转Hu-IFN-α基因草鱼雌核发育F1的遗传配型分析

The Hu-IFN-α gene, which was transducted into downstream promoter of β-actin gene of common carp (Cyprinus carpio), was recombined by DNA recombination technology. These recombined genes were injected into 1-2 cell fertilized eggs of grass carp (Ctenophatyngodon idellus) by microinjection technology, we gained transgenetic fish by molecular detection methods. In order to analyse the genetic expression of tranHu-IFN-α gene gynogensis F₁, which male individualization were gained by raising methyltestosterone, molecular genetic marker technology was used. In our research, 30 random primers were picked out from 48 and were used into RAPD-PCR, the result indicated that 1 169 clear, steady and repeated DNA finger printing bands were achieved. On the basis of gentic distance matrix among tranHu-IFN-α gene gynogenesis F₁ group, the genetic relationship of gynogenesis F₁ were analysed by UPGMA, the results showed the genetic patterns are close between the 3# male gynogenesis F₁ and the the 23# female of gynogenesis F₁, 5# and 27#, 2# and 28#, 2# and 30#. The data indicated that these group could be served as parent of tranHu-IFN-α grass carp (Ctenophatyngodon idellus) pure line.     

水产养殖动物基因组研究的现状及其应用前景

The genomic resources from human and several model organisms have been increased very fast since 1990. The techniques for developing genomic resources have already been very advanced and smart. These could make scientists see and improve organism in genomic level. For Chinese aquaculture scientists and aquatic industry, developing genomic resources and genetic tools for the native species are most important in the genomic era. The genomic resources and genetic tools for several aquatic species have been developed and some of them have been used in the marker based selection and other researches. The genome research work on aquaculture species was reviewed in this paper, especially a USDA genome project was focused. Some functional genomic research for aquatic animal was also discussed here. The importance and necessity of China aquaculture species genome project were discussed. Common carp and other cultured fishes in Cyprinidae such as grass carp, silver carp, bighead carp etc were recommended as the candidate species for genome research, because the output of all carps is almost up to 1/3 of total fisheries output in China. Common carp with another virtue for genome research is that there are much more families and strains in common carp than those in other cultured species in China, and those families and strains are the basis for genome research and mapping quantitative trait loci associated with important economic trait. Although the first linkage map of common carp made by Sun needs to be added with more markers for mapping QTL and Type I markers ,it has laid the groundwork for QTL mapping and markerassisted selection in common carp. Because the model organism zebrafish and common carp, grass carp and other carps cultured in China all belong to Cyprinidae, the China carp genome research will obtain a lot of useful information from zebrafish genome research. How the China carp genome program will be conducted and what kinds of strategy involved in this program were all suggested. How the results of the genome research of aquaculture species will be used in the aquaculture industry was reviewed and analyzed here.     

彭泽鲫的分子遗传分析及其与方正银鲫A系的比较

Genetic homogeneity between Pengze crucian carp and strain A of silver crucian carp was studied by using transferrin,isozyme and RAPD markers.The studied individuals of Pengze crucian carp showed transferring patterns were the same of silver crucian carp A strain while distinct from those of other crucian carp populations.As far as isozyme is concerned,the MDH,LDH and EST are all of the same with only slight differences in SOD between them.The RPAD patterns clearly indicated high homogeneity among 16 individuals (6 sampled from individuals of two years old and the others aged one) from crucian carp of Pengze and 5 individuals from strain A of silver crucain carp.Nearly indentical banding patterns were observed among all individuals.Average genetic distance within all the individuals is only 0.011,suggesting crucian carp of Pengze might possess indentical genetic background with strain A of silver crucian carp.     

海洋贝类雌核发育研究进展和展望

The paper summarized the progresses and achievements of the studies on the gynogenesis of the marine mollusks at home and abroad in recent years,and pointed out the study direction on gynogenetic shellfishes and 6-DMAP used as an ideal medicine for raising the induced rate of marine shellfish gynogenesis.     

罗非鱼杂交F1代与亲本的遗传关系及其杂种优势的利用

Genetic relationships between the parental fishes Nil e tilapia,blue tilapia and their hybrids were assayed with RAPD analysis using 1 6 primers, of which 11 generated polymorphic products. Genetic similarity indice s demonstrated that the offsprings of Nile tilapia♀×blue tilapia♂were genetic ally intermediate between the parental fishes,while the offsprings of blue tilap ia♀×Nile tilapia♂were extremely similar to blue tilapia.This suggested that hybrids F₁ have high possibility to develop heterosis if they are intermediate between the parental fishes in genetic characteristics. In addition, products O PA07₁₉₀₀,OPA07₉₆₀,OPZ14₇₂₀ and OPZ14₆₀₀ were found to be molecular markers for identifying blue tilapia, Nile tilapia and the offspring of Nile tilapia♀×blue tilapia♂.     

