27 resultados para 16SrRNA


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形态和分子系统学的研究结果均表明,鲤科中的鲃系是一个单系类群,由鲃亚科、鲤亚科、裂腹鱼亚科和野鲮亚科组成,但它们各自的单系性及相互关系还没有系统发育证据.采用PCR方法获得了27种鱼类的16SrRNA基因序列,其中包括18种野鲮亚科鱼类,代表了中国野鲮亚科13个属.应用邻接法、最大简约法、最大似然法和Bayesian法分别构建其分支系统图.结果表明,野鲮亚科鱼类形成了一个单系类群,并且包括两个主要分支类群:其中一支包含鲮属、缨鱼属和墨头鱼属;另一个支系包括野鲮属、拟缨鱼属、异华鲮属、卷口鱼属、唇鲮鱼属、泉

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O objetivo principal deste estudo foi investigar a interação de 24 cepas de E. faecalis isoladas de infecções endodônticas primárias às proteínas de matriz dentinária, como também a moléculas de matriz presentes em lesões de endocardites. A análise desta interação foi feita através de técnica enzimática, com confirmação pela técnica de fluorescência. Além disto, foi realizada a confirmação do isolamento da espécie E. faecalis, através da técnica de PCR para o gene 16SrRNA e a análise da presença de genes de virulência da referida espécie microbiana para aderência às supostas proteínas de matriz incluindo às de ligação ao colágeno (ace, gelE, esp, agg e efaA). O maior padrão de interação das cepas ocorreu com a fibronectina (83,4%), seguido pelo fibrinogênio (62,5%) e colágeno humano tipo I (52%). Curiosamente, a aderência observada para o colágeno do tipo I, foi de pequena magnitude, quando comparado com a amostra padrão da ATCC 29212. As cepas ATCC 29212, A1, A43 e A68 interagiram com todas as proteínas de matriz utilizadas neste estudo. Um percentual expressivo das cepas testadas apresentou amplificação para efaA (86,9%) e para ace (73,9%). Paralelamente, todas as cepas apresentaram amplificação para gelE e foram negativas para os genes agg e esp. Adicionalmente, não houve correlação entre a detecção dos genes de virulência e a interação às proteínas de matriz, evidenciando que, mesmo com a detecção dos genes nas amostras, se faz necessário avaliar a expressão gênica por qPCR.

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The pantherine lineage of the cat family Felidae (order: Carnivora) includes five big cats of genus Panthera and a great many midsized cats known worldwide. Presumably because of their recent and rapid radiation, the evolutionary relationship among pantherines remains ambiguous. We provide an independent assessment of the evolutionary history of pantherine lineage using two complete mitochondrial (mt) genes (ND2 and ND4) and the nuclear beta-fibrmogen intron 7 gene, whose utility in carnivoran phylogeny was first explored. The available four mt (ND5, cytb, 12S, and 16SrRNA) and two nuclear (IRBP and TTR) sequence loci were also combined to reconstruct phylogeny of 14 closely related cat species. Our analyses of combined mt data (six genes; approximate to 3750 bp) and combined mt and nuclear data (nine genes; approximate to 6500 bp) obtained identical tree topologies, which were well-resolved and strongly supported for almost all nodes. Monophyly of Panthera genus in pantherine lineage was confirmed and interspecific affinities within this genus revealed a novel branching pattern, with P. tigris diverging first in Panthera genus, followed by P. onca, P. leo, and last two sister species P. pardus and P. uncia. In addition, close association of Neofelis nebulosa to Panthera, the phylogenetic redefinition of Otocolobus manil within the domestic cat group, and the relatedness of Acinonyx jubatus and Puma concolor were all important findings in the resulting phylogenies. The potential utilities of nine different genes for phylogenetic resolution of closely related pantherine species were also evaluated, with special interest in that of the novel nuclear beta-fibrinogen intron 7. (c) 2005 Elsevier Inc. All rights reserved.

