Cytogenetic mechanism for the aneuploidy and mosaicism found in tetraploid Pacific oyster Crassostrea gigas (Thunberg)

Chromosome constitution was investigated in adult tetraploid Pacific oyster produced by blocking the first polar body of triploid eggs which were fertilized with haploid sperms. A high incidence of aneuploid and heteroploid mosaics were found among the offspring. Of 20 individuals identified, only 9 (45%) were eutetraploid which contained 40 chromosomes; 2 (10%) were aneuploid (hypotetraploid), which contained 39 and 38 chromosomes, respectively; and 9 (45%) were heteroploid mosaics. One mosaic was consisted of cells containing 40 and 39 chromosomes, respectiovely (1:1 in cell number), while the other 8 were consisted of cells containing chromosomes varying between tetraploid and triploid. It was also interesting to note that 3 mosaics even contained some diploid cells with 20 chromosomes. A certain number of cells of 2 tetraploids and 8 mosaics spread with 32–37 well-scattered and some clumped chromosomes at metaphase. The percentage of aneuploid cells with chromosomes varying between triploid and tetraploid correlated significantly with that of heteroploid mosaics cells with clumping chromosomes (P<0.05). Our findings suggested that reversion existed in both tetraploid and triploid oyster and chromosome clumping may underline the chromosome elimination in tetraploid oyster. It seems that the reversing cells, at least some of them, continuously eliminate their chromosomes until the most stable diploid state is established.     

Polyploidy induction by hydrostatic pressure shock and embryo development of sea cucumber Apostichopus japonicus

The manipulation of the chromosome set for commercially valuable marine animals is important for enhancing aquacultural production. In this study, triploid and tetraploid sea cucumber Apostichopus japonicus were induced by hydrostatic pressure shock, and the conditions of appropriate induction were tested with different starting times, and hydrostatic pressure intensities and durations. The highest rate of triploid induction reached 20% and that of tetraploid was 60%. In consideration of the survival rate and hatch rate, the appropriate treatment for triploid was 55 Mpa of hydrostatic pressure for 5 min at 55 min after fertilization (a.f.), while for tetraploid it was 60 Mpa for 5 min at 61 min a.f. The triploid of the sea cucumber could survive through the pelagic larval stage and attachment stage, and develop like the control group of the experiment. The tetraploid, however, could not survive the attachment stage.     

Preliminary study on chromosomes of induced triploid in the red sea bream,Pagrosomus major

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Chromosome behavior of heat shock induced triploid in Fenneropenaeus chinensis

Triploidy was induced in Chinese shrimpFenneropenaeus chinensis by 30±0.5°C heat shock treatment (initiated at 20 min after fertilization) for 10 min to inhibit the release of PB2 at 18.0°C. The highest triploid rate obtained was 84.5% in nauplius stage. The effect of heat shock treatment on meiosis and cleavage of eggs was investigated in this work aimed to establish efficient procedures for triploid induction and to gain understanding of the mechanism of triploid production. Three pronuclei that could be observed in the treated eggs under fluorescence microscope developed into triploid embryos. Some abnormal chromosome behavior was observed in heat shocked eggs. Contribution No. 4407 from the Institute of Oceanology, Chinese Academy of Sciences. This work was supported by Project under Major State Basic Development Program (G1999012009) and Knowledge Innovation Program of the Chinese Academy of Sciences (KZCX 2-211)     

Chromosome Behavior of Heat Shock Induced Triploid in Fenneropenaeus Chinensis

INTRODUCTIONTriploidanimalsareusefulbecausetheyaresterile ,characterizedbypositivegrowthinthere productiveseason ,usefulfortesting ,forintroductionofnon nativespecies ,andforprotectionofde velopedstrains.Theinductionoftriploidyhadbeenreportedinmanyaquaculturespecies .Triploidshavebeensuccessfullyinducedinshrimpusingtemperatureshockorchemicalshock (CB ,6 DMAP)in 4species :Sicyoniaingentis (Xiangetal.,1 991 ) ,Fenneropenaeuschinensis (Xiangetal.,1 992 ;Daietal.,1 993;Baoetal.,1 993;Li…     

Comparison of taste components between triploid and diploid oyster

The main taste components of triploid and diploid oyster (Crassostrea gigas) were compared. Free amino acids, inosine monophosphate, succinate, trimethylamine oxide and betaine in fresh and boiled extractives were analyzed. The protein, lipids, glycogen, moisture and ash, which may affect the flavour, were evaluated. In boiled extractives, the amino acids were 394.1 mg(100 g)⁻¹ in diploid and 183.5 mg(100 g)⁻¹ in triploid. However, in fresh oyster extractives, they were 320.0 mg(100g)⁻¹ and 147.3 mg(100g)⁻¹ respectively. The inosine monophosphate in triploid was 44% more than that in diploid, and a little difference existed in the content of trimethylamine oxide between them. The contents of these taste components were the basis for taste flavour pattern determination.     