海湾扇贝个体间单向人工授精的分子生物学验证

Hybrid family of Argopecten irradians irradians was created by fertilization between two northern bay scallop individuals. Two families were analyzed in this study. The first family, P_a-P_b, is a pair mating between two scallops named P_a and P_b, while the second one crossed by individuals of Y₁ and P₀. Marker inheritance and segregation were studied in the 10 progenies of each family by randomly amplified polymorphic DNA (RAPD) analysis. 102 RAPD primers were first screened by parental animals of both families. Only the primers with polymorphisms between the two parental animals in each family were select ed for further analysis. In both families, parents and 10 progeny were nalyzed with selected primers. In family P_a-P_b, a total of 122 bands generated from 12 selected primers. 37 of them were polymorphic between two parents. The maternal P_a of this family had 17 molecular markers while paternal Pb had 20 markers. In Y₁-P₀，95 bands were produced by 10 selected primers. 32 bands were polymorphic between maternal Y₁ and paternal P₀, who had 17 and 15 molecular markers respectively. In both families, each progeny analyzed in this study had at least 8 maternal markers and 5 paternal markers. Based on segregation patterns at all markers analyzed, we concluded that none of the progeny analyzed were from self-fertilization, and oneway hybridization between two individuals was successful in both of the two bay scallop families.     

建鲤与异育银鲫生长、生化组成和饲料利用的比较

In order to investigate the difference in utilization of diets with different quality in Jian carp（Cyprinus carpio var. Jian）and allogynogenetic silver curcian carp(Carassius auratus gibelio), a 55 d growth trial was conducted and low quality diet (LQ-diet) and high quality diet (HQ diet) were tested. LQ-diet contained 33.91% dietary protein which is mainly from soybean meal while HQ-diet contained 45.59% dietary protein which is mainly from fish meal. The initial average body weights were from 5.58 g to 5.82 g for two fish strains. The trial was carried out in a system consisting of 12 self circulation 320 L tanks. During the experiment, the fish were fed to satiation twice a day (at 9: 00 and 15:00), and uneaten feed was collected 1 h after feeding and dried. Feces were collected twice a day (at 11:00 and 16:45) from the fecal traps and dried at 70 ℃. The results show that feed intake was higher in Jian carp than in allogynogenetic silver curcian carp when fed LQ diet, while there was no significant difference between weight gain, feed conversion efficiency, protein efficiency rate and apparent digestibility. When fed HQ diet, Jian carp showed a lower feed intake, but higher feed conversion efficiency and protein efficiency rate than allogynogenetic silver curcian carp while there was no significant difference in the weight gain and apparent digestibility of both species. For Jian carp, feed intake and protein efficiency rate for the fish fed HQ diet and LQ diet were not significantly different, while the fish fed HQ diet showed higher weight gain, feed conve rsion efficiency and apparent digestibility. For allogynogenetic silver curcian carp, the fish fed HQ diet showed significantly higher feed intake, apparent digestibility, weight gain, feed conversion efficiency and lower protein efficiency rate. For Jian carp, body contents of dry matter, protein, lipid and energy for the fish fed HQ diet and LQ diet were significantly higher. For allogynogenetic silver curcian carp, body contents of dry matter and protein was significantly higher, while body contents of lipid and energy were affected by diet qualities. Compared to Jian carp, allogynogenetic silver curcian carp showed better utilization when fed LQ diet while poorer utilization when fed HQ diet.     

