Abstract:The oogenesis and ovarian development cycle of Sepiella maindroni, artificial breeding, are observed on histological and cytological characteristic by using routine histological techniques. According to the size of oocytes, the morphology of nucleus and the relationship between oocytes and follicle cells, the oogenesis can be divided into four stages as follows: oogonium stage, oocytes stage, maturation stage and resorbing oocytes stage. Moreover, the histological characteristic and variational disciplinarian of oocytes at each stage is described in detail. Basis of the observation of oogenesis, the results illuminate that this cuttfish’s oogenesis are provided with particularity. About twelve days after hatching, there are oogonium in the ovary, and then the oogonium proliferate quickly. Examined with a light microscope, the oocytes is follicular type, and their development are not at the same developmental stage in the ovary. According to the oogenesis, the eggs have no primary membrane, there are only secondary membrane and tertiary membrane.The secondary membrane is excreted by follicle cells; and the tertiary membrane is made by the secretion of albumen gland、nidamental gland and ink sac. Basis of the morphological characteristic and variational disciplinarian through histological observation, the process of ovarial development of breeding Sepiella maindroni can be divided into 6 stages.

Abstract:In order to identify the strain of Streptococcus iniae(S.iniae) from the infected tilapia at molecule level, the 16S rRNA gene was amplified by using the universal primers, and the PCR product was cloned and sequenced．A sequence of 1447 base pair(bp) was obtained from the bacteria．The sequence was compared to that derived from GenBank using BLASTn，and the closest sequence was the 16S rRNA gene of S.iniae SCCF5L strain，with a similarity of 99.4% and we name it S.iniaeCGX strain. The phylogenetic analysis was based on the 16S rRNA genes sequences of S.iniaeCGX strain and other related organisms，including the representative strains of S.iniae, S.difficilis and S.agalactiae species in GenBank database. The result showed that all strains of S.iniae specie formed a separate and distinct phylogenetic branch, the strains of S.difficilis and S.agalactiae species formed another two separate and distinct phylogenetic branches. The results above showed that the strain of Streptococcus from the tilapia brain was S.iniae species.

Abstract:Scales were used for the age determination of high-finned grouper (Cromileptes altivelis)．The relationships between body length and age were described by four growth models in order to find out the best one．The results showed that the annual-ring feature belongs to the loose-close．The false ring and spawning check can be seen occasionally．According to the scale radius，body length and body weight，the growth condition of C. altivelis was analyzed．The radius of scale was linearly and directly correlative to body length．The relationship between body length ( L) and body weight ( W) was expressed as Wt＝2.3×10-5Lt3.0245．Four growth functions provided a high goodness of fit and preferably described the growth of C. altivelis．Among them, the Gomperz growth function was especially suited to describe the growth patterns of under 10 years fish．The growth of body length and body weight can be described by the von Bertalanffy growth equation Lt=566.0139[1-e-0.1549(t+1.0988)]；Wt=4531.0076[1-e-0.1641(t+0.76635)]3.0245．The age of 5.98 years was the turning point of body growth with body weight 1346.26 g．The fish grew faster before the age of 6．

Abstract:Experiments in this study on the effect of water temperature upon the development of pelagospheric larvae of peanut worm Sipunculus nudus were conducted over larvae from swimming period to setting stage, in which, parents worms used for larvaeproducing were collected from native sea waters, and larvae were obtained by manmade induction to spawning and hatching. The results showed that the development durations of larval S.nudus from swimming to setting stage were closely related to the change in water temperature with a relationship in response to Logistic curve, indicating an optimum temperature range of 27.5-32.0 ℃ for the development of the larvae at swimming stage, a critical range of 27.5-28.0 ℃ and an optimum temperature of 30.0-34.0 ℃ for the larvae at metamorphosing, with the evidence that at 30 ℃ or above larvae at metamorphosing could arrive at obviously higher rates of both metamorphosis and postmetamorphosed survival, and the survival rate of postmetamorphosis being above 20%. Discussions were also given in this study to explore the possibility that there may be a close relationship between the distribution of natural resources of S. nudus and the distribution of water temperatures along the China Sea, assuming that the increase in number of the natural resource distribution of the worm from the North to the South might result from the change in water temperature.

