中国绿水螅分子系统发生地位的初步探讨

目的:初步探讨中国绿水螅(Hydra sinensis)分子系统发生地位以及水螅属内部各类群系统发生关系。方法:采用酚-氯仿法提取中国绿水螅总DNA,扩增线粒体COI和16S r RNA基因片段并进行DNA序列测定,再利用Clustal及MEGA等生物信息学分析软件进行系统发生分析。结果:在本研究重建的所有系统发生树中,中国绿水螅始终与绿水螅Hydra viridissima的不同种群一起构成绿水螅单系群。同时,棕色水螅群的单系性被基于COI基因的NJ树以及基于16S r RNA基因的NJ树和ML树支持,唯独基于COI基因的ML树不支持棕色水螅群的单系发生。在基于COI基因的ML树中纤弱水螅族在系统树的基部独立为一支系,而绿水螅群和其他棕色水螅群水螅一起组成另一支系,提示纤弱水螅族水螅的系统发生地位值得进一步探讨。值得注意的是,根据本文的结果,棕色水螅群内3族的划分仍然有一定疑问。基于COI基因的NJ树和ML树支持普通水螅族、寡水螅族和纤弱水螅族各自族内的单系发生,但16S r RNA基因的NJ树和ML树中仅普通水螅族水螅聚为单系群,而寡水螅族和纤弱水螅族水螅各自并非单系发生。结论:把水螅属划分为绿水螅群及棕色水螅群有一定的合理性,但棕色水螅群内寡水螅族、普通水螅族和纤弱水螅族3族的划分还有待商榷。     

大乳头水螅RACE cDNA文库的构建

目的:为筛选和克隆大乳头水螅发育调控相关基因的全长cDNA,构建大乳头水螅RACE cDNA文库.方法:提取大乳头水螅总RNA后从其中分离mRNA,运用SMART技术构建RACE cDNA文库.为鉴定所构建文库的质量,根据GenBank中大乳头水螅actin基因cDNA序列设计5'RACE和3'RACE的引物及用于扩增actin基因编码区全长序列的引物.结果:琼脂糖凝胶电泳结果表明,RACE cDNA文库中全长cDNA的长度集中在500-2 000bp之间.5'RACE、3'RACE PCR及扩增actin基因编码区全长序列时均以本文构建的大乳头水螅RACE cDNA文库为模板,这3个PCR反应均能扩增出产物,产物大小与目标片段预计大小相似.PCR产物分别经T/A克隆及测序后证明为大乳头水螅actin基因cDNA的相应序列.结论:RACE cDNA文库的成功构建为通过RACE方法获得大乳头水螅功能基因cDNA全长序列奠定了基础.     

大乳头水螅在中国的首次发现

淡水水螅有效物种已记述36种。史新柏等(1987)对哈尔滨的强壮水螅Hydra robustaIto,1947作过报道;范学铭报道了中国水螅4个新种,即哈尔滨水螅Hydra harbinensisFan、漠河水螅Hydra mohensisFan,1999、大庆水螅Hydra daqin-gensisFan,2000和北京水螅Hydra beijingensisFan,2003,     

磁化水对水螅出芽生殖及创伤恢复后水螅出芽生殖的影响

通过经 H型磁化杯 (磁场为 4 0 m T的非均匀场 )磁化水培养正常生活的水螅和横切恢复后的水螅出芽生殖的观察 ,探索磁化水对具有两个胚层的低等水生动物细胞增殖的影响。1　材料方法1.1　材料　采用黑龙江省广泛分布的雌雄异体、实验室培养近 2年的强壮水螅 ,(H ydra robusta) ,培养液为储放两周以上的自来水 ;每天定时喂食 1次 ,食物为裸腹氵蚤 (Moina) ,摄氏 2 2～ 2 6℃条件下培养。1.2　方法1)取刚脱离亲体的水螅 2 8个 ,分成实验组 (2 1个水螅 )和对照组 (7个水螅 ) ,分别以单个水螅培养在磁化水和非磁化水培养液中。每天定时检查产…     

异种嫁接的水螅畸形体初步观察

摘要：把强壮水螅（Hydra robusta）和普通水螅（H.vulgaris）除头后进行异种切口嫁接实验,共获得3例畸形体,对其进行280 d的培养,获得78只畸形子代。追踪观察发现：①畸形体及其无性生殖子代均具有2种钩刺丝囊（强壮水媳钩刺丝囊和普通水螅钩刺丝囊）;②其身体伸展时最长达52 mm;③无性出芽生殖的平均...     