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通过改进裂解温度和延长裂解时间并增加苯酚/氯仿洗脱次数的DNA提取方法获得南京玄武湖底泥中的DNA,通过PCR法来扩增微囊藻的16SrRNA基因.结果表明在所有采样点中均得到微囊藻基因组DNA,并且纯度较高,OD_(260)/ OD_(280)均高于1.54,最高值达到1.89.PCR的扩增结果显示所有样点的DNA都得到212 bp大小的微囊藻16SrRNA基因片断,表明这种方法可以有效的从底泥中提取微囊藻的DNA,从而为研究底泥微囊藻生理生态及其越冬、上浮、形成水华的机理提供更有利的方法.

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对从青岚湖采集的 10属 14种河蚌的 19个样本进行了 16SrRNA的序列测定 ,并同GenBank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 .0 2 74— 0 .2 2 90 ,平均为 0 .132 5 ,种内遗传距离在0 .0 0 34— 0 .0 0 6 8之间 ,平均为 0 .0 0 4 5 ,种间遗传距离远大于种内距离。表明以 16SrR

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石爬属鱼类的青石爬和黄石爬的物种界限一直不清楚。采用形态判别和线粒体 16SrRNA基因序列分析相结合的方法 ,分别研究分析了青石爬和黄石爬的物种划分、地理分化及遗传多态性。结果表明 :(1)区别青石爬和黄石爬的重要特征腹鳍起点至臀鳍起点的距离是否大于至鳃孔下角的距离 ,腹鳍相对位置 ,头部相对大小与上颌须的须状延长部分等相互之间有一定的相关性 ,但是在研究的样本中没有截然的界限 ,而是有较大的重叠 ,难以区分 ;(2 )从地埋分布看 ,金沙江不同支流的样本在上述特征上有一定的区别 ,但是没

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首次测定了中国淡水贝类———蚌科 (Unionidae) 1 3个属代表种类的线粒体 1 6SrRNA部分序列。用Clustal X排序软件进行 1 6SrRNA序列的对位排列 ,序列总长度为 30 5— 32 0bp。通过Mega 2 0软件对所得线粒体 1 6SrRNA片段序列进行比较 ,共发现 1 0 8个碱基存在变异 ,其中包括 77个简约信息位点 ,并用“Pairwisedistance”计算了各属间的相对遗传距离。以贻贝为外类群 ,采用Mega 2 0软件中的“Neighbore Joinin

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在红鲤、镜鲤和野鲤中进行线粒体 1 6SrRNA和Cytb基因片段的序列测定。结果发现 ,3个鲤品种在多达 859个碱基长度上无变异 ,但是与已报道的鲤全序列中的同源序列则有约 1 5%的趋异。这个结果提示 3个鲤品种可能在起源中是独立的一支 ,并且分化极低

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近年的研究表明,海绵微生物是某些海绵天然产物的真正产生者。因此,人们将海绵微生物作为开发海绵天然产物的重要来源之一。采用琼脂块法和液体扩散法,从分离自中国黄渤海大连海域的海绵优势种—繁茂膜海绵的28株放线菌中筛选到4株具有抗菌活性的放线菌,并对它们进行生物学鉴定。采用经典和现代分类鉴定方法,对4株具有抗菌活性的繁茂膜海绵放线菌的形态特征、培养特征、生理生化特征、细胞壁化学组分和16SrRNA序列进行了研究,得出种水平的鉴定结果:Hmp-S14为西唐氏链霉菌Streptomycessetonii;Hmp-S19为灰色链霉菌Streptomyces griseus;Hmp-S24为桔橙小单抱菌Micromonospora aurantiaca;Hmp-S26为生二素链霉菌Streptomyces ambifaciens。在四株具有抗菌活性的繁茂膜海绵放线菌中,菌株Hmp-S19的抗菌活性优于其它三株,并且与已报道的20多种灰色链霉菌菌株有不同的生理生化特性,故进一步优化其发酵条件并初步研究了S19抗菌素的理化性质。通过单因子和均匀设计实验,优化菌株Hmp-S19摇瓶发酵条件。确定最佳发酵培养基:玉米粉0.6%,葡萄糖0.1%,豆饼粉0.5%,NaCl 0.3%,KH2PO40.08%,CaCO30.08%,MgSO40.02%;最佳发酵条件:接种龄30h,接种量5%,初始pH7.0,发酵时间96h,装液量100ml/50ml,培养温度28 ℃。应用二剂量法测定519一抗菌素的相对效价,为5154μ/ml,较原始发酵培养基和发酵条件(3364μ/ml)提高了53%。通过pH纸层析和捷克八溶剂系统纸层析试验,初步判定519抗菌素为两性、非水溶性I型抗菌素。Hmp-S19发酵液经预处理、萃取、硅胶柱层析、制备薄层层析等步骤,对S19抗菌素进行分离纯化得粗制品,并进行了液 相色谱一质谱检测。

