The c-myc coding DNA sequences of cyprinids (Teleostei : Cypriniformes): Implications for phylogeny

The family Cyprinidae is one of the largest fish families in the world, which is widely distributed in East Asian, with obvious difference in characteristic size among species. The phylogenetic analysis of cyprinid taxa based on the functionally important genes can help to understand the speciation and functional divergench of the Cyprinidae. The c-myc gene is an important gene regulating individual growth. In the present study, the sequence variations of the cyprinid c-myc gene and their phylogenetic significance were analyzed. The 41 complete sequences of the c-myc gene were obtained from cyprinids and outgroups through PCR amplification and clone. The coding DNA sequences of the c-myc gene were used to infer molecular phylogenetic relationships within the Cyprinidae. Myxocyprinus asiaticus (Catostomidae), Misgurnus anguillicaudatus (Cobitidae) and Hemimyzon sinensis (Homalopteridae) were assigned to the outgroup taxa. Phylogenetic analyses using maximum parsimony (MP), maximum likelihood (ML), and Bayesian retrieved similar topology. Within the Cyprinidae, Leuciscini and Barbini formed the monophyletic lineage respectively with high nodal supports. Leuciscini comprises Xeno-cyprinae, Cultrinae, East Asian species of Leuciscinae and Danioninae, Gobioninae and Acheilognathinae, and Barbini contains Schizothoracinae, Barbinae, Cyprininae and Labeoninae. Danio rerio, D. myersi and Rasbora trilineata were supposed to separate from Leuciscinae and Barbini and to form another lineage, The positions of some Danioninae species were still unresolved. Analyses of both amino acid variation with parsimony information and two high variation regions indicated that there is no correlation between variations of single amino acid or high variation regions and characteristic size of cyprinids. In,addition, the species with smaller size were usually found to be basal within clades in the tree, which might be the results of the adaptation to the primitive ecology and survival pressure.

Occurrence of Camallanus cotti in greatly diverse fish species from Danjiangkou Reservoir in central China

Two thousand four hundred fifty-eight fish comprised of 53 species were captured in the Danjiangkou Reservoir, in the northwestern part of Hubei Province, central China during 2004, to examine Camallanus cotti infections. We found that 19 cypriniform, 3 siluriforme, and 4 perciforme fishes were infected by the nematode. Our study revealed the species, Hemiculter bleekeri bleekeri, Culter oxycephaloide, Pseudolaubuca sinensis, Acanthobrama simony, Mylopharyngodon piceus, Ctenopharyngodon idella, Gnathopogon imberbis, G. argentatus, Saurogobio dabryi, S. dumerili, Gobiobotia ichangensis, Liobagrus marginatoides, and Ctenogobius shennongensis as new hosts of the worm. The number and range of fish host species found in this survey were much greater than any of the previous investigations. The mean prevalence, prevalence, mean abundance, and intensity of infection varied in different fish species, indicating a possible host preference. Moreover, we suggest that this nematode is a native parasite of cypriniform fishes in China, perhaps initially in the reaches of the Yangtze River.

Molecular and morphological data suggest that Spinibarbus caldwelli (Nichols) (Teleostei : Cyprinidae) is a valid species

The taxonomic problem of the cyprinid species of genus Spinibarbus, occurring in southern China and northern Vietnam, was resolved on the basis of molecular and morphological analyses. Spinibarbus caldwelli and Spinibarbus hollandi have a smooth posterior edge of the last unbranched dorsal fin ray among species in the genus. Spinibarbus caldwelli is currently regarded as a junior synonym of S. hollandi because of ambiguities in diagnostic characters. In this article, 11 mtDNA cytochrome b sequences of Spinibarbus specimens were analyzed together with Barbodes gonionotus and Puntius conchonius as outgroups. Our results showed that specimens identified as S. hollandi from Taiwan were different from those from the Asian mainland at a high level of genetic divergence (0.097-0.112), which is higher than that between the two valid species, S. sinensis and S. yunnanensis ( 0.089), and suggested that Taiwan specimens should be considered as a different species from the Asian mainland one. In a molecular phylogenetic analysis, the sister-group relationship between Taiwan specimens and the Asian mainland specimens was supported strongly by a high confidence level ( 100% in bootstrap value). Further analysis of morphological characters showed that overlap of diagnostic characters is much weaker than previously suggested. Taiwan specimens had 8 branched rays in the dorsal fin, whereas those from the mainland had almost 9-10. The molecular and morphological differences suggest S. caldwelli to be valid. The molecular divergence shows the genetic speciation of S. hollandi and S. caldwelli might have occurred 5.6-4.9 million years ago; the former could be a relict species in Taiwan, and the latter dispersed in the Asian mainland.

