丁鱥雌核发育鱼的异源精子诱导及其与亲本的RAPD比较分析

Gynogenesis is thought to be a useful method to generate fully inbred line and to initiate monosex culture in teleost fish. This article presents results of a study of gynogenesis of tench, Tinca tinca L. from Eltrix River of Xinjiang, China, induced by the common carp ( Cyprinus carpio ) sperm inactivated with UV irradiation. When cold shocking method was used to prevent exmasion of the second polar body in order to produce gynogenetic diploids, the optimum treated parameters were screened as cold shocking for 20min at 4℃, 5min after insemination. On this condition, 10.23% of the treated eggs survived at feeding stage. A comparative study between the gynogenesis and their parents was made by RAPD technique. 82 random primers of 10 nucleotide long sequence were used in RAPD analysis. The results showed that each primer gives 3- 12 fragments for each sample with the fragment length 330 - 2 460 bp, and the genetic similarity between gynogenesis and their mother was 97.4 %, while the genetic similarity between gynogenesis and their father was 27.4%, and there were no bands only shared by gynogenesis and their father, the common carp. It revealed that the chromosome of their father, the common carp, did not penetrate into the construction of gynogenetic tench.     

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.     

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.     

复合有益菌剂对工厂化大菱鲆育苗水净化效果研究

On the basis of purifying effect of single strain probiotic bacteria on water of intensive turbot breeding, the purifying effect of multistrain bacteria product was studied. The results of experiments showed that the free multistrain mixed bacteria had better purifying effect on turbot breeding water than that of any singlestrain bacteria. this phenomenon can be explained with the principle of “ biopump”. The immobilized multistrain probiotics product, biofilm, also showed excellent purifying effect on wate r of turbot breeding. The removal rates of COD and ammonia in 5 days were up to 73.1% and 80.0%, respectively. The concentration of nitrite kept low, which indicated most ammonia had been transformed into nitrate and accumulation of harmful intermediate productnitrite was decreased. This paper supplied theoretical and technical foundation for saving sea water through biopurification of wastewater from marine fish culture.     

渔业统计——渔业管理者和科学家的一个重要工具

FISHERIES STATISTICS： AN IMPORTANT TOOL FOR MANAGERS AND SCIENTISTS This presentation focuses on the type of information collected through fisheries statistics that could be useful to scientists and fisheries managers. In Norway the use of fisheries statistics as an analyzing tool for marine scientists can be dated back to around 1973. At that time the Directorate of Fisheries started to computerize information from logbooks of the trawl fleet. The fisheries managers started to use fisheries statistics as an analyzing tool by the late 1970's. Once again the computerization of sales notes information together with additional statistical information, enabled the manager to regroup the information according to new problems to be analyzed and new questions to be answered.     

动植物蛋白源替代鱼粉研究进展

With the fast development of aquaculture, fish meal needs increased in recent years, however the quantity of fish catching decreases gradually. Fishmeal is a limited feed resource, and serious concem exists on the future availability of this feedstuff for incorporation in fish diets. Undoubtedly, fish meal is well recognized as the best dietary protein source for most marine carnivorous fishes which required high dietary protein levels compared to omnivorous or herbivorous fish. Fishmeal is known for their high content of essential amino acids and fatty acids, low carbohydrates, high digestibility, low levels of anti-nutritional factors (for fresh fish meal) and is a very good source of minerals and is highly palatable. Thus fish meal is in high demand as the protein source for many formulated diets. However, production of fish meal consumes approximately 35 % of the total global fish catch, and the increasing price and potentially unstable supply in the market could be limiting factors for marine fish culture. There have been strong efforts to define and develop cost-effective protein sources that can, at least partly, substitute for expensive high-quality fish meals in least-cost feed formulations. The search for fish meal substitutes and altemative dietary protein sources is an international research priority that could be of considerable economic advantages. Therefore it is urgent task to find animal and plant protein sources in place of fish meal. Among these, plant feedstuffs have received most attention in recent years, but due to their amino acid unbalances, .presence of anti-nutritional factors and low palatability, a high level of replacement of fish meal with plant feedstuffs in omnivorous fish is generally not well accepted. This paper reviews the research status for other protein sources replacing fish meal based on available information in the literature. Animal and plant protein sources nutrient values are evaluated from the aspect of digestibility, antinutrients, physiological status and suitable supplementation.     