中国水螅属一新种(螅形目,水螅科)

记述水螅科水螅属1新种：哈尔滨水螅Hydra harbinensis sp.nov.。本种与其它已报告的雌雄同体水螅有明显区别,只与美洲的海曼水螅较为相似,但也有不同,其与海曼水螅的区别为：生活于静水,能直立并可横向附着生活,实验室培养用储放的自来水可正常生活及生殖。粘刺丝囊小型。胚鞘的鞘膜表面有网格,每根圆锥形的棘刺发自每一网格中央,棘刺无结节,不分叉。精巢数目为2～6个,卵为4～6个。模式标本保存于哈尔滨师范大学生命与环境科学学院生物学系无脊椎动物研究室。     

直饮水对水螅生长与再生影响的比较

检测了30种直饮水对水螅生长与再生的影响。每种水使用切割的水螅组织10个和水螅个体30条,本实验共使用水螅1200条。结果表明：1）分别在30种直饮水中浸泡水螅组织96h,有17％的直饮水致其解体;浸泡水螅个体24h及48h,分别有37％和43％的直饮水致其解体。2）在市售的七大类直饮水中,矿泉水的安全性最高,可直接用于水螅的培养、再生等实验。人工配制的矿物质水不宜作为水螅的实验用水。     

中国水螅属一新种(螅形目,水螅科)

记述采自北京地区水螅属1新种：北京水螅Hydra beijingensis sp．nov．。新种的主要鉴别特征是出现体部与柄的分化;钩刺丝囊内刺丝缠绕具有2种形态：大部分钩刺丝囊(占2／3左右)内顶端基部刺丝呈4～5圈横向弹簧状缠绕,而占所观察刺丝囊总数1／3左右的囊内刺丝为不规则缠绕和纵向缠绕。标本保存在哈尔滨师范大学生命与环境科学学院生物学系无脊椎动物研究室。     

双头无基盘水螅的初步观察

自然形成的双头无基盘畸形体水螅极为罕见,为探索其形成原因,人工嫁接水螅14例,获得5例相同的双头水螅。对其分别进行连续观察发现:嫁接的双头水螅的双头可同时摄食,食物集聚于1个胃区。随个体生长其体柱延长,逐渐发育为2个胃区,在2个胃区间发生2~6个芽体,所有子代发育正常。出芽位置至触手环的距离同正常水螅。之后分化为2个出芽区,出芽区之间体柱较透明,同茎区组织。第21天开始发生基盘,发生过程异常缓慢。     

水螅的快速繁殖方法

本文介绍了水草附着法,藻苔贴壁法和苔丝附着法。作者利用18—25℃的高水温,充足的水蚤,喂食后及时换水和藻类的光合作用等几个条件,将水螅人工地附着于水草、藻苔和着生绿藻的玻璃绳上,高速、省工、适时地繁殖水螅,十分成功。夏季室温(20—25℃左右)较高,水蚤较易得刭,又是教学使用水螅的旺季.利用水螅的出芽繁殖,几年来为几百所学校提供了数万条活水螅用于教学。冬季保留少数螅种喂水蚯蚓越冬,亚甲基蓝和(口了)啶黄水溶液可防冶水螅的车轮虫病。     

Genomic fingerprints in the tribe Triticeae produced by PCR using a tRNA consensus primer

Forty-one accessions belonging to ten genera of the tribe Triticeae representing both wild and cultivated species were analyzed by polymerase chain reaction (PCR). Of two consensus tRNA primers tested, one primer revealed characteristic amplification products for all of the species. A total of 35 tRNA markers were scored across all accessions. Five genus-specific and three species-specific markers were obtained. Genomic fingerprints were largely conserved within a genus. The phylogram obtained using parsimony has separated most of the accessions into their prevailing taxonomic species and genus groups. The phylogram showed close association among the three genera Secale, Triticum and Hordeum as expected. The Triticum-Secale relationship was closer than the Triticum-Hordeum or the Secale-Hordeum relationships. The tree also reflected the close associations among the forage grass species belonging to Leymus and Elymus. Thus tDNA-PCR helped to identify species and genera. This revised version was published online in July 2006 with corrections to the Cover Date.     

