东海原甲藻修订及与相关原甲藻的分类学比较

分析了东海原甲藻(Prorocentrum donghaiense Lu)显微结构,并与具齿原甲藻(Prorocentrum dentatum Stein)模式种和Schiller的钝头原甲藻的描述等进行了比较,结果表明,它们之间的形态结构和个体大小具有很大的差别,这些差异远超出了同种个体因环境不同所造成的形态变化范围．从细胞形态及其表面结构可以判断,日本、韩国海区所记录并报道的“P．dentatum”与我国东海的东海原甲藻应属同一种．因此可以认为,我国东海赤潮高发区以及在韩国、日本海区的出现的高生物量(high biomass bloom—forming species)赤潮原甲藻不是Stein所发表的具齿原甲藻。而是东海原甲藻,并对其进行了进一步修订,其种名应为东海原甲藻Prorocentrum donghaiense Lu．     

东海具齿原甲藻的扫描电子显微结构

在光学显微镜分析鉴定的基础上,通过高分辨率的扫描电子显微镜,对东海赤潮原甲藻标本和美国国家海洋藻种中心(CCMP)的具齿原甲藻(Prorocentrum dentatum)藻株(CCMPl517)的细胞表面结构进行观察和比较,认为两者为同一种类．在分析大量文献资料的基础上,可以认为发生在我国东海水域的赤潮原甲藻种类是具齿原甲藻(Prorocentrum dentatum),而非新种．     

慢原甲藻——南海热带附生甲藻新记录

从海南陵水新村港分离出1株附生原甲藻,在光学显微镜和扫描电子显微镜下进行形态学特征分析,并结合核糖体18S rRNA基因序列分析,鉴定为慢原甲藻(Prorocentrum rhathymum),为国内首次报道.国内曾将该种与墨西哥原甲藻(Prorocentrum mexicanum)归为同种异名.     

慢原甲藻——南海热带附生甲藻新记录

从海南陵水新村港分离出1株附生原甲藻,在光学显微镜和扫描电子显微镜下进行形态学特征分析,并结合核糖体18S rRNA基因序列分析,鉴定为慢原甲藻(Prorocentrum rhathymum),为国内首次报道。国内曾将该种与墨西哥原甲藻(Prorocentrum mexicanum)归为同种异名。     

我国东海赤潮原甲藻应属哪种？

阐述并讨论了中国东海长江口外经常发生的原甲藻赤潮原因种的种类认知问题．论述了自从Stein提出具齿原甲藻(Prorocendrum dentatum Stein)新种后迄今中外专家报告该种的描述及分布,并对比了具齿原甲藻与长江口的原甲藻的异同．研究证实我国东海的原甲藻与具齿原甲藻有较大的差别,主要表现在体积大小、藻体末端形态以及藻体前端突起的大小和形态等方面．本文汇集了国内外对具齿原甲藻与我国原甲藻的研究成果,对比认为。我国长江口外经常发生赤潮的原甲藻为东海原甲藻(Prorocendrum donghaiense Lu)．文中还讨论了由陆斗定发表的东海原甲藻需要补充修正的观点．在过去的数年中,我国东海长江口海域频繁爆发大面积的原甲藻赤潮,但对于赤潮原因种原甲藻(Prorocentrum)的定种问题存在两种不同的观点,一种观点认为本种是具齿原甲藻,另一种观点认为是东海原甲藻．为了阐明此种原甲藻的命名问题,于2002年11月在深圳召开了题为“我国东海赤潮原因种原甲藻分类学国际研讨会”．认为是P．dentatum的依据是长期以来国际上通常把类似我国东海的原甲藻定为P．dentatum,其根据是它的前端有突起等．而持不同观点的专家认为我国东海的这种原甲藻在细胞长度上与Stein描述的原P．dentatum差异很大,并且除少数样品末端尖伸外,大多数细胞末端是钝圆的．专家们还认为在日本、韩国等亚洲海域分布的被报告为P．dentatum的原甲藻与我国东海的此种原甲藻为同一种．综观各种观点,作者认为东海本种原甲藻应为东海原甲藻.     

北部湾海洋原甲藻的形态特征及其系统进化分析

为探讨广西北部湾海洋原甲藻(Prorocentrum micans)的形态特征及其系统进化意义,利用光学显微镜、分子生物学方法,对海洋原甲藻BBW-01 藻株的形态特征进行了描述,并分析了其分子系统进化关系.结果表明,各地理株系的海洋原甲藻的形态特征相似,仅在细胞大小上存在差异.海洋原甲藻BBW-01 与采自于广东大亚湾的海洋原甲藻形态特征最为接近,其壳板后端的7 个呈“V”字形对称排列的大孔可作为海洋原甲藻鉴定的重要指标.18S rDNA 序列同源检索和系统进化分析表明,海洋原甲藻BBW-01 与源自中国南海的海洋原甲藻的亲缘关系最近,并与其他2 株海洋原甲藻聚成一支,属于浮游、兼性浮游类原甲藻.因此,对赤潮原因种的准确识别有助于预防和减轻赤潮对海洋环境和人类带来的危害.     