鱼类热忍耐温度研究进展

The paper is a review of the studies on the temperature of the thermal tolerance in fishes. It was described from four aspect: (1) The key parameters of temperature of thermal tolerance in fishes; (2) The influence factors of temperature of thermal tolerance in fishes; (3) The research methods of thermal effect of fishes; (4) The evaluation of effects of thermal pollution on fishes, especially, with the coastal power station will be‘built in China recent years, the effects of thermal discharge water on the marine fisheries resource wered iscussed.     

我国赤潮频发现象分析与海藻栽培生物修复作用

In this paper, the history, main events and present status of red tide (HAB, harmful algal blooms) along China coast in recent years were reviewed and presented. It showed that the HAB's frequency and scale, number of HAB spec ies, percentage of toxic HAB events and the degree of damages to marine environment and economy have sharply increased in China since 1960's. Eutrophication was key factor for high occurrence of red tide. In this paper, main causes of frequent HAB occurrence along China coast was discussed. Many factors might influence the occurrence of red tide, which included weather, climate, coastal current, tidal current, water temperature, salinity, hydrodynamic and nutrient conditions, trace metals and the variation of biological environment. Numerous evidences from all over the world revealed the linkage between the increases in nutrient loading and the occurrences of high biomass blooms. Eutrophication was one of the important causes that involved in high occurrence of HAB. The main sources of nutrients potentially stimulating HABs included terrestrial runoff, aquaculture selfpollution, atmospheric deposition, sea projects and other pollution events in the ocean. Studies showed that the input from land contaminations and the selfpollution of marine aquaculture accelerated eutrophication in coastal waters and were also important impact factors on red tide. Researches suggested that nutrient composition could affect the species composition of phytoplankton as well as the development of some HABs. The changes in nutrient supply ratios, primarily N∶P, often resulted in shifts in red tide species composition. The correlation between cysts and formation of HAB was discussed from the viewpoi nt of transformation of cyst and vegetative cell, the effects of trace elements and other organic substances on the occurrence of HAB were presented also. It indicated that the nutrient control could be an effective way to reduce the risk of red tide occurrence. Seaweed would play an important role for decreasing marine eutrophication. Among the different methods of red tide controlling studied, seaweed biomass has received much attention due to the cost saving, low sensitivity to environmental and impurity factors, the possible contaminant recovery from the biomaterial and its elevated adsorption capacity. Cultivated seaweeds have very high rates of productivity higher than that of seaweed in its natural habits and grow well in water bodies with higher nitrogen and other nutrients. Seaweeds are able to absorb large quantities of nitrogen, phosphorus and carbon dioxide, produce large quanti ties of oxygen, and have excellent effect on decreasing eutrophication. Large amounts of C, N and P are accumulated into seaweed tissues as they accumulate considerable biomass over a period of months or years depending on the cultivation season. When seaweeds are harvested, nutrients are removed from the sea area. An investigation was carried out for inorganic nitrogen and inorganic phosphorus concentration at Lusi Coast, Qidong County, Jiangsu Province in China, where there were about 270 hm2 for Porphyra yezoensis cultivation with eutrophic sea water in recent years. While during Porphyra yezoensis cultivation, from Sep 2003 to May 2004, the concentration of ammonium nitrogen declined form 0.511-0.778 mg·L^-1 to 0.006-0.057 mg·L^-1, nitrite nitrogen concentration declined from 0.010-0.040 mg·L^-1 to 0.001-0.009 mg·L^-1, and nitrate nitrogen concentration declined from 0.466-0.549 mg·L^-1 to 0.286-0.0568 mg·L^-1, the average concentration of inorganic phosphorus declined from 0.024 mg·L^-1 to 0.019 mg·L^-1. Furthermore, during five hours, the concentration of ammonium nitrogen in the seawater declined form 220.88 μmol·L^-1 to 8.59 μmol·L^-1 by cultivated Gracilaria lemanaiformis, and the concentration of ammonium nitrogen declined form 213.84 μmol·L^-1 to zero by cultivated Enteromorpha clathrata. Other bioremediation mechanisms of seaweed inhibiting the red tide microalgae such as nutrients competition and allelopathic effects were also discussed.     

