新疆3种雅罗鱼线粒体DNA控制区序列的差异和系统进化关系

对分布在新疆的准噶尔雅罗鱼(Leuciscus merzbacheri)、贝加尔雅罗鱼(Leuciscus leuciscus baicalensis)和高体雅罗鱼(Leuciscus idus)3个鱼种共24尾个体的线粒体DNA D-loop控制区核苷酸序列进行了测定,获得24条长度为667—669bp的同源基因序列。3种雅罗鱼之间的序列差异在6．39％—9．89％之间,贝加尔雅罗鱼与高体雅罗鱼种间序列同源性高,变异程度小;贝加尔雅罗鱼与准噶尔雅罗鱼种间序列同源性最低,变异程度最大。所采集的贝加尔雅罗鱼两个地理群体(赛里木湖和额尔齐斯河)内mtDNA的平均核苷酸碱基序列差异为1．07％和1．08％;两群体间的序列差异为1．07％,显示两个地理群体间无明显分化。DNA序列数据显示,这3种鱼类线粒体DNAD-loop序列变异丰富,24尾个体呈现独自的单倍型。同源基因序列平均含AT碱基64．1％,GC碱基35．9％,显示准噶尔雅罗鱼、贝加尔雅罗鱼、高体雅罗鱼的线粒体DNAD-loop区核苷酸组成的不均一性。分子系统树提示,贝加尔雅罗鱼与高体雅罗鱼亲缘关系较近,准噶尔雅罗鱼是3种雅罗鱼中较古老的鱼种。     

新疆3种雅罗鱼线粒体DNA细胞色素b序列的差异与系统进化

利用鲤科鱼类线粒体DNA细胞色素b通用引物对分布在新疆的准噶尔雅罗鱼(Leuciscusmerzbacheri)、贝加尔雅罗鱼(L .baicalensis)和高体雅罗鱼(L .idus) 3个鱼种共1 5尾个体的线粒体DNACytb部分核苷酸序列进行了测定,获得1 5条长度为41 3bp的同源基因序列。同源基因序列分析显示,在3种雅罗鱼1 5条mtDNACytb基因片段中,A、T、C、G碱基的平均含量分别是:2 7 4%、2 6 7%、1 7 2 %、2 8 7% ,其中A T含量(5 4 1 % )高于G C含量(4 5 9% ) ;共检测到5 2个突变位点;转换比颠换发生频率高,转换颠换比值(R)是3 5。mtDNACytb的进化速率在3种雅罗鱼间表现出不一致性,贝加尔与高体雅罗鱼、贝加尔与准噶尔雅罗鱼种间遗传距离分别是0 1 2 5 1和0 1 2 61 ,保守性较低;高体与准噶尔雅罗鱼种间的遗传距离是0 0 0 0 7,高度保守。mtDNACytb分子系统树显示,在3种雅罗鱼中贝加尔雅罗鱼独立成一枝,高体雅罗鱼和准噶尔雅罗鱼聚成一类。提示了,在mtDNACytb分子水平上,高体雅罗鱼和准噶尔雅罗鱼的亲缘关系十分相近,贝加尔雅罗鱼与准噶尔雅罗鱼的亲缘关系相距较远。我们对准噶尔、贝加尔和高体雅罗鱼mtDNACytb方面的研究与陈星玉对其骨骼类型的研究结果相吻合。     

准噶尔雅罗鱼组织同工酶表达特性

使用不连续PAGE法,分析了10尾准噶尔雅罗鱼(Leuciscus merzbacheri)眼睛、鳃、皮、背部肌肉、鳍和肝胰脏6种组织的10种同工酶(LDH,CCO,EST,CAT,POD,ME,MDH,G6PD,GDH,ADH)的差异表达,并对部分同工酶基因位点及表达酶谱表型进行了分析,以期为其种质资源保护和开发以及遗传育种等方面的研究提供基础资料。结果显示,10种同工酶中9种在6种组织中出现了明显的组织差异性,仅CCO在6种组织中的差异性较小。在对准噶尔雅罗鱼的10种同工酶的遗传多样性分析中,共记录到了21个基因位点,其中Est-1、Me-B、s-MDH、G6pd-A、G6pd-B和Adh-A为多态性基因位点。多态位点比例为:P=6/21=28.57%。     

