鱼类杂交育种及其在鲌属鱼类中的应用

对中国鲌鱼杂交研究的现状及其问题进行了介绍.重点阐述了鲌属鱼类杂交育种的研究现状、细胞遗传学、生化遗传特征及分子标记技术的应用和进展.     

“石斑鱼杂交育种技术”首次批量培育出石斑鱼新品种

2009年12月29日,由广东省大亚湾水产试验中心承担的广东省科技计划项目“石斑鱼杂交育种技术”项目顺利通过由广东省海洋与渔业局组织的专家验收。     

石斑鱼类的人工繁育技术

石斑鱼（Epinephalus）属鲈形目能科,是广泛分布于印度洋和太平洋的暖水性鱼类,在中国近海有31种,其中,南海有30种,东海8～9种,黄渤海仅1种。目前我国大陆沿海养殖的种类主要有：赤点石斑鱼、青石斑鱼、点带石斑鱼、斜带石斑鱼、鲑点石斑鱼、云纹石斑鱼、鞍带石斑鱼等。     

鱼类杂交育种的原理和应用

所谓杂交育种是人们将遗传性不同的生物体相互交配(或结合)使之产生优良杂种的过程。它已作为培育鱼类新品种的基本方法之一，即使在选择育种的许多情况下也往往要应用杂交育种的一些基本原理才能凑效。     

杂交育种技术在海水养殖贝类中的应用及研究进展

我国是世界上最大的水产养殖大国,同时也是海水养殖大国。2001年海水养殖产量达1.13×108t,其中贝类养殖产量占海水养殖总量的80%以上[1],贝类养殖在整个海水养殖业中占有十分重要的地位。但是长期以来,我国贝类只是进行单一的野生品种的养殖,很多养殖品种一直处于野生状态。长期近亲繁育、累代养殖,导致了近交衰退,种质逐年下降,表现为生长缓慢、经济性状下降、抗逆性降低等等。再加上病害的发生、养殖海区环境恶化等因素,致使近些年几种重要的养殖贝类均发生了大规模的死亡事件,造成巨大经济损失,其中仅1997、1998年的扇贝死亡,山东省…     

人工增殖放流石斑鱼类的标识技术研究

本文介绍了对平均体长5cm～8cm的石斑鱼苗进行背部挂线、尾部挂线、剪背鳍、剪尾鳍和尾部烙印5种标识手术和术后40d对比饲养观察结果。研究表明：①尾部烙印标志法具有对石斑鱼幼鱼的行为习性、生长、存活影响较小,标识脱标率低、保留时间长、标志醒目,操作简单的优点,是五种标识方法中结果最好的标识技术②标识操作的鱼苗规格以6．0cm～8．0cm为宜;③鱼苗在标识后暂养期以14d为宜。     

石斑鱼类养殖技术体系的创建和石斑鱼养殖产业持续发展的思考

石斑鱼是驰名世界的名贵海产鱼类,是中国南方沿海广东、海南、福建、广西等海水养殖业的主要对象,经济价值巨大。中国石斑鱼类养殖兴起于20世纪80、90年代;进入21世纪,中山大学的研究团队围绕石斑鱼苗种繁育和养殖产业化的理论和技术进行了一系列研究,对其生殖调控、生长发育、营养需求、病害防治、种质分析、养殖技术和养殖模式等方面进行集成创新,建立苗种繁育和成鱼养殖各个环节的技术规范和管理措施,实现了石斑鱼苗种规模化繁育和自养自足,从而带动石斑鱼养殖产业发展到相当的规模。石斑鱼类养殖产业化持续健康发展的总体思路是：以建设资源高效利用、改善生态环境、产品优质安全的现代渔业基本内涵为目标,改革和提升传统养殖技术和养殖模式,转变产业发展方式,不断优化和提升产业链上的各项技术,包括：（1）优化苗种人工繁育技术,提高苗种成活率和苗种质量;（2）培育抗逆、抗病、优质、高产的优良品种;（3）配制高效、优质的人工配合饲料;（4）建立病害检测和防控新技术和研制主要疾病的特异性疫苗;（5）建立高效、低碳、节能的养殖技术,因地制宜地采用先进的养殖模式;（6）加强石斑鱼类的生物学基础研究以及生物科学和工程技术的交叉融合、协同发展,为石斑鱼养殖产业的技术创新和集约化与工程化奠定基础。     

