Development and Characterization of Microsatellite Markers for the Pacific Abalone （Haliotis discus） via EST Database Mining

The EST database of the Pacific abalone (Haliotis discus) was mined for developing mierosatellite markers. A total of 1476 EST sequences were registered in GenBank when data mining was performed. Fifty sequences (approximately 3.4%) were found to contain one or more mierosatellites. Based on the length and GC content of the flanking regions, duster analysis and BLASTN, 13 microsatellite-containing ESTs were selected for PCR primer design. The results showed that 10 out of 13 primer pairs could amplify seorable PCR products and showed polymorphism. The number of alleles ranged from 2 to 13 and the values of Hoand He varied from 0.1222 to 0.8611 and 0.2449 to 0.9311, respectively. No significant linkage disequilibrium (LD) between any pairs of these loci was found, and 6 of 10 loci conformed to the Hardy-Weinberg equilibrium (HWE). These EST-SSRs are therefore potential tools for studies of intraspecies variation and hybrid identification.     

The relationships between rheological properties and structural changes of chilled abalone meat

The quantitative correlation between rheological properties and structural characteristic values of chilled abalone meat was studied. Structural changes were observed, and these values were enumerated using image processing and analysis technique. Structural changes in the myofibrils and collagen fibrils were the greatest in chilling for 24 h. After chilling for 48h, similar structures of vertical and cross sections were observed. For chilling from 0h to 72h, the instantaneous modulus E0 of the both section meat decreases gradually with time, but no significant differences were observed after chilling for 48h. The relaxation time and viscosity of both sections attained the same values for the same chilling time, but increased gradually with increasing chilling time. Meanwhile, a negative correlation between the structural characteristic values (Dm, Am, Rvm), and rheological properties (E1, τ1 , η1) clearly exists. Some logarithmic expressions have been obtained for these negative correlations. These r     

In Silico Screening for Microsatellite Markers from Expressed Sequence Tags of Porphyra yezoensis （Bangiales,Rhodophyta）

The genomic resources of Porphyra yezoensis expressed sequence tags （ESTs） were utilized to identify simple sequence repeats （SSRs）, or microsatellites. This method took the advantage of using ESTs and microsatellites either for the establishment of gene identities or for the acquisition of high polymorphism. The microsatellites can be used as gene markers when microsatellites are tagged to genes. Revealed by bioinformatics analysis, 1162 out of 21954 ESTs contained microsatellites and cluster analysis indicated that 984 of these ESTs fell into 112 contigs, while the other 178 ESTs were singletons. A total of 290 unique SSR-containing genes were identified. The AAC SSRs were the most populous type of microsatellites. GC-rich microsatellites were predominant among all the microsatellites.     

CYTOLOGICAL COMPARISON OF GAMETOGENESIS OF SCALLOPS, ARGOPECTEN IRRADIANS AND CHLAMYS FARRERI

Histological characteristics of gametogenesis of two kinds of scallops, gonochoric Chinese scallop, Chlamys farreri and hermaphroditic bay scallop, Argopecten irradians were investigated in this study. Spermatogenesis in C. farreri has different developmental stages: spermatogonia, primary spermatocyte, second spermatocyte, spermatid and spermatozoon. A large number of same developmental stage spermatic cells converge at a definite area of the testis. Premeiotic, previtellogenic and vitellogenic oocytes can be found during oogenesis in C. farreri, where oocyte distribution is obviously irregular. The A. irradians gonad consists of two different parts in one individual: one part functions as testis, the other as ovary. Between these two parts is a special appearance area, where a large number of spermatic cells are bound with two layers of acellular substance with many oocytes in it.     