Phylogenetic Analysis of Dipterocarps Using Random Amplified Polymorphic DNA Markers

The phylogenetic relationships among 12 species belonging tothree different genera (Shorea, HopeaandAnisoptera) of Dipterocarpaceaewere studied using random amplified polymorphic DNA (RAPD) markers.A modified CTAB DNA extraction protocol was used to obtain tannin-and polysaccharide-free genomic DNA from mature leaves. Clusteranalysis of data from six random primers placed the 12 speciesin three groups corresponding to their respective genera. Fourdistinct nodes ofShoreaspp. and two ofHopeaspp. could be identified.Anisopteramegistocarpaserved as an outgroup, and was unique when comparedto the other genera examined. RAPD profiles of five individualsofH. odoratawith six random primers were identical, suggestingthat there is little intraspecific variation in this species.The RAPD technique can thus be successfully applied for thestudy of phylogenetic relationships of this important groupof tropical timber trees.Copyright 1998 Annals of Botany Company Dipterocarpaceae,Anisopteraspp.,Hopeaspp.,Shoreaspp., RAPD markers, tropical timber trees.     

五种索科线虫RAPD亲缘关系分析

采用RAPD技术构建了索科线虫4属5种的指纹图谱。从47个引物中筛选出12个稳定性好、多态性高的引物,共扩增出161条谱带,其中150条谱带具遗传多态性,占93·17%。所获片段长度大小为200~3200bp,单个引物扩增的条带数在11~16之间,平均为13·42条。采用RAPDistance软件及MEGA程序,计算Nei氏相似系数和遗传距离,建立UPGMA和NJ聚类图,两个聚类图拓扑结构相同,将5种索科线虫分为两大分支:同属于蚊幼寄生罗索属线虫的食蚊罗索线虫(Romanomermisculicivorax)与武昌罗索线虫(R.wuchangensis)亲缘关系最近,先聚在一起,再与同翅目(Homoptera)寄生长沙多索线虫(Agamermischang-shaensis)聚为一支;鳞翅目(Lepidoptera)寄生中华卵索线虫(Ovomermissinensis)和同翅目寄生两索属线虫(Amphimermissp·)亲缘关系较近,两者聚为一支。5种索线虫属内种间的遗传距离较小,食蚊罗索线虫与武昌罗索线虫之间遗传距离仅为0·1789;而属间遗传距离较大,在0·4471~0·5488之间。上述结果表明:RAPD技术可以应用于索科线虫亲缘关系的分析,能够反映出不同线虫间的遗传差距,从而成功地进行属、种的分类及进化问题研究。     

利用RAPD技术对三种伪角毛虫及五种全列虫(原生动物,纤毛虫)的属间及属内关系探讨

利用11种随机引物对腹毛类纤毛虫中两个相近属内8个海洋种(3种伪角毛虫和5种全列虫)进行了种间及种内关系的RAPD分析,所得清晰条带显示种间均具有明显的差异.计算和比较了种群间、同属种间及不同属间的相似性指数.利用RAPDistance软件构建的邻接(NJ)树表明:伪角毛虫属与全列虫属之间能很好地分开,而不同种群间表现了很好的同源性.本工作表明RAPD技术可以很可靠的用来区分形态相似种,同时也表明RAPD分析在纤毛虫属间水平的系统关系推定中存在一定的适用性.     

玉米品种遗传多态性与根系内生细菌种群的相互关系

对辽宁省14个玉米主栽品种根系内主要细菌种群的分析结果表明,芽孢杆菌属（Bucillus ssp．）为玉米内生细菌的主要种群,其它种属包括肠杆菌属、沙雷氏杆菌属、假单孢菌属、黄单孢菌属和棍状杆菌属。内生细菌在不同玉米品种和不同生育期之间存在程度不同的差异。从90条随机引物中筛选出条带清晰、重复性好的13个引物,对14个玉米主栽品种进行遗传多态性的RAPD分析,共扩增出条带139条,其中多态性条带101条,多态性比率72．7％。利用SPSS统计分析软件对14个玉米品种在DNA水平和内生细菌种群水平上进行聚类分析,结果表明,品种亲缘关系在这两个水平上表现基本一致,即品种内生细菌的种类及其数量在很大程度上受品种的遗传背景控制。     

Phylogenetic relationships in Elymus (Poaceae: Triticeae) based on the nuclear ribosomal internal transcribed spacer and chloroplast trnL-F sequences

To estimate the phylogenetic relationship of polyploid Elymus in Triticeae, nuclear ribosomal internal transcribed spacer (ITS) and chloroplast trnL-F sequences of 45 Elymus accessions containing various genomes were analysed with those of five Pseudoroegneria (St), two Hordeum (H), three Agropyron (P) and two Australopyrum (W) accessions. The ITS sequences revealed a close phylogenetic relationship between the polyploid Elymus and species from the other genera. The ITS and trnL-F trees indicated considerable differentiation of the StY genome species. The trnL-F sequences revealed an especially close relationship of Pseudoroegneria to all Elymus species included. Both the ITS and trnL-F trees suggested multiple origins and recurrent hybridization of Elymus species. The results suggested that: the St, H, P, and W genomes in polyploid Elymus were donated by Pseudoroegneria, Hordeum, Agropyron and Australopyrum, respectively, and the St and Y genomes may have originated from the same ancestor; Pseudoroegneria was the maternal donor of the polyploid Elymus; and some Elymus species showed multiple origin and experienced recurrent hybridization.     

