真鲷与黑鲷杂交与多倍体育种系列研究-Ⅰ

真鲷属于鲷科,真鲷属,分布于我国四大海域。黑鲷属于鲷科,鲷属,我国沿海均有分布。真鲷与黑鲷杂交属于属间远缘杂交。本研究利用春季繁殖的真鲷种群与春季繁殖的黑鲷种群进行交配。用HCG与LHRH-A_2催产,网箱内自然受精。正交以真鲷为♀,黑鲷为♂;反交以黑鲷为♀,真鲷为♂。两种杂交组合的雌雄性比均为1:1.5。正交组合的受精率为73％,反交组合为67％。正交组合的孵化率为85％,反交组合为87％。该研究证明海水鱼属间远缘杂交可以成功,并为海水鱼杂交育种推广应用奠定了基础。     

黑鲷与灰鳍鲷杂交育种

黑鲷属于鲷科鲷属，广泛分布于我国沿海，灰鳍鲷同属于鲷科鲷属，在我国仅分布于南海。黑鲷与灰鳍鲷杂交属于种间杂交。本研究利用科季繁殖的黑鲷种群与灰鳍鲷种群进行酱。用HCG与LHRH－A2催产，网相内自然受精。正交以黑鲷为♀，灰鳍鲷为♂；反交以黑鲷为♂，灰鳍鲷为♀。两种杂交组合的雌雄比均为1：1.5。正交组合的受精率为83％，反交组合为86％，正交组合的孵化率为91％，反交为89％，该研究证明海水鲁种间杂交可以成功，并为海水鱼杂交育种推广应用奠定了基础。     

鲷科于类属间远缘杂交的发育和生长

鲷科鱼类的肉味鲜美 ,营养价值高 ,生长快 ,其主要养殖品种真鲷 (Ｐａｇｒｏｓｏｍｕｓｍａｊｏｒ)、黑鲷 (Ｓｐａｒｕｓｍａｃｒｏ ｃｅｐｈａｌｕｓ)、黄鳍鲷 (Ｓｐａｒｕｓｌａｔｕｓ)和平鲷 (Ｒｈａｂｄｏｓａｒｇｕｓｓａｒ ｂａ)的人工繁殖及种苗生产已获得成功[1,2 ] ,但未见其杂交育种成功的研究报道。这 4种鲷分属同科不同属。笔者从1 989年开始进行这 4种鲷的杂交育种生产试验 ,均成功孵化出杂种仔鱼 ,并获得全长 6 6～ 86ｍｍ的平鲷与黑鲷杂交后代幼鱼 6 .0万尾。为海水鱼的属间远缘杂交研究及杂交育种生产提供了理论和技术依据1…     

真鲷与黑鲷杂交繁育技术研究

通过对真鲷黑鲷杂交繁育,培育出一种能结合父母本性状,具有生长速度快、抗逆性强的杂交子一代。2006年3月从海区选购真鲷（Pagrosomusmajor）、黑鲷（Spraus macrocephalus）亲鱼,通过强化培育,性腺促熟,于4月进行干法人工授精。通过对真鲷♀×黑鲷8及黑鲷♀×真鲷♂获得杂交受精卵,结果真鲷♀×黑鲷♂受精率为87．4％,孵化率为85％,出苗率14．28％;黑鲷♀×真鲷♂受精率为92．1％,孵化率为90％,出苗率7．2％,该批次共获得正交反交杂交鱼苗32000尾。从杂交实际结果看,不管正交还是反交都比自交的结果差,说明它们之间存在生殖隔离,杂交繁育难度相当大,要达到规模生产。需要继续研究、试验。     

真鲷与黑鲷杂交繁育技术研究

2006年3月从海区选购真鲷、黑鲷亲鱼,通过强化培育,性腺促熟,于4月开展真鲷♀×黑鲷♂及黑鲷♀×真鲷♂,结果真鲷♀×黑鲷♂受精率为87.4%,孵化率为85%,出苗率14.28%;黑鲷♀×真鲷♂受精率为92.1%,孵化率为90%,出苗率7.2%,共获得正交反交杂交鱼苗32 000尾。     