鲍遗传育种研究进展

Abalones are important farming species with a high economic value. They have already been farmed for more than 50 years. As problems and new requirements rose continuously in culture industry of abalone, studies on genetic and breeding techniques are needed to improve characteristics and to gain new traits. This review concentrates on advances in genetics and breeding techniques in abalone. As for genetic studies, karyological analyses, allozyme, DNA markers and genetic diversity were reviewed. So far, karyological analyses in abalone have been performed in 12 species that can be divided into three groups according to the chromosome number. In some economically important species, loci of allozymes and. microsatellites have been isolated and applied to investigate the genetic structure of natural and hatchery populations and to identify the result of chromosome set manipulation, but the related reports are only a few yet. The resultsof investigation with DNA markers and allozymes showed that the genetic structure of natural populations presents two characteristics: excessive homozygosity and subdivision. Advances of various breeding techniques, including introduction, selection,hybridization, polyploidy, gynogenesis and gene manipulation, were reviewed in the other part. Although Haliotis discus discus, introduced from Japan, has become one of the most important culture species in China, the economic, social and environmental effects of introduction have been rarely studied. Selection is one of the most important and basic breeding techniques, but the studies on selection are only a few and preliminary, referring to the relations between genetic characteristics and the traits of growth and resistance, genetic diversity and heritability of quantitative traits, and the effect of selection. Interspecific hybridization was the first breeding program carded out in abalone. Experimental hybridization have been carded out for about 20 crosses. Heterosis,such as faster growth and high survival rate, has been observed in some crosses. Triploids have been successfully induced in many species of abalone with physical or chemical shock, e.g.H, discus hannai, H. rufescens, H.diversicolor diversicolor and H. rnidae. Field experiments were conducted in some species of triploid abalone. In comparison with triploid, the research on tetraploid is still in quest stage. The progress of induction of gynogenesis in abalone is quite slow. Conditions of sperm inactivation, diploid restoration and nuclear behavior of gynogenetically activated eggs have been researched on in H. discus han nai. Notwithstanding the gene transfer technology in abalone is in the quest stage, the research have already involved preparation of exogenic DNA,means of gene transfer, identification integration and expression of target gene, etc. Three research directions in these topics were proposed : to investigate the germplasm resources of abalone deeply and widely, to make use of traditional breeding methods and modem biotechnique synthetically, and to combine the science research with production practice.     

大菱鲆胚胎的玻璃化冷冻保存

Vitrification solutions were examined for their suitability of cryopreservation of turbot (Seophthulnms maximus) embryos. PMPI gave higher survival rate (75.68%) than other vitrification solutions and was considered to be suitable liar the cryopreservation of turbot embryos. The freezing point of the vitrification solutions consisted of PG and MeOH in the proportion of 3:2 was measured when the vitrification solutions were cryopreserved. It showed that the vitrification solutions with a cryoprotectant concentration of over 41% have no freezing point and the freezing points of vitrification solutions containing 33.33%-40% cryprotectants were between -32.4℃ and -65.4℃, The freezing point decreased with the increase of cryoprotectant concentration. The resistance of turbot embryos at different stages to the cryoprotectants was studied, It was found that the turbot embryos from 4-5 pairs somite to tail bud were more resistant to cryoprotectants and suitable to vitrify. A live embryo was obtained after cryopreservation in liquid nitrogen for 14h and was hatched out。     

水产品中三聚氰胺残留的测定研究

method for determination of melamine residue in fishery products by liquid chromatography-mass spectrum （LC-MS/MS）was established. The conditions of LC-MS/MS was optimized. The sample was ultrasonically extracted with 1% triehloroactic acid and ethylamine, and cleaned by the solid phase extraction （MCX） cartridge. The analyte was identified by electrospray ionization （ESI） in positive mode using multiple reaction monitoring （MRM）. The determination ion pairs are 127/85 and 127/68,and the confirmation ion pairs are 127/85. The method was validated and good results were obtained with respect to spiked recoveries. The linear range was from 10 to 400 ng/mL,the average recoveries were between 80.5%and 94.1% in the spiked rang of 0.01 -10.0 mg/ kg. LOQ was 0.01 mg/kg.     

Stabilization of myosin by ionic compounds as affected by F-actin

ABSTRACT:   Suppressive effects of non-ionic (sorbitol, maltose, and trehalose) and ionic (Na-glutamate, Na-acetate, Na-sulfate, and ammonium sulfate) compounds on the thermal inactivation of myosin subframgent-1 (S-1) and myofibril Ca²⁺-ATPase were compared. All compounds suppressed S-1 denaturation. When myofibrils were used (at 0.1 M KCl), sugars and sugar alcohol (non-ionic compounds) suppressed denaturation similar to S-1, while Na-glutamate, Na-acetate, and Na-sulfate weakly suppressed them. Ammonium sulfate accelerated denaturation, but suppressed denaturation when heated in 2 M KCl, at which myosin lost protection by F-actin. It was thus concluded that ionic compounds affected the denaturation of myofibrils in two ways; suppression as established with S-1, and acceleration as a result of loss of protection by F-actin caused by increase in ionic strength.     

Freshwater fish community structured more by dispersal limitation than by environmental heterogeneity

Abstract –  We examined the relative contribution of environmental heterogeneity and dispersal limitation on freshwater fish community composition in 18 Greek protected lakes and reservoirs. Environmental heterogeneity was measured by spatial pattern metrics (calculated by digital map processing, e.g., patch richness density, area-weighted mean patch area), altitude, maximum lake depth and trophic status. Dispersal limitation was measured by straight-line distances among lake centres. Ecosystems were clustered according to species composition. We examined the correlation of similarity in species composition among ecosystems with that of environmental heterogeneity and with straight-line distances, for the entire dataset, as well as for the occurring clusters. Fish species richness increased with ecosystem area and decreased with altitude. The clusters identified (aquatic ecosystems of Northern vs. ecosystems of Western Greece), implied an underlying biogeographical pattern as defined, with Pindus range acting as a natural barrier. Between ecosystems similarity, based on fish species composition, showed a weak to insignificant correlation with environmental heterogeneity, but was significantly correlated to dispersal limitation for the entire dataset as well as within each occurring cluster. Thus, natural barriers, species biogeography and dispersal limitation played a more significant role in shaping freshwater fish communities than environmental heterogeneity.     

