Intraspecific diversity of Aureobasidium pullulans strains from different marine environments

Totally more than 500 yeast strains were isolated from seawater, sea sediments, mud of sea salterns, marine fish guts and marine algae. The results of routine and molecular biology identification methods show that nine strains among these marine yeasts belong to Aureobasidium pullulans, although the morphologies of their colonies are very different. The marine yeasts isolated from different marine environments indicate that A. pullulans is widely distributed in different environmental conditions. These Aureobasidium pullulans strains include A. pullulans 4#2, A. pullulans N13d, A. pullulans HN3-11, A. pullulans HN2-3, A. pullulans JHSc, A. pullulans HN4.7, A. pullulans HN5.3, A. pullulans HN6.2 and A. pullulans W13a. A. pullulans 4#2 could produce cellulase and single cell protein. A. pullulans N13d could produce protease, lipase, amylase and cellulase. Both A. pullulans HN3-11 and A. pullulans HN2-3 were able to produce protease, lipase and cellulase. A. pullulans JHSc could secrete cellulase and killer toxin. Both A. pullulans HN4.7 and A. pullulans HN5.3 could yield lipase and cellulase. A. pullulans W13a was able to secrete extracellular amylase and cellulase while A. pullulans HN4.7 and A. pullulans N13d could produce siderophores. This means that different A. pullulans strains from different marine environments have different physiological characteristics, which may be applied in many different biotechnological industries.     

Two new species of the genus anthenoides (Echinodermata: Asteroidea) from southern China

Two new species of the genusAnthenoides from the southern China are described.A. laevigatus sp. nov. shows affinities withA. epixanthus. A. tenuis sp. nov. is related toA. cristatus (Sladen). Contribution No. 1472 from the Institute of Oceanology, Academia Sinica     

Effect of Alexandrium tamarense on three bloom-forming algae

We investigated the allelopathic properties of Alexandrium tamarense (Laboar) Balech on the growth of Prorocentrum donghaiense Lu, Chattonella marina (Subrahmanyan) Hara et Chihara and Heterosigma akashiwo (Hada) Hada in a laboratory experiment. We examined the growth of A. tamarense, C. marina, P. donghaiense and H. Akashiwo in co-cultures and the effect of filtrates from A. tamarense cultures in various growth phases, on the three harmful algal bloom (HAB)-forming algae. In co-cultures with A. tamarense, both C. marina and H. akashiwo were dramatically suppressed at high cell densities; in contrast, the growth of P. donghaiense varied in different inoculative ratios of A. tamarense and P. donghaiense. When the ratio was 1:1 (P. donghaiense: A. tamarense), growth of P. donghaiense was inhibited considerably, while the growth of P. donghaiense was almost the same as that of the control when the ratio was 9:1. The growth difference of P. donghaiense, C. marina and H. akashiwo when co-cultured with A. tamarense indicated that the allelopathic effect may be one of the important factors in algal competition and phytoplankton succession involving A. tamarense. In addition, the filtrate from A. tamarense culture had negative impacts on these three HAB algae, and such inhibition varied with different growth phases of A. tamarense in parallel with reported values of PSP toxin content in Alexandrium cells. This implied that PSP toxin was possibly involved in allelopathy of A. tamarense. However, the rapid decomposition and inactivation of PSP toxin above pH 7 weakened this possibility. Further studies on the allelochemicals responsible for the allelopathy of A. tamarense need to be carried out in future.     

