寡毛双眉虫Diophrys oligothrix形态和形态发生的研究

报告了寡毛双眉虫(Diophrys. oligothrix) 的形态学特征以及它在无性分裂时期形态发生的全过程。观察了前仔虫口围带的形成过程,并将其与亲代的口围带进行比较,认为前仔虫口围带是经过重建的。从口器、大核等的情况反映出寡毛双眉虫与游仆虫、盾纤虫以及尾刺虫在进化上的关系。并认为前、后仔虫形成后仍继续恢复其典型的纤毛模式。     

一种腹柱虫包囊形成和解脱过程中核的行为研究

腹柱虫Ｇａｓｔｏｓｔｙｌａ ｓｐ．成包囊过程中,其纤毛器的分化不论是从细胞前端或是细胞后端开始,其大、小核的变化是相同的。首先,两个大核先出现复制带,然后两个大核融合成一个圆球形融合大核。另外也观察到二核包囊和四核包囊,融合大核在包囊进入休眠之前出现复制带,改组后的融合大核在休眠包囊中分裂为二。在成包囊过程中,两个小核中的一个被吸收。出包囊时,小核分裂为二,融合大核行无丝分裂成两个大核。     

包囊游仆虫休眠细胞中大核和线粒体DNA的多态性

为了探索纤毛虫休眠细胞中两套遗传系统的作用特征，对包囊游仆虫（Euplotes encysticus）休眠细胞与营养细胞大核DNA和线粒体DNA进行了RAPD比较.结果显示，在所选用的35条随机引物中，包囊游仆虫大核DNA共扩增出220条片段，其中以休眠细胞大核DNA为模板扩增出18条特有片段，以营养细胞大核DNA为模板扩增出44条特有片段，两者存在28%的差异.在所选用的32条随机引物中，线粒体DNA共扩增出154条片段，其中以休眠细胞线粒体DNA为模板扩增出19条特有片段，以营养细胞线粒体DNA为模板扩增出25条特有片段，两者有29%的差异.这些结果表明，包囊游仆虫休眠细胞与营养细胞的大核DNA结构存着一定的差异；两者的线粒体DNA结构也存在差异.因此，包囊游仆虫在休眠细胞形成过程中，大核DNA、线粒体DNA结构可能都发生了一定的变化，并且这些变化可能与休眠细胞形成过程中的形态结构和代谢活动等剧烈变化以及休眠状态下的生理生化变化密切相关.所得结果为揭示纤毛虫细胞结构的分化与细胞遗传物质的作用关系提供了基础资料.     

RNA-mediated epigenetic programming of a genome-rearrangement pathway

Genome-wide DNA rearrangements occur in many eukaryotes but are most exaggerated in ciliates, making them ideal model systems for epigenetic phenomena. During development of the somatic macronucleus, Oxytricha trifallax destroys 95% of its germ line, severely fragmenting its chromosomes, and then unscrambles hundreds of thousands of remaining fragments by permutation or inversion. Here we demonstrate that DNA or RNA templates can orchestrate these genome rearrangements in Oxytricha, supporting an epigenetic model for sequence-dependent comparison between germline and somatic genomes. A complete RNA cache of the maternal somatic genome may be available at a specific stage during development to provide a template for correct and precise DNA rearrangement. We show the existence of maternal RNA templates that could guide DNA assembly, and that disruption of specific RNA molecules disables rearrangement of the corresponding gene. Injection of artificial templates reprogrammes the DNA rearrangement pathway, suggesting that RNA molecules guide genome rearrangement.     

茄子雄性不育株和可育株的胞质DNA和核DNA差异分析

以茄子(Solanum melongenaL)雄性不育株“正兴1号”(S)和可育株(F)的总DNA为模板,对60个随机引物进行了筛选,找到5个其RAPD(Random amplified polymorphic DNA,RAPD)扩增产物在茄子雄性不育系和对照材料间存在稳定差异的引物．将该5个引物同时扩增总DNA、核DNA和线粒体DNA(mitochondrial DNA,mtDNA)．以总DNA为模板时得到多态性片段9个,以核DNA为模板时得到5个,以mtDNA为模板时得到9个．以总DNA为模板时得到的9个扩增片段中,有5个在总DNA和mtDNA中同时出现,而在核中没有出现,即认为来自mtDNA,这5个片段中,有4个来自可育株,1个来自不育株,说明不育株和可育株在mtDNA上存在差异;有4个片段同时出现在以总DNA和核DNA为模板的扩增中,但在以mtDNA为模板的扩增中却没有出现,认为是来自核DNA,这4个片段中,有3个来自不育株,一个来自可育株,说明可育株和不育株在核DNA上也存在差异．结果初步表明：新发现的茄子雄性不育可能是核质相互作用所引起．     