鲤鱼性状遗传及其在杂交育种上的应用

<正> 我们在鲤鱼杂交育种过程中作过一些遗传研究工作,现将我们积累的资料及其在选种上的分析应用介绍如下: 一鲤鱼体色和鳞被性状遗传(一)体色遗传观察与分析体色是划分鲤鱼品种和检验其纯度的一个重要指标。细胞学观察,不同鲤品种的体色是由于鱼体鳞片和皮肤含有不同的色素细胞及其数量分布差异性所致。如鲤、镜鲤含有大量的黑色素和黄色素细胞,而红色素细胞、银色反光组织较少,呈现一般鲤的体色     

杂交石斑鱼(斜带石斑鱼♀×赤点石斑鱼♂)与其亲本形态性状比较研究

研究了杂交石斑鱼[斜带石斑鱼(Epinephelus coioides)♀×赤点石斑鱼(E.akaara)♂]与其亲本的生长差异。采用形态学测量方法,分别测定了3种石斑鱼体质量和7个形态性状,对各形态性状对体质量影响进行通径分析,并分析杂交子代杂种优势。结果显示,3种石斑鱼各形态性状与体质量之间均呈极显著相关(P0.01),斜带石斑鱼、赤点石斑鱼体质量与全长的相关性最高,相关系数分别为0.946、0.932,杂交子代体质量与尾柄高的相关性最高(0.955),与全长的相关系数为0.927;形态性状与体质量的多元线性回归方程中,斜带石斑鱼保留体长、体高、尾柄高和体厚,赤点石斑鱼保留全长、体高和尾柄高,杂交子代保留全长、体高、尾柄高和体厚。对杂交子代杂种优势进行分析发现,体质量平均杂种优势为3.43%,体长平均杂种优势为3.02%,杂交子代同时表现出正向超亲优势和负向超亲优势。该研究表明杂交子代(斜带石斑鱼♀×赤点石斑鱼♂)与斜带石斑鱼的差异要小于赤点石斑鱼,杂交子代形态性状更偏向于母本。     

Temporal genetic heterogeneity of juvenile orange-spotted grouper (Epinephelus coioides, Pisces: Serranidae)

Juveniles of orange-spotted grouper ( Epinephelus coioides ), a tropical serranid species, are heavily harvested for aquaculture seeds from nursing grounds in several Southeast Asian countries. Because juveniles of similar sizes are present in a nursery area throughout the year, we aimed to determine whether more than one genetically distinct population contributes to juvenile aggregations. We examined the temporal genetic heterogeneity of juvenile aggregations collected at four different times of the year at a nursery area in coastal waters of the Andaman Sea in Trang province, Thailand. Also, we examined the differences between these temporal samples and an outgroup collected from the Gulf of Thailand (Chantaburi). The genetic variation at six polymorphic microsatellite loci within each sample was moderate, with observed heterozygosities across all loci ranging from 0.551 to 0.629 and number of alleles per locus ranging from 7.0 to 8.33. Results indicated substantial genetic differences between the two geographically distant samples, Trang and Chantaburi (F_st=0.040–0.050, P <0.005), and between the July sample and the remaining samples from Trang (F_st=0.096–0.106, P <0.005). The observed temporal genetic heterogeneity of E. coioides juveniles may reflect high variability in the reproductive success of each spawning event and the existence of spatially isolated groups of spawners.     