Molecular identification and culture observation on <Emphasis Type="Italic">Acrochaete leptochaete</Emphasis> (Chaetophoraceae,Chlorophyta) from China

Acrochaete leptochaete (Huber) Nielsen (Chaetophoraceae, Chlorophyta) was isolated from the macroalgae Chaetomorpha collected from intertidal pools in Rongcheng, Shandong, China. 18S rDNA combined with ITS regions were used to ascertain the morphological identification of the isolated material. Based on the unialgal culture, asexual reproduction and growth characteristics of A. leptochaete were investigated over wide ranges of temperature and irradiance. Results revealed that asexual reproduction of A. leptochaete could be realized by biflagellate zoospores. The zoospores germinated directly to give self- replicating generations. Zoospore germination was bipolar. A temperature range from 13-21°C and a lower irradiance of 36 μmol/(m2·s) were most favorable for the growth of A. leptochaete. Thallus organization, an important taxonomic criterion for the genus Acrochaete, was affected markedly by temperature and irradiance. Our results extend the knowledge about the species’ general biology and its morphological plasticity. For classification and identification of a simple microphytic algae like A. leptochaete, which are traditionally placed in the class Chaetophoraceae, we propose that molecular tools associated with culture observations are applied.     

Preliminary identification of three new isolates in genus Alexandrium (Dinophyceae) from China sea area

The 5.8S ribosomal DNA sequences (5.8S rDNA) and their flanking regions, internal transcribed spacer 1 and spacer 2 (ITS1 and ITS2) of three new isolates in genus Alexandrium (Alexandrium sp. qd1, Alexandrium sp. qd2, Alexandrium sp. gz) from China were amplified, sequenced, and subjected to phylogenetic analysis. Alexandrium sp. gz and Alexandrium sp. qd1 were grouped with high bootstrap values with four strains/species, i.e., A. catenella South Korea strain, A. catenella Japan strain, and two from China, Alexandrium sp. AC03 and Alexandrium sp. AN01 being proposed to be A. catenalla in a previous study. Then Alexandrium sp. gz and Alexandrium sp. qd1 were identified as Alexandrium catenella. As A. catenella was isolated from Qingdao and Guangzhou sea areas, it supposedly distributed at least in these two areas and was genetically different. Alexandrium sp. qd2 differed greatly from species in Alexandrium. It clustered with Symbiodinium californium, Symbiodinium sp. G15 and Gymnodinium sp. Zhao 01 with 100% bootstrap value; so Alexandrium sp. qd2 affiliates to genus Symbiodinium, and is probably a free-living Symbiodinium species.     

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

Although both Astasia longa and Euglena gracdlis belong to different genera, they share many morphological characters except that A. longa has no chloroplast. In the 1940‘ s, on the basis of the finding that in darkness or upon addition of scrne chemicals, E. gmcilis would fade reversibly or irreversibly, scrne scholars hypothesised that A. longa evolved from E. gracilis by losing chloroplast.The authors‘ use of RAPD and cladistic analyses in a study on the evolutionary relationship between A .longa and E. gmcilis showed that the A. longa ‘ s relationship with E. gmcilis was closer than that with other green euglenoids. This proves the hypothesis that A. longa evolved from E. gmcills is reasonable. The results d this study suggest that saprophytic colorless euglenoids were transformed fromgreen euglenoids by losing their choroplasts.     

Microsatellite variation between Japanese eel （Anguillajaponica） and European eel （Anguilla anguilla）

Allelic variation in a total of 7 microsatellites was examined between elvers of freshwater eels （Anguillajaponica and Anguilla anguilla）. The number of alleles at these loci ranged from 8 to 26. A single test of each locus revealed significant deficits of heterozygotes （P〈0.01）. Significant departure from expectations of Hardy-Weinberg equilibrium （HWE） was found for all loci within four subpopulations of A. japonica, which opposes the panmixia hypothesis of Schmidt. Also exact tests of population differentiation based on allelic frequency distribution disagree the hypothesis of random distribution of individuals among populations. Population structure among four populations of A. japonica was revealed with FST value of 0.009 8 （P=-0.000 48; 10 000 iteration）. Pairwise matrixes of FST and RST showed a significant difference between two distantly related species-A, japonica and A. anguilla. Divergent time of the two species calculated by Goldstein method is over 2 million years. The results may challenge the Schmidt＇s theory about the distribution of freshwater eels.     

