ANALYSIS OF EVOLUTIONARY RELATIONSHIP BETWEEN ASTASIA LONGA AND EUGLENA GRACILIS BY USING RAPD TECHNIQUE AND CLADISTIC ANALYSIS

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Two New Species of the Genus finocladia Kylin （Chordaria-les, Phaeophyta） from Yellow Sea Coasts of China

Two new species of the genus Tinocladia Kylin, T. gracilis sp. nov. and T. microsporangii sp. nov,, are described. T. gracilis is characterized by its slender frond, variable size, infrequently branched medullary filaments, and short assimilating filaments with 2 - 7 cells. T. microsporangii is characterized by its medullary cells which are slightly larger than sub-cortical ones; narrow sub-cortical layer, usually consisting of 4- 6 cells and very small unilocular sporangia. The holotypes are deposited at the Herbarium of the Institute of Oceanology, Chinese Academy of Sciences, Qingdao, China.     

On a new species ofPavlova (Prymnesiophyceae) from China

Pavlova viridis sp. nov. is described on the basis of light and electron microscope observations. The material was collected from the coast of haiyang county of Shandong, China. It is characterized by 1) yellowish green to green chloroplast, 2) rudimentary short flagellum 0.3 μm long and hook-shaped, 3) long flagellum with small spherical knob-scales, 4) absence of pyrenoid and stigma, and 5) marine habitat. This new species andP. salina seem to be most closely related to each other but their colour, the insertion of their three appendages, the shape of their knob scales, and their cell periplasts are quite different from one another. Contribution No. 1879 from the Institute of Oceanology, Academia Sinica     

Proteocephalus sinocyclocheili n. sp. (Cestoda: Proteocephalidae) in a fishSinocyclocheilus grahami tingi Fang from Yunnan, China

A new species ofProteocephalus from the fishSinocyclocheilus grahami tingi, a subspecies endemic to Fuxian Lake in Southwest China,is described. This is the first report of cestode from this host. The new species differs fromP. exiguus, P. Longicollis, P. parasiluri andP. torulosus, the only other species ofProteocephalus currently reported from China, by lacking an apical sucker, fromP. exiguus by having 45–108 rather than 24–54 testes per proglottis, and a maximum of 15 rather than 28 uterine diverticula; fromP.longicollis by having an average of 45–108 rather than 75–115 testes per proglottis and testes being in a single layer instead of 2 layers; and fromP. torulosus by having 45–108 instead of 150 testes per proglottis and testes being in 3–4 layers; and fromP. parasiluri by having 45–108 rather than 180 testes per proglottis. It resemblesP. percae, P. pollanicola, andP. thymalli in number of testes and uterine diverticula, but differs by lacking an apical sucker.Proteocephalus thymalli, like the new species, has a vagina that opens dorsally to the cirrus sac, butP. thymalli slao has postovarian testes, which the new species lacks. The possible significance of parasitological examinations of the other two subspecies ofS. grahami in two lakes close to Fuxian Lake is discussed in relation to host speciation and coevolution of the host-parasite system.     

PHYLOGENETIC RELATIONSHIPS AMONG PUFFERFISH OF GENUS TAKIFUGU BY RAPD ANALYSIS

AmpLifications with 300 10-base primers under predetermined optimal reaction yielded 2762 reproducible amplified fragments ranging from 200 to 3000 bp. Genetic distances among five species of Takifugu and Lagocephalus spadiceus as outgreup were calculated based on the presence/absence of the amplified fragments. Topological phylogenie trees of the 5 species of Takgfugu and the outgroup were gen-erated by Parsimony and Neighbor-Joining analysis based on RAPD data set. The genetic distance between T. rubripes and T. pseudommus was almost the same as that between individuals within each species, but was much smaller than that among T. rubripes, T. pseudommus and the other species. The molecu-lar data from RAPD analysis convincingly showed that T. rubripes and T. pseudommus should be the same species.     