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本研究从磷霉素(FOM)生产车间和储存仓库附近土壤中筛选手性环氧化顺式丙基磷酸(cPA)合成FOM的高效生物转化细菌。采用两种筛选模型,从土壤样品中分离到了183株待选细菌,其中147株为FOM抗性细菌,36株为cPA利用细菌。采用改进的琼脂块培养法,初筛获得了22株生物转化合成FOM菌株;再经振荡培养法,复筛获得3株生物转化活力较高的菌株,分别为FB3212、FB4211和FB7912。通过培养特征、生理生化特征性试验和16SrRNA序列分析鉴定,确定FB3212为赤红球菌(Rhodococcusruber),FB4211属于De沂ia菌属,FB7912为梭形芽抱杆菌(Bacillusfusformis),皆为新发现的生物转化合成FOM菌种。通过转化特性试验,FB3211、FB42n和FB7912的FOM转化合成均属于VO3-依赖型。甘油与Co2+对转化有明显的促进作用。在分批培养实验中FoM浓度在72h内随时间延一长而增高。底物cPA浓度对转化的影响最大,在三株菌中,FB7912的底物的抑制效应最小,转化活性最高,当cPA浓度为10mg/mL时,转化效果最佳,磷霉素转化产量为1.15mg/mL,转化率为10.17%。

