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).     

PHYLOGENETIC RELATIONSHIPS AMONG PUFFERFISH OF GENUS TAKIFUGU BY RAPD ANALYSIS

ＩＮＴＲＯＤＵＣＴＩＯＮＰｕｆｆｅｒｆｉｓｈｅｓｏｆｔｈｅｇｅｎｕｓＴａｋｉｆｕｇｕ (Ｔｅｔｒａｏｄｏｎｔｉｆｏｒｍｅｓ,Ｔｅｔｒａｏｄｏｎｔｉｄａｅ)ａｒｅｍａｉｎｌｙｄｉｓｔｒｉｂｕｔｅｄａｌｏｎｇｔｈｅｃｏａｓｔａｌｒｅｇｉｏｎｓｏｆＣｈｉｎａ ,ＪａｐａｎａｎｄＫｏｒｅａａｎｄａｒｅｃｏｍｍｅｒｃｉａｌｌｙｉｍｐｏｒｔａｎｔｔｏｔｈｅｓｅｃｏｕｎ ｔｒｉｅｓ .Ｏｖｅｒ 2 0ｓｐｅｃｉｅｓｈａｖｅｂｅｅｎｒｅｃｏｒｄｅｄｉｎｔｈｅＹｅｌｌｏｗＳｅａａｎｄｔｈｅＥａｓｔＣｈｉｎａＳｅａ (Ｃｈｅｎｇ…     

Use of rapd for detecting and identifyingPorphyra (Bangiales, Rhodophyta)

Randomly amplified polymorphic DNA (RAPD) technique was applied to assess the genetic variations and phylogenetic relationships in 4 species ofPorphyra. The samples were collected from the coast of Canada, Vietnam, Zhoushan, Fujian and Qingdao in China. Amplifications with 20 primers were carried out under predetermined optimal reaction conditions (samples were first heated at 94°C for 5 min. and followed by 45 cycles of 1 min at 94°C, 1 min at 36°C, and 2 min at 72°C, then held at 72°C for 10 min). The amplified products were scored as present (1) or absent (0) for each DNA sample and an index of genetic similarity (F) was calculated by using Nei & Li’s matching coefficient method (1979). The value of (1−F) was used to quantify the genetic distances between species and construct a phylogenetic tree. The relationship indicated by the UPGMA and NJ cluster analysis on the values of the genetic distance is in good overall agreement with classical taxonomy. The obvious differences between natural and cultivated population ofP. haitanensi suggest that variation or hybridization with other species occurred during the culture. Contribution No. 100 from the Experimental Marine Biology Laboratory, IOCAS. Contribution No. 3289 from the Institute of Oceanology, Chinese Academy of Sciences. The research supported by the Foundation of the Chinese Academy of Sciences.     

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.     

Invasion and spreading of Cabomba caroliniana revealed by RAPD markers

Applying randomly amplified polymorphic DNA (RAPD), the genetic variation ofCabomba caroliniana Gray (cabomba or fanwort), a new alien plant in China, was analyzed in this paper. Total 143 bands, including 47 polymorphic bands, were amplified from 23 primers in 20 samples. The sampling distance was large, but its genetic diversity was low. The main results were that: (1) Cabomba, which grew and dispersed mainly in fragment, was an abundant and dominant species in freshwater, and its main dispersal mechanism was vegetative reproduction (2) Cabomba was originally introduced into China as an aquarium submerged plant. Somehow, those discarded cabomba became invasive species in the areas of Hangzhou, Shanghai, and Meicheng, and other places. (3) Although the level of genetic diversity in cabomba was low, their rapid dispersion and propagation could seriously harm to local aquatic community. Therefore, specific measure should be used to control cabomba from uncontrolled spreading and damage to local vegetation communities. Supported by National Natural Science Foundation of China (No. 39970145) and Major State Basic Research Development Program of China (No. G2000046803).     

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)     

Hybrids between olive flounder Paralichthys olivaceus and stone flounder Kareius bicoloratus： karyotype, allozyme and RAPD analyses

The hybrid between olive flounder Paralichthys olivaceus and stone flounder Kareius bicoloratus was produced by artificial insemination of olive flounder eggs with stone flounder sperm. Sinistral and dextral are two types of hybrid progeny after metamorphosis. Karyotypes of both hybrid flounders are the same as those of the two parental species. Of the 22 loci examined from 12 allozymes, 12 confirmed hybridization of the paternal and maternal loci in hybrids and no difference was found in allozyme patterns of sinistral and dextral hybrid fishes. RAPD patterns of these specimens were also studied with 38 primers selected from 104 tested. Among them, the PCR products of 30 primers showed hybridization of the paternal and maternal bands. Genetic variation between hybrids and their parental stocks was analyzed by RAPD using 10 of the above 38 primers. The average heterozygosity and genetic distance were calculated. The results suggested that the filial generation could inherit a little more genetic materials from paternal fish than that from maternal fish. Supported by National Natural Science Foundation of China (No. 30571445), National High-Technology Research and Development Program (863 Program, No. 2006AA10A404), and Project from the Ministry of Science and Technology of China (No. 2006DKA30470-017)     