Abstract:Studies on techniques of parent fish nursing and artificial propagation of Elopichthys bambusa were carried out in Jinshan fishery farm of Huzhou city in Zhejiang province from 2003 to 2006. The results revealed that the two-year-old wild fish from Taihu lake after cultivating intensively in ponds for three years, the gonad maturity rate of the female and male fish was up to 100%. When the sexual mature parent fish were induced by both hormone injection and water flow, their average rates of spawning, fertilization and hatching were 100%, 86.5% and 88.3% respectively. Furthermore, the process of embryonic development was also recorded. The fertilization eggs hatch under 21~23℃, and the effective accumulated temperature is 2325.34（℃•h）

Abstract:Lipovitellin（Lv）was purified from ovaries of mature female swordtai1 fish（Xiphophorus helleri）by gel filtration with Sephacryl S300 HR and anion exchange chromatography with HiTrap Q. The results indicated that the purified Lv appeared approximately 530 kD in native polyacrylamide gel electrophoresis(PAGE).Lv was characterized as a phospholipoglycoprotein by NativePAGE and staining of gels for carbohydrates, lipids and phosphorus. Polyclonal antibodies against the purified Lv from swordtai1 fish were produced in the mice. Double immunodiffusion showed that the determination of the titre for antiLv sera was l∶32. Westernblotting analyses demonstrated that antiLv sera have specificity and Lv has common antigenicity with vitellogenin(Vtg) while purified Vtg and Lv had crossreacted obviously with the antiLv sera. Double immunodiffusion showed that antiLv sera have sexspecificity and speciesspecificity.

Abstract:In order to study the metabolism and elimination of norfloxacin (NFLX) in Pseudosciaena polyactis, single oral administration(10 mg/kg b.w) of NFLX has been investigated in healthy Pseudosciaena polyactis at (22±2) ℃. The medicine was extracted by methanol from plasma and perchloric acid (6%) from tissues. NFLX was determined by high performance liquid chromatography(HPLC). The mobile phase was methanol, 0.05 mol/L phosphate buffer and 0.05 mol/L terabutyl ammonium bromide (24.5/73.5/2, V/V/V) adjusted to pH 2.0 with phosphoric acid. The ZORAX SBC18 column was used and ultraviolet detector was 280 nm. The mean recoveries were all higher than 88.73％ for samples and interday CV for plasma and tissues were all under 9.75 %. The sensitivity was 0.01 μg/mL. The result demonstrated: Plasma concentrationtime data of NFLX were best fitted using a twocompartmental open model, absorption and distribution were fast but elimination was slow, with absorption, distribution and elimination halflife of 0.703 0, 2.092 6 and 154.326 5 h, respectively. The maximal plasma concentration was 0.886 4 μg/mL, peaking at 2.091 4 h after dosing. Area under the plasma drug concentrationtime curve from 0 to ∞ was 97.803 8 μg/h·mL. The concentration of NFLX was higher than other tisssues and all tissues’ concentration is higher than plasma at the same time. The speed from fast to slow is kidney, liver, muscle with elimination halflife of 135.88,173.25,223.55 h, respectively. As an edible tissue, muscle had a slowest depletion, so muscle was selected as target tissue in this experiment. According to the maximum residue limit(MRL) of 50 μg/kg in muscle, the withdrawl period should not be less than 23 d under this experiment condition; then the scheme of NFLX is established to fish bacterial disease, that is，the dose orally is l0 mg/kg, at the intervals of 24 h for 2～3 times．This study had theory value to application of norfloxacin (NFLX) to disease prevention and cure for aquatic animals and consumer’s health safety.