黑鲷和真鲷自交及正反交子代生长性状和肌肉营养成分的分析比较

以黑鲷(Acanthopagrus schlegelii)和真鲷(Pagrus major)为繁育亲本,采用完全杂交配组方法,分别对黑鲷(BB)、真鲷(RR)自交与正反交后代(RB,Pagrus major♀×Acanthopagrus schlegelii♂;BR,Acanthopagrus schlegelii♀×Pagrus major♂)的生长性状和肌肉营养成分进行了分析比较。结果显示:养殖180 d时,黑鲷、真鲷自交及正反交后代的体质量绝对增长率的排序为正交>真鲷自交>反交>黑鲷自交,体质量特定增长率各组间无显著差异(P>0.05);养殖240 d时,正交组的体质量绝对增长率显著高于其他组(P<0.05),真鲷自交组与反交组无显著差异(P>0.05),黑鲷自交组最低(P<0.05)。4个群体鱼的粗蛋白质量分数在19.3%~22.0%,粗脂肪质量分数在0.9%~2.6%,其中真鲷自交组最低(0.9%),正交组最高(2.6%);4个群体鱼的必需氨基酸含量占氨基酸总量的40.16%~41.69%,必需氨基酸指数在82. 17%~87.36%,鲜味氨基酸占氨基酸总量的38.1%~39.0%,杂交组均高于自交组。研究表明:正交组生长速度最快,杂交后代的生长速度均快于黑鲷;4个群体鱼肌肉的蛋白质含量均较高,氨基酸种类齐全,就蛋白质营养水平而言,反交子代与黑鲷、真鲷相当,高于正交子代;就蛋白质的氨基酸水平而言,反交子代优于黑鲷和正交子代;杂交后代的鲜味氨基酸含量均优于黑鲷和真鲷,表现出一定的杂交优势。研究结果证明黑鲷、真鲷杂交能够提高后代的生长性能,改良肌肉品质,增强选育的综合效果。     

真鲷与黑鲷杂交繁育技术研究

2006年3月从海区选购真鲷、黑鲷亲鱼,通过强化培育,性腺促熟,于4月开展杂交试验,结果真鲷♀×黑鲷♂受精率为87.4%,孵化率为85%,出苗率14.28%;黑鲷♀×真鲷♂受精率为92.1%,孵化率为90%,出苗率7.2%,共获得正交反交杂交鱼苗32 000尾。     

用RAPD技术探讨5种鲷科鱼类的亲缘关系

用RAPD技术对鲷科鱼类的黑鲷、真鲷、二长棘鲷、平鲷和黄鳍鲷进行亲缘关系的研究。在事先优化的反应条件下，在使用的60个随机引物中，有29个引物扩增出清晰稳定的片段，共计200条，大小在200～2500 bp之间。5种鲷均有其特异性扩增片段，可作为种类鉴别的依据。根据MEGA2.1软件的UPGMA和NJ程序构建的分子系统树显示：同属鲷属的黄鳍鲷和黑 鲷的亲缘关系最近，其次是二长棘鲷和真鲷，平鲷与二长棘鲷和真鲷的亲缘关系较近，与黄鳍鲷和黑鲷较远,黑鲷和二长棘鲷的亲缘关系最远。两种聚类的结果相一致。研究结果与借助形态学和生化特征进行的传统系统分析具有一致性。在40个个体之间遗传距离矩阵中，最大值达到0.8916。多数的遗传距离指数在0.50～0.85之间。通过分析，认为RAPD技术对分析属级间的亲缘关系具有一定的适用性。     

4种鲷科鱼类的线粒体细胞色素b基因序列及分子系统学分析

从鲷科鱼类中黄猪鲷(Sparus latus)、黑鲷(S.macrocephalusM)、真鲷(Pagrosomus major)和平鲷(Rhabdosargus sarba)的线粒体DNA扩增出约430bp的细胞色素b(Cytb)基因，经Clastal X同源排序后得408bp序列，对该序列进行分析并结合GeneBank中黄鲷(Taius turnifrons)、灰鳍鲷(S.berda)、犁齿鲷(Evynnis japonica)和高背四长棘鲷(Argyrops spinifer)的该区段DNA序列比较，共发现126个核苷酸位点存在变异(30．7％)，序列中的转换大于颠换，碱基替换多发生在密码子的第3位点。黑鲷和灰鳍鲷亲缘关系最近，序列差异为3.8％；而黄鲷和平鲷的亲缘关系最远，序列差异为21．7％。用MEGA2．1软件中的NJ法构建的分子系统树，将上述8种类分为4组，其中黄鲷属、平鲷属和鲷属分类的结果与形态分类学的观点一致；而传统分类上把犁齿鲷、真鲷和高背四长棘鲷归为3个不同属，在本研究分子系统树中，这3个属却聚成一支。本研究目的在于从分子水平上阐明鲷科鱼类的分类与系统进化关系。     

真鲷与黑鲷及杂交F1耐受温度、生长比较

本文通过对真鲷、黑鲷及真鲷♀×黑鲷♂杂交的F1耐高温、低温的比较,杂交F1具有父母本优良性状,能安全度夏和过冬;通过二三者之间生长测量,真鲷和杂交F1的生长速度明显快于黑鲷,从体重增长比较杂交F．增重速度比黑鲷快30％以上,但低于真鲷。     

Preliminary Results on Rearing of Sparus Aurata × Pagrus Pagrus Hybrids. Performance Comparison With the Parental Species