Preparation and characterization of agar,agarose, and agaropectin from the red alga Ahnfeltia plicata

Agar, agarose, and agaropectin were extracted from the red alga Ahnfeltia plicata, and their properties and structures were characterized. Agar was extracted by a comparatively low alkaline consumption of 1.2%. It exhibited a gel strength of 1 152.50±74.25 g/cm~2 and a sulfate content of 0.55%±0.08%. The yield of agar from A. plicata was 24.53%, which is higher than those of other agarophytes commonly used in China. Three kinds of the method were compared for the purification of agarose, and the physicochemical properties of agarose that was prepared under the optimal condition were identical to those of commercially available agarose. Furthermore, agaropectin was purified from A. plicata and characterized by GC, HPLC,UV-spectrum, and FI-IR to understand its composition and structure. It was the first time to comprehensively study the agar and its fractions from the red alga of A. plicata. This research provided an eco-friendly agar extraction method from A. plicata and revealed its potential application for the production of agar, agarose,and agaropectin.     

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.     

Occurrence and diversity of <Emphasis Type="Italic">Pichia</Emphasis> spp. in marine environments

A total of 328 yeast strains from seawater, sediments, mud of salterns, the guts of marine fish and marine algae were obtained. The results of routine identification and molecular methods show that five yeast strains obtained in this study belonged to Pichia spp., including Pichia guilliermondii 1uv-small, Pichia ohmeri YF04d, Pichia fermentans YF12b, Pichia burtonii YF11A and Pichia anomala YF07b. Further studies revealed that Pichia anomala YF07b could produce killer toxin against pathogenic yeasts in crabs while Pichia guilliermondii 1uv-small could produce high activity of extracellular inulinase. It is advisable to test if Pichia ohmeri YF04d obtained in this study is related to central-venous-catheter-associated infection.     

Two new species of the genusDorylaimopsis Ditlevsen, 1918 (Nematoda: Adenophora,Comesomatidae) from the Bohai Sea,China

Two nematode species of the genusDorylaimopsis (Family Comesometidae) from the Bohai Sea are described.Dorylaimopsis rabalaisi n. sp. similar toD. punctata Ditlevsen, 1918 from which it can be separated by the small size, short and simple spicules without ventral apophyses or joint line.Dorylaimopsis turneri n. sp. differs from the closest speciesDorylaimopsis angelae (Inglis, 1967) by its higher values of de Man ratio ‘a’, much shorter spicules and higher ratio of gubernacular to spicules length. A key to species of the genus based on the key by Jensen (1979) is given. This project was supported by the National Science Foundation, No: 3860804     

Chlamydomonas sajao nov. sp. (Chlorophyta,Volvocales)

A new species ofChlamydomonas, namely,C. sajao nov. sp. of the Volvocales, Chlorophyta was isolated from a duckweed growing near a ricefield in the vicinity of Guangzhou, China. This interesting unicellular green alga, similar toC. mexicana from Mexico, secretes quantities of extracellular mucilaginous polysaccharides, and may be employed in improving soil quality. The new species resemblesC. waldenburgensis Moewus in most characteristics but differs in three important features.     

Age and growth characteristics of crimson sea bream <Emphasis Type="Italic">Paragyrops edita</Emphasis> Tanaka in Beibu Gulf

Abstact  Age and growth characteristics of crimson sea bream Paragyrops edita Tanaka in Beibu Gulf were studied through bottom trawling and gillnet fleets fishing from July 2006 to December 2007. A total number of 1155 individuals, ranging from 49 to 249 mm in standard length was examined. The age of the fish was determined from sagittal otoliths. One year growth was made up of one translucent and one opaque zone. A maximum likelihood estimation procedure was used to fit the Von Bertalanffy, Logistic and Gompertz growth functions to the length-at-age data. ARSS indicated that there were no significant differences in growth between sexes in the three growth models (P>0.05), and the Von Bertalanffy growth function L _t =292.8{1−exp [−0.167(t+1.116)]} was selected as the most appropriate growth model according to Akaike’s information criterion (AIC).     