滩羊体大品系遗传标记的研究

采用RAPD(Rondom Amplified Polymorphic DNA)技术,利用混合基因池(DNA pool)法,对滩羊体大品系、普通品系进行了DNA多态性分析,从100种具有10个碱基的随机引物中,筛选出84种引物在滩羊群体基因组中共扩增出358条带,其中22种引物的扩增产物表现为多态(占22％),且扩增出32条有差异的条带,占总带数的8.94％;62种引物的扩增产物表现为单态(占62％)。滩羊体大品系的特异性条带有5条,而普通品系的特异性条带有7条,这些特异标记可以用来鉴定滩羊的两个品系;滩羊体大品系与普通品系间的遗传距离为0.136±0.087,表明两品系之间的亲缘关系很近。     

Phylogenetic positions of two marine ciliates, Metanophrys similis and Pseudocohnilembus hargisi (Protozoa, Ciliophora,Scuticociliatia), inferred from complete small subunit rRNA gene sequences

The small subunit rRNA (SSrRNA) gene was sequenced for two marine scuticociliates Metanophrys similis and Pseudocohnilembus hargisi. The results show that this gene comprises 1763 and 1753 nucleotides in the two marine ciliates respectively. Metanophrys similis is phylogenetically closely related to the clade containing Mesanophrys carcini and Anophyroides haemophila, which branches basally to other species within the order Philasterida. Pseudocohnilembus hargisi groups with its congener, P. marinus, with strong bootstrap support. Paranophrys magna groups with the clade including Cohnilembus and Uronema, representing a sister clade to that containing the two Pseudocohnilembus species.     

Phylogenetic positions of two marine ciliates, Metanophrys similis and Pseudocohnilembus hargisi (Protozoa, Ciliophora, Scuticociliatia), inferred from complete small subunit rRNA gene sequences

The small subunit rRNA (SSrRNA) gene was sequenced for two marine scuticociliates Metanophrys similis and Pseudocohnilembus hargisi. The results show that this gene comprises 1763 and 1753 nucleotides in the two marine ciliates respectively. Metanophrys similis is phylogenetically closely related to the clade containing Mesanophrys carcini and Anophyroides haemophila, which branches basally to other species within the order Philasterida. Pseudocohnilembus hargisi groups with its congener, P. marinus, with strong bootstrap support. Paranophrys magna groups with the clade including Cohnilembus and Uronema, representing a sister clade to that containing the two Pseudocohnilembus species.     

Phylogenetic positions of two marine ciliates, Metanophrys similis and Pseudocohnilembus hargisi (Protozoa, Ciliophora, Scuticociliatia), inferred from complete small subunit rRNA gene sequences

The small subunit rRNA (SSrRNA) gene was sequenced for two marine scuticociliates Metanophrys similis and Pseudocohnilembus hargisi. The results show that this gene comprises 1763 and 1753 nucleotides in the two marine ciliates respectively. Metanophrys similis is phylogenetically closely related to the clade containing Mesanophrys carcini and Anophyroides haemophila, which branches basally to other species within the order Philasterida. Pseudocohnilembus hargisi groups with its congener, P. marinus, with strong bootstrap support. Paranophrys magna groups with the clade including Cohnilembus and Uronema, representing a sister clade to that containing the two Pseudocohnilembus species.     

中心蛋白3定位于游仆虫细胞中形成棘毛的基体上

中心蛋白与中心体(或基体)有着密切关系,作为一种细胞骨架蛋白,形成原生动物巨大的细胞骨架网络;为了研究从八肋游仆虫细胞中克隆到的一种中心蛋白基因(EoCen3)的功能,我们构建了八肋游仆虫细胞大核人工染色体pBTub-Tel,定位分析该基因所表达的中心蛋白在细胞中的分布和行为.该人工染色体利用密码子优化的增强型绿色荧光蛋白基因作为标记,在八肋游仆虫细胞中实现高表达,避免用抗生素作为标记对该细胞的喂养产生影响.定位结果显示,该类中心蛋白特异定位于游仆虫形态发生过程中的基体上.在八肋游仆虫无性生殖细胞分裂的起始阶段基体开始发育,首先形成第一个基体,Eo-Cen3定位于新形成的基体中;随后基体开始复制,EoCen3定位于正在复制的基体中.在细胞发育过程中绿色荧光标记的基体成倍增加,说明EoCen3与参与棘毛形成的基体的复制有一定的关系.     