Mitochondrial 16S rRNA sequence variations and phylogeny of the Chinese sisorid catfishes

Partial sequences of mitochondrial 16S rRNA gene were obtained by PCR amplification for comparisons among nine species of glyptosternoid fishes and six species of non-glyptosternoids representing 10 sisorid genera. There are compositional biases in the A-rich impaired regions and G-rich paired regions. A-G transitions are primarily responsible for the Ts/Tv bias in impaired regions. The overall substitution rate in impaired regions is almost two times higher than that in the paired regions. Saturation plots at comparable levels of sequence divergence demonstrate no saturation effects. Phylogenetic analyses using both maximum likelihood and Bayesian methods support the monophyly of Sisoridae. Chinese sisorid catfishes are composed of two major lineages, one represented by (Gagata (Bagarius, Glyptothorax)) and the other by "glyptosternoids + Pseudecheneis". The glyptosternoids may not be a monophyletic group. A previous hypothesis referring to Pseudecheneis as the sister group of monophyletic glyptosternoids, based on morphological evidence, is not supported by the molecular data. Pseudecheneis is shown to be a sister taxon of Glaridoglanis. Pareuchiloglanis might be paraphyletic with Pseudexostoma and Euchiloglanis. Our results also support the hypothesis that Pareuchiloglanis anteanalis might be considered as the synonyms of Pareuchiloglanis sinensis, and genus Euchiloglanis might have only one valid species, Euchiloglanis davidi.

Phylogenetic relationships of glyptosternoid fishes (Siluriformes : Sisoridae) inferred from mitochondrial cytochrome b gene sequences

To explore phylogenetic relationships among glyptosternoid fishes, we determined nucleotide sequences of the complete mitochondrial cytochrome b gene region (1138 base pair). Thirteen species of glyptosternoid fishes and six species of non-glyptosternoids represent 10 sisorid genera were examined. Molecular phylogenetic trees were constructed using the maximum parsimony, minimum evolution, maximum likelihood, and Bayesian methods. Bayesian and maximum likelihood analyses support the monophyly of glyptosternoids, but our hypothesis of internal relationships differs from previous hypothesis. Results indicated that glyptosternoid is a monophyletic group and genera Glyptosternum and Exostoma are two basal species having a primitive position among it. Genera Euchiloglanis and Pareuchiloglanis form a sister-group. Then they form a sister-group with Pseudexostoma plus Oreoglanis. Our result also found that Pareuchiloglanis anteanalis might be considered as the synonyms of Parechiloglanis sinensis, and genus Euchiloglanis might have only one valid species, Euchiloglanis davidi. (C) 2003 Elsevier Inc. All rights reserved.

Some nematodes of fishes from central China, with the redescription of Procamallanus (Spirocamallanus)fulvidraconis (Camallanidae)

The present paper comprises a systematic survey of nematodes based on helminthological examinations of 176 specimens of freshwater fishes, belonging to 22 species, from central China (mostly lakes in Hubei Province) collected during the autumn of 2001. The following six species were recorded: Procamallanus (Spirocamallanus) fulvidraconis Li, 1935, Camallanus cotti Fujita, 1927, Dentiphilometra monopteri Moravec et Wang, 2002, Pingus sinensis Hsu, 1933, Proleptinae gen. sp. larv., and Eustrongylides sp. larv. Data on their morphology, morphological variability, host range, prevalence, intensity and distribution are provided. SEM studies of P. fulvidraconis and larval Physalopterinae, used for the first time in these species, revealed some additional morphological details and made it possible to redescribe the former. In contrast to the existing description of P. fulvidraconis, this species was found to possess two spicules and a V-shaped gubernaculum with unequal arms (originally mistaken for the left spicule), as well as deirids, whose location can be considered an important taxonomic feature. Larvae of the Physalopterinae have not previously been reported from fishes in China. The finding of larval Eustrongylides in Paramisgurnus dabryanus represents a new host record. All but one nematode species from this zoogeographically interesting region are briefly described and illustrated.