Molecular identification based on ITS sequences for Kappaphycus and Eucheuma cultivated in China

The systematic classification of the Eucheumatoideae is difficult because of their variable morphology and interpretation of reproductive structures. Kappaphycus and Eucheuma specimens cultivated on the Hainan and Fujian coast of China were introduced from Vietnam, the Philippines and Indonesia. Combined with morphological characteristics, all Kappaphycus and Eucheuma cultivated strains were identified by internal transcribed spacer (ITS) sequences. The phylogenetic tree was constructed using neighbor-joining and maximum likelihood methods. The results indicate that different ITS sequence lengths occurred in the different genera and species. An obvious difference in morphology could be found in the protuberance shape between Kappaphycus and Eucheuma. The protuberance in Eucheuma was thorn-like and in Kappaphycus was wartlike or papillate. Their ITS sequence lengths differed significantly in nucleotide variation rates up to 58.55%-63.90%. All nucleotide variations occurred in the ITS1 and ITS2 regions except for five nucleotide transversions in the 5.8S rDNA region. In addition, the difference was at the branches among congeneric species. Kappaphycus sp. had branches with small buds, while K. alvarezii did not have such a feature. The nucleotide variation rates varied from 7.02% to 7.48% among species; within the same species of the clades it was <1.20%. Eucheumatoideae algae cultivated in China consisted of three clades, K. alvarezii, Kappaphycus sp., and E. denticulatum. The results indicate that ITS sequence analysis was an effective way for identification of interspecies and intraspecies phylogenetic relationships and might provide a clue for molecular identification of algal Eucheumatoideae.     

Chromosomal localization of 5S rDNA in Chinese shrimp (Fenneropenaeus chinensis): a chromosome-specific marker for chromosome identification

Chinese shrimp (Fenneropenaeus chinensis) is an economically important aquaculture species in China. However, cytogenetic and genomic data is limited in the organism partly because the chromosomes are difficult to isolate and analyze. In this study, fluorescence in-situ hybridization (FISH) was used to identify the chromosomes of F. chinensis. The 5S ribosomal RNA gene (rDNA) of F. chinensis was isolated, cloned and then used as a hybridization probe. The results show that the 5S rDNA was located on one pair of homologous chromosomes in F. chinensis. In addition, triploid shrimp were used to evaluate the feasibility of chromosome identification using FISH and to validate the method. It was confirmed that 5S rDNA can be used as a chromosome-specific probe for chromosome identification in F. chinensis. The successful application of FISH in F. chinensis shows that chromosome-specific probes can be developed and this finding will facilitate further research on the chromosomes of penaeid shrimps.     

Triploidy induction in Pacific oysterCrassostrea gigas by caffeine with thermal shock

Zygotes of Pacific oyster (Crassostrea gigas) were treated for triploid induction with caffeine (10 mmol/L, 15mmol/L and 20mmol/L) in combination with thermal shocks (at 40 minute post-fertilization) lasting for 5 and 10 minutes. The highest yield of triploids, 41.5%, was obtained from the treatment with 20 mmol/L caffeine at 34°C lasting 10 minutes. The triploid levels were less than 30.0% in other treatments. Triploid induction was more effective in treatment with 15–20 mmol/L caffeine at 34–38 °C than with lower concentrations of caffeine at temperatures below 32 °C. Our results suggest that triploid induction with caffeine in combination with thermal shocks is less efficient than some other methods reported previously.     

Temporal variation in biodeposit organic content and sinking velocity in long-line shellfish culture

We measured the organic content and sinking velocities of biodeposits from two scallop species (Chlamys farreri, Patinopecten yessoensis) and abalone (Haliotis discus hannai) that were cultured on suspended long-lines. Measurements were conducted every two months from April 2010 to February 2011. The shellfish were divided into three size groups (small, middle, and big sizes). At each sample point, we assessed biodeposit organic content, average sinking velocity, the frequency distribution of sinking velocities, and the correlation between organic content and sinking velocity. The organic content of biodeposits varied significantly among months (P<0.05) and the pattern of change varied among species. Sinking velocities varied significantly, ranging from <0.5 cm/s to >1.9 cm/s. The sinking velocities of biodeposits from C. farreri and P. yessoensis were 0.5–1.5 cm/s and from H. discus hannai were <0.7 cm/s. The organic content was significantly negatively correlated to the sinking velocity of biodeposits in C. farreri (P<0.001) and P. yessoensis (P<0.05).     