用RAPD技术对特化等级裂腹鱼类亲缘关系的探讨

为确定特化等级裂腹鱼类的遗传分化关系 ,对 7种 2 2个裂腹鱼个体进行随机扩增多态DNA(RAPD)研究。从所使用的 40个随机引物中 ,选择了 37个扩增带谱清晰的引物进行分析。根据构建的分子系统树显示 ,特化等级裂腹鱼类划分为 3个属级分类单元较为合适 ,这与采用形态学特征进行系统分析所得出的结论一致。在 2 2个个体之间遗传距离矩阵中 ,最大的遗传距离指数达到了 90 38% ,此外 ,还有较多的遗传距离指数也在 6 0 %～ 90 %之间。通过分析 ,认为用RAPD标记技术来分析属级或属级以上分类单元的亲缘关系 ,具有一定的适用性。对于一些遗传差异较大的类群 (如裂腹鱼类 ) ,应用属级或属级以上分类单元的亲缘关系 ,很可能会出现个体之间遗传距离接近或达到 10 0 %的情况 ,从而无法探讨其相互之间的亲缘关系。     

Confirmation of relationships among Boesenbergia (Zingiberaceae) and related genera by RAPD

The relationships among 19 accessions of Zingiberaceae belonging to 11 species of Boesenbergia, six species of Kaempferia, and two species of Scaphochlamys from Southern Thailand were studied using random amplified polymorphic DNA (RAPD) profiles from leaf tissue samples. The RAPD was carried out using 10 random decamer arbitrary primers. Amplification occurred in five out of 10 tested primers (OPAM-01, OPAM-03, OPAM-12, OPB-14, OPZ-03). Total of 53 amplified bands were observed. Data obtained from the RAPD fingerprints from the samples clarified some doubts in morphological classification. The data were analyzed for the Nei and Li's Dice similarity coefficient for pair-wise comparison between individual samples and the distance matrix. The dendrogram resulting from cluster analysis, UPGMA and a principal component analysis of the RAPD result confirms a higher degree of relationship between Boesenbergia and Scaphochlamys than between Boesenbergia and Kaempferia.     

水龙骨亚科植物(水龙骨科)的形态解剖学研究

对水龙骨科水龙骨亚科5属16种植物的孢子形态、叶表皮结构、叶柄和根状茎的横切、根状茎上的鳞片以及叶脉特征进行了详细的观察和分析,探讨了该亚科植物的系统位置和亲缘关系。结果表明,在该亚科植物类群中,主要分布于亚洲亚热带地区的篦齿蕨属、拟水龙骨属和水龙骨属的亲缘关系较近,而主要分布于亚洲热带地区的棱脉蕨属和亚洲温带地区的多足蕨属的系统位置则分别较孤立。     

Molecular characterization of five medicinally important species of Typhonium (Araceae) through random amplified polymorphic DNA (RAPD)

The interrelationship of five medicinally important species of Typhonium (Araceae) including T. venosum, which was previously placed under the genus Sauromatum, was inferred by analysis of random amplified polymorphic DNA (RAPD). DNA from pooled leaf samples was isolated and RAPD analysis was performed using 20 decamer oligonucleotide primers. Out of a total of 245 bands amplified, 12 were found to be monomorphic while 233 bands were polymorphic including 86 species-specific bands. The genetic similarities were analyzed from the dendrogram constructed by the pooled RAPD data using a similarity index. The dendrogram showed two distinct clades, one containing T. roxburghii, T. trilobatum and T. venosum and the other containing the remainder two species, i.e. T. diversifolium and T. flagelliforme. Both the clusters shared a common node approx. at 23.7% level of similarity. The maximum similarity of 31.2% was observed between T. venosum and T. trilobatum. In view of its close genetic similarity with other members of Typhonium, transfer of Sauromatum venosum to the genus Typhonium and merger of the two genera was supported.