The precocious Chinese mitten crab: Changes of gonad, survival rate, and life span in a freshwater lake

Changes of the gonad. survival rate, and life span of the precocious (i.e., sexually mature in their first October-November, at the age of 5 or 6 months) Chinese mitten crab (Eriocheir sinensis) were studied in a small, shallow, macrophytic, freshwater lake along the middle reaches of the Changjiang River, China. The gonosomatic index (GSI) reached peak during the next March when the female GSI was 12.32 +/- 1.75 SD% and the male GSI A as 4.24 +/- 0.19%. The sexual glands degraded from then on. In the first ten-day period of the following July, there was no complete ovum in the ovary. and the sperm became thin and lost adhesion. The population declined sharply from November to July, and the last one (a female) died in the middle of July, which indicated that the life span of the precocious crab was about 12 months (from larval hatching in June to death in July of next year). The survival curve might be expressed as Y = 1.09exp(-0.018x) (Y survival rate; x: days) for the precocious crabs stocked in experimental cages.

Taxonomic studies of parasitic nyctotherans from chinese Anura amphibians IV. Spirocytopharynxa gen. nov and Macrocytopharynxa gen. nov.

Lian-Xiang U, Jian-Guo Wang and Wu-Han Xiao (2002) Taxonomic studies of parasitic nyctotherans from Chinese Anura amphibians IV. Spirocytopharynxa gen. nov. and Macrocytopharynxa gen. nov. Zoological Studies 41(1): 77-84. This paper describes 2 new genera and 5 new species of nyctotherans from Anura amphibians distributed in southern China. Based on the composition of the upper and bottom flaps, the number and position of sutural lines, the length of the oral groove, and the morphology and position of the end of the cytopharynx, the adoral zone of the membranelle (AZM), and the macronucleus, two new genera were established. The 3 new species, Spirocytopharynxa sinensis, S. guangxiensis, and S. quadranus, belong to the new genus Spirocytopharynxa, Another 2 species, Macrocytopharynxa (Nyctotheroidae Nie, 1932) pyriformis n. comb. and M. lingchuanensis n. sp., belong to the new genus Macrocytopharynxa. The characteristics for diagnosis of the 2 new genera and 5 new species are described in this paper.

RAPD and AFLP techniques for the analysis of genetic relationships in two genera of Decapoda

RAPD was used fur analysing three (sub-)species of mitten crabs (Eriocheir sinensis, E. japonicus, and E. japonicus hepuensis) and three populations of E. sinensis. The results show that their relationships on DNA level are similar to the classical taxonomic hypotheses (Dai, 1991). No diagnostic RAPD marker could be found, but there were statistically significant genetic differences among these taxa (P < 0.001) or populations (P < 0.001). That is, the intraspecific similarities were larger than the interspecific similarities; the intrasubspecific similarities were larger than the intraspecific similarities; and the intrapopulational similarities were larger than the interpopulational similarities. In AFLP analysis, no significant genetic difference has been found between E. sinensis and E. japonicus, but AFLP markers among four species of Macrobrachium (M. rosenbergii. M. nipponense, M. hainanense, and M. asperulum) were found. The DNA similarities among these four species of Macrobrachium are in accordance with morphological similarities.

Molecular phylogenetic systematics of twelve species of acipenseriformes based on mtDNA ND4L-ND4 gene sequence analysis

Acipenseriformes is an endangered primitive fish group, which occupies a special place in the history of ideas concerning fish evolution, even in vertebrate evolution. However, the classification and evolution of the fishes have been debated. The mitochondrial DNA (mtDNA) ND4L and partial ND4 genes were first sequenced in twelve species of the order Acipenseriformes, including endemic Chinese species. The following points were drawn from DNA sequences analysis: (i) the two species of Huso can be ascribed to Acipenser; (ii) A. dabryanus is the mostly closely related to A. sinensis, and most likely the landlocked form of A. sinensis; (iii) genus Acipenser in trans-Pacific region might have a common origin; (iv) mtDNA ND4L and ND4 genes are the ideal genetic markers for phylogenetic analysis of the order Acipenseriformes.