Comparative studies of inorganic carbon utilization inEmiliania huxleyi and some non-calcifying marine microalgae

Inorganic carbon utilization in the non-calcifying marine microalgae,Nannochloropsis oculata, Phaeodactylum tricornutum andPorphyridium purpureum was compared with high- and low-calcifying strains ofEmiliania huxleyi grown in artificial seawater medium aerated with either air (0.03% V/V CO₂) or CO₂-free air. For high-calcifying strain ofE. oculata andP. tricornutem, similar growth patterns were observed in air-and CO₂-free air-grown cultures.P. purpureum showed a less final cell density in CO₂-free air than in air-grown culture. However, low-calcifying strain ofE. huxleyi was able to grow only in air-grown culture, but not in CO₂-free air-grown culture. Measurements of alkalinity, pH, concentration of dissolved inorganic carbon (DIC) and free CO₂ showed different patterns of DIC utilization. WithN. oculata, P. tricornutum andP. purpureum the pattern of DIC utilization was characterized by an increase of pH and a decrease of DIC but a constant alkalinity in the cultures aerated with air or CO₂-free air, suggesting that bicarbonate utilization was concomitant with an efflux of OH⁻. Both alkalinity and pH were maintained rather constant in air-grown culture of low-calcifying strain ofE. huxleyi, suggesting that diffusive entry of CO₂ could meet the requirement of DIC for its photosynthesis and growth. High-calcifying strain ofE. huxleyi, however, showed a pattern of decrease of alkalinity and DIC but an almost constant pH, indicating that bicarbonate was the major form of inorganic carbon utilised by this organism and bicarbonate uptake is unlikely to be accompanied by an efflux of OH⁻. The final pH values reached byN. oculata, P. tricornutum andP. purpureum in a closed system were 10.75, 10.60 and 9.85 respectively, showing that bicarbonate utilisation is concomitant with an efflux of OH⁻. While the final pH of 8.4 in high-calcifyingE. huxleyi suggests that bicarbonate utilization was not accompanied by an efflux of OH⁻. Contribution No. 3557 from the Institue of Oceanology, Chinese Academy of Sciences. This work was supported by NERC grant GRE3/7853 U. K. and partly supported by Bio-Engineering Center, SSTC 96-C01-05-01.     

Comparative studies on structural feature of agar polysaccharides fromPorphyra haitanensis grown in south and north China

The structural feature of agar polysaccharides fromPorphyra haitanensis grown in south China and transplanted to the north was investigated by fractionation on DEAE—Sephadex A 50, chemical analysis, and infrared and¹³C-NMR spectroscopy. The agars composed mainly of charged molecules were eluted from DEAE—Sephadex A 50 with 1.0 mol/L NaCl solution from the southernP. haitanensis and with 0.5 mol/L NaCl from the northern one. The¹³C-NMR spectra showed that agarobiose [(1→3)-β-D-galactopyranosyl-(1→4)-3,6-anhydro-α-L-galactopyranose] and the biological precursor of agarobiose [(1→3)-β-D-galactopyra nosyl-(1→4)-6-sulfate-α-L-galactopyranose] were the major disaccharide repeating units in the charged fractions. The content of the biological precursor in the agar polysaccharides from southernP. haitanensis was higher than that in the northern one, the content of the biological precursor extracted from cold water was higher than that from hot water, and the content of 6-OMe-D-galactose in the southernP. haitanensis polysaccharides was higher than in the northern one. This distinct difference will be of significance for further study of the physiological characters ofP. haitanensis. Contribution No. 1849 from the Institute of Oceanology, Academia Sinica.     

Redescription and taxonomic reconsideration ofNemertopsis gracilis var.bullocki Coe, 1940

Specimens ofNemertopsis gracilis var.bullocki Coe, 1940 collected from Huian, Fujian Province, China, are described. The variety is upgraded to specific level on the basis of: the dorsal surface having a pair of blackish brown longitudinal stripes meeting anteriorly with a broad transverse band of the same color; the proboscis having 11–13 distinct proboscis nerves in its longitudinal muscle layer. Project 39170111 and 39470097 supported by NSFC.     

Report of Pontoniinae Shrimps （Crustacea： Decapoda） Collected by Joint Chinese-German Marine Biology Expeditions to Hainan Island, South China Sea. Ⅲ. PericHmenes

The present paper reports 12 species of genus Periclimenes, subfamily Pontoniinae, collected from Hainan Island, South China Sea, by the Joint Chinese-German Marine Biology Expeditions, in which, Periclimenes consobrinus and P. tenuipes are new records from Chinese waters, and Periclimenes amymone, P. holthuisi, P. soror, and P. toloensis are recorded for the first time from Hainan Island.     