额尔齐斯河高体雅罗鱼指环虫种群动态及中国一新记录种（英文）

2009年4月到2012年7月对额尔齐斯河(中国段)高体雅罗鱼鳃部寄生的指环虫进行了取样调查,以期了解该鱼指环虫的种群生态学特点。结果表明,高体雅罗鱼共感染了三种指环虫,分别为号筒指环虫、小刺指环虫、大翼指环虫,该鱼的总感染率为28.47%,平均感染强度为7.39(1—42)。感染率、感染强度在不同体长组的宿主中表现出不同的变化趋势,指环虫的感染率在30 cm以上的宿主中达到最大,为50%,而在L10范围内未发现感染;在不同体长组宿主中指环虫种群的分布类型主要为聚集分布,另外,根据形态学特征鉴定大翼指环虫(Dactylogyrus alatus f.major Linsyow,1878)为我国指环虫属一新记录种,论文对其主要形态特征进行了描述并绘制了特征图,同时与寄生于高体雅罗鱼鳃上的国内已记述指环虫属虫种的主要形态特征作了比较。     

鲤科四种鱼肠道菌群的相似性及其与系统进化的关系

在同一背景条件下,对草鱼、白鲢、团头鲂和鲤鱼等4种鱼的肠道菌群中好氧性菌或兼性厌氧菌进行分析鉴定,得到4种鱼之间的肠道菌群相似值.用UPGMA和NJ法构建鲤科的雅罗鱼亚科、鲢亚科、鳊亚科和鲤亚科等4种亚科的系统演化树.不同亚科的鱼间菌群分布有较大差异,但雅罗鱼亚科和鳊亚科在菌群分布方面有极显著的正相关性.哈夫尼亚菌属、致病杆菌属、气单胞菌属、柠檬酸菌属、假单胞菌属等在4种鱼肠道中均有分布,链球菌属、拉恩氏菌属、芽孢杆菌属、葡萄球菌属等属于共生菌.雅罗鱼亚科和鳊亚科相邻邻接为一个共同接点,该接点与鲤亚科连接为一个接点,鲤亚科可能是最原始的类型。 Abstract:This paper is an analysis and appraisal of aerobe and facultative anaerobe among intestinal bacterium of the following fishes:Ctenophar yngodomidellus,Hypophtha lmichthys molitrix,Megalorama ambly cephala Yih and Cyprinus carpio haematopterus under same surroundings.After that,We contain similarity values of intestinal bacteria were obtained between every two kinds of the four fishes.We constructed the revolutionary trees of in Cyprinidate,Leuciscinae,Abramidinae,Cyprininae and Hypophthalmi by NJ method and UPGMA method according to the similarity values.     

六种鲤科鱼类核仁组织者区的研究

采用银染及荧光染色技术,对6种鲤科鱼的NORs进行了研究。结果表明：这6种鲤科鱼中瓦氏雅罗鱼、花 、麦穗鱼、白甲鱼具有2个NORs,长春鳊、墨头鱼具有4个NORs。根据实验研究结果对鲤科鱼类NORs多态性及演化等进行了讨论。 The NORs were examined in 6 species of Cyprinidae by both silver nitrate and chromomycin A3,which led to the detections of 2 NORs in 4 species(Leuciscus waleckii,Hemibarbus maculates,Pseudorasbora parva,and Varicorhinus simus)and of 4 NORs in 2 species (Parabramis pekinensis,and Garra pingi pingi).Based on our results,the variation and evolution of fish NORs were discussed.     

云南保山猪线粒体DNA D-loop区序列初步分析 Primary Analysis of Mitochondrial DNA D-loop Region Sequence in Baoshan Pig

摘要：为了解云南保山猪(Baoshan pig)的遗传多样性及其遗传背景,我们测定了19个个体线粒体DNA D-loop高变区I 15 363～15 801片段序列438 bp。检测到10种单倍型,包括8个多态位点,其中5次T/C转换、1次G/A转换、1次G/C颠换和1次A/T颠换,其A、T、G、C碱基的平均含量分别为35.4%、26.9%、13.2%和24.5%,A+T含量(62.3%)明显高于G+C含量(37.7%)。对于保山猪的保种及其持续利用有着重要的理论指导意义。 Abstract:To investigate the genetic diversity and genetic data of Baoshan pig in Yunnan province,the mitochondrial DNA D-loop hypervariable segment I sequences 15 363～15 801 (438 bp) in 19 individuals of Baoshan pig were sequenced.Ten mitochondrial haplotypes were identified in the samples,with 8 sites showing polymorphism,which were 5 T/C and 1 G/A transitions,1 G/C and 1 A/T transversions.The contents of A,T,G and C were 35.4%,269%,13.2% and 24.5%,respectively.The content of A+T (62.3%) was significantly higher than that of G+C (37.3%).It will be of importance to conservation and sustainable utilization in Baoshan pig.     