Two new species of Actinolaimidae thorne, 1939 (Nemata : Dorylaimida) from China

Two new species of Actinolaimidae are described from China. Trachactinolaimus brevicaudatus n. sp. is 3.4-4.4 mm long; a = 40-60, b = 3.5-5.2, c = 20-21 in female and 34-39 in male; odontostyle length is 29-33 mu m; spicules are 67-77 mu m long; and the stoma has four onchia with numerous mural denticles. The female has a longitudinal vulva, and the male has 20 to 23 contiguous ventromedian supplements. Egtitus sinensis n. sp. is 1.7-2.2 mm long; a = 24-33, b = 3.1-3.9, c = 16-19 in female and 0.7-0.9 in male; odontostyle length is 25-29 mu m; spicules are 55-56 mu m long; and the prerectum 53-77 pm long. The cardia is short and blunt conoid, 13-19 mu m long. The male has 12 to 13 ventromedian supplements at intervals of 2-3 mu m.

Diversity of intestinal helminth communities of carp from six lakes in the flood plain of the Yangtze river, China

The infra- and component communities of intestinal helminths of carp Cyprinus carpio were investigated in six lakes in the flood plain of the lower and middle reaches of the Yangtze River, China. Eight species of helminth parasites were recorded. The intestinal helminth communities were species rich in Niushan and Tonghu lakes where the digenean Asymphylodora japonica was the dominant species, whereas in Qinggang and Yanglan lakes a species-poor helminth community had only one species, Khawia sinensis. The degree of similarity within localities was highest in Qinggang and Yanglan lakes, and was high between communities where K. sinensis was the dominant species. The rich composition of these helminth communities may be because China is the heartland for carp while the poor helminth composition of those in Qinggang and Yanglan lakes may reflect the poor fauna there. It is suggested that species compositions of intestinal helminth communities of carp may be diversified in lakes in the hood plain of the Yangtze River. (C) 1999 The Fisheries Society of the British Isles.

日光温室豇豆产量和品质对水分和氮素水平的响应

依据在日光温室条件下的田间试验,研究了水分和氮素水平对豇豆产量和品质的影响,以期筛选较佳水分和氮素控制指标,为豇豆优质高产提供依据。结果表明:同一氮肥处理下,豇豆产量随土壤水分含量增加而显著增加,达到土壤相对含水量(RH)75%~85%时产量最高,豇豆高产较佳的水分和养分组合为:氮肥120 kg/hm2,壤RH为60%~85%。水分对豇豆鲜菜可溶性糖含量影响不明显;在水分充足条件下氮肥能提高豇豆的可溶性糖含量;水分有助于提高豇豆的Vc含量,水分、氮肥适宜豇豆可获得较高的Vc含量。豇豆硝酸盐含随施氮量增加显著增大。在氮素水平为60~120 kg/hm2条件下,水分对豇豆硝酸盐含量影响不明显,在氮素水平为180kg/hm2条件下,水分明显增加豇豆硝酸盐含量。按照蔬菜中硝酸盐含量安全标准,在氮水平为60~120 kg/hm2条件下豇豆的硝酸盐含量均在250 mg/kg以下,达到一级安全标准。氮水平120 kg/hm2是较佳的豇豆安全施肥量。综合考虑水分和氮肥对豇豆安全品质、营养品质以及产量的效应,建议豇豆生产中氮肥用量为120 kg/hm2左右,田间持水量保持在60%~85%。