海参养殖池中毛蚶生物沉积作用的研究

对毛蚶的生物沉积作用以及对养殖水体中物质的运输的影响进行了研究.实验结果表明,毛蚶能够加速养殖池中悬浮颗粒物的沉积,小个体(壳长2.60～2.85 cm)的生物沉积率为45.9～89.5 mg ·ind-1·d-1;中等个体(壳长3.77～3.97 cm)为101.0～167.0 mg·ind-1·d-1;大个体(壳长...     

Diagnosis of bacterial kidney disease by detection of Renibacterium salmoninarum by real-time PCR

Bacterial kidney disease (BKD), caused by Renibacterium salmoninarum (Rs), is a serious threat to salmon in aquaculture as well as to wild populations. We have developed a real-time polymerase chain reaction (PCR) for detection of Rs in kidney samples. The PCR is based on detection of unique parts of the 16S rRNA gene of Rs and DNA equivalent to 1-10 Rs genomes was detected per reaction. No cross-reactivity with other fish pathogenic or related bacteria could be demonstrated. Analysis of individual kidney samples collected from BKD classified populations identified 39.9% of the fish as positive by real-time PCR compared with 28.0% by polyclonal enzyme-linked immunosorbent assay (ELISA). The real-time PCR assay was found to be well suited for complementary use with ELISA for diagnosis of BKD, with the ability to detect clinical as well as covert Rs infections. The infection level determined by the polyclonal ELISA and by real-time PCR was significantly correlated.     

刺参蛋白胰蛋白酶解产物生物活性的研究

用胰蛋白酶对刺参蛋白进行水解,研究其酶解液所具有的抗氧化活性及其对肠道菌群的作用。通过测定刺参蛋白酶解液清除DPPH(二苯基苦基苯肼)自由基、Feton体系产生的羟基自由基和邻苯三酚自氧化体系产生的超氧阴离子自由基的能力。同时,研究了刺参蛋白酶解液体外对肠道菌群生长的影响。结果表明,刺参蛋白胰蛋白酶解液具有抗氧化活性和促进乳酸杆菌生长、抑制大肠杆菌和产气杆菌生长的作用。     

栉孔扇贝生物沉积作用的研究

2001年3～9月,在自然养殖状态下对栉孔扇贝(Chlamys farreri)的生物沉积及其对物质输运的影响进行研究.结果表明,栉孔扇贝能加速海洋中颗粒物质的沉积,生物沉积率分别为小个体(壳长30～40 mm)72.31～109.85 mg·ind-1·day-1、中等个体(壳长50～60 mm)103.49～207.77 mg·ind-1·day-1和大个体(壳长60～70 mm)120.05～237.65 mg·ind-1·day-1.栉孔扇贝的生物沉积与其壳长呈正相关线性关系,与其干组织重呈正相关的指数关系,而单位重量的生物沉积则与壳长和干组织重分别为负相关的线性和幂指数关系.海水温度和环境中饵料数量是影响栉孔扇贝的生物沉积的重要因子.     

丁雌核发育鱼的异源精子诱导及其与亲本的RAPD比较分析

鱼类人工雌核发育的研究始于20世纪50年代,在随后的50年里发展迅速。雌核发育可以为纯系建立、性别控制、基因定位等研究提供一种有效途径,乃鱼类遗传育种研究工作活跃领域之一[1-3]。丁(Tincatinca)隶属于鲤科、雅罗鱼亚科、丁属,是一种广温性鱼类(0～37℃)。主要分布于欧洲,在我国仅分布于新疆额尔齐斯河流域[4]。由于丁雌鱼生长比雄鱼快,Linhart等[5]对其人工诱导雌核发育和雌核发育子代性逆转进行了研究。但到目前为止,与其他大多数鱼类雌核发育研究工作一样,尚未获得功能性雄鱼。本研究旨在对丁人工雌核发育条件进行探索,…     

丁(鱼岁)雌核发育鱼的异源精子诱导及其与亲本的RAPD比较分析

鱼类人工雌核发育的研究始于20世纪50年代,在随后的50年里发展迅速.雌核发育可以为纯系建立、性别控制、基因定位等研究提供一种有效途径,乃鱼类遗传育种研究工作活跃领域之一[1-3].