ABSTRACTS AND KEY WORDS

Influence of Sudden Changes of Temperature and Salinity on the Survival Rate of Larval Prawns Released into the Sea
Wang Yun-zhong et al. （1）
Abstract The experiments on the influence of sudden changes of temperature and salinity on the survival rate of larval Penaeus chinensis and Penaeus japonicus were carried out. The results indicated that the survival rate during 48 hours of P. chinensis was 100% when the salinity suddenly decreased 4 or increased 2 and the temperature decreased 6~C or increased 3 ~C, respectively; the 48 hours survival rate of P.japonicus was 100% when the salinity decreased 2 or increased 2and the temperature decreased 3 ~C or increased 6~C, respectively. The tolerance on the sudden change of salinity of P. chinensis was better than that ofP. japonicus; when the temperature decreased suddenly, the tolerance of P. chinensis was better than P. japonicus; it was just the reverse when the temperature increased suddenly.     

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.     

牙鲆雌核发育研究进展

牙鲆(Paralichthys olivaceus)是一种重要的海水经济鱼类,其雌性生长速度显著快于雄性。因此,生产全雌牙鲆对提高养殖产量和经济效益具有重要意义。雌核发育是一种特殊的有性生殖方式,在这种生殖方式中,灭活的精子进入卵子,激活胚胎发育,但精核不与卵核融合形成合子,所以雌核发育的个体只具有母本的遗传信息。目前,牙鲆已成为雌核发育技术应用于产业的代表性鱼类。本文对牙鲆人工诱导雌核发育的研究进展进行综述,介绍牙鲆减数分裂雌核发育和有丝分裂雌核发育的诱导方法,以及雌核发育技术在性别控制、克隆制备等育种研究中的应用。最后,作者提出"雌核发育育种体系"构想,并对其在鱼类育种和遗传研究中的应用前景进行展望。     

牙鲆三个同源纯系的生长和遗传性状比较

利用同一牙鲆(Paralichthys olivaceus)家系F09119的母本,建立了减数雌核发育系(F1324)、卵裂雌核发育系(F1346)及近交家系(F1313)3个纯系,对3个纯系群体的生长和遗传性状进行比较分析。在受精后85~388 d对3个家系的体长、体宽、体重及成活率进行测定比较,结果显示:F1346生长最快,388 d时体长、体宽和体重分别为(28.89±2.77)cm、(10.00±1.21)cm和(254.91±83.11)g,F1313次之,F1324最慢(P0.05)。388 d时F1324成活率最高(46.50%),F1313次之(38.00%),F1346最低(26.80%)。F1346、F1324和F1313平均等位基因数(Na)分别为2.0、2.0和2.35,平均有效等位基因数(Ne)分别为1.87、1.95和2.1,多态信息含量(PIC)分别为0.35、0.36和0.4,平均观测杂合度分别为0、0.8155和0.6366;卵裂雌核发育系和减数雌核发育系的遗传多样性明显低于近交系。平均期望纯合度F1313(0.503)F1324(0.5105)F1346(0.532);平均期望杂合度正好相反,F1313(0.4967)F1324(0.4894)F1346(0.4679)。卵裂雌核发育系纯合度是减数雌核发育系的1.0421倍,是近交系的1.0577倍;减数雌核发育系纯合度是近交系的1.0149倍。F1346与F1313之间的遗传距离最大(0.1568),遗传相似度最小(0.8549);F1346与F1324之间的遗传距离最小(0.0406),遗传相似度最大(0.9602)。本研究结果为牙鲆纯系构建及解析卵裂雌核发育系、减数雌核发育系和近交系的遗传区别提供了理论依据。     