营养盐限制条件下塔玛亚历山大藻对东海原甲藻的化感作用研究

为明确塔玛亚历山大藻(Alexandrium tamarense)对东海原甲藻(Prorocentrum donghaiense)生长的化感作用,研究了在N、P限制及正常营养盐条件下(又称富营养)塔玛亚历山大藻无细胞滤液对东海原甲藻生长的影响,并探讨了3种不同营养盐条件下两种藻共培养时的生长状况。结果表明,半连续培养时,营养盐限制下,塔玛亚历山大藻无细胞滤液对东海原甲藻的生长均有一定影响。N限制下,5 d后东海原甲藻藻密度显著低于未加滤液的对照组,藻密度为1.02×107 cells L-1,对照组为1.7×107 cells L-1;P限制下,东海原甲藻藻密度与对照组差异不显著,5 d后藻密度为1.44×107 cells L-1;富营养条件下,东海原甲藻藻密度与对照组无明显区别。共培养时,塔玛亚历山大藻对东海原甲藻生长的抑制作用更为显著,N、P限制下,4 d后东海原甲藻全部死亡,且聚集成团形成沉淀;富营养条件下,仍有少量东海原甲藻存活(藻密度3.3×104 cells L-1)。这表明,塔玛亚历山大藻对东海原甲藻的生长有一定的化感作用。营养盐限制可促进塔玛亚历山大藻化感物质的合成和释放,化感作用是塔玛亚历山大藻抑制东海原甲藻生长的原因之一。     

米氏凯伦藻与东海原甲藻共培养条件下的种群竞争

米氏凯伦藻(Karenia mikimotoi)与东海原甲藻(Prorocentrum donghaiense)为有害赤潮生物,两者经常形成复合型赤潮.该文设置东海原甲藻的起始密度为400 cells·mL-1,米氏凯伦藻分别为200 cells·mL-1、400 cells·mL-1和800 cells·mL-1,通过共培养实验,初步研究米氏凯伦藻和东海原甲藻的种间关系.结果表明:共培养条件下,受到东海原甲藻的影响,米氏凯伦藻的生长受到抑制.米氏凯伦藻不同的起始密度对东海原甲藻的生长有不同的影响,较低的起始密度(200 cells·mL-1、400cells·mL-1)促进东海原甲藻的生长,使其增长率提高,生长曲线达到拐点的时间提前;高的起始密度(800 cells·mL-1)对东海原甲藻的生长有一定的抑制作用,使其增长率降低,生长曲线达到拐点的时间推迟.     

东海原甲藻对不同磷源的利用特征

采用单因子实验,以NaNO3作为唯一氮源,固定初始氮浓度为40μmol.L-1,设置六个初始磷浓度梯度:0.1μmol.L-1、0.5μmol.L-1、1.0μmol.L-1、2.0μmol.L-1、5.0μmol.L-1、10.0μmol.L-1,对比研究了东海原甲藻(Prorocentrum donghaiense)对4种不同磷源的利用特征及其这4种不同磷源对东海原甲藻的营养价值。结果表明:东海原甲藻既可以直接吸收利用无机磷——磷酸二氢钠(NaH2PO4),又可以不同程度地利用3种小分子有机磷:三磷酸腺苷二钠盐(ATP)、D-葡萄糖-6-磷酸钠(G-6-P)和甘油磷酸钠(G-P);东海原甲藻在以NaH2PO4、G-6-P和ATP分别作为磷源时的最大比生长率(μmax)值相差不大,3种磷源对最大生物量的贡献也基本相近;但是以G-P作为磷源时则明显有最低的μmax值,其对最大生物量的贡献也最低。东海原甲藻对这4种磷源的利用特征说明:长江口自然海水中与三种小分子有机磷结构相似的溶解有机磷在一定程度上也可以为东海原甲藻赤潮的爆发和维持提供磷源。     