Abstract:To investigate the protective role of three antioxidantsvitamin C, vitamin E and Zn（Zn2S4•7H2O）in CdCl2induced Eriocheir sinensis sperm DNA damage, single cell gel electrophoresis was used to detect the relationship between the contents of antioxidants and DNA damage. Four parameters, L tail (Tail length in arbitrary units), Tail DNA (relative % of DNA in the comet tail), TM (tail moment means integrated value of DNA density multiplied by the migration distance，L tail×Tail DNA) and OTM(Olive tail moment means the product of the distance between the center of gravity of the head and the center of gravity of the tail and percent tail DNA), were used to evaluate the DNA damage of the Eriocheir sinensis sperms which were treated by 5 mmol/ L CdCl2. The result showed that the four parameters all increased remarkably and the sperm DNA was seriously damaged by CdCl2. After that the protective effects of 3 kinds of antioxidants on CdCl2induced DNA damage were investigated. The contents of VC were 400 μmol/L,1 mmol/L, 3 mmol/L and 5 mmol/L；that of VE were 50 μmol/L,100 μmol/L,200 μmol/L and 400 μmol/L；and that of Zn were 400 μg•Ml/L,1 mg/L,2 mg/L and 4 mg/mL, respectively. The result showed that all these three antioxidants can protect 〖WTBX〗Eriocheir sinensis sperm DNA from oxidizing. The bigger the content of VE and Zn, the stronger the protective effect was. When the content of VC was 3 mmol/L, the protective effect was the best and the ratio of cell drag was comparatively low. But when the content of that was 5 mmol/L, the ratio of cell drag increased and the DNA damage became more serious. So it was concluded that the content of VC which could protect the sperm DNA from damaging was limited. Whereas the protective effect of VE and Zn had no limit of the content. Furthermore, by contrast to VC and Zn, VE appeared better protective effects with concentration of 400 μmol/L. Vitamin E and Zn possessed antagonism to CdCl2 induced DNA damage but vitamin C possessed dual nature. Vitamin E was a sort of perfect antioxidants in antagonism to CdCl2induced DNA damage. In addition, CASP is a good software to analyse comet image externally better than other analysis methods such as routine method. This research can provide methods for the research of Eriocheir sinensis feed additive, species protection, and sperm quality assessment.

Abstract:The comparative analysis on number, morphology, and distribution of intermuscular bones in lower teleosts nonacanthomorph such as Osteoglossomorpha, Elopomorpha, Clupeomorpha, Ostariophysi, was conducted. Pike eels (Muraenesox cinereus), estuarine tapertail anchovies (Coilia ectenes) yellow catfish (Pelteobagrus fulvidraco) and arowanas (Osteoglossum bicirrhosum) owned 409, 221, 70 and 8 intermuscular bones, respectively. The numbers of intermuscular bones in the fishes of Cyprinidae fell down on the range from 99 to 133. The morphology of intermuscular bones here was classified into seven types based on the bone complexity. From Osteoglossomorpha to Ostariophysi, the bones were evolved from simple type to various complexity types, then, retrogressed to simple types. Arowanas only owned simplest “I”type epineurals, another type oneendbifork epineurals was found in pike eel, meanwhile several simpletype epipleurals appeared. In herrings (Tenualosa reevesii) and estuarine tapertail anchovies (Coilia ectenes), a little more complexity epineurals like oneendmultifork type were found. As to Xenocypridini and Cyprinidae, most complexity type tree type was evolved. From Leuciscini, the epineurals initially retrogressed to a little simple type, finally to Siluriformes, the epineurals totally was disappeared, and the epipleurals also retrogressed to most simple type. Comparing with epineurals, the epipleurals emerged a little later, and formed a litter simpler. The complexity degree of various epineurals with rostrocaudal arrangment decreased gradually. However, the decreasing trend of the complexity degree of epipleurals was not evident. On the whole, Consideration taken together with lower teleostean phyologenetic relation, the morphology and arrangement of intermuscular bones, two paths of intermuscular bone evolution in Cyprinidae were postulated. One is from “I” type (nofork type) to “卜” type (oneendunequalbifork type), then to “Y” type (oneendequalbifork type), finally to oneendmutifork type, another is from “I” type to “卜” type, then to “Y” type, then to twoendbifork type, then to twoendmutifork type, finally to tree type. The research results provided here will help us to understand the evolution of intermuscular bones.