Bacteriological analysis of the digestive tube of the mud snail (Bullacta exarata Philippi) and its rearing shoal

The bacterial flora in the digestive tube of Bullacta exarata Philippi and its rearing shoal were investigated. A total of 157 strains of heterotrophic bacteria, isolated from crop, stomach intestine and other parts of the digestive tube, mainly belong to the genera Photobacterium, Bacillus, Pseudomonas Vibrio and some genera of the family Enterobacteriaceae. There are significantly more varieties of bacteria in crop than in stomach and intestine. A total of 173 strains of bacteria were isolated from the rearing shoal, belonging to 13 genera. The 5 predominant genera, such as Bacillus and Photobacterium, are the same as those in the digestive tube, but greatly differ in percentages. The number of heterotrophic bacteria and Vibrio in rearing shoal change in line with the alteration of the temperature, and are significantly affected by the use of pesticides.     

Mapping the resting cysts of dinoflagellate Alexandrium catenella along the coast of Qinhuangdao,China

Dinoflagellate Alexandrium catenella is a cosmopolitan bloom-forming species with complex life cycle, the formation and germination of resting cysts are critical for its bloom dynamics. In the coastal waters of Qinhuangdao, A. catenella has been identified as the major causative agent for paralytic shellfish poisoning, but there is little knowledge concerning its resting cysts in this region. In this study, three surveys were carried out along the coast of Qinhuangdao from 2020 to 2021 to map the distribution of A. catenella resting cysts, using a quantitative PCR (qPCR) assay specific for A. catenella. The resting cysts were detected in surface sediments during all the three surveys, and their distribution patterns were similar. High abundance of resting cysts (maximum 1 300 cysts/g sediment (wet weight)) were found in a region (119.62°E–119.99°E, 39.67°N–39.98°N) northeast to the coastal waters of Qinhuangdao, where surface sediments were mainly composed of clay and silt (percentage above 50%). Prior to the formation of the A. catenella bloom in March 2021, the abundance of A. catenella vegetative cells in seawater had extremely significant positive correlation with the abundance of resting cysts in surface sediments, reflecting the important role of resting cysts in the initiation of A. catenella blooms. As far as we know, this is the first report on the distribution of A. catenella cysts along the coast of Qinhuangdao. The results will offer a sound basis for the future monitoring and mitigation of toxic A. catenella blooms and paralytic shellfish poisoning events in this region.

     

Effects of co-existing microalgae and grazers on the production of hemolytic toxins in Karenia mikimotoi

Karenia mikimotoi (Miyake & Kominami ex Oda) Hansen & Moestrup is associated with harmful algal blooms in temperate and subtropical zones of the world. The hemolytic substances produced by K. mikimotoi are thought to cause mortality in fishes and invertebrates. We evaluated the composition of the hemolytic toxin produced by K. mikimotoi cultured in the laboratory using thin-layer chromatography.In addition, we evaluated the effect of co-occuring algae (Prorocentrum donghaiense and Alexandrium tamarense) and the cladoceran grazer Moina mongolica on hemolytic toxin production in K. mikimotoi. The hemolytic toxins from K. mikimotoi were a mixture of 2 liposaccharides and 1 lipid. Waterborne clues from P. donghaiense and A. tamarense inhibited the growth of K. mikimotoi but increased the production of hemolytic toxins. Conversely, K. mikimotoi strongly inhibited the growth of caged P. donghaiense and A. tamarense. In addition, the ingestion of K. mikimotoi by M. mongolica induced the production of hemolytic toxins in K. mikimotoi. Taken together, our results suggest that the presence of other microalgae and grazers may be as important as environmental factors for controlling the production of hemolytic substances. K. mikimotoi secreted allelochemicals other than unstable fatty acids with hemolytic activity. The production of hemolytic toxins in dinoflagellates was not only dependent on resource availability, but also on the risk of predation. Hemolytic toxins likely play an important role as chemical deterrents secreted by K. mikimotoi.