Estimation of age,growth and maturation of purpleback flying squid, <Emphasis Type="Italic">Sthenoteuthis oualaniensis</Emphasis>, in Bashi Channel,central Pacific Ocean

The age, growth and maturation of Sthenoteuthis oualaniensis were determined with statolith data collected with a light purse seine from the Bashi Channel of central Pacific Ocean. The estimated longevity of the squid was no more than 6 months for females, and no more than 5 months for males. Growth in mantle length (ML) was best described by logistic models for both females and males, while growth in body weight (BW) was best fitted by power curves. The maximum absolute growth rate (AGR) and instantaneous growth rate (IGR) in ML or BW both occurred at 91–105 days for females and 76–90 days for males. Back calculated hatching dates were from October to January, with a peak in December, although the short duration of sampling date might have had an influence on the result. The lower percentage of mature females (37.2%) suggested that the study area during the sampling date was not a spawning ground for the species. Size and age at first maturity were 183 mm ML and 136 days for females, whereas they were 156 mm ML and 85 days for males.     

虎纹蛙性腺发育的细胞学研究

通过组织切片对虎纹蛙性腺发育过程进行研究，根据其细胞学特征，雌性虎纹蛙的性腺发育过程分为卵原细胞期、初级卵泡期、生长卵泡期和成熟卵泡期；在第一个性周期内，从卵原细胞到成熟卵泡整个发育阶段需要2a。雄性虎纹蛙的生殖细胞成簇排列，性腺发育分为精原细胞、初级精母细胞、次级精母细胞、精子细胞以及精子形成5个阶段；性成熟期为1a。并报道了虎纹蛙的镶嵌型性腺。     

Characterization,expression and function analysis of DAX1 gene of scallop (Chlamys farreri jones and preston 1904) during its gametogenesis

DAX1, a member of nuclear receptor superfamily, has a function in the sex determination and gonadal differentiation of several vertebrate species. However, little information about DAX1 of invertebrates is available. Here we cloned a homolog of scallop (Chlamys farreri Jones and Preston 1904) dax1, Cf-dax1, and determined its expression characteristics at mRNA and protein levels. The cDNA sequence of Cf-dax1 was 2093 bp in length, including 1404 bp open reading frame (ORF) encoding 467 amino acids. Unlike those of vertebrates, no conserved LXXLL-related motif was found in the putative DNA binding region of Cf-DAX1. Fluorescence in situ hybridization showed that Cf-dax1 located on the short arm of a pair of subtelocentric chromosomes. Tissue distribution analysis using semi-quantitative RT-PCR revealed that Cf-dax1 expressed widely in adult scallop tissues, with the highest expression level found in adductor muscle, moderate level in mantle, gill and testis, and low level in kidney, ovary and hepatopancreas. The result of quantitative real-time PCR indicated that the expression of Cf-dax1 was significantly higher (P<0.05) in testis than in ovary at the same stage, showing a sex-dimorphic expression pattern. Furthermore, immunohistochemical detection found that Cf-DAX1 mainly located in spermatogonia and spermatocytes of testis and in oogonia and oocytes of ovary, implying that DAX1 may involve in gametogenesis of bivalves.     

An evaluation on using soil aggregate stability as the indicator of interrill erodibility

Aggregate stability is a very important predictor of soil structure and strength,which influences soil erodibility.Several aggregate stability indices were selected for estimating interrill erodibility of four soil types with contrasting properties from temperate and subtropical regions of China.This study was conducted to investigate how closely the soil interrill erodibility factor in the Water Erosion Prediction Project(WEPP) model relates to soil aggregate stability.The mass fractal dimension(FD),geometric mean diameter(GMD),mean weight diameter(MWD),and aggregate stability index(ASI) of soil aggregates were calculated.A rainfall simulator with a drainable flume(3.0 m long × 1.0 m wide × 0.5 m deep) was used at four slope gradients(5°,10°,15° and 20°),and four rainfall intensities(0.6,1.1,1.7 and 2.5 mm/min).Results indicated that the interrill erodibility(Ki) values were significantly correlated to the indices of ASI,MWD,GMD,and FD computed from the aggregate wet-sieve data.The Ki had a strong positive correlation with FD,as well as a strong negative correlation with ASI,GMD,and MWD.Soils with a higher aggregate stability and lower fractal dimension have smaller Ki values.Stable soils were characterized by a high percentage of large aggregates and the erodible soils by a high percentage of smaller aggregates.The correlation coefficients of Ki with ASI and GMD were greater than those with FDand MWD,implying that both the ASI and GMD may be better alternative parameters for empirically predicting the soil Ki factor.ASI and GMD are more reasonable in interrill soil erodibility estimation,compared with Ki calculation in original WEPP model equation.Results demonstrate the validation of soil aggregation characterization as an appropriate indicator of soil susceptibility to erosion in contrasting soil types in China.     