Genetic polymorphism of mitochondrial DNA HVS-I and HVS-Ⅱ of Chinese Tu ethnic minority group

We analyzed the two hypervariable segments HVS-Ⅰ and HVS-Ⅱ of 108 Chinese Tu ethnic minority group samples for forensic and population genetics purposes.Comparing with Anderson sequence,79 polymorphic loci in HVS-Ⅰ and 40 in HVS-Ⅱ were found in Chi-nese Tu ethnic minority group mtDNA sequences,and 90 and 64 haplotypes were then defined.Haplotype diversity and the mean pair-wise differences were 0.9903±0.0013 and 5.7785 in HVS-Ⅰ,and 0.9777±0.0013 and 3.5819 in HVS-Ⅱ,respectively.By analyzing the hypervariable domain from nucleotide 1,6180 to 1,6193 in HVS-Ⅰ,we defined some new types of sequence variations.We also compared the relationship between Tu population and other populations using mtDNA HVS-Ⅰ sequences.According to Rst genetic distances,the phylogenetic tree showed that the Tu population,the Xi'an Han population,the Chinese Korean,and the Mongol ethnic group were in a clade.This indicated a close genetic relationship between them.There were far relations between the Tu population and other Chinese southern Han populations,Siberian,European,African,and other foreign populations.The results suggest that Tu population has a multi-origin and has also merged with other local populations.     

广州地区106例汉族人群MICA和MICB微卫星基因座 单体型研究

利用聚合酶链反应和荧光（6－FAM）自动化检测技术对广东地区汉族106例无亲缘关系样本进行MICA基因外显子5和MICB基因内含子1微卫星基因座多态性及其单体型分布调查。根据群体资料估算两者间的单体型频率、连锁不平衡参数、相对连锁不平衡参数。结果显示,广州地区汉族人群MICA和MICB微卫星基因座基因型分布符合Hardy-Weinberg平衡法则,共检出MICA微卫星基因座 5个等位基因, MICB微卫星基因座14个等位基因。其中MICA A5基因频率最高（0.2877）,A4基因频率最低（0.1321）。MICB CA14等位基因频率最高（0.3255）,CA19、CA28等位基因频率最低（0.0047）,未检出CA27。21种MICA－MICB单体型频率大于1%(连锁不平衡参数>0), 其中单体型A5－CA14 (16.73%), A5.1－CA18 (8.75%), A4－CA26(3.76%),A9－CA15(3.66%)和A6－CA21(2.61%)为强连锁常见单体型（χ2>3.84, P<0.05）。广州地区汉族人群MICA和MICB微卫星基因座多态性和单体型分布有其自身特点,MICA和MICB微卫星基因座适合做为遗传标志,用于人类学、遗传疾病基因连锁分析、法医学亲子鉴定和个体识别等研究领域。Abstract: This study is to investigate genetic polymorphisms and haplotypes of microsatellite locus in the exon 5 of the MICA gene and intron 1 of the MICB gene based on 106 samples of Guangzhou Han Population by polymerase chain reaction and fluorescent technique (6-FAM). The corresponding haplotype frequencies, linkage disequilibria values and relative linkage disequilibria values were estimated based on population data. The results show that the genotype distributions of MICA and MICB microsatellite meet Hardy-Weinberg equilibrium in Guangdong Han population. In total, 5 alleles of MICA microsatellite locus and 14 alleles of MICB microsatellite locus were observed. MICA A5 was the most common allele (0.2877), whereas A4 was the least popular one (0.1321). MICB CA14 was the most common allele (0.3255), and CA19 and CA28 were the least popular ones (0.0047). CA27 was not observed. Twenty-one kinds of MICA-MICB haplotypes occurred at frequencies of more than 1% (linkage disequilibria value>0). The common MICA-MICB haplotypes were A5-CA14（16.73％）, A5.1- CA18 (8.75%), A4- CA26(3.76%),A9-CA15(3.66%) and A6-CA21(2.61%)（χ2>3.84, P<0.05）, and they were strong linkage disequilibria. The polymorphisms and haplotypes distributions of MICA and MICB microsatellite locus in Guangzhou Han population have their own genetic characteristics. The microsatellite locus of the exon5 of the MICA gene and intron 1 of the MICB gene could be used as the genetic markers in the studies of anthropology, linkage analysis of genetic disease genes, individual identification and paternity test in forensic medicine.     