Observation of female gamete (Carpogonia) germination of Porphyra yezoensis and P. oligospermatangia

Several papers have reported that part or whole leafy thallus seemingly consisting of zygotospores can give rise to both blades and conchocelis in the same culture ofPorphyra. Study on samples of wild and cultivatedPorphyra yezoensis andP. oligospermatangia were conducted to clarify the origination of the young blades in the culture. It is confirmed that single cells on the blade of both species, which normally intermixed with zygotospores, germinated into young blades. TEM and SEM observation has shown that the single cells ofPorphyra yezoensis had typical features of female gamete (carpogonia) but archeospore. Therefore, the female gametes are responsible in developing leafy thalli. This project was sponsored by the NSFC (No. C-0205-05-39770593)     

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.     

DISTRIBUTION AND POPULATION DYNAMICS OF PARACALANUS PARVUS,PRACALANUS CRASSIROSTRIS,AND ACARTIA BIFILOSA（COPEPODA,CALANOIDA）IN THE BOHAI SEA

Paracalanus parvus,Paracalanus crassirostris,and Acartia bifilosa are dominant and widely distributed in the Bohai Sea,and comprise an important part of zooplankton in terms of biomass as well as production rate.In order to understand their seasonal distribution and population dynamics,their stage-specific abundance in different months of the year were analyzed based on the never analyzed yet samples collected in 1959.The three species showed clear and remarkable seasonal variation in abundance,which maximized in spring and summer,when they formed high biomass patches or concentrations in the nearshore area.For Paracalanus parvus,two peaks were observed in the annual circle,one in June and the other in September,For Paracalanus crassirostris,one peak occurred in summer and a small one in December.The seasonal pattern of Acartia bifilosa was different in different regions.In Bohai Bay it had a two-peak pattern,with the first large peak occurring in May and the second one in October.In Laizhou Bay,a winter peak in December and January could be observed besides the spring one.The number of generations during the reproductive season for the three species was estimated based on the annual cycle in abundance and ambient temperature.     

Morphological and toxicity characteristics of Pseudo-nitzschia pungens strain PP0201-01 isolated from the East China Sea

The morphology and toxicity ofPseudo-nitzschia strain PP0201-01, isolated near the estuary of Changjiang (Yangtze) River in Year 2002 and tentatively identified asPseudo-nitzschia pungens, were studied. The strain was identified morphologically asP. pungens using scanning electronic microscope (SEM). The analytical results of LC-PDA (liquid chromatography with photodiode array detector), LC-MS (liquid chromatography with mass detector), and the toxicity data from mouse bioassay, showed that no domoic acid was produced by the strain. However, the mechanism of domoic acid production byPseudo-nitzschia species is complicated. More works are needed to assess the toxic species and to evaluate the potential risk of amnesic shellfish poisoning (ASP) in China. Supported by the projects of Ministry of Science and Technology (No. 2001CB409704 and No. 2001BA804A20), and the innovation project of the Institute of Oceanology, CAS (No. 200223107).     

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.     

Genetic differentiation withinEriochoir sinensis (milne, edwards) revealed by allozyme and RAPD

We analyzed 17 allozymes, and 20 primers in order to detect the genetic differentiation between commercial populations (Changjiang River, Liaohe River) ofEriochoir sinensis. Ten allozymes (LDH, MDH, ME, IDH, EST, ALP, AAT, CTL, POD, SOD) showed 21 loci by vertically discontinuos buffer system polyacrylamide gel electrophoresis. RAPD profiles generated by 12 ten-base primers showed 63 loci. The percentage of polymorphic loci and the expected heterozygosity obtained by using allozyme analysis were lower than those obtained by RAPD. The index of similarity between these two populations were 0.955 and 0.932 as revealed by allozyme analysis and RAPD technology. There was gene flow between the above populations. Contribution No. 4025 from the Institute of Oceanology and Limnology, Chinese Academy of Sciences. Project 39470141 supported by NSFC. Work also supported by Climbing Program B (PDB 6-5-3) and No. 863 High-technology Program 863-819-01-05.     

A genetic linkage map of marine shrimp Penaeus (Fenneropenaeus) chinensis based on AFLP, SSR, and RAPD markers

The Chinese shrimp Penaeus (Fenneropaeneus) chinensis is an important species in marine fishery and aquaculture in China. A female Chinese shrimp Penaeus (Fenneropaeneus) chinensis was captured from west coast of the Korean peninsula and mated with a “Yellow Sea No. 1” male to produce the first filial generation (F₁) 100 F₂ full-sib progeny from brother-sister crosses between F₁ families was used for the mapping study. A genetic linkage map of the Chinese shrimp was constructed, based on 354 markers, including 300 amplified fragment length polymorphism (AFLP) markers, 42 microsatellite (SSR) markers, and 12 randomly amplified polymorphism (RAPD) markers. Forty-seven linkage groups (LGs) were identified. The total map length was 4 580.5 cM, with an average spacing of 11.3 cM, covering 75.8% of the estimated genome size. The construction of this genetic linkage map was part of a genetic breeding program. This linkage map will contribute to the discovery of genes and quantitative trait loci (QTLs) in Chinese shrimp.     