EFFECTS OF CO—CULTURE AND SALINITY ON THE GROWTH AND AGAR YIELD OF GRACILARIA TENUISTIPITATA VAR LIUI ZHANG ET XIA

Gracilaria tenuistipitata var Liui were mono-cultivated and co-cultivated with Pinctada martensii under high(33) and low(21) salinity conditions in laboratory.The daily growth rate of the alga was determined.Tissue carbon and nitrogen contents,the yield and fractional composition of agar were analyzed.Results showed that:1.Gracilaria grew better under low salinity conditions,the daily growth rate was twice that under high salinity conditions.Co-cultivated algae grew faster than mono-culti-vated algae under low salinity conditions,the daily growth rate was about 37.6?higher.2.Compared with mono-cultivated algae,tissue nitrogen contents of co-cultivated algae were higher,while the C:N ratios were much lower.3.The agar yields of co-cultivated algae were much lower than those of mono-culti-vated algae.Agar yield was found to be negatively correlated to the tissue nitrogen contents,and positively correlated to the C:N ratios.4.The highest fractional yields obtained from co-cultivated algae were extracted with 40?ethanol,while from mono-cultivated algae,the highest fractional yields obtained were extracted with distilled water at room temperature.     

AGE STRUCTURES OF MODULES OF CLONAL PEATLAND SEDGE Carex middendorffii

Age structure of a plant population carries important information on population dynamics. The traditional age classification of individuals by development phases could not explain the generation relationship neither between individuals nor between modules, and it could not accurately predict the future of population or the tendency of peatland evolution. In a peatland of the Xiao Hinggan Mountains, China, at the middle of the growth season, the age structures of 3 modules, ramets, active buds and rhizomes of a Carex middendorffii clonal population were investigated, with the method of classifying age classes of ramets and active buds by counting generation quantity of tiller nodes, and classifying age classes of rhizomes by their real survival time. The quantity of vegetative ramets was dominant. Tiller nodes of ramets can propagate vegetatively for a maximum of 3 generations. The population of ramets consisted of 3 age classes of ramets at the middle of the growth season, and showed a stable age structure. In the two sampling events, there was no significant difference between quantities and age structure of the population. The maximum age of an excavated rhizome was 12 years old. Rhizomes were classified in 8 age classes, and age classes 4-6 contributed most to the total biomass. There was no significant difference in total length and total biomass per unit area, or in biomass per unit length in rhizomes between the two samplings. Four age classes of active buds were recognized, and their number increased from July to August. The Carex middendorffii clonal population achieved regeneration by budding from the tiller nodes of ramets. The age structures of the 3 modules suggested that the Carex middendorffii clonal population could persist in the early development phase of the oligotrophic peatland in the Xiao Hinggan Mountains, but it could not be dominant. It also faces the risk to disappear from the community as the peatland develops further.     

Temperature Influences on the Expression of GFP Promoted by the Upstream Sequence of cpcB from Arthrospiraplatensis

In order to investigate the regulation mechanism of the phycocyanin gene,a series of functional analyses of the upstream sequence of cpcB gene from Arthrospira platensis were conducted in E. coli with green fluorescent protein encoding gene(gfp) as the reporter. Results showed that the gfp gene could express at a high level under the promotion of the upstream sequence,suggesting the existence of some strong promoter elements in it. The expression of GFP was influenced by temperature. Higher temperature led to higher expression level. The bioinformatics analyses followed by mutation analyses on the secondary structure of translation ini-tiation region(TIR) revealed that RNA thermosensor might account for the temperature regulation.     

In vitro acute cytotoxicity of neonicotinoid insecticide imidacloprid to gill cell line of flounder Paralichthy olivaceus

In vitro acute cytotoxicity of neonicotinoid insecticide imidacloprid (IMI) to the gill cell line of flounder (FG) that collected in the gill of Paralichthys olivaceus, was examined by 3 widely used endpoint bioassays: NR (neutral red), MTT (3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide) and TCP (total cell protein). The result shows that the IMI increased at concentrations ≥0.5 μg/ml. The IC50 value of NR, MTT, and TCP was 41.86, 38.46, and 39.08 μg/ml, respectively. The ultrastructural observation revealed that the mitochondria of the cells exposed to 60 μg/ml IMI for 48 h were severely damaged, swollen or disrupted, while their nuclei and rough endoplasmic reticulum (RER) remained normal. This would suggest that the mitochondria are probably the primary target of IMI.     

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.