中国三种实验用小型猪线粒体DNAD-loop多态性分析

分析中国三种实验用小型猪线粒体DNA（mtDNA）D-loop的多态性,建立各品种品系猪的遗传标记,为各品种、品系猪的鉴别提供依据。应用PCR技术分别对西双版纳近交系小耳猪、广西巴马小型猪、贵州小型香猪和长白猪的血液总DNA样品中mtDNAD-loop进行扩增,用23种限制性内切酶消化,观察其酶切多态。PCR扩增其mtDNAD-loop5′端227bp高变区域,应用PCR?SSCP和PCR直接测序分析,观察其单链构象多态和序列多态。结果显示：三种小型猪之间未见酶切长度多态、单链构象多态和序列多态。与长白猪之间表现出单链构象多态和序列多态。本研究认为：三种实验用小型猪之间mtDNA多态性贫乏,证明其亲源关系很近,在母系起源和进化上有一致性,应用PCR RFLP、PCR SSCP和PCR直接测序分析,尚不能作为三种实验用小型猪品种、品系鉴定的依据,但与长白猪等欧系猪比较有一定差异。 Abstract：the present study is to analyze the polymorphism of the mtDNA D-loop in three breeds of laboratory miniature pigs in China , and to establish its cytoplasmic DNA markers to distinguish among them . The polymorphism of mtDNA D-loop and its 5′-end high variable regions were detected by PCR-RFLP , PCR-SSCP and PCR-direct sequencing on Xishuangbanna Small-ear inbred pig, Guizhou miniature Xiang pig , Guangxi Bama miniature pig and Landrace.There was no polymorphism obtained among or within three breeds of Chinese laboratory miniature pigs besides Landrace. It is concluded that the polymorphism of mtDNA D-loop within the three breeds of Chinese laboratory miniature pigs is poor , These methods cannot be used to distinguish among them , but it can be used to distinguish them from Landrace.     

准噶尔雅罗鱼β-肌动蛋白基因启动子克隆及序列分析

利用PCR方法克隆了准噶尔雅罗鱼(Leuciscus merzbacheri)的β-actin基因启动子片段SZ21,大小是2398bp。对克隆的启动子序列进行了转录调控元件的生物信息学预测分析,同时,基于启动子中包含的开放阅读框和内含子序列,探讨了准噶尔雅罗鱼与鲤鱼(Cyprinus carpio)、草鱼(Ctenopharyngodon idella)、青鱼(Mylopharyngodon piceus)、团头鲂(Megalobrama amblycephala)、泥鳅(Misgurnus mizolepis)间的系统进化关系。结果显示,该启动子序列的3个核心启动子转录元件:CAAT-box、CArGmotif和TATA-box分别在转录起始位点(+1)上游的-89、-59、-26处,序列中还含有MEF2、SATB、CHRF、INRE、MTEN、E-box、RU49、ZBPF、CREB、Enhance region、CEBP位点等多种转录调控元件。在剪接体内含子中,剪接位点遵循GT…AG法则。启动子SZ21序列含有3个内含子和155个氨基酸。内含子1、内含子2、内含子3的系统发育分析表明,团头鲂与草鱼和青鱼的亲缘关系要比与准噶尔雅罗鱼的更近一些,这与传统分类中的亲缘关系显示不一致,其原因尚需探讨。     

A comparative study of neuropeptides in the intestine of two stomachless teleosts (Poecilia reticulata,Leuciscus idus melanotus) under conditions of feeding and starvation