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对隆肛蛙属的物种构成进行了订正,建立新属肛刺蛙属Yerana gen. nov.;订正后的隆肛蛙属现仅隶2种, 即隆肛蛙F. quadrana和太行隆肛蛙F. taihangnicus。运用形态学分析探讨了隆肛蛙属物种及种群的形态差异和分类关系,通过分子系统学研究探讨了隆肛蛙属物种及种群的分类和系统发育关系,运用动物地理学方法结合系统发育关系探讨了隆肛蛙属种群的地理分布格局成因与历史过程。主要结果和推论如下: 1.隆肛蛙属物种构成的订正及一新属建立 建立新属肛刺蛙属,将隆肛蛙属中的原叶氏隆肛蛙F. yei归隶新属肛刺蛙属并更名为叶氏肛刺蛙Y. yei,,新属建立的主要依据为:(1)雄性肛部隆起,肛孔下方有两个布满黑刺的大的白色球形隆起,具单咽下内声囊, 第一指具婚刺;(2)形态量度分析表明叶氏肛刺蛙与隆肛蛙和太行隆肛蛙的形态差异远大于后两者之间的差异;(3)叶氏肛刺蛙的分布区与隆肛蛙和太行隆肛蛙的分布区距离较远且呈隔离状态;(4)分子系统学研究资料(Jiang et al.,2005)证明叶氏肛刺蛙与隆肛蛙和太行隆肛蛙非单系发生;叶氏肛刺蛙在第二支中位于基部。因此,隆肛蛙属现仅隶2种,即隆肛蛙和太行隆肛蛙。 2.隆肛蛙属种群形态学研究 对隆肛蛙属中隆肛蛙和太行隆肛蛙的15个地理种群565只标本的28项形态性状进行了测量,运用典型判别分析法对其分析的结果表明:(1)太行隆肛蛙与隆肛蛙形态差异明显,支持其为不同的物种;(2)原隆肛蛙河南伏牛山种群和山西中条山种群应为太行隆肛蛙的地理种群;(3)隆肛蛙不同地理种群之间形态差异明显,其中四川安县种群、陕西周至种群和湖北利川种群与模式产地重庆巫山种群的差异可能达到了亚种或亚种以上分化水平。对隆肛蛙属量度分析的15个种群进行定性形态分析表明其分为三种形态型,对应隆肛蛙、过渡型和太行隆肛蛙,其变异特征主要为内跗褶、雄性肛部隆起及疣粒分布、第五趾外侧缘膜等,这与量度分析结果相似。 3.隆肛蛙属种群分子系统学研究 测定隆肛蛙属Feirana的2种19种群的线粒体12S rRNA和16S rRNA基因片段、ND2基因的DNA序列,比对后共计1953bps。(1)遗传多样性与距离分析:结果表明,隆肛蛙属种群具很高的遗传多样性,19个种群样品表现出19种单倍型(遗传多样性指数Hd=1.0); ND2基因的进化信息含量远高于12SrRNA和16SrRNA。隆肛蛙属2种群组内的种群间的遗传距离远小于两种群组间的距离,种群在不同基因上的遗传距离表现的关系与对应的系统树一致。(2)系统发育关系分析:结果表明,不同基因片断基于不同方法构建的隆肛蛙属种群系统发育树结构基本一致,基本表明隆肛蛙属种群为单系发生;它们在系统树中分为两大支,分别对应于隆肛蛙和太行隆肛蛙;支持中条山种群(沁水、历山和济源种群)和伏牛山种群(栾川和内乡种群)为太行隆肛蛙的地理种群,而原隆肛蛙秦岭中东段的部分种群(柞水、宁陕、长安大坝沟种群)也应为太行隆肛蛙的地理种群。(3)亚种分化分析:根据遗传距离分析和系统发育关系分析结果,并考虑形态上的差异情况以及地理分布信息,隆肛蛙所隶种群组可分为2亚种,即隆肛蛙指名亚种F. quadrana quadrana包括四川盆地东缘大巴山东段-巫山-武陵山北麓种群和秦岭中段(周至板房子和长安广货街)种群,他们在系统关系树上聚为一支;安县亚种F. quadrana anxianensis包括四川盆地西缘岷山东麓-龙门山-大巴山和秦岭西段的种群(安县、青川、文县、南江和凤县种群),他们在系统关系树上聚为一支。太行隆肛蛙所隶种群组也可分为2亚种,即太行隆肛蛙指名亚种F. taihangnicus taihangnicus包括中条山的种群(沁水、历山和济源种群)和中东秦岭的部分种群(柞水、长安大坝沟和宁陕种群),他们在系统关系树上聚为一支;太行隆肛蛙伏牛亚种F. taihangnicus funiuensis,为伏牛山地区的种群(栾川和内乡种群),他们在系统关系树上聚为一支。 4.隆肛蛙属种群动物地理学研究 隆肛蛙属19种群的分歧年代分析: 以长江巫山段和黄河三门峡段的形成历史时期为参考点,根据已测隆肛蛙属19种群及其外群包括N. pleski、P. yunnanesis、P. robertingeri、F. limnocharis的1953bps DNA序列构建分子钟,获得各支系的分歧年代。结果表明:①棘蛙族在70Ma左右开始其独立演化历程,这与Roelants et al.(2004)的分析结果~60±15Ma左右开始分化基本一致,后者印证了本文的分子钟。②隆肛蛙属的起始分化年代较早,隆肛蛙和太行隆肛蛙两种群组的最近祖先种群大概在46Ma~50Ma左右;隆肛蛙和太行隆肛蛙种群组内的种群分化年代相对两种群组间晚得多, 隆肛蛙种群组内两亚种分化起始年代约为10Ma左右,而太行隆肛蛙种群组内两亚种分化起始年代约为6Ma。 隆肛蛙属种群分布格局形成过程分析: ①隆肛蛙属的系统关系与地理分布格局密切相关,大部分系统分支分级与地理距离成正比;②隆肛蛙属最近祖先种群的分化中心可能位于秦岭中部地区, 隆肛蛙属的种群分布格局的形成表现为隔离分化与扩散相结合的机制,由隔离分化产生的隆肛蛙祖先种群主要从秦岭中部向西南方向扩散,后隔离分化为两亚种;太行隆肛蛙祖先种群向东北方向扩散也分化为两亚种。 隆肛蛙属种群分布区域地质历史的探讨:本文所建分子钟和种群分化方式印证了该区域的几次主要地质事件,包括岷山-龙门山-西秦岭等地区的快速差异隆起、第四纪冰期等。 The specific composition of the genus Feirana should be revised. A new genus Yerana gen. nov.(Ranidae:Dicroglossinae)was established based on morphological data-set and molecular phylogeny, as a result, only two species F. quadrana and F. taihangnicus are classified into Feirana now. Morphological differences and taxonomy of populations of Feirana were investigated based on morphological and morphometric data; phylogenetic relationships and taxonomy of populations of Feirana were elucidated using molecular data, and then the proceeding of the distribution pattern of populations of Feirana were discussed. The main results and conclusions and proposals were presented as following: 1. Revising of the specific composition of the genus Feirana and establishment of a new genus The new genus Yerana, only containing the type species Y. yei, was established based on the following evidences: (1) In adult male, distinct up-heaved circular vesicle presents around the anal, and under anal there are two white balls on which black spines exist, black horny spines scatter on the upper side of first finger, and internal single subgular vocal sac presents; (2) there is obvious morphometric differences between Yerana and Feirana; (3) Yerana is distributed far from Feirana; (4) evidences of molecular phylogeny(Jiang et al.,2005)suggested that Yerana take a special phylogenetic clade which is different from other genus included in the tribe Paini. As a result, there are only two species in Feirana, i.e., F. quadrana and F. taihangnicus. 2. Morphological research of populations of Feirana Twenty-eight characters of 565 individuals of 15 populations of the genus Feirana were measured, the results of Canonical Discriminant analysis of the morphometric data-set indicated that: (1) there are very prominent differences between the two species F. quadrana and F. taihangnicus. The validity of species F. taihangnicus was approved here; (2) Mt. Funiu population and Mt. Zhongtiao population should belong to the species F. taihangnicus; (3) Obvious differences exist among 12 populations of F. quadrana, the differentiation among Zhouzhi population, Anxian population, Lichuan population, and Wushan population together with the others probably reach sub-specific or specific level. Result of morphological comparison between 15 different populations show that 3 morphological types are recogenized in according with F. quadrana, F. taihangnicus and intergradation, this result conform to the result of morphometric analysis. 