Competition Among Dinoflagellate Alexandrium Tam—arense, Raphidophyte Heterosigma Carterae and Diatom Skeletonema Costatum under Combinations ofTwo Temperatures and Five Salinities

Competition among HAB (Harmful Algal Bloom) species Dinoflagellate Alexandrium tamarense, Raphidophyte Heterosigma carterae, and Diatom Skeletonema costatum was studied in the laboratory. Experiments with these three major HAB species under combinations of different salinities (10, 18, 25, 30, 35) and temperatures (19℃, 25℃) were carried out. The results showed that S. costatum successfully competed with the other two species at salinities of 18, 25,30, and 35 at temperatures of 19℃ and 25℃. However, H. carterae showed its advantage at low salinity of 10 and became the single dominant species at salinity 10 and 25℃. A. tamarense could not compete successfully with the other two species especially at low salinities. However, it could remain at low density in the presence of higher densities of other algae.     

REPORT ON PONTONIINE SHRIMPS（CRUSTACEA：DECAPODA）COLLECTED BY JOINT CHINESE—GERMAN MARINE BIOLOGY EXPEDITIONS TO HAINAN ISLAND,SOUTH CHINA SEA i.ANCHISTUS,CONCHODYTES,CORALLIOCARIS

The present paper reports 7 species of pontoniine shrimps collected from Hainan Island,South China Sea,by the Joint Chinese-German Marine Biology Expeditions,in which,Conchodytes nipponensis(De Haan,1844)and Coralliocaris brevirostris Borradaile,1898 are new records from Chinese waters.     

Three new species of Batrachospermum Roth （Batrachospermaceae, Rhodophyta） in China

Three new species of Batrachospermum Roth (Batrachospermales, Rhodophyta) from China are described in this paper. B. yunnanense sp. nov. has long-cylindrical trichogynes with long stalks and is diagnostic of section Virescentia. Within this section, B. yunnanense is similar to B. helminthosum Bory emend. Sheath et al., but it is dioecious and has curved carpogonial branches, while the latter is monoecious and has straight carpogonial branches. It is also similar to B. transtaganum Reis, but it differs from the latter in long carpogonia,big carposporophytes and carposporangia. It is considered that B. nothocladoideum sp. nov. is assigned to section Contorta, subsection Kushiroense, because its carpogonial branches are twisted and gonimoblast filaments are loosely agglomerated. This new species similar to B. iriomotense Kumano, but with short fascicles, long-ovoid or subpyriform cells, numerous terminal hairs, long-ellipsoid trichogynes, big carposporophytes and small carposporangia. The plant is quite tough and cartilaginous and similar to Nothocladus in gross morphology, but its carposporophytes are compact instead of diffuse. This shows that it may be a transitional species between section Contorta and genus Nothocladus. So, B. transitorium sp. nov. should belong to section Contorta, subsection Kushiroense, because of its curved or twisted carpogonial branches and loosely agglomerated gonimoblast filaments, with globose or subglobose cells in fascicles similar to B. spermatiophorum Vis et Sheath, but no colourless spermatiophores. In terms of small and numerous carposporophytes, B. transitorium sp. nov. is similar to some species of section Batrachospermum. However, their other features are unique, indicating its transitional nature between section Contorta and Batrachospermum.     

A Fast and Indriect Fluorescent Antibody Assay for the Vibro in Large Yeelow Croaker Pseudosciaena Crocea (Richardson)

A fast and indirect fluorescnet antibody assay for the Vibrio alginolyticus and V.Parahaemolyticus infecting the large yellow croaker has been developed.The specific antisera for the two strains of vibrio were prepared with New Zealand rabbit and the antiserum and cross-reactive efficacy was tested by coagulation in tube.It showed that the goat anti-rabbit IgG had been labeled by fuorescence isothiocyanate(FITC).The results showed that positive reactions were 100% for the large yellow croaker Pseudosciaena crocea with typical symptom of vibrio infection,while the positive reaction to the pathogen in healthy yellow croakers reached 40%,but seemed negative for aquaculter water.The results demonstrated that this fast and indirect fluorescent antibody assay can be used not only to test the vibrio pathogen in diseased yellow croaker but also in infected animals with no symptom.     