Possible application of mibolerone for induced sex inversion of grouper Epinephelus coioides

SUMMARY: Thirty immature juvenile grouper Epinephelus coioides (19–168 g bodyweight, BW) were randomly stocked in four units 6 t tanks to determine if mibolerone can be used to induce sex inversion in groupers. After acclimatization and weaning to artificial feed, the feed given daily (4% BW/day) was supplemented with 0, 50, 100, and 200 μg mibolerone/kg feed for about 18 weeks. Thereafter, the hormone treatment was withdrawn and the experiment was terminated at Week 24. Ten fish were killed for gonad histology at stocking to serve as an initial control while about three to five fish were killed every 8 weeks. In general, ovaries of initial controls showed the presence of moderate stromal cells and gonia and few primary oocytes. At Weeks 8 and 16, ovaries of the control fish (0 μg/kg) were similar to that of the initial control except that primary oocytes increased at Week 24. Gonads of fish fed diets containing 100 and 200 μg/kg had none to moderate spermatocytes and few spermatids at Week 8 and 16, although spermatozoa were not observed, indicating that the fish were undergoing spermatogenesis. Spermatogenesis at 50 μg/kg was not as advanced since only few spermatocytes occurred at Weeks 8 followed by moderate gonia and no spermatocytes and spermatids at Week 16. However, the presence of few primary oocytes was observed when mibolerone was withdrawn suggesting that sex-inversed fish reverted back to a female condition. These results show that sex inversion in juvenile grouper can be induced by oral administration of mibolerone and may have possible application on mature females to produce functional males.     

Establishment and characterization of a new cell line derived from kidney of grouper, Epinephelus akaara (Temminck & Schlegel), susceptible to Singapore grouper iridovirus (SGIV)

A marine fish cell line derived from the kidney of red-spotted grouper, Epinephelus akaara, designated as EAGK was established and characterized. The EAGK cells multiplied well in Leibovitz's L-15 medium containing 10% foetal bovine serum at 25 °C and have been subcultured for more than 90 passages. Karyotyping, chromosomal typing and ribosomal RNA (rRNA) genotyping analysis revealed that EAGK had a modal diploid chromosome number of 82 and was a fibroblast cell line originated from grouper. A severe cytopathic effect was observed in EAGK cells incubated with Singapore grouper iridovirus (SGIV), but not with soft-shelled turtle iridovirus, viral nervous necrosis virus or spring viraemia of carp virus. SGIV replication was further confirmed by immunofluorescence, electron microscopy and virus titre determination. Bright fluorescence was observed after transfection with fluorescent protein reporter plasmids, indicating that EAGK cells can be used to identify gene functions in vitro. In addition, the cell organelles including mitochondria and endoplasm reticulum changed and aggregated around virus factories after SGIV infection, suggested that the EAGK cell line could be an important tool for investigation of iridovirus-host interactions.     

Utilization of glucose and starch by the grouper Epinephelus malabaricus at 23°C

ABSTRACT : The aim of the present study was to investigate carbohydrate utilization by the grouper Epinephelus malabaricus reared at 23°C. Two isoenergetic semipurified diets were prepared with two carbohydrate sources (glucose and starch). Each diet was fed to triplicate groups of grouper in a recirculating rearing system for 8 weeks. Water temperature was held constant by a thermal controller at 23 ± 1°C. Weight gain, feed efficiency (FE) and protein efficiency ratio (PER) of fish fed the starch diet were significantly higher ( P  < 0.05) than those of fish fed the glucose diet. Body lipid content of the starch-fed group of fish was higher than that of glucose-fed group of fish. Hepatic hexokinase and glucose-6-phosphate dehydrogenase activities were higher in fish fed the starch diet than fish fed the glucose diet. Fish fed the glucose diet had higher hepatic glucose-6-phosphatase activity than fish fed the starch diet. These results suggest that starch is better utilized by grouper than glucose when the water temperature is 23°C.     