3. Molecular phylogenetic study on populaions of Feirana Fragment of 12SrRNA and 16SrRNA genes, and ND2 gene of 19 populations of two species of Feirana were sequenced and aligned, from which 1953 bps were received. (1) analyses of genetic distance and hereditary diversity indicated that: genetic distance between populations in each group were less than distance between two groups of Feirana, 19 haplotypes were recognized from 19 samples of 19 populations, so the hereditary diversity of populations of Feirana was very high (Hd=1.0), phylogenetic information in ND2 gene is more than fragment sequence of 12SrRNA and 16SrRNA genes. (2) Result of molecular phylogeny indicate that the phylogenetic trees constructed using different methods based on different sequence data sets showed the revised genus Feirana is monophyletic since the 19 populations of Feirana were firstly clustered together as one large clade, which was further clustered into two major clades, corresponding to F. quadrana(GroupⅠ) and F. taihangnicus(GroupⅡ), respectively. So populations of Qinshui and Lishan in Mt. Zhongtiao, populations of Luanchuan and Neixiang in Mt. Funiu, and populations of Zhashui, Dabagou of Chang’an and Ningshan in eastern Mt. Qinling should belong to the species F. taihangnicus; (3) Subspecific differentiation. on the basis of genetic distance, phylogenetic trees and geographical distribution, F. quadrana should have two subspecies, i.e., F. quadrana qudadrana, consisting of the populations Guanghuojie of Chang’an and Zhouzhi in Mid-Mt. Qinling, populations in Wushan area and northern Mt. Wuling (Lichuan), and F. qudadrana anxianensis, consisting of the populations in eastern Mt. Ming shan-Mt. Longmen-western Mt. Daba-western Mt. Qinling (Anxian, Qingchuan, Wenxian, Nanjiang and Fengxian); F. taihangnicus should also has two subspecies, i.e., F. taihangnicus taihangnicus, consisting of the populations in Mt. Zhongtiao and eastern Mt. Qinling, and F. taihangnicus funiuensis, consisting of the populations in Mt. Funiu. 4. Zoogeography of populaions of Feirana Analysis for divergent time of 19 populations of Feirana: Using the dates of run-through of Wushan segment of Changjiang River as the time when the population of Lichuan started differentiated from the populations of Wushan and Shennongjia, and the dates of Sanmenxia segment of Yellow River as the time when the populations in Mt. Zhongtiao started differentiated from the population of Dabagou in Chang’an, molecular clock was established using sequences with 1953 bps of 19 populations of Feirana and outgroup including N. pleski, P. yunnanesis, P. robertingeri, F. limnocharis in order to estimate divergent time of all clades. Result of that indicated that: ① the tribe Paini started to evolve independently at about 70Ma when is in consistent with that estimated by Roelants et al.(2004)with result of about ~60±15Ma, they were corroborated by each other, this confirms the validity of this molecular clock; ② divergent time for speciation of Feriana is early, ancestral populations of F. quadrana and F. taihangnicus were found about 46Ma~50Ma; differentiation of populations within species is greatly late to the divergence of the two species, divergent time for F. quadrana is 10Ma and divergent time for F. taihangnicus is 6Ma. Proceeding of distribution pattern of Feirana. Phylogenetic relationships of populations of Feirana matched quite with distribution pattern of them, the relationships among clades showed in phylogenetic trees is direct ratio to geographical distance of them; the estimated date of speciation between two species of Feirana was as early as speciation of Paa yunnanesis and Nanara pleski; middle part of Mt. Qinling is the center of speciation of Feirana, combination of mult-events of dispersal and vicariance are probably the mechanism of speciation of Feirana, F. quadrana colonized the mid-Mt. Qinling and then differentiated into two subspecies in southwest direction, ancestral population of F. taihangnicus colonized the mid-Mt. Qinling and then differentiated into two subspecies in northeast direction. On geological history of the distribution of Feirana. According to molecular clock and speciation model of populations of Feirana, some geological events are confirmed, including special rise of Mt. Minshan- Mt. Longmen-western Mt. Qinling, glacial age.