鰋鮡鱼类(Glyptosternoid fishes)的系统进化和动物地理学研究

鰋鮡鱼类共包括9 属43 种（亚种），隶属于鲇形目（Siluriformes）鮡科 （Sisoridae）鰋鮡亚科（Glyptosternae）中鰋鮡族（Tribe Glyptosternini）的鰋鮡 亚族（Subtribe Glyptosternina）。鰋鮡鱼类形态上的共同特征为：无胸吸着器，胸、 腹鳍水平展开，第一根鳍条完全分节或在外缘生出许多软骨细条，被外表皮所裹， 在腹面看到的是许多与分节或软骨细条大致对应的横纹皱褶。鰋鮡鱼类集中分布 于青藏高原周边的水系中，部分属种向西分布到了中亚地区的阿姆河上游，是适 应山区急流环境的一群鱼类。近年来一系列的研究表明，鰋鮡鱼类为一单系类群， 它的起源和演化与青藏高原的隆升有着直接的关系。鰋鮡鱼类的系统发育研究， 可以为青藏高原隆升的年代、幅度和形式提供间接的证据，其系统发育树的拓扑 结构也直接反映了东喜马拉雅地区诸水系的形成与演变。 本研究对鰋鮡鱼类9 属进行了系统整理。1. 发现并描述了异齿鰋属 （Oreoglanis）两新种，分别为分布于景东无量山（澜沧江水系）的景东异齿鰋 （O. jingdongensis）和分布于怒江水系南景河和南滚河的无斑异齿鰋（O. immaculatus）。认为分布于中国的异齿鰋属鱼类均属于尖须异齿鰋种组（O. siamensis species group）。指出区分异齿鰋属两个种组：尖须异齿鰋种组和细尾 异齿鰋种组（O. delacouri species group）的特征是下唇中部是否具有中央缺刻， 尾型的差别（新月型尾或凹型尾）不能用来区分两个种组，给出了异齿鰋属的检 索表，并绘制了异齿鰋属鱼类分布图。2. 对鮡属（Pareuchiloglanis）鱼类进行了 系统整理，通过外部形态度量性状的比较，认为分布于澜沧江水系的，曾经被鉴 定为扁头鮡（P. kamengensis）的标本，应属于大鳍鮡（P. macropterus），扁头鮡 和大鳍鮡之间的最明显的差别在于腹鳍前长的不同。前者的腹鳍前长为体长的 53.2-64.9%，后者的腹鳍前长均不达体长的50%。3. 通过比较金沙江水系分布的 5 种鮡属鱼类，中华鮡（P. sinensis）、前臀鮡（P. anteanalis）、壮体鮡（P. robusta）、 四川鮡（P. sichuanensis）和天全鮡（P. tianquanensis），认为天全鮡和四川鮡之 间分布水系重叠，外部形态亦无差别，天全鮡很可能为四川鮡的同物异名。给出 了鮡属鱼类的系统检索表，并绘制了鮡属鱼类分布图。利用分子系统学的原理和方法对鰋鮡鱼类进行系统发育研究。测定了6 属 15 种鰋鮡鱼类和鮡科中非鰋鮡鱼类4 属7 种共28 个体的线粒体Cyt b 基因部分 片段和全序列（1138 bp），结合从GenBank 下载的相关类群相同的基因序列，以 魾属（Bagarius）的巨魾（B. yarrelli），纹胸鮡属（Glyptothorax）的穴形纹胸鮡 （G. cavia）、亮背纹胸鮡（G. dorsalis）、扎那纹胸鮡（G. zainaensis），福建纹胸 鮡（G. fukiensis fukiensis）、海南纹胸鮡（G. fukiensis hainanensis），黑鮡属（Gagata） 的长丝黑鮡（G. dolichonema）以及褶鮡属（Pseudecheneis）的黄斑褶鮡（P. sulcatus） 和无斑褶鮡（P. immaculatus）作为外类群，采用贝叶斯法（Bayesian）、最简约 法（maximum pasimony, MP）和邻接法（neighbour-joining, NJ）构建系统发育树。 结果显示： 1. 鰋鮡鱼类为一单系类群，并且与褶鮡属互为姐妹群关系； 2. 原鮡属、鰋属和凿齿鮡属是鰋鮡鱼类的三个基部类群； 3. 异齿鰋属构成为一个单系群，大鳍异齿鰋最早从该属的基部分化出来； 4. 石爬鮡属构成一个单系，并与分布于金沙江水系的中华鮡＋前臀鮡构成姐妹 群，黄石爬鮡和青石爬鮡的单倍型相互交错；显示两个物种的分类是不合适的， 而是同一水系不同支流种群之间梯度变异的例子，依据本次研究所得出的三个分 支图，结合青石爬鮡自西至东分布于金沙江、雅砻江、大渡河、青衣江、岷江等 的分布格局以及形态特征的分布，显示形态特征的分布变化有以下趋势：自西至 东，腹鳍位置逐渐前移；颌须渐趋缩短；胸鳍趋向发达、伸达腹鳍起点。这些变 化趋势是同一水系不同支流种群之间梯度变异的极好例子； 5. 分布于澜沧江以西（包括澜沧江）水系的鮡属鱼类（扁头鮡、细尾鮡、短鳍 鮡）与分布于怒江和伊洛瓦底江水系的拟鰋属鱼类构成为一支，并且二者共同与 分布于元江上游（红河水系）的大孔鮡构成为单系。其中，短体拟鰋和拟鰋互为 姐妹种；而扁头鮡和短鳍鮡也互为姐妹种；细尾鮡的系统地位则尚不能确定； 6. 无论在MP 树、贝叶斯树还是NJ 树中，石爬鮡属、鮡属、拟鰋属和异齿鰋属 构成一个单系，并且支持率达到了100％。 7. 鰋属位于鰋鮡鱼类的基部，是较早就从鰋鮡鱼类祖先中演化出来的一个类群， 鰋类的口吸盘是一个趋同性状，是在急流环境条件下形成的一种适应性性状，在 鰋类各属中不是同源特征； 8. 鮡属鱼类不是一个单系类群，分布于金沙江流域的鮡属鱼类与同流域分布的石爬鮡属鱼类聚成一支；澜沧江及其以西水系分布的鮡属鱼类与同流域分布的拟 鰋属鱼类聚成一支，之后这两支又同异齿鰋属共同构成为一个大支。 结合鰋鮡鱼类的系统发育分支图对其动物地理学进行研究。鰋鮡鱼类是由类 似纹胸鮡属（Glyptothorax）鱼类的祖先演化而来，鰋属（Exostoma）和凿齿鮡 属（Glaridoglanis）是较早就从从类似原鮡的祖先中演化出来，鮡属、石爬鮡属、 拟鰋属和异齿鰋属这一大支由类似原鮡的祖先演化而来。在青藏高原强烈隆起等 重大地理隔离事件发生之前，类似于现生的原鮡属鱼类已经广布喜马拉雅山脉东 西两侧。在青藏高原强烈隆起等重大地理隔离事件发生后，鰋鮡鱼类在相同的地 理隔离下独自演化为现在的分布格局。