利玛原甲藻中聚酮合酶基因克隆与分析

为探讨聚酮合酶 (polyketide synthase, PKS)基因与藻毒素合成的关系,揭示PKS基因在赤潮毒素合成中的作用,采用兼并引物,通过PCR技术获得利玛原甲藻(Prorocentrum lima)可能存在的I型PKS基因;并对所获得PKS基因的同源性进行了分析,构建了基于PKS氨基酸序列的系统进化树;采用RT-PCR技术分析了PKS基因在利玛原甲藻中的表达状况;并通过多聚腺苷酸RNA的扩增、细菌的分离鉴定、限制性内切酶酶切、Southern blotting等技术对PKS基因进行了分析.结果表明,利玛原甲藻中PKS基因与海洋原甲藻聚为一支,在利玛原甲藻中有显著表达;以Oligo(T)引物进行RT-PCR扩增时,可出现18S rRNA和PKS基因相应条带;限制性内切酶酶切和Southern blotting结果显示,该基因中存在明显的甲基化;16S rRNA基因序列分析显示,从利玛原甲藻培养液中分离到的细菌与海洋放线菌假诺卡氏菌属(Pseudonocardia)基因序列同源性达到99%,该菌株中并不存在PKS基因.结果显示,所获得的PKS基因是利玛原甲藻聚酮合酶基因,基因序列已提交GenBank (EF521601);PKS可能在腹泻性贝毒合成中起着关键作用.     

Cytotoxicity in the marine dinoflagellate Prorocentrum mexicanum from Brazil

The microscopic algae in the oceans are crucial food for filter feeding bivalve shellfish (oysters, mussels, scallops, clams, etc.) as well as for the larvae of commercially important crustaceans. Some species of microalgae have the capacity to produce potent toxins, such as saxitoxins and ciguatoxins, which may intoxicate humans. Among the marine phytoplankton, the dinoflagellates are the main toxin producers. Studies on the marine phytoplankton from the S?o Sebasti?o Channel, southeastern coast of Brazil, showed a great diversity of dinoflagellates. Some species were collected and cultured at the Marine Biology Center of the S?o Paulo University (USP). The polar (PEs) (aqueous) and apolar (AEs) (methylene chloride) extracts of the cultivated dinoflagellate species were tested on different stages of the sea urchin development, on mouse erythrocytes and on microfilaments organization in a neuroblastoma cell line. Prorocentrum mexicanum PE and AE induced cells anomalies and cell division inhibition of sea urchin eggs at EC50 of 78.75 microg/mL (95% CI from 32.56 to 190.50) and 22.50 microg/mL (95% CI from 2.96 to 170.80) respectively (n=3). Both AE and PE of P. mexicanum induced hemolysis with EC50 of 65.07 microg/mL (95% CI from 27.40 to 154.60) and 84.29 microg/mL (95% CI from 53.26 to 133.40 microg/mL), respectively. P. mexicanum PE was tested in immunofluorescence for actin filaments organization in neuroblastoma cultured cell.     

Type studies in the Polyporaceae 11. Species described by J. F. C. Montagne, either alone or with other authors

Of the 93 polypores described by Montagne, either alone or with other mycologists, 32 are accepted, 53 are synonyms, 3 names are invalid, 2 species are of unknown affinities, 2 types have not been found while 1 name is rejected. The following three combinations are proposed: Phylloporia capucina (Mont.) Ryv. comb, nov., Coltricia fragilissima (Mont.) Ryv. comb, nov., and Gloeophyllum mexicanum (Mont.) Ryv. comb. nov.     

Ribosomal 5S genes in relation to C-value in amphibians.

We have measured the amount of 5S-ribosomal DNA in the genomes of Xenopus laevis, Triturus cristatus carnifex and Ambystoma mexicanum, three species of Amphibians which have widely different C-values. Our best estimate is that these organisms have about 24,000, 32,000 and 61,000 5S-genes per haploid genome respectively. A trend to increasing 5S gene copynumber with increasing C-values in amphibians is apparent, probably linked to the need to supply more ribosomes to the larger cells which are associated with larger genomes, particularly during the critical phases of oogenesis and embryonic cleavage. The correlation between the two is poor however, and whilst C-value may determine a minimum gene copy-number, there appears to be little constraint on exceeding this minimum in some species. Certain problems encountered in measuring gene copy-numbers, i.e. the criterion dependance of such numbers and the effect of having pseudogenes, are highlighted.     

