形菌门丰度较高,芽孢杆菌属(Bacillus)为绝对优势菌群。另外,由于火山物质的同源性,导致各沉积物中细菌的代谢通路多与C、N、S、Fe等元素的生化循环相关,群落结构及功能具有一定程度的相似性。但由于各研究区域在沉积组分、环境因素及地质演化进程上的差异,致使芽孢杆菌属、地杆菌属(Geobacter)、酸杆菌属(Acidobacterium)、嘉利翁氏菌属(Gallionella)、Blastocatella等部分种群在不同火山沉积物中呈现差异化分布,具有典型的地域适应特性。【结论】五大连池火山沉积物中含有较为丰富的细菌资源,...     

16S rDNA克隆文库法探索转基因香石竹对土壤细菌群落的影响

【目的】通过研究转基因香石竹对土壤细菌群落的影响,为转基因香石竹的环境安全性评价提供依据。【方法】通过构建16S rDNA克隆文库,分析种植转基因和非转基因香石竹的土壤中细菌的群落结构组成。【结果】转基因和非转基因香石竹土壤中,共有的菌群有变形菌门(Proteobacteria)、浮霉菌门(Planctomycetes)、酸杆菌门(Acidobacteria),其中α-变形菌门、β-变形菌门、浮霉菌门为优势菌群;而在放线菌门(Actinobacteria)、疣微菌门(Verrucomicrobia)及未培养菌(Uncultured bacterium clone)等菌群存在部分差异。【结论】通过16S rDNA克隆文库方法揭示了转基因香石竹的土壤中细菌多样性十分丰富,其栽培对土壤细菌群落结构影响有限。     

高通量测序分析小头裸裂尻鱼皮肤和肠道的微生物多样性

【目的】解析小头裸裂尻鱼不同部位的微生物群落结构、物种组成、多样性特征以及菌群功能差异。【方法】通过Illumina MiSeq扩增子高通量测序,分析小头裸裂尻鱼皮肤黏膜、肠道黏膜和肠道内容物3个部位微生物菌群组成差异,并通过Tax4Fun预测菌群潜在功能。【结果】皮肤黏膜微生物α多样性最高,其Shannon指数显著高于肠道黏膜(P<0.05)和肠道内容物(P<0.001)。主坐标分析表明,皮肤黏膜微生物显著区别于其他2个部位。在门水平小头裸裂尻鱼3个部位相对丰度前五的微生物类群均为放线菌门(Actinobacteria)、变形菌门(Proteobacteria)、厚壁菌门(Firmicutes)、绿弯菌门(Chloroflexi)和蓝藻门(Cyanobacteria),其中肠道内容物中放线菌门相对丰度(46.53%)显著高于肠道黏膜(29.23%,P<0.05)和皮肤黏膜(25.83%,P<0.01);肠道黏膜中变形菌门的相对丰度(40.33%)显著高于肠道内容物(26.10%,P<0.05)。对各部位相对丰度前10的菌群进行分析发现,小头裸裂尻鱼皮肤黏...     

放牧对贝加尔针茅草原土壤细菌群落结构的影响

利用变性梯度凝胶电泳(PCR-DGGE)技术,研究了放牧对贝加尔针茅(Stipa baicalensis)草原土壤细菌群落结构的影响。结果表明,放牧条件下,贝加尔针茅草原0～20cm土壤中细菌类群包括变形菌门(Proteobacteria)的α、β、γ、δ变形菌纲,放线菌门(Actinobacteria)、芽单胞菌门(Gemmatimonadetes)、绿弯菌门(Chloroflexi)、厚壁菌门(Firmicutes)、酸杆菌门(Acidobacteria)、疣微菌门(Verrucomicrobia)、拟杆菌门(Bacteroidetes)、硝化螺旋菌门(Nitrospirae)等,其中变形菌门、放线菌门属优势类群,疣微菌门细菌是中度放牧样地仅有的类群;典型对应分析表明,土壤细菌群落与放牧强度、土壤理化性状密切相关。轻度放牧样地土壤细菌群落与有机碳、C/N比有较为密切的关系,中度放牧样地土壤细菌群落则与全N有较密切的关系,重度放牧样地土壤细菌群落与土壤pH有关。     

心脑血管疾病大额住院消费统计分析

通过对医院2004-2006年心脑血管疾病大额住院消费(消费大于50000元,以下简称大额消费)病例发病率高的前五种疾病的构成情况、药费、材料费消耗情况进行分析,认为加强大额病例中发病率高的病种的重点管理,是降低医疗费用的有效途径。建议制定常见病大额病种预定额付费方案和审查报销制度;采用适宜技术;控制药费,防止过度医疗,有效地遏制医疗费用的过快增长。     

Evidence that the C-terminus of OprM is involved in the assembly of the VceAB-OprM efflux pump

Although the architecture of tripartite multiple drug resistance (MDR) efflux pumps of Gram-negative bacteria has been well characterized, the means by which the components recognize each other and assemble into a functional pump remains obscure. In this study we present evidence that the C-terminal domain of the Pseudomonas aeruginosa OprM and the α-helical hairpin domain of Vibrio cholerae VceA play an important role in the recognition/specificity/recruitment step in the assembly of a functional, VceAB-OprM chimeric efflux pump. To our knowledge, this is the first evidence directly linking the C-terminal domain of an outer membrane efflux protein to its recruitment during the assembly of a tripartite efflux pump.     