Embryonic and larval development of honeycomb grouper Epinephelus merra Bloch

Spawning and successful rearing of larvae of honeycomb grouper Epinephelus merra Bloch 1793 upto juvenile stage was accomplished at the finfish hatchery of Mandapam Regional Centre of Central Marine Fisheries Research Institute during 2004. The fertilized eggs were free, spherical and buoyant with size ranging from 710 to 730 μm. Complete early embryonic development took place within 24–27 h and hatching occurred. The hatchlings measured 1.5 mm. Mouth opening (115 μm) appeared at 72 h when the larvae were 2.2 mm in size. Pectoral fin developed on the fifth day. Complete metamorphosis took place and by the 60th day the larvae transformed into juveniles (45 mm) and attained skin colouration and honeycomb pattern.     

Evaluation of genetic diversity of eight grouper species <Emphasis Type="Italic">Epinephelus</Emphasis> spp. based on microsatellite variations

ABSTRACT:   Genetic diversity among eight species of grouper, Epinephelus bleekeri , E. coioides , E. malabaricus , E. ongus , E. akaara , E. maculatus , E. merra and E. fuscoguttatus , was studied using six microsatellite loci, Em-01 *, Em-03 *, Em-07 *, Em-08 *, Em-10 *, and CA-07 *, with the aim of exploring the feasibility of using microsatellite data for species identification. The results showed high levels of genetic differentiation among species ( F _ST  = 0.4403 and R _ST  = 0.4954). Species identification based on fixed allelic differences was possible between E. coioides , E. ongus , and E. fuscoguttatus at Em-01 * and between E. fuscoguttatus and E. ongus at Em-08 *. Private alleles were found in all species, except for E. ongus . Pairwise F _ST ranged 0.238–0.578 ( P  < 0.008 Bonferroni correction), and Nei's genetic distance ranged 0.433–2.710. Size homoplasy was observed at Em-03 * 157 allele, which was characterized by a T–C transition at the 119th nucleotide site of PCR products. The genetic assignment test unambiguously assigned each individual to the correct species. Thus, this test can be used for species identification of unknown individuals when the multilocus genotypes of the six microsatellite loci are available. The phylogenetic (neighbor-joining) tree, which was constructed based on the genetic distance matrix, separated the eight grouper species into two main groups.     

Precocious sex change and spermatogenesis in the underyearling Malabar grouper Epinephelus malabaricus by androgen treatment

The Malabar grouper Epinephelus malabaricus is an important candidate species for commercial aquaculture in tropical and subtropical areas. In nature, this species requires more than 10 years to change sex from female to male and have active spermatogenic tissues in the testis. Thus, it is essential to find a means of producing sperm for seed production. This is the first report of artificial sex change in underyearling E. malabaricus . Female E. malabaricus with immature ovaries at 144 days post-hatch (DPH) were fed a diet with 17α-methyltestosterone (MT) at 50 μg g⁻¹ diet for 6 months. Sex change occurred in most of the treated fish, which had testis with all stages of spermatogenic germ cells including spermatozoa. In contrast, most of the control fish had immature ovaries. These results, which reveal that germ cells in the underyearling grouper have the ability to produce spermatozoa in response to exogenous androgen, demonstrate that sex change can be artificially induced during ovarian development.     

Requirements of vitamin C (L-ascorbyl-2-monophosphate-Mg and L-ascorbyl-2-monophosphate-Na) and its effects on immune responses of grouper, Epinephelus malabaricus