3种染色体组加倍方法诱导黄河鲇雌核发育的比较

用经紫外线照射后遗传失活的黄河鲇精子、刺激性成熟的黄河鲇卵子, 进行雌核发育, 采用冷休克、热休克和秋水仙素浸泡等3种染色体组加倍方法诱导黄河鲇卵进行二倍体雌核发育。实验结果表明: 在人工诱导黄河鲇二倍体雌核发育过程中, 以冷休克处理组的效果最好, 胚胎孵化率达到8. 1%; 其次为秋水仙素处理组, 胚胎孵化率达到6. 3%; 热休克处理组诱导效果较差, 胚胎孵化率仅为4. 0%。     

Fast development of genetically uniform strains by gynogenesis in Japanese flounder (Paralichthys olivaceus)

The Japanese flounder is a major species in both aquaculture and research. Inbred strains of Japanese flounder were developed efficiently in our laboratory by meiotic and mitotic gynogenetic reproduction techniques. To determine the induction efficiency of gynogenesis and the rate at which full homozygosity is produced, six meiotic gynogenetic females (G1–G6) that had experienced meiotic gynogenesis once and three common females (C1–C3) were selected for production of mitotic gynogenesis in our experiments. Of the nine adult females, all six gynogenetic fish successfully produced viable offspring. However, only one of the three common fish did. Using microsatellite markers, we estimated the homozygosity of gynogenetic fish induced by mitotic gynogenesis. We found that the homozygosity of seven lines (C1, G1–G6) increased quickly, individually from 43%, 72%, 69%, 71%, 78%, 69% and 58% to 100%, 100%, 100%, 90%, 100%, 100% and 90% offspring complete homozygosity. Under mitotic gynogenesis, individuals with higher homozygosity had a higher induction rate. The G4 line showed the highest induction rate, achieving 44.59% convert hatching rate and 29.28% convert normality rate. We conclude that meiotic gynogenesis may be a feasible method to produce DH genetic material in Japanese flounder.     

Chromosome set manipulation and sex control in common carp: a review

The development of techniques for production of gynogenetic, androgenetic, polyploid, and monosex progenies in common carp (Cyprinus carpio L.) is described from a chronological perspective. Gynogenetic progenies were obtained either by suppression of the second meiotic division in eggs (meiotic gynogenesis) or by suppression of the first mitotic division in haploid embryos (mitotic gynogenesis). As a rule, gynogenetic progenies of common carp were all-female, revealing female homogamety (females—XX, males—XY) in this species. Induced gynogenesis results in increased homozygosity; the rate of increase depends on the type of gynogenesis. Inbreeding coefficient (F) for one generation of meiotic gynogenesis in common carp is about 0.6, while diploids obtained by mitotic gynogenesis are homozygous for all genes (F = 1.0). Mitotic gynogenesis was used for production of clones in common carp. In androgenetic progenies of common carp, YY males were identified, that after crossing with normal females (XX) produced all-male progenies. Triploids of common carp are characterized by a significant reduction in gonad development (especially ovaries). However, the reduction in gonad development did not result in an increase of somatic growth rate of fish. The procedure for androgen treatment to induce phenotypic sex reversal in genotypic females (XX) was elaborated. All-female progenies of common carp were produced on a large scale by crossing normal females (XX) with hormonally sex-reversed males (XX). Rearing of all-female progenies in conditions when fish normally reach sexual maturity before reaching of market size increased production yield by 7–8%. In a few cases distant hybridization resulted in polyploidy of fish without application of any physical treatment. The ability of hybrid females between crucian carp (Carassius auratus) and common carp to produce diploid (with unreduced chromosome number) gametes resulted in opportunities to produce triploid and tetraploid hybrid progenies.     