Seasonal fluctuation of Myxobolus gibelioi（Myxosporea） plasmodia in the gills of the farmed allogyno—genetic gibel carp in China

The seasonal fluctuation of the plasmodia ofMyxobolus gibelioi Wu et Wang, 1982 in the gill filaments of the allogynogenetic gibel carpCarassius auratus gibelio (Bloch) in a fish pond in Hubei Province of China was investigated from August 1999 to July 2000. A total of 445 fish was examined; the overall prevalence of the plasmodium infection in the fish was 64.94% and mean abundance of plasmodia was 11.65±27.85 (mean±SD). Significant seasonal changes in prevalence and mean abundance, with higher levels of the plasmodia infection from late spring to autumn, were observed. This research was financially supported by the Projects for Knowledge Innovation of the Chinese Academy of Sciences (No. KSCX2-1-04 and No. 1999051) and the Science & Technology Commision of Wuhan City (No. 995004095G).     

Effect of nitrogen and phosphorus on the growth of a red tide dinoflagellateScrippsiella trochoidea (Stein) Loeblich III

Scrippsiella trochoidea (Stein) Loeblich III was grown in a nitrogen-or phosphorus-limited batch culture system in laboratory. Growth rates and cellular Chl-a were measured as functions of nitrate and phosphate concentrations. Growth rates were hyperbolic with both nitrate and phosphate concentration and fit the Monod equation. The minimum cell quota of nitrogen and phosphorus and then the optimum N∶P ratio ofS. trochoidea were estimated in this study. Measurement of phosphate concentration in Jiaozhou Bay suggest that phosphorus is the limiting factor ofS. trochoidea growth. Contribution No. 3679 from the Institute of Oceanology, Chinese Academy of Sciences. Project 39790110 supported by NSFC, the study aslo supported by the National Climbing Project B (PDBG-7-2) and partly supported by an MF grant from Hong Kong University of Science and Technology.     

Reproductive morphology ofEucheumagelatinae (Esper) J. Agardh (Solieriaceae,Gigartinales, rhodophyta)

Eucheuma gelatinae (Esper) J. Agardh has vegetative and reproductive features distinguishing it from other species ofEucheuma. The occurrence of reproductive nemathecia containing carpogonial branches, auxiliary cells and post-fertilization stages including gonimoblast and pericarp initiation, fusion cell formation stages and carposporophyte development are described and ilustrated for the first time for this species. Contribution No. 2380 of the Institute of Oceanology, Chinese Academy of Sciences.     

A Taxonomic Study of the Genus Tinocladia （Chordariales,Phaeophyta） in China Coasts I. Two new species, T. eudesmoides sp.nov, and T. zhangii sp.nov.

Two new species of the genusTinocladia Kylin (1940)T. eudesmoides Ding et Lu sp. nov. andTinocladia zhangii Ding et Lu sp. nov., are described.T. eudesmoides is mainly characterized by its longitudinally and transversely produced sub-cortical filaments from the medulla, its short uniseriate assimilating filaments, usually composed of 6–10 cells with slightly swollen and narrow ultimate cells, and its unilocular sporangia (60–115)×55–70μm, variable in size.T. zhangii is mainly characterized by its very solid frond, not rip apart by hands, its evident longer primary branches, its short and slender secondary branches, with very evident sub-cortical layer, and filaments divided repeatedly, and its large and small unilocular sporangia, with the latter borne on an evidently swollen pedicel. Project B80981816 supported by the NSFC. Contribution No. 4443 from the Institute of Oceanology, Chinese Academy of Sciences, Qingdao, China.     

Light and electron microscope observations onNephroselmis gaoae sp. nov. (Prasinophyceae)

Nephroselmis gaoae sp. nov. is described on the basis of light and electron microscope observations of cultured material originally collected and isolated from seawater of Jiaozhou Bay, Qingdao, China. The periplasts on the cell body and flagella are covered by five types of scales, two types on the flagella and three on the body. Among these, the morphology and the number of spines of large stellate body scales differ remarkably from those of previously described species ofNephroselmis. Apart from these, the unusual fine structure of the eyespot (stigma) is very characteristic. As in the other species ofNephroselmis, the eyespot lies immediately under the two-membraned chloroplast envelope; unlike the others, however, it is not composed of a number of osmiophilic globules, but consists of about 14 curved rod-shaped osmiophilic bodies arranged loosely and randomly. This feature distinguishes the present new species not only from the other species ofNephroselmis but also from the other motile algal species, the eyespots structure of which had been previously described. Contribution No. 2316 from the Institute of Oceanology, Chinese Academy of Sciences. This paper was presented at the XIVth International Seaweed Symposium held in Brittany, France, 1992.