Abstract:After 21 days’ exposure of adult male goldfishes(Carassius auratus) to monocrotophos(0.01、0.10、1.00 mg•L-1), we measured activities of lactate dehydrogenase(LDH), acid phosphatase(ACP), alkaline phosphatase(ALP) in testis and observed micro-, ultrastructure of testis by light and electron microscopy separately. We found that the gonado-somatic(GSI) decreased significantly, ground membrane dissolved, leydig’s cell swelled and interstitial tissue enlarged, as a result of exposure to monocrotophos at the concentration of 1.00 mg•L-1. Monocrotophos induced dissolution of nuclear membrane of the Sertoli’s cell, and the increase of lipid droplet and myelin-like figure number in Sertoli’s cell. The activities of LDH, ACP and ALP in testis decreased gradually, as the exposure concentration of monocrotophos increased. Exposure to monocrotophos at concentrations of 0.10 and 1.00 mg•L-1 resulted in the decrease of LDH and ACP activities in testis. However, the ALP activity did not decrease significantly at the exposure concentrations of all. It is supposed that the energy metabolism was interrupted as a result of the changes of micro-, ultrastructure and the decrease of LDH, ACP, ALP activities in goldfish testis, which caused the reproductive toxicity effect.

Abstract:Ruditapes philippinarum, which is naturally distributed along the coast of the Pacific and India Oceans, is one of the most popular shellfish cultured in China and many parts of the world. China is the world’s largest producer of Ruditapes philippinarum. Some inappropriate ways of aquaculture, however, have resulted in the worseness of environment, which would restrict and endanger the development of aquaculture. The study of carrying capacity may resolve the problem of ecosystem by overstocking, so the study on carrying capacity of cultured tidal shellfish is very necessary. The carrying capacity for shellfish Ruditapes philippinarum that cultured in the Rushan Bay was estimated by calculating the demand for and the supply of organic carbon produced by primary production. Ingestion rate of filter-ingestion animals and other field data were used for model validation. The results obtained showed that the highest carrying capacity of the bay for the Ruditapes philippinarum occurred in October and the lowest in August in tidal cultured areas among different shell length. The average carrying capacity of the bay for the Ruditapes philippinarum with the shell length of 1.5-2.5cm was 2692ind/m2 and still in suitable opportunity, the shell length of 2.5-3.5cm was 1157 ind/m2 and near the actual culture density, and the shell length above 3.5cm was 2692 ind/m2 and over the best culture density.

Abstract:Nowadays, the selective Germany mirror carp (Cyprinus carpio L. mirror) is one of the excellent breeds adapting to pond condition in the North area of China. In china, however, the information on their genetic variability is unavailable. Here, to evaluate the genetic diversities and relationships among 3 populations (BL, BZ and TJ) located at Suihua, Harbin and Tianjin of China respectively, 10 polymorphism microsatellite markers were used. The results demonstrated that 3 populations showed high polymorphism in 9 loci (PIC>0.5), and the average heterozygosity degree per locus was 0.6293. Meantime, 144 alleles were detected in all examined loci. Among them, BL and TJ populations had higher genetic variation than BZ population. The analyses of Nei’s DA genetic distance indicated the closest relationship was between BL and TJ, and the largest distance was observed between BL and BZ. Only little genetic differentiation was detected among the 3 populations (Fst=0.0315), and the internal variation was 96.85% of the total variation. Furthermore, the own alleles frequencies of each population were commonly lower (0.0125 ~ 0.1500), but the shared alleles frequencies of total populations were relatively higher. Therefore, we proposed that the selective populations of Germany mirror carp in different regions of China still kept high genetic diversity level, and the genetic differentiation among populations was not significant.