两个猪苓菌株生长速度及酯酶同工酶比较研究

对不同地理分布的猪苓纯培养菌株进行了种性和酯酶同工酶的比较研究,结果表明,鸡爪苓（Z）纯培养菌株和猪屎苓（ZJ）纯培养菌株的种性有很大不同,两个纯培养菌株的酯酶同工酶酶带类型差异较大,亲缘关系较远。     

Synthesis of modified fragments of fibrinogen and their effect on the activity of proteolytic enzymes

New analogues of the Gly-Pro-Arg and Arg-Gly-Asp fragments of fibrinogen were synthesized: Gly-Pro-Arg-Pro (I), Gly-Pro-Arg-Pro-Met-OMe (II), Gly-Pro-Arg-Pro-Phe (III), Gly-Pro-Arg-Pro-Asp (IV), Gly-Pro-Arg-Pro-Glu (V), and Arg-Asn-Trp-Asp (VI). Their effect on the activity of proteases of various types was studied with the method of lysis of fibrin plates. All the peptides were found to inhibit plasmin activity (by 60–85%) and the γ-subunit of nerve growth factor (by 55–93%). Tetrapeptide (VI) proved to be an effective inhibitor of tissue activator of plasminogen and the γ-subunit of nerve growth factor (by 96 and 93%, respectively). The peptides exerted practically no effect on the activity of urokinase and moderately inhibited the activity of streptokinase [(III), IV), and (VI)], papain [(I), (II), IV), and (VI)], subtilisin [(V) and (VI)], α-chymotrypsin [(III), (V), and VI)], and Bacillus subtilis metalloprotease (VI). They inhibit trypsin [except for (I) and (III)] when applied on fibrin plates at a concentration of 1 × 10^?2 M, while, at the concentration of 1 × 10^?3 M, (I) and (II) induced an increase in proteolytic activity by 35 and 47%, respectively.     

Studies on the energy coupling sites of photophosphorylation IV. The relation of proton fluxes to the electron transport and ATP formation associated with Photosystem II

1. By using dibromothymoquinone as the electron acceptor, it is possible to isolate functionally that segment of the chloroplast electron transport chain which includes only Photosystem II and only one of the two energy conservation sites coupled to the complete chain (Coupling Site II, observed P/e₂ = 0.3–0.4). A light-dependent, reversible proton translocation reaction is associated with the electron transport pathway: H₂O → Photosystem II → dibromothymoquinone. We have studied the characteristics of this proton uptake reaction and its relationship to the electron transport and ATP formation associated with Coupling Site II.

2. The initial phase of H⁺ uptake, analyzed by a flash-yield technique, exhibits linear kinetics (0–3 s) with no sign of transient phenomena such as the very rapid initial uptake (“pH gush”) encountered in the overall Hill reaction with methylviologen. Thus the initial rate of H⁺ uptake obtained by the flash-yield method is in good agreement with the initial rate estimated from a pH change tracing obtained under continuous illumination.

3. Dibromothymoquinone reduction, observed as O₂ evolution by a similar flash-yield technique, is also linear for at least the first 5 s, the rate of O₂ evolution agreeing well with the steady-state rate observed under continuous illumination.

4. Such measurements of the initial rates of O₂ evolution and H⁺ uptake yield an H⁺/e⁻ ratio close to 0.5 for the Photosystem II partial reaction regardless of pH from 6 to 8. (Parallel experiments for the methylviologen Hill reaction yield an H⁺/e⁻ ratio of 1.7 at pH 7.6.)

5. When dibromothymoquinone is being reduced, concurrent phosphorylation (or arsenylation) markedly lowers the extent of H⁺ uptake (by 40–60%). These data, unlike earlier data obtained using the overall Hill reaction, lend themselves to an unequivocal interpretation since phosphorylation does not alter the rate of electron transport in the Photosystem II partial reaction. ADP, P_i and hexokinase, when added individually, have no effect on proton uptake in this system.