Growth and Grazing Rates of the Marine Planktonic Ciliate Strombidinopsis sp. on Red-Tide and Toxic Dinoflagellates

ABSTRACT We investigated growth and grazing rates of Strombidinopsis sp. when feeding on several species of red-tide and/or toxic dinoflagellates. Strombidinopsis sp. one of the largest aloricate choreotrichs so far reported, grew well on Lingulodinium polyedrum, Gymnodinium sanguineum, Scrippsiella trochoidea, Cochlodinium polykrikoides , and Prorocentrum minimum , but failed to grow on Amphidinium carterae. Specific growth rates of Strombidinopsis sp. increased rapidly with increasing prey density up to ca. 100 ng C ml^-1, but were saturated or increased slightly at higher concentrations. Maximum specific growth rates of Strombidinopsis sp. on various prey species were 1.38 day^-1 for C. polykrikoides , 1.27 for G. sanguineum , 1.06 for P. minimum , 0.83 for L. polyedrum , and 0.67 for S. trochoidea. Threshold prey concentrations (where net growth = 0) were 12–38 ng C ml^-1. Maximum ingestion and clearance rates of Strombidinopsis sp. were 353 ng C grazer^-1 day^-1 and 110 μ, l grazer^-1 h^-1, respectively. Strombidinopsis sp. exhibited higher maximum growth, ingestion, and clearance rates than the mixotrophic dinoflagellate Fragilidium cf. mexicanum or the heterotrophic dinoflagellates Protoperidinium cf. divergens and P. crassipes , when grown on the same prey species. In addition, the sequence of prey species arranged according to growth response of Strombidinopsis sp. differed considerably from those of Fragilidium cf. mexicanum, Protoperidinium cf. divergens , and P. crassipes.     

CULTIVATION STUDIES OF GIGARTINA SKOTTSBERGII IN SOUTHERN CHILE

Increases in population and agriculture in coastal areas can result in increased nutrient inputs and alterations in the ratios of organic to inorganic nutrients in coastal waters. Such changes in coastal nutrient regimes can affect phytoplankton community structure by creating conditions favorable for growth and dominance of algae that were not dominant before. The effect that changes in ratios and concentrations of nutrients have on toxicity of harmful algal species is not well known. There seems to be a relationship; however, between nutrient stress and toxin production among harmful phytoplankton producing low-N toxins, e.g. Diarrhetic Shellfish Poisoning (DSP) toxins. Even less is known about the relationship between organic nutrient uptake and toxin production. Benthic species and species in coastal areas are probably exposed to greater fluxes of dissolved organic nitrogen (DON). In this study, benthic and planktonic species of Prorocentrum were grown on L1 media with the sole N-source varying among treatments as nitrate, ammonium, urea, L-glutamic acid, and high molecular weight natural DON. An ELISA specific to the DSP toxins, okadaic acid and 35-methylokadaic acid, was used to determine toxin production by each species when grown on the different N sources. Preliminary results indicate that some organic forms of N support growth as well as inorganic forms for Prorocentrum minimum , P. mexicanum , and P. hoffmannianum.     

Predation and dispersal of large and small seeds of a tropical palm

Seed size may vary greatly among individuals within plant species. What effects the extremes of this variation have for seeds taken by small mammals are poorly understood. Not all seeds removed by small mammals are necessarily eaten. Small rodents are common seed predators, but they may disperse a significant proportion of seeds by scatter hoarding them via burial. Size-dependent predation and dispersal of seeds has not been directly tested within a plant species for tropical rodents. This study tested whether or not large and small nuts of Astrocaryum mexicanum (Palmae) differed in their fates due to handling by the spiny pocket mouse Heteromys desmarestianus (Heteromyidae). Exclosures were used to give small rodents exclusive access to A. mexicanum nuts. H. desmarestianus preferentially consumed large over small A. mexicanum nuts, but cached (in burrows and by scatter hoarding) similar proportions of these nuts by size. Small nuts tended to be buried farther away from exclosures than large nuts. Although sample sizes of buried nuts were small, the rodents retrieved all buried large nuts, but 30% of the small nuts remained buried long enough to germinate. I also examined predispersal predation by insects and found that insects appear to have no size preference for A. mexicanum nuts, but insect predation appears to hinder nut development. Thus, nuts attacked by insects develop to be significantly smaller, with a low proportion of undamaged endosperm, than uninfested nuts. It is hypothesized that the preferential predation of large A. mexicanum nuts by H. desmarestianus is a response by these rodents to insect predation.     

Detection of a malaria parasite (Plasmodium mexicanum) in ectoparasites (mites and ticks), and possible significance for transmission

Two species of sandflies (Lutzomyia) are competent vectors of Plasmodium mexicanum, a malaria parasite of lizards. The very patchy distribution of sites with high P. mexicanum prevalence in the lizards, and often low or even nil sandfly density at such sites, provoked an evaluation of 2 common lizard ectoparasites, the tick Ixodes pacificus and the mite Geckobiella occidentalis, as potential passive vectors. Plasmodium sp.-specific polymerase chain primers were used to amplify a long segment of the mitochondrial cytochrome b gene that is unlikely to survive intact if the parasite cells are killed within a blood-feeding arthropod. The segment was strongly amplified from sandflies (the positive control for the method) from 1 to 96 hr postfeeding on an infected lizard. For ticks, the gene fragment was poorly amplified at 0 hr postfeed, and not amplified after 2 hr. In contrast, strong amplification of the parasite DNA was observed from mites from 0 to 20 hr postfeed, and weak amplification even at 96 hr.     