Abstract:Argopecten irradians concentricus Say, introduced from America, has been extensively cultured in recent years in Beibu bay in Zhanjiang, but the research about its morphological growth is not found to have been reported, this makes the culture of Argopecten irradians concentricus Say lack of the guiding of relevant theory. The morphological growth of Mexican bay scallop(Argopecten irradians concentricus Say) in Zhanjiang Beibu bay sea area was investigated in this paper by using model fitting procedure under such conditions as seawater temperature ranging from 18.2℃ to 30.8℃, salinity ranging from 27.8 to 30.4, limpidity ranging from 1.5m to 5.7m, water depth at 9.7m, average planktonic plant cell abundance at 4.3104cellL-1 and the autumu-winter-spring culture mode in which the time span of the investigation was exactly equivalent to one culture cycle of Mexican bay scallop. Results showed that the growth of morphological traits of Mexican bay scallop conformed to Logistic model, with squared multiple correlation coefficient R2 all exceeding 0.98 (P<0.001) and root mean square error (RMSE) averaging out at 0.897. All Logistic models of varying morphological traits were statistically significant (ANOVA, P<0.001). Growth parameters contained in Logistic model, estimated by Levenberg-Marquardt iteration method, could be employed in establishing the growth equations of varying traits of interest. Growth limits and confidence intervals of varying morphological traits of Mexican bay scallop in Zhanjiang sea area were, shell length 53.8mm, shell height 54.6mm, shell breadth 27.1mm,hinge length 28.4mm, adductor length 26.1mm, adductor diameter 16.2mm, respectively; the fastest growth intervals and growth inflection points of varying morphological traits were, shell length 1.5-5.7 months and 3.6 months, shell height 1.4-5.9 months and 3.6 months, shell breadth 2-6.4 months and 4.2 months, hinge length 0.7-4.8 months and 2.8 months, adductor length 1.9-6.5 months and 4.2 months, adductor diameter 1.6-5.5 months and 3.5 months, respectively. Growth delay with shell width and adductor length took place. In terms of the absolute growth rate, there was this sequence, viz., shell length>shell height>shell breadth>adductor length>adductor diameter, the relative growth rate were different among traits of interest and gradually tended to 0. It was found in the study that delay of the growth of shell width and adductor length traits occurred, there existed significant difference in terms of morphological growth between varying geographic populations, the growth of morphological traits were controled by both temperature and growth phase. New viewpoints of opportune harvesting according to the growth of shell width and adductor length traits and of arranging the fast growth period into most favorable season so as to maxically harness growth potential were proposed. Results obtained can serve as theoretical guidelines for predictions of future changes in culture ecology, selection breeding and production of Mexican bay scallop.

Abstract:Based on the observation and comparative analysis, overall and systematic development of M．latissimum in the stages from conjugation of gamete to discharge of zoospore, on the sporophyte stage during the life history of M．latissimum is entirely expatiated in this article. Meanwhile, on the basis of the results of the experiment, we divided the sporophyte stage of M．latissimum into five periods : conjugation of gametes（gamete 7.5 μm×2.0 μm）, zygocyte （4-20 μm）, sporange （18-40 μm）, zoospore formation （sporange 35-55 μm） and discharge of zoospore （zoospore 9.4 μm×3 μm）. In this paper it may be in China to find the systemic acknowledgement on the division of the sporange stage during the life history of Monostroma as well as to the seedling cultivation in practice. 

Abstract:The effects of 6-DMAP on assembly of microtubules and pronuclear migration between normal and gynogenetic fertilized eggs ofChlamys farreri were studied carefully by direct immunofluorescence with antitubulin. According to observation, chromosomes were segregated by spindle, the first and the second polar bodies were emitted and the female and the male pronuclei fused into zygotonucleus in normal eggs. Treatment of eggs at metaphaseanaphase stages caused drastic modifications of the spindle organization. The antitubulin staining revealed that soon after addition of 60μg/L l6-DMAP, outward astral fibers were no longer detected, whereas the spindle showed elongation and flattening. Finally 6-DMAP induced a significant disorganization of the spindle at the metaphase of meiosis II, the formation of the second polar body was inhibited effectively. The fertilized eggs raised in the presence of 6DMAP did not show any further pronuclear movements. when the treated eggs were washed and further maintained in fresh water ,migration resumed at a slower rate than in normal eggs.