An 8‐week feeding trial was conducted to evaluate two vitamin C derivatives, L‐ascorbyl‐2‐monophosphate‐Mg (C2MP‐Mg) and L‐ascorbyl‐2‐monophosphate‐Na (C2MP‐Na), to satisfy the vitamin C requirement and to test their effects on the immune responses of juvenile grouper, Epinephelus malabaricus. C2MP‐Mg and C2MP‐Na were each supplemented at 20, 50, 80, 150, 250, and 400 mg kg^?1 diet in the basal diet providing of 7, 18, 31, 51, 93, 145 mg ascorbic acid (AA) equivalent of C2MP‐Mg kg^?1 diet and 4, 10, 17, 31, 47, 77 mg ascorbic acid (AA) equivalent of C2MP‐Na kg^?1 diet, respectively. Basal diet without AA supplementation was included as control. Each diet was fed to triplicate groups of grouper (mean initial weight 3.20 ± 0.05 g). Fish fed diets supplemented with either C2MP‐Mg or C2MP‐Na had significantly (P < 0.05) greater weight gain (WG), feed efficiency and survival than those fed the unsupplemented control diet. Liver ascorbate concentrations in fish generally increased as dietary C2MP‐Mg or C2MP‐Na supplementation level increased. Haemolytic complement activity was higher in fish fed diets supplemented with 92 mg AA equivalent of C2MP‐Mg kg^?1 or 10–17 mg AA equivalent of C2MP‐Na kg^?1 than fish fed the unsupplemented control diet. Lysozyme activity was higher in fish fed ≥51 mg AA equivalent of C2MP‐Mg kg^?1 or ≥47 mg AA equivalent of C2MP‐Na kg^?1 than fish fed the unsupplemented control diet. Analysis by broken‐line regression of WG indicated that the adequate dietary vitamin C concentration from each vitamin C derivative in growing grouper is 17.9 mg AA equivalent of 2MP‐Mg kg^?1 and 8.3 mg AA equivalent of C2MP‐Na kg^?1, and it also indicated that C2MP‐Mg is about 46% as effective as C2MP‐Na in meeting the vitamin C requirement of grouper.     

Requirements of vitamin C (l-ascorbyl-2-sulphate and l-ascorbyl-2-polyphosphate) and its effects on non-specific immune responses of grouper, Epinephelus malabaricus

An 8‐week feeding trial was conducted to determine two vitamin C derivatives, l ‐ascorbyl‐2‐sulphate (C2S) and l ‐ascorbyl‐2‐polyphosphate (C2PP), to satisfy vitamin C requirement and to test their effects on the non‐specific humoral immune responses of juvenile grouper, Epinephelus malabaricus. C2S and C2PP were each supplemented at 20, 50, 80, 150, 250 and 400 mg kg^?1 diet in the semi‐purified basal diet providing of 7, 16, 28, 55, 86, 142 mg ascorbic acid (AA) equivalent of C2S kg^?1 diet and 4, 9, 15, 31, 49, 75 mg AA equivalent of C2PP kg^?1 diet, respectively. Basal diet without AA supplemented was included as a control. Each diet was fed to triplicate groups of grouper (mean initial weight: 6.69 ± 0.07 g). Fish fed diets with ≥28 mg AA equivalent of C2S or ≥4 mg AA equivalent of C2PP kg^?1 had significantly (P < 0.05) greater weight gain (WG) than fish fed the unsupplemented control diet. Liver AA concentrations were higher in fish fed diets with ≥16 mg AA equivalent of C2S or ≥9 mg AA equivalent of C2PP kg^?1 than fish fed the control diet. Alternative pathway of complement activation (ACP) was higher in fish fed diets with ≥55 mg AA equivalent of C2S or ≥15 mg AA equivalent of C2PP kg^?1 than fish fed the control diet. Lysozyme activity was higher in fish fed ≥86 mg AA equivalent of C2S or ≥15 mg AA equivalent of C2PP kg^?1 than fish fed the control diet. Analysis by broken‐line regression of WG indicated that the adequate dietary vitamin C concentration from each vitamin C derivative in growing grouper is 46.2 mg AA equivalent of C2S kg^?1 diet and 17.8 mg AA equivalent of C2PP kg^?1 diet, and it also indicated that C2S is approximately 39% as effective as C2PP in meeting the vitamin C requirement for grouper. The data suggest that both C2S and C2PP supplementation support non‐specific immune responses of grouper.