Two new species of Echinobothrium (Cestoidea: Diphyllidea) from myliobatiform elasmobranchs in the Gulf of California, México

Two new species of diphyllidean cestodes, Echinobothrium mexicanum n. sp., and Echinobothrium fautleyae n. sp., are described, the former from the spiral intestines of the bat rays Myliobatis californicus and Myliobatis longirostris and the latter from the spiral intestine of the cownose ray Rhinoptera steindachneri, all caught in the Gulf of California, México. Echinobothrium mexicanum is most easily distinguished from the majority of the species of the genus by the number of large apical hooks in each dorsoventral group (23) and from the remainder in the number of lateral hooklets in each group on the scolex. Echinobothrium fautleyae differs conspicuously from all other members of the genus in that the first and last lateral hooklets in each group are distinctly larger than the other hooklets in each group. Serial sections of the scoleces of these 2 new species confirm that the attachment structures on the scolex are bothria, rather than bothridia, as has been described in other species in the genus, suggesting that the attachment structures of the scolex should be examined in more detail in the genus in general. Palmate microtriches of various forms cover the proximal bothrial surfaces of both new species and are also found on the dorsal bothrial surfaces of E. fautleyae. The distal bothrial surfaces of E. mexicanum are covered with slender spiniform and short filiform microtriches. The cephalic peduncles of both species possess short filiform microtriches only. Comparison of these 2 new species with the other 3 species in this genus that have been examined with SEM suggests that the various forms and distributions of these microtriches is unique in each of these 2 species. We believe these features will prove to be taxonomically useful.     

Breeding relationships in the genus Brachypodium (Poaceae: Pooideae)

Results of hybridisation experiments involving five perennial European species of Brachypodium ( B. sylvaticum, B. glaucovirens, B. pinnatum, B. phoenicoides and B. retusum ), the annual B. distachyon , and the C. American perennial B. mexicanum are presented. Four of the European perennials are largely self-incompatible, whereas B. sylvaticum, B. distachyon and B. mexicanum are highly self-compatible. Despite their avowed ancient and relict status, the European species have mostly retained the ability to interbreed and produce viable F₁ hybrids. The F₁ hybrids obtained between the five European perennials all exhibit some degree of fertility. Hybrids between B. distachyon and at least some of the European perennials are vigorous and long-lived, but show no evidence of fertility. No hybrids involving B. mexicanum could be raised. Success in raising hybrids, and the degree of fertility of such hybrids, partly agrees with closeness of taxonomic relationships as measured by morphological, anatomical, biochemical and molecular characteristics, but also with closeness in chromosome number irrespective of taxonomic affinity. Hence some interspecific hybrids are more easily produced and are more fertile than certain intraspecific hybrids involving parents of different chromosome number. All hybrids form good numbers of bivalents, but meiosis varies from almost regular in hybrids between European perennials with the same chromosome number to very irregular in hybrids between B. distachyon and any perennial species.     

Male gametocyte fecundity and sex ratio of a malaria parasite, Plasmodium mexicanum

Evolutionary theory predicts that the sex ratio of Plasmodium gametocytes will be determined by the number of gametes produced per male gametocyte (male fecundity), parasite clonal diversity and any factor that reduces male gametes' ability to find and combine with female gametes. Despite the importance of male gametocyte fecundity for sex ratio theory as applied to malaria parasites, few data are available on gamete production by male gametocytes. In this study, exflagellating gametes, a measure of male fecundity, were counted for 866 gametocytes from 26 natural infections of the lizard malaria parasite, Plasmodium mexicanum. The maximum male fecundity observed was 8, but most gametocytes produced 2-3 gametes, a value consistent with the typical sex ratio observed for P. mexicanum. Male gametocytes in infections with higher gametocytaemia had lower fecundity. Male fecundity was not correlated with gametocyte size, but differed among infections, suggesting genetic variation for fecundity. Fecundity and sex ratio were correlated (more female gametocytes with higher fecundity) as predicted by theory. Results agree with evolutionary theory, but also suggest a possible tradeoff between production time and fecundity, which could explain the low fecundity of this species, the variation among infections, and the correlation with gametocytaemia.