Abstract:Apostichopus japonicus was reared under different temperatures, salinities and light cycles so as to analyze the effects of temperature, salinities and light cycle on the growth and behavior of it. A. japonicus were sampled from Dakoujing Shrimp Cultural Cage, Lüshun City, which were 1yearold, and were accustomed to the experimental conditions for two days before each experiment. The average initial body wet weight of the experimental sea cucumber was 36.89 g (25.24-59.11 g), and the average initial body length was 6.68 cm (5.12-8.74 cm). In the temperature experiment, the sea cucumbers were reared in six plastic barrels (the cubage of each was about 60 L and one brick in each barrel) at six temperatures (9, 12, 15, 18, 21 and 24 ℃). The results showed that the body wet weight increased under 9-21 ℃ and 15 ℃ was the optimum temperature to the growth of A. japonicus. When the temperature was beyond 22 ℃, A. japonicus began to aestivate. As soon as the temperature below 20 ℃, aestivation was over. In the salinity experiment, the sea cucumbers were reared in the same plastic barrels under six salinities (23, 26, 29, 32, 35 and 38). The salinity for growth of the A. japonicus was 29-33, and the optimum salinity was 32, the reason of which was that the sea cucumbers had been accustomed to the range of salinity in the ocean for a long time. The A. japonicus were reared in four plastic barrels for the light cycle experiment, and the conditions of the barrels were fully light (24hourlight), fully dark (24hourdark), half light dark (12hourlight and 12hourdark) and natural light. The growth of the sea cucumbers was best under the half light dark condition, and worst under the fully dark condition. In addition, the correlation analyses were given in each experiment.

Abstract:Halfsmooth tonguesole (Cynoglossus semilaevis Günther) is a cultured marine fish exploited recently in China. The female individuals of tongue sole grow 1-2 times faster than male individuals. Thus, the development of allfemale stock would be of significant benefit for aquaculture. Sexrelated molecular marker is a useful tool for studying sex determination mechnism and controling fish sex. In order to screen sex specific molecular markers, AFLP analysis technique was firstly developed in halfsmooth tonguesole. DNA extraction from liver tissues was carried out according to stardand procedures. Phenotypic sex of the fish was determined by histological sectioning and staining. 64 AFLP primercombinations were used to screen the genome DNA of halfsmooth tonguesoles. 4 primercombinations amplified 7 femalespecific markers that were female specific in halfsmooth tonguesole. The 7 femalespecific markers were named CseF382, CseF575, CseF783, CseF464, CseF136, CseF618 and CseF305, respectively. One femalespecific AFLP marker (CseF382) was amplified, recovered from the gels, cloned, and sequenced. This femalespecific AFLP marker was converted into single locus PCR marker of a sequencecharacterized amplified region (SCAR). A simple PCR method of using the specific primers was developed for identifying genetic sex of halfsmooth tongue sole. This PCR method will be more efficient and less expensive than with AFLP markers. The isolation of sexspecific molecular markers lays a basis for elucidation of sex determination mechanism, and provides a tool for sex control in halfsmooth tongue sole.