Seasonal variation of the glycogen enzyme activity in diploid and triploid Pacific oyster gonad during sexual maturation

The glycogen content and the activities of two key enzymes in glycogen metabolism, glycogen phosphorylase and gly- cogen synthetase, in the gonad of diploid and triploid Pacific oysters (Crassostrea gigas) were compared during maturation. The glycogen content in the gonad of diploids decreased with gametogenesis (by 85.7%), but the glycogen content in the gonad of trip- loids did not vary significantly. Activity of glycogen phosphorylase (GP) in the gonad of diploids decreased with gametogenesis (by 55.5%), while GP activity of triploids did not vary significantly during maturation. Activity of glycogen synthetase (GS) in the gonad of diploids increased slightly with gametogenesis, reaching a peak in June. Activity of GS declined sharply from June to July, which might be due to gonad spawning. GS activity of triploid oysters in spawning time (July and August) was significantly higher than that in other months, which might be explained with a ‘compensating’ mechanism for the higher glycogen content in triploids.     

Seasonal changes in Porteresia coarctata (Tateoka) beds along a subtropical coast

Shoot density, standing crop (above- and below-ground biomass) and habitat of salt marsh grass Porteresia coarctata were investigated along the coast of Bakkhali estuary, Cox’s Bazar, Bangladesh from January to December 2006. Shoot density of P. coarctata was influenced by season and was found to be higher (>2 500 shoots/m 2 ) in post-monsoon and minimal in monsoon season; plants were particularly active in vegetative propagation during pre-monsoon. Above-ground biomass was greater along the protected coast compared with the exposed one in this estuary. Below-ground biomass was higher (7.75-269.53 g DW/m 2 ) than that above ground (2.20-114.75 g DW/m 2 ). Standing crops of P. coarctata showed a negative relationship (R=-0.77; P<0.05) with sedimentation rate, while seasonal activity influenced sedimentation. The recorded sedimentation rate was lower (6.09 mg/(cm 2 ·d)) in pre-monsoon and highest (14.55 mg/(cm 2 ·d)) in monsoon season. The mean value of pore water salinity was higher (34.25±5.05) during post-monsoon and lowest (18.0±3.71) in monsoon season. The soil was sandy clay in this P. coarctata bed; it consisted of 86% sand, 13% clay and 1% silt. Soil organic matter dropped during the monsoon season (0.78%-0.67%) and was highest ((2.17±1.42)%-(2.3±1.47)%) during post-monsoon, probably owing to accumulation of decomposed peat on the marsh surface. The mean pore water NH 4 -N concentration ranged from 2.44±1.65 to 3.33±1.82 μg/L, with a minimum air temperature of 22.09°C in post-monsoon and a maximum of 31.16°C in pre-monsoon. Variations of physico-chemical parameters in the soil, water, and climate governed biological parameters of P. coarctata in the Bakkhali estuary, and were comparable with estuarine environments elsewhere.     

Effects of different organic residues on rice yield and soil quality

Calcaric regosols are a valuable land resource, distributed widely across western China. Soil quality has deteriorated considerably in recent years owing to the blind pursuit of economic benefits. A 2-year field experiment was carried out to evaluate the effects of using spent mushroom compost, leguminous plant (Vicia sepium L.) compost, and a combination of the two (at a 1:1 and 2:1 ratio), on rice yield and soil quality in a suburb of China. Vicia sepium L. composted with spent mushroom compost at a 1:1 ratio produced the highest grain and stover yield, grain and stover phosphorus concentration, and phosphorus uptake of rice; they were 56.5%, 93.2%, 89.3%, 198.6% and 22.2% greater than control soil, respectively. The 2:1 ratio (Vicia sepium L.: spent mushroom compost) produced the highest grain N concentration, stover N concentration, and N uptake; they were 31.6%, 31.4%, and 40.7% higher than control, respectively. Soil physical, chemical, and environmental properties were improved with the application of Vicia sepium L. composted with spent mushroom compost at a 2:1 ratio. In particular, soil water-stable aggregates, organic carbon, particulate organic carbon, total nitrogen, available potassium, and cation exchange capacity increased, whereas bulk density, pH, and phytoavailable heavy metals decreased. This organic treatment is beneficial to improve soil quality indicators, and contribute to soil restoration.     