翘嘴鳜人工雌核发育的初步研究

采用紫外线照射灭活精子,经冷休克诱导染色体加倍的方法对翘嘴鳜(Siniperca chuatsi)进行人工诱导雌核发育育种。灭活翘嘴鳜精子,紫外线照射剂量80 mJ/cm2是最佳的辐射剂量。冷休克结果表明,在0～3℃冰水下,冷休克起始时间为授精后3～7 min均有效,以5 min为最适;冷休克持续时间10-50 min均有效,以30 min最佳。综合以上两项因素,授精5 min后,休克时间持续30 min组的原肠胚存活率和孵化率均为最高,与其他处理组差异显著(P<0.05),此时雌核发育率最高,达到4.72%。     

异源精子诱导犬齿牙鲆的雌核发育

用冷冻保存的花鲈精子作为异源精子,采用紫外线(UV)对精子进行照射使其遗传物质失活,然后与卵子进行"授精",可以刺激犬齿牙鲆鱼卵进行雌核发育,在受精后一定时间采用冷休克处理"授精"卵,抑制第二极体排出成功获得了犬齿牙鲆雌核发育二倍体鱼苗.实验表明:犬齿牙鲆同源精子经80 mJ/cm2的紫外线照射可以完全失活,冻存花鲈精子经紫外线照射后也同样具有诱导犬齿牙鲆雌核发育的能力.无论同源精子还是异源精子,最佳诱导条件为在18℃条件下,精子经80 mJ/cm2的紫外线照射,受精后4～5 min将受精卵放在3℃海水中进行冷休克处理,持续时间为45 min,同源精子和异源精子二倍体诱导分别为32.66%±7.03%和28.00%±6.48%.采用形态学、流式细胞仪DNA含量分析和微卫星标记技术对雌核发育鱼苗进行了分析,证明了雌核发育鱼苗为雌核发育二倍体.     

快速近交对团头鲂遗传结构的影响和近交效应的估算

利用RAPD技术定量分析了快速近交(全同胞近交,雌核发育)对团头鲂养育群体遗传结构的影响。经群体遗传学分析软件ARLEQUIN对RAPD图谱计算,在实测171个位点中,系统选育7代群体(F7)、全同胞近交4代群体(S4)及雌核发育2代群体(G2)的多态位点个数从大到小依次为F7(133)>S4(117)>G2(106);位点基因多样性从大到小依次为F7(0.30)>S4(0.28)>G2(0.24);根据这两个指标,一代雌核发育对基因纯合的影响力度为一代全同胞交配的3.3倍和6倍。3群体的群体内方差百分比为83.10%,群体间方差百分比为16.90%,固定性系数Fst(0.169)的p<0.01,遗传差异极显著,证明同选育群体相比,近交群体的基因型发生了明显异化。通过试验估算的近交系数值低于理论上的近交系数值,应是环境影响的存在和作用所致。     

七彩鲑雌核发育及其倍性研究

利用紫外线照射对七彩鲑(Salvelinus fontinalis)精子进行灭活处理,使其遗传物质失活,作为同源精子诱导源,与七彩鲑卵子进行受精,采用冷休克方法诱导雌核发育。结果表明:同源精子可以诱导七彩鲑的雌核发育,紫外照射强度为72 m J/cm2;距离为8 cm;冷休克温度为20℃。七彩鲑减数分裂雌核发育倍性测定以鸡(Gallus sp.)红细胞DNA含量(2.5 pg/N)为标准、七彩鲑普通育苗子一代为参照,采用流式细胞术和微卫星标记技术对30尾七彩鲑减数分裂雌核发育鱼苗进行分析,证明了雌核发育鱼苗为雌核发育二倍体。     

Artificial and natural cross breeding between Atlantic salmon and salmonids currently present in Japan

We evaluated the potential for hybridization between Atlantic salmon and salmonids currently present in Japan. We performed artificial and natural crosses and tested for hybridization using short interspersed elements. We documented the occurrence of hybrids and gynogenesis following artificial crossing. However, the survival rate of these offspring was low (<2 % to the fry stage). We were unable to obtain viable offspring from natural crosses of Atlantic salmon and chum salmon or masu salmon. Our results suggest that there is little risk of hybridization between Atlantic salmon and salmonids that are currently present in the wild in Japan. Furthermore, our results suggest that gynogenesis may occur naturally.