中国蜜环菌属系统学研究

对中国已经发表的13个异宗配合的蜜环菌生物种的200多号标本用形态学和解剖学的方法进行了系统学研究，对其进行了详细的形态结构描述和重要性状的显微结构绘图，讨论了它们和近缘种间的区别，总结了这些种类的世界分布，国内分布范围和生境，给出了中国蜜环菌属的分种检索表，在此基础上报道新种5个：中国蜜环菌（CBS F）A. sinensis H.C. Wang，G.F. Qin & J. Zhao，褐囊蜜环菌（CBS H）A. bruneocystidia H.C. Wang，G.F. Qin & J. Zhao，秦氏蜜环菌（CBS J）A. qinii H.C. Wang & J. Zhao，杏孢蜜环菌（CBS L）A. amygdaliformospora H.C. Wang，G.F. Qin & J. Zhao，紫盖蜜环菌（CBS N）A. violacea H.C. Wang，G.F. Qin & J. Zhao。 用性亲和性实验对我国的同宗配合蜜环菌进行了研究，改进了用于同宗配合蜜环菌性亲和性实验的实验方法，将我国的同宗配合蜜环菌划分为8个新生物种，澄清了过去对它们的模糊认识，同时探讨了它们和欧洲及北美异宗配合的蜜环菌A. mellea的关系。 选取世界各地同宗配合的蜜环菌和我国部分存在疑问的异宗配合的蜜环菌生物种菌株进行了部分β-Tubulin序列测定，在此基础上进行了系统发育分析，探讨了各生物种间的系统发育关系，对一些疑难问题进行了讨论。