Development of three multiplex PCR primer sets for ark shell (<Emphasis Type="Italic">Scapharca broughtonii</Emphasis>) and their validation in parentage assignment

Scapharca broughtonii is a commercially important and over-exploited species. In order to investigate its genetic diversity and population structure, 43 novel polymorphic microsatellites were isolated and characterized. The number of alleles per locus ranged from 3 to 22 with an average of 6.93, and the observed and expected heterozygosities varied between 0.233 and 1.000, and 0.250 and 0.953, with an average of 0.614 and 0.707, respectively. Three highly informative multiplex PCRs were developed from nine of those microsatellites for S. broughtonii. We evaluated and validated these multiplex PCRs in 8 full-sib families. The average polymorphism information content (PIC) was 0.539. The frequency of null alleles was estimated as 3.13% of all the alleles segregation based on a within-family analysis of Mendelian segregation patterns. Parentage analysis of real offspring demonstrated that 100% of all offspring were unambiguously allocated to a pair of parents based on 3 multiplex sets. Those 43 microsatellite loci with high variability will be helpful for the analysis of population genetics and conservation of wild stock of S. broughtonii. The 3 sets of multiplex PCRs could be an important tool of pedigree reconstruction, population genetic analysis and brood stock management.     

Distribution patterns of chlorophyll <Emphasis Type="Italic">a</Emphasis> in spring and autumn in association with hydrological features in the southern Yellow Sea and northern East China Sea

Two field studies were conducted to measure pigments in the Southern Yellow Sea (SYS) and the northern East China Sea (NECS) in April (spring) and September (autumn) to evaluate the distribution pattern of phytoplankton stock (Chl a concentration) and the impact of hydrological features such as water mass, mixing and tidal front on these patterns. The results indicated that the Chl a concentration was 2.43±2.64 (Mean ± SD) mg m^?3 in April (range, 0.35 to 17.02 mg m^?3) and 1.75±3.10 mg m^?3 in September (from 0.07 to 36.54 mg m^?3) in 2003. Additionally, four areas with higher Chl a concentrations were observed in the surface water in April, while two were observed in September, and these areas were located within or near the point at which different water masses converged (temperature front area). The distribution pattern of Chl a was generally consistent between onshore and offshore stations at different depths in April and September. Specifically, higher Chl a concentrations were observed along the coastal line in September, which consisted of a mixing area and a tidal front area, although the distributional pattern of Chl a concentrations varied along transects in April. The maximum Chl a concentration at each station was observed in the surface and subsurface layer (0–10 m) for onshore stations and the thermocline layer (10–30 m) for offshore stations in September, while the greatest concentrations were generally observed in surface and subsurface water (0–10 m) in April. The formation of the Chl a distributional pattern in the SYS and NECS and its relationship with possible influencing factors is also discussed. Although physical forces had a close relationship with Chl a distribution, more data are required to clearly and comprehensively elucidate the spatial pattern dynamics of Chl a in the SYS and NECS.     

Effects of feeding and stocking density on digestion of cultured Atlantic salmon Salmo salar L.

The combined effects of feeding rate （0.8%, 1.0%, and 1.2% initial body weight/day）, feeding frequency （two, three, and four times/day） and stocking density （10, 15, and 20 kg/m3） in recirculating aquaculture systems （RAS） on growth performance, digestion and waste generation of Atlantic salmon （Salmo salar L.） were investigated in an 8-week orthogonal experiment （L9（3）3） with a constant daily water renewal at 7.50% of total volume. No mortality occurred during the experimental period. Feed conversion ratio （FCR） varied from 0.90 to 1.13 and specific growth rate （SGR） ranged from 0.48% to 0.69%/day. SGR, thermal growth coefficient （TGC） and FCR were not significantly （P〉0.05） affected by the three factors, while net protein utilization （NPU） was significantly （P〈0.05） affected. Apparent digestibility coefficients （ADC） of dry matter in the present study were in the range 66.12%-73.55%. ADC in protein, lipid and energy were statistically different among all treatments and in the range of 90.07%-93.67%, 81.54%-89.15%, and 67.55%-71.87%, respectively. The proportion of mean total ammonia nitrogen excreted ranged from 1.37% to 1.64% of feed nitrogen at steady state, and the concentration of nitrogenous and phosphorus compounds were differently correlated to the three factors. The results will provide valuable reference data for culture management decisions in the Atlantic salmon farming industry.