Abstract:So far studies have focused on bacteria isolation aimed at establishing causes of fish diseases and medication methods. Now, more and more attention has been given to the composition of the microflora, its variations in time, and effect on whiteleg shrimp (Litopenaeus vannamei). This is why the problem of bacterial flora in water ought to be studied. The major role of ammonifying bacteria is to break down nitrogenous organic matter to ammoniac nitrogen. The research of ammonifers in the pond water of Litopenaeus vannamei is increasing with the demand for environment friendly aquaculture. Four bacterial strains (No. zjs01，zjs02，zjs03 and zjs04), isolated from the pond water of Litopenaeus vannamei in Jinshan district of Shanghai, were cultured in ammonifying bacteria rich medium and identified by two methods. One is the sequence analysis of 16S rDNA, the other is bacteria identification system. First, sequence analysis of the 16S rDNA was done. Genomic DNA of four strains was isolated respectively, then their full length of the 16S rDNA were amplified by PCR respectively, using universal primers to the 16S rDNA. After purification by gel extraction, the PCR products were cloned and subsequently sequenced by Shanghai Invitrogen Biotechnology Company (SIBC). The phylogenetic trees were constructed, based on the result of online alignment. At the same time, the four sequences of 16S rDNA were submitted to NCBI（http://www.ncbi.nlm.nih.gov） in order to obtain accession number of strains of zjs01, zjs02, zjs03 and zjs04. Then the API 2000 Bacteria Identification System (Biomerieux Company) was applied in assessment the identification result of zjs01, zjs02, zjs03 and zjs04 by molecular method. Finally the 4 strains were identified as, zjs01: Brevundimonas diminuta, zjs02 and zjs03: Alcaligenes faecalis, zjs04: Enterobacter aerogenes. And the accession number of the strains (zjs01, zjs02, zjs03 and zjs04) is DQ857897, DQ857898, DQ857895, DQ857896 respectively. The method of detecting bacterial 16S rDNA using PCR technique is specific，sensitive，rapid and accurate in detecting bacterium in culture pond. Also an interesting thing should be indicated, it is absolutely necessary that the 16S rDNA PCR products should be cloned before DNA sequencing. This paper will establish theory groundwork for application of ammonifers microbiological preparation to bioremediation of the polluted culture water.

Abstract:The genetic structure and segregation of molecular markers in F2 of Chinese shrimp ( Fenneropenaeus, chinensis) were studied by random amplified polymorphic DNA (RAPD) technology to explore its usefulness in linkage mapping construction. Two hereditable types of DNA markers, the non-segregating type and segregating type, were identified from the amplification results of 22 random primers. The non-segregating marks in both parents and progenies accounted for 54.1% of all markers, reflecting one genotypic combinations of G♀ and G♂ (AA×AA). There were three kinds of segregation, including Mendelian segregation, deviated segregation and abnormal segregation, which accounted for 45.9% of all marks. The standard Mendelian segregating markers were divided in 3:1 ratio and 1:1ratio, the former accounted for 14.7% of all segregated markers respectively and the latter accounted for 64.7%. The 3:1-segregated markers in the F2 represented four genotypic combinations of G♀ and G♂ (AA×aa、Aa×aa、Aa×Aa and Aa×AA), and one genotypic combinations of F1♀ and F1♂ (Aa×Aa); the 1:1-segregated markers in the F2 represented two genotypic combinations of G♀ and G♂ (Aa×aa and Aa×Aa), and one genotypic combinations of F1♀ and F1♂(Aa×aa); non-parental heteroduplex, so far, had not be conjectured their genotypic combinations, which were segregated 1:1 ratio in the F2 progenies. Non-standard Mendelian segregating marks and abnormal segregating marks accounted respectively for 11.7% and 8.9% of all segregated markers respectively. Totally, 76.5% segregating markers, explained by two way pseudo-testcross, could be used to construct genetic maps of F. chinensis. Accordingly, the result would lay a theoretical basis for genetic linkage map construction using RAPD markers and F2 population．

Abstract:A high performance liquid chromatography method had been established for the determination of malachite green and leucomalachite green in aquatic water. Malachite green and leucomalachite green were extracted with dichloromethane, then concentrated, and dissolved with acetonitrile. Samples were chromatographed using an acetonitrile-ammonium acetate buffer mobile phase on a C１8 column in line with a post column reactor filled with lead (IV) oxide and celite. The liner range was from 0.01μg•mL-1 to 1μg•mL-1 with r2 of 0.9999. The average recoveries of MG were 84.00%, 86.40%, 93.50%, and LMG were 74.00%, 89.00%, 91.80% when samples were spiked with MG and LMG of 0.1, 0.5, 1.0μg•mL-1 respectively. The detection limits of MG and LMG were 0.003μg•mL-1 and 0.004μg•mL-1 respectively. The RSDs were 4.278%-6.250% and 2.179%-6.757% respectively. The method is sensitive, accurate and suitable for the determination of trace malachite green and leucomalachite green in aquatic water. Key words: malachite green ; leucomalachite green; high performance liquid chromatography; aquatic water