6. The involvement of a proton uptake reaction with an H⁺/e⁻ ratio of 0.5 in the Photosystem II partial reaction H₂O → Photosystem II → dibromothymoquinone strongly suggests that at least 50% of the protons produced by the oxidation of water are released to the inside of the thylakoid, thereby leading to an internal acidification. It is pointed out that the observed efficiencies for ATP formation (P/e₂) and proton uptake (H⁺/e⁻) associated with Coupling Site II can be most easily explained by the chemiosmotic hypothesis of energy coupling.     

林带阻力系数与透风系数关系的理论分析

根据冲量定理分析了林带对气流的阻力,首次得到了林带阻力系数的估算模式（Ｃｄ＝（１．８＋０．２α）（１－α）ｓｉｎ２ω,并利用有关文献发表的资料进行了验证．文章还对来流平行林带时林带对气流的阻力进行了讨论,指出来流平行林带时林带对气流的阻力仅为来流垂直林带时林带对气流阻力的０．７—１．１％,可以不予考虑．     

2种嵩草属植物形态特征对生境干旱化的响应

该研究于西藏自治区东南部的东达山高山草甸沿生境干旱化梯度设置10个样地,采用线性回归方法分析优势种高山嵩草(Kobresia pygmaea)和矮生嵩草(K.humilis)叶片数量与其它构件数值(分株数量、茎基直径、根系数量和根系长度)之间的关系,并采用线性回归斜率测度分株功效、叶片萌生能力、根系分生功效和根系伸长功效,以探讨嵩草的分株能力、茎基生长和根系生长对生境干旱化过程的响应机制。结果显示:(1)2种嵩草植物的叶片数量与4种构件数值均为显著线性正相关关系。(2)随着生境干旱化程度增加,高山嵩草分株数量增加,矮生嵩草分株受干旱抑制程度高,分株数量呈下降趋势;2种嵩草分株功效下降,即单株叶片数量因干旱化程度增加而减少。(3)2种嵩草的茎基直径、叶片萌生能力随着生境干旱化程度增加而下降;高山嵩草叶片萌生能力的变化与生境干旱化梯度一致,具有连续性;矮生嵩草叶片萌生能力对生境干旱化的适应性弱,干旱到一定程度发生骤降。(4)随着生境干旱化,高山嵩草的根系数量和长度均增加,矮生嵩草根系长度增加,但数量却无规律变化;2种嵩草的根系分生功效和根系伸长功效均下降。研究表明:嵩草属植物分布的最适宜生境为表面稍有积水的沼生生境。为适应生境干旱化,高山嵩草降低叶片萌生率以减少蒸腾作用,增加分株数量以增强对空间的占有能力,并增加根系数量和长度来提高对土壤水分的吸收能力,因此对干旱有较强适应性且分布范围广;矮生嵩草只通过降低叶片萌生率,增加根系长度响应干旱化生境,其分布范围较窄。     

The detection of salivary enzymes of phytophagous Hemiptera: a compilation of methods

The role of Hemipteran saliva and salivary enzymes is central to an understanding of the etiology of damage that these insects cause to plants. The dilute nature of the salivary secretions and the minute quantities in which they are often obtainable have made analysis and detection of salivary components very difficult. Such investigations in this laboratory have led us to formalise the techniques involved and we believe that the compilation of these methods presented herein may be useful to other research workers in this area. Methods are described for acid and alkaline phosphatase, esterase, /S-glucosidase, carbohydrases, invertase, amylase, proteinase, pectinase, cellulase, catalase, peroxidase, catechol oxidase, superoxide dismutase and ascorbic oxidase.     

民勤绿洲边缘地下水位变化对植物种群生态位的影响

在民勤绿洲边缘,利用空间区域不同地下水位(湖区8～12m,泉山区15～17m,坝区20～23m)和时间序列(1984～1992年)民勤沙井子地区地下水位下降(7.45～11.65m)梯度,研究地下水位下降对荒漠植物种群生态位的影响。结果表明:空间区域地下水位下降,植物种群生态位宽度均减小,种群退化;时间序列地下水位下降,白刺种群在扩展,其它植物种群在退化。白刺种群生态位宽度在民勤绿洲边缘荒漠植物群落中最大,是该区的建群种。由于白刺种群在地下水位7.45～11.65m范围内扩展,民勤绿洲生态环境治理中地下水位达到该范围是一个主要目标。     

The Method of Unweighted Means in Unbalanced Analysis of Variance

For factorial experiments with unbalanced data the method of unweighted means is an alternate method of analysis which is computationally simpler than likelihood ratio based F-tests. The quality of the null approximation to the F-distribution of the statistic for the method of unweighted means and the power of this test relative to the likelihood ratio test are discussed. Recommendations are made of when in the course of data analysis the superior operating characteristics of the likelihood ratio test may not outweigh the computational simplicity of the method of unweighted means test.