Summary Endocrine cells containing bombesin-, enkephalin-, gastrin/CCK-, 5-HT-, and substance P-like material were demonstrated in the alimentary tract of Poecilia reticulata and Leuciscus idus melanotus. Endocrine cells with neuropeptide-Y-like immunoreactivity were found only in P. reticulata, those with VIP-like immunoreactivity only in L. idus melanotus. Gut nerves showing bombesin-, G/CCK-5-HT-, neurotensin-, substance P-and VIP-like immunoreactivity were observed in both species investigated, enkephalin- and neuropeptide Y-like immunoreactivity in P. reticulata alone. The distribution and amount of endocrine cells and nerves along the gut as visualized with the appropriate antisera varied in both teleosts. Histologically, the intestinal tract of these stomachless fish can be divided into three regions. A large number of endocrine cells with VIP-like immunoreactivity was noted in the rectum of L. idus melanotus. Endocrine cells containing bombesin-, enkepha-lin- and substance P-like material were found only in intestinal parts I and II in L. idus melanotus. Neuropeptide Y-like immunoreactivity was absent from intestinal part I of P. reticulata. The influence of starvation on the immunoreactivity of nerves and enteroendocrine cells in the teleost intestine was examined. After a starvation period of more than 6 weeks, no alterations were observed either in the appearance or amount of nerve and endocrine cell immunoreactivity.     

Freshwater sponge silicateins: comparison of sequences and exon-intron structure of genes

Siliceous sponge spicules contain silicateins--proteins taking part in biogenic silica precipitation and determination of the spicule morphological features. The exon-intron structure of four silicatein-alpha isoforms: -alpha1,-alpha2, -alpha3 and -alpha4 from endemic baikalian sponge Lubomirskia baicalensis was studied. For eight sponge species, including both cosmopolitan (Spongilla lacustris, Ephydatia muelleri, E. fluviatilis) and Baikal endemic (L. baicalensis, L. incrustans, Baikalospongia intermedia, B. fungiformis, Sw. papyracea) species, seventeen gene fragment sequences of different silicatein isoforms were determined. It was shown that cosmopolitan and endemic Baikalian sponges differ from each other by gene structure (have different length ofintrons). Among Baikalian sponges silicatein-alpha1 has the most variable intron length, and silicatein-alpha4 is the most conservative. Phylogenetic analysis of amino-acid silicatein sequences allow identify different silicatein isoforms, which authentically differ form four clusters on phylogenetic tree. Phylogenetic analysis of exon-intron sequences gives the possibility to separate different sponge species in the clusters.     

Use of the chromosomal co-location of the minor 5S and the major 28S rDNA as a cytogenetic marker within the genus Leuciscus (Pisces, Cyprinidae)

The chromosomes of three species from the genus Leuciscus (the ide L. idus, the European chub L. cephalus and the common dace L. leuciscus) were examined with the FISH technique for 5S and 28S rDNA probes. The analysis showed that among the three examined species, 5S rDNA signals were located on two large and four small subtelocentric chromosomes in L. leuciscus, on one large and five small subtelocentric chromosomes in L. idus, while in L. cephalus the probe signals were found on two metacentric chromosomes and one large and one small subtelocentric chromosome pairs. In all analysed species, the 28S rDNA probe signals were placed on only one chromosome pair, subtelocentric in the common dace and the European chub, and submetacentric in the ide. The three species differed in the number of sites in which both probe signals were present. In conclusion, the co-location of the 5S and 28S rDNA proved to be a useful cytogenetic marker among the studied fishes. Moreover, this marker could be adapted to other cyprinids.     

准噶尔雅罗鱼染色体核型及带型的初步研究

以肾细胞作材料,采用秋水仙素-低渗-空气干燥法、Ag-NORs、C-带和G-带显带技术对准噶尔雅罗鱼(Leuciscus merzbacheri)染色体进行了研究。结果表明:(1)准噶尔雅罗鱼2n=50,核型组成为18m+14sm+6st+12t,NF=82,没有异型性染色体分化。(2)Ag-NORs的数目在不同的细胞中表现出多态性,数目为1～2个,出现1个Ag-NORs的频率最低(10%),出现2个的频率最高(70%);Ag-NORs主要出现在m1对和m4对同源染色体上;未发现有Ag-NORs联合的现象。(3)准噶尔雅罗鱼的染色体均呈现C-带阳性,可分为着丝粒C-带和端粒C-带。(4)同源染色体上G-带带纹基本一致,其带纹在每对染色体上的数目及分布具有明显特征性。     

DNA polymorphism, haplotype structure and balancing selection in the Leavenworthia PgiC locus.