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采用PCR-DGGE方法,对我国东北地区长期石油和重金属污染农田土壤中的假单胞菌多样性及其种群结构进行研究.结果表明:石油污染区土壤中假单胞菌多样性指数显著高于重金属污染区;石油污染区旱田土壤假单胞菌多样性接近于对照清洁土壤,但低于相似污染程度的石油污染区水田土壤,表明污染物类型与耕作管理方式是影响土壤中假单胞菌多样性的主要因素.经16SrRNA的V6/V7区测序,Pseudomonas mendocina、P.stutzeri和P.aerugi-nosa是该石油和重金属污染区土壤中的优势类群,说明在长期污染胁迫下,这3种假单胞菌分别得到了不同程度的富集,推测与石油烃的自然降解及假单胞菌的重金属抗性有关.

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放线菌Frankia与放线菌根植物在根瘤中共生的现象发生在8科24属被子植物上.系统发育分析表明:Frankia可分为4个族,其菌株对寄主植物有明显的偏好性.用大理旱冬瓜根瘤接种昆明和武定旱冬瓜幼苗,收集幼苗上的根瘤,再对幼苗接种数次.用武定旱冬瓜和川滇桤木幼苗做同样实验,最后对Frankia的16SrRNA基因进行ARDRA分析,发现接种幼苗根瘤中Frankia与接种根瘤中Frankia产生相同的16SrRNA基因ARDRA带型.

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从南海红树林木榄(Bruguiera gymnorrhizo)根际土壤中分离到一株具有拮抗杉木致害菌尖孢镰刀菌萎蔫专化型SF2(Fusariumoxysporum f.sp.vasinfectum)活性的海洋细菌3728菌株,对分离菌株的形态特征、培养特征、生理生化特征和16S rRNA基因序列进行了系统的研究。发现细菌3728与枯草芽孢杆菌(Bacillus subtilis)序列相似性最高,达到100%,在系统进化树中与枯草芽孢杆菌(Bacillus subtilis AJ276351)处于同一分支上,结合形态和生理生化分析结果,将其鉴定为枯草芽孢杆菌。