Assessing the germplasm of Laminaria (phaeophyceae) with random amplified polymorphic DNA (RAPD) method

Eighteen gametophytes includingL. japonica, L. ochotensis andL. longissima, were verified with random amplified polymorphic DNA (RAPD) technique. Eighteen ten-base primers were chosen from 100 primers selected for final amplification test. Among the total of 205 bands amplified, 181 (88.3%) were polymorphic. The genetic distance among different strains ranged from 0.072 to 0.391. The dendrogram constructed by unweighted pair-group method with arithmetic (UPGMA) method showed that the female and male gametophytes of the same cell lines could be grouped in pairs respectively. It indicated that RAPD analysis could be used not only to distinguish different strains ofLaminaria, but also to distinguish male and female gametophyte within the same cell lines. There is ambiguous systematic relationship if judged merely by the present data. It seems that the use of RAPD marker is limited to elucidation of the phylogenetic relationship among the species ofLaminaria. Contribution No. 4389 from the Institute of Oceanology, Chinese Academy of Sciences. This research was supported by the State's High-tech Project of China, Marine 863 project (No. 819-03-02) and (2001AA621090).     

Molecular identification of Prorocentrum (Dinophyceae) species

A fragment of a large sub-unit ribosomal DNA (LrDNA) of 12 strains ofProrocentrum species was amplified by polymerase chain reaction (PCR). The PCR products were digested by 3 restriction endonucleases (Cfo I, Hae III, and RSA I) and then resolved in agarose gels. Results show that different species had different RFLP patterns, except forP. arcuatum (ME 131), which had the same pattern toP. micans (ME160 and 04). The same fragment of 19 strains of the genus was also amplified and subjected to denaturing gradient gel electrophoresis (DGGE). 11 different patterns were resolved. Different cultures of a same species had the same pattern. The results of RFLP and DGGE analyses showed that eight newly isolated epibenthicProrocentrum species were different from each other, and also from other cultured ones examined in this study.P arcuatum (ME132) could not be differentiated fromP. micans (ME160 and 04), it was probably mis-identified, since they are quite different morphologically.P. redfieldii (ME138) could also not be distinguished formP. triestinium (ME132), it should be regarded as a synonym ofP. triestinium. Unexpectedly, a restriction site was found inP. micans, compared with previous sequence data. Project supported by National Basic Research Priorities Program (2001CB409701, 2001CB409710) and supported by NSFC (40376040, 40025614)     

Evalution on nutritive value ofPortunus trituberculatus

This study on the nutritive indexes (total nitrogen, amino acids, crude fats, inorganic elements, unsaturated fatty acids) of meat, male and female reproductive gland ofPortunus trituberculatus showed that their nutritive value is in the order meat>female reproductive gland>male reproductive gland and that they do not raise the total cholesterol (TC) and triglyceride (TG) contents of the serum of the animals which eat them but increase the contents of high density lipoprotein cholesterol (HDL-C) in the animal's blood. These findings implied that people who have high blood lipid and aortic atheroma can safely use them as food. This study showed thatP. trituberculatus has high nutritive value.     

A new radiolarian species Clathrocorys gracilis from northern South China Sea

A new species,Clathrocorys gracilis nov. spec. of Tripocyrtidae (Radiolarians: Nassellaria) from surface water in northern South China Sea, is described in this paper. The new species has four radial beams arising from the cephalic base centre, each foot with cup-shaped structure and ramified bar in the proximal section (near cephalic base), and each wing with a large mesh in the middle, a medium mesh and a small mesh on each side respectively. Project supported by the NSFC (No. 40276044) and the Chinese Academy of Sciences (No. KSCX2-SW-101B).     

Two new species of Ophidiasterid sea-stars from the vicinity of Diaoyudao,East China Sea

Two new species of the asteroid family Ophidiasteridae are described from two specimens collected by commercial fishery vessels of Qingdao from the vicinity of Diaoyudao (25°45′N, 123°15′E), East China Sea.Hacelia tuberculata sp. nov. shows affinities withH. tyloplax (H. L. Clark, 1914) andLinckia gracilis sp. nov. is related toL. laevigata (Linnaeus). Contribution No. 1163 from the Institute of Oceanology, Academia Sinica.     

Studies on extraction procedure and antioxidative activity of phlorotannins fromSargassum kjellmanianum

Known only in phaeophyceae, phlorotannins (brown algal polyphenols) are natural products with potential uses in pharmacology. This study yielded an extraction procedure to obtain high purity, high molecular weight phlorotannins fromSargassum kjellmanianum and revealed the characteristics of their infrared and flourescence spectra. The antioxidative activity of phlorotannins, which was about 2.6 times as strong as that of 0.02% BHT (tert-butyl-4-hydroxytoluene), showed potential for preventing oil rancidification. Contribution No. 2918 from the Institute of Oceanology, Chinese Academy of Sciences. The project was supported by The State Ninth-five Key Program of China. (No. 96-916-04-01).