A study of DNA polymorphism and divergence was conducted for the cytosolic phosphoglucose isomerase (PGI:E.C.5.3.1.9) gene of five species of the mustard genus Leavenworthia: Leavenworthia stylosa, L. alabamica, L. crassa, L. uniflora, and L. torulosa. Sequences of an internal 2.3-kb PgiC gene region spanning exons 6-16 were obtained from 14 L. stylosa plants from two natural populations and from one to several plants for each of the other species. The level of nucleotide polymorphism in L. stylosa PgiC gene was quite high (pi = 0.051, theta = 0.052). Although recombination is estimated to be high in this locus, extensive haplotype structure was observed for the entire 2.3-kb region. The L. stylosa sequences fall into at least two groups, distinguished by the presence of several indels and nucleotide substitutions, and one of the three charge change nucleotide replacements within the region sequenced correlates with the haplotypes. The differences between the haplotypes are older than between the species, and the haplotypes are still segregating in at least two of five species studied. There is no evidence of recent or ancient population subdivision that could maintain distinct haplotypes. The age of the haplotypes and the results of Kelly's Z(nS) and Wall's B and Q tests with recombination suggest that the haplotypes are maintained due to balancing selection at or near this locus.     

松嫩平原针茅草原的特征及其生态地理规律的探讨

 经考察,松嫩平原的针茅草原主要有4个群落类型：贝加尔针茅(Stipa baicalensis)群落、贝加尔针茅+线叶菊(Filifolium sibiricum)群落、大针茅(S．grandis)+贝加尔针茅群落和大针茅群落。通过对针茅草原各类型基本结构特征和生态地理分布规律的分析,并根据它们所反映的水土条件的差异,认为贝加尔针茅群落为该地区的地带性植被。在中国温带草原区划中的位置属于草甸草原,应与内蒙古高原典型草原分开,成为一个独立的分区     

Comparison of calcium storage between a Baikalian gastropod and holarctic relatives

In Lake Baikal, extremely thin shells are reported as a typical feature of endemic gastropods. This statement derived only from observations; no experimental data were available up to now. Therefore, we quantitatively investigated the calcium distribution in the endemic prosobranch gastropod Benedictia baicalensis and compared the results with those of Lithoglyphus naticoides, a near relative, non-endemic, palaearctic species. The shell of the endemic mollusc B. baicalensis consists of 94.9+/-26.0 microg Ca(2+)/microl animal volume (n=43), and in L. naticoides 865.0+/-271.5 microg Ca(2+)/microl (n=10). Calcium contents in the tissue of B. baicalensis vary between different sampling stations and different sampling dates (from 9.4+/-5.1 (n=33) to 20.5+/-8.4 microg Ca(2+)/mg dry weight DW (n=16)) and are only 1/5-1/10 compared to L. naticoides (88.5+/-39.1 microg Ca(2+)/mg DW (n=9)). But the values for hemolymph calcium concentration and osmolality in both species are identical (B. baicalensis: osmolality: 84.4+/-5.3 mosm/kg (n=40); hemolymph calcium concentration: 4.6+/-1.7 mmol/l (n=40). L. naticoides: osmolality: 85.0+/-2.0 mosm/kg (n=8); hemolymph calcium concentration: 5.2+/-5.0 mmol/l (n=40).). This is the first experimental study demonstrating, that - besides a similar hemolymph ionic composition - the Baikalian species is characterized by significantly lower calcium storage in shell and tissue than the nearly related non-endemic species.     

Factors influencing movement behaviour and home range size in ide Leuciscus idus

Seventeen individuals of ide Leuciscus idus were radio-tracked weekly from September 2003 to September 2004 in the River Elbe, Czech Republic, to examine migration patterns and the influence of environmental factors on their diurnal behaviour. Of the 10 environmental factors measured, L. idus were significantly influenced by turbidity, which increased diurnal movement and the home range size of the species. The peak of longitudinal movement occurred in the spring, indicating pre-spawning migration. Migrating fish moved downstream and later returned upstream to the vicinity of their original locations, displaying a homing behaviour.