Evolutionary relationship and species separation of four morphologically similar stichotrichous ciliates （Protozoa, Ciliophora）

Phylogenetic and taxonomic studies on ciliate protists using molecular approaches have been demonstrated to be very reliable to form strong conclusions and results. In the present work, species separation of some morphologically similar stichotrichous ciliates, two species of Pseudokeronopsis and two species of Apokeronopsis, was reexamined using amplified ribosomal DNA restriction analysis (PCRRFLP). Five of 10 restriction enzymes revealed species-specific polymorphic patterns, of which four similar stichotrichs could be significantly separated and identified. Among them, EcoR I offered almost no significantly different restriction fragment patterns, but the four species could be separated from one another and identified with Hae III. Distinctly different restriction digestion haplotypes and similarity indices separated the species, and were used to construct a phylogeny. Phylogenies based on ITS2 nucleotide sequences and ITS2 secondary structures supported the separation of Pseudokeronopsis and Apokeronopsis using RFLP analysis, although three Pseudokeronopsis carnea populations did not cluster together. In addition, phylogenetic analyses using multiple algorithms confirmed that these two genera formed two distinct groups within the urostylids.     

云南鼠疫菌随机引物扩增DNA多态性指纹图谱分析

目的：分析云南不同地区、不同年代分离鼠疫菌的基因表型。方法：采用随机引物扩增技术（RAPD）对菌株进行分析。结果：共检测了28株鼠疫菌，除一株外，27株均显示有13条带型，并与假结核菌和小肠结肠炎菌有明显的区别。结论：云南家、野两型鼠疫菌株可能具有相同的随机引物扩增基因表征。     

DNA typing from single hairs

The characterization of genetic variation at the DNA level has generated significant advances in gene and disease mapping, and in the forensic identification of individuals. The most common method of DNA analysis, that of restriction fragment length polymorphism (RFLP), requires microgram amounts of relatively undegraded DNA for multi-locus typing, and hundreds of nanograms for single-locus comparisons. Such DNA frequently cannot be obtained from forensic samples such as single hairs and blood stains, or from anthropological, genetic or zoological samples collected in the field. To detect polymorphic DNA sequences from single human hairs, we have used the polymerase chain reaction (PCR), in which specific short regions of a gene can be greatly amplified in vitro from as little as a single molecule of DNA. We have detected genetically variable mitochondrial and nuclear DNA sequences from the root region of shed, as well as freshly-plucked, single hairs; mitochondrial DNA (mtDNA) sequences have been detected in a sample from a single hair shaft. We have used three different means of DNA typing on these samples: the determination of amplified DNA fragment length differences, hybridization with allele-specific oligonucleotide probes, and direct DNA sequencing.     

兰州市雁儿湾污水处理厂活性污泥中纤毛虫春季群落特征

从2006年2月至2006年4月,对兰州市雁儿湾污水处理厂曝气池活性污泥中的纤毛虫群落进行了调查研究.共记录到纤毛虫57种,隶属于11目21科27属.缘毛目为优势类群.其中优势种为：小口钟虫（Vorticella microstoma）、沟钟虫（Vorticella convallaria）、八钟虫（Vorticella octava）、卑怯管叶虫（Trachelophyllum pusillum）、彩盖虫（Opercularia phryganeae）、有肋楯纤虫（Aspidisca costata）、片状漫游虫（Litonotus fasciola）.同时记录到10种常见种.与两对照样点相比较,活性污泥中纤毛虫种类数、丰度都大于两对照样点.结果表明,缘毛目优势种可以作为污水处理效果的指示生物.     

油蒿种群遗传分化的RAPD分析

采用随机扩增多态性DNA(RAPD)方法对油蒿(Artemlsia ordosica Krasch．)的5个种群进行了研究．用15个随机引物扩增出297条清晰谱带，其中285条为多态性谱带．利用POPGENE 32软件对数据进行处理，结果如下：(1)油蒿有着丰富的遗传多态性，多态位点百分率达96％，各种群多态位点百分比在84．6％～90．9％之间．(2)油蒿的种群间分化较小Gat=0．1364，86．36％的遗传变异存在于种群内，各种群的遗传一致度都在96％以上(3)聚类分析显示，地理分布近的种群被聚到了一起，反映了油蒿种群的遗传分化和地理距离有着一定的相关性．     

ISSR 和 RAPD PCR技术对东北地区主推玉米品种的鉴别

运用现代分子生物学简单重复区间序列扩增多态性（ISSR）和随机扩增片段多态性DNA（RAPD）两种分子标记技术， 对吉林省主推6个玉米杂交种及其父本、 母本共计18份基因图谱进行鉴别. 以ISSR为主要检测方法、 RAPD为验证方法，分别从30条ISSR引物中选出4条、 从30条RAPD引物中筛选出2条扩增效果明显、 特异性高的引物. 结果表明， 运用ISSR和RAPD-PCR技术提高了玉米品种鉴定和纯度鉴定的准确性.     

Analysis of RAPD and mitochondrial 16S rRNA gene sequences from Trichiurus lepturus and Eupleurogrammus muticus in the Yellow Sea

Random amplified polymorphic DNA (RAPD) technique is applied to 12 individuals from each species of the hairtail fishes Trichiurus lepturus and Eupleurogrammus muticus in the Yellow Sea. The percentage of polymorphic sites, degree of genetic polymorphism and genetic distance are compared and the phylogenetic tree is constructed by Neighbor-joining method. The partial mitochondrial 16S rRNA gene is amplified by polymerase chain reaction (PCR) and the PCR products are directly sequenced after being purified. These sequences, together with the homologous sequences of another Trichiuridae species Lepidopus caudatus obtained from GenBank, are used to analyze nucleotide difference and to construct a UPGMA phylogenetic tree by means of biological informatics. Analysis shows: (1) the RAPD technique is a highly sensitive method for investigating genetic diversity in T. lepturus, and E. muticus. T. lepturus exhibits a lower polymorphism and genetic diversity than E. muticus; (2) according to the analysis of the partial mitochondrial 16S rRNA gene sequences, a very low intraspecific variation and considerably high divergence among species were found, which reveals a dual nature of conservatism and variability in mitochondrial 16S rRNA gene; (3) five primers generate the species-specific RAPD sites and these sites can be served as the molecular markers for species identification and (4) it can be proved at DNA variation level that T. lepturus and E. muticus are of two species respectively pertaining to different genera, which supports the Nelson taxonomic conclusion.     

水稻苯达松敏感致死基因的RAPD标记的克隆及测序

用3S Spin DNA Agarose Gel Purification Kit试剂盒将与水稻苯达松敏感致死基因相连锁的RAPD遗传标记S20—420和S316—600回收纯化，连接于pGEM—T载体并克隆测序，得到了S20—420和S316—600的全序列，其长度分别为423bp、606bp．将两端序列设计特异PCR扩增引物可用于检测水稻苯达松敏感致死基因和标记辅助育种．     

濒危植物连香树遗传多样性研究

以我国特有的濒危被子植物连香树为材料,采用RAPD标记对分布于浙江、安徽、湖南、湖北、四川、陕西、河南的11 个天然居群进行了检测。以20个引物共扩增出691条DNA片段,其中多态性条带328条,占总条带数的475 %。AMOVA分析表明,连香树的基因分化系数为0479 7,居群内变异占总变异的5203 %。由Nei基因分化系数估计的基因流仅为0542 4,表明居群间基因交流困难。UPGMA聚类分析表明,安徽歙县居群与河南济源居群的关系最远。遗传漂变等因素可能是连香树目前遗传结构的主要成因。保护现有生境、通过培育实生苗并扩大繁殖栽培的范围是目前对连香树比较好的保护措施。     

草鱼基因组随机扩增多态性引物及多态性位点的筛选

为了建立草鱼基因组DNA多态性分析的指标体系,应用RAPD技术,从180条10碱基随机引物中筛选出了31条能扩增出多态性DNA片段的引物。用这31条多态性引物共扩增出了327条重复性好、带型清晰、分辨率高的谱带。扩增产物的片段大小范围在400—2000bp之间。单一引物扩增条带为5—14条。用这些多态性引物在草鱼基因组DNA中检测到了93个多态性位点,并对这些多态性位点上的等位基因频率进行了统计分析。这31条多态性引物和所检测到的93个多态性位点初步为草鱼基因组的多态性分析提供了可靠的分析指标体系。