Hybridization between gilthead sea bream, Sparus aurata, and red porgy, Pagrus pagrus, was undertaken and viable hybrids were produced by an artificial hormonal-induced cross. The hybrids, from the cross of female red porgy and male gilthead sea bream (Pp × Sa), survived through the yolk-sac larval stage. The hybrids produced from the cross of female gilthead sea bream and male red porgy (Sa × Pp) presented higher mortality in the larval stage (98.4%) compared to gilthead sea bream (80.0%) and red porgy (92.5%). Afterwards, the Sa × Pp hybrid was compared to the offspring of its parental species under the same rearing conditions (tanks of 500 l capacity, natural light, ambient temperature, self-feeding with commercial feed) in a 40–week experiment. In this comparative study, the Sa × Pp hybrid had intermediate mortality rate (6%), compared to gilthead sea bream (0%) and red porgy (21%). The growth profile of this hybrid resembled the slow growing phases of its parents (red porgy in warm period and gilthead sea bream in cold period). The feed conversion ratio between fish forms was not significantly different (1.54–1.84 in the Sa × Pp hybrid, 1.53–1.56 in the maternal species and 1.33–1.40 in the paternal one). The body weight variation in the Sa × Pp hybrid was quite high (31.7–37.9%), compared to that in gilthead sea bream (19.7–24.7%) and red porgy (18.0–31.3%). With respect to body coloration, the Sa × Pp hybrid was distinguishable and classified into groups of red porgy-like (55.1%), gilthead sea bream-like (40.6%), and intermediate forms (4.3%). The preliminary results of this study proved that the produced Sa × Pp hybrid had too low a performance for aquaculture use, but the interesting consequent hereditary potential could be a useful tool in fish hybridization practice.     

Prothrombin time, activated partial thromboplastin time and fibrinogen values in Mediterranean marine teleosts

Coagulation profiles for marine fish were determined with the standard global screening assays, the prothrombin time (PT) and activated partial thromboplastin time (APTT) using homologous tissue thromboplastin and phospholipid extracts to investigate species-specificity of brain thromboplastins, and to establish a base-line coagulation profile for marine fish of economical importance for Mediterranean aquaculture (sea bream, Sparus aurata, sea bass, Dicentrarchus labrax, red porgy, Pagrus pagrus, common dentex, Dentex dentex). Sea bass and sea bream displayed the shortest PT (9.0–15.0 s) and APTT (22.1–35.8 s) in comparison to the other species (PT: 24.8–37.2 s, APTT: 39.6–66.4 s). Red porgy displayed the lowest fibrinogen value (1.2–2.3 g l^–1), while no differences were found among the other species (2.2–2.8 g l^–1). It is concluded that the interaction of the tissue factor and plasma clotting factor is relatively species specific and that the use of homologous brain extracts is preferable for haemostasis study in marine fish.     

Comparison of iodine and glutaraldehyde as surface disinfectants for red porgy (Pagrus pagrus) and white sea bream (Diplodus sargus sargus) eggs

The efficacy of iodine and glutaraldehyde as fish egg surface disinfectants were assessed in red porgy (Pagrus pagrus) and white sea bream (Diplodus sargus sargus) eggs, two species of interest for Mediterranean aquaculture. Iodine was effective in reducing the bacterial load of the 1-day-old eggs when applied at 50 mg L⁻¹ for 5 min. The same concentration did not cause any significant change in hatching success or survival of the larvae for the first 5 days. Glutaraldehyde failed to reduce the bacterial load of the fish eggs at concentrations that were safe for the eggs (100 mg L⁻¹ for 5 min), as it had a significant effect in preventing hatching of the developed embryo. Disinfecting 0-day-old eggs with iodine resulted in a significant reduction of hatching percentage, while larval survival thereafter was unaffected. The results of the present study suggest that iodine may be an appropriate egg disinfectant for both red porgy and white sea bream.     

Self-feeding activity patterns in gilthead sea bream (Sparus aurata), red porgy (Pagrus pagrus) and their reciprocal hybrids

This study was conducted to monitor feeding activity of gilthead sea bream (Sa) and red porgy (Pp) and to investigate the hereditary effects on feeding in their hybrids (female Sa×male Pp, female Pp×male Sa), obtained after artificial crosses. All fish groups were held in 500-l tanks under ambient temperature (15–25°C) and light conditions (daylength from 9 h 30 min to 14 h 48 min). Fish (initial body weight 100–165 g) were fed by means of self-feeders, which were connected to a computer, recording fish feeding activity for a year. Results showed that there was a 24-h periodicity in all fish types over the experiment. In their daily feeding patterns, both parental species were diurnal: red porgies had a stable daily pattern over the experiment, while gilthead sea bream shifted their activity from a general daylight fluctuation in warm periods to an afternoon peak in cold periods. The Sa×Pp hybrids showed intermediate feeding patterns compared to their parents in warm periods and a pattern that was similar to the red porgy in cold periods, while the Pp×Sa hybrids had rather unchangeable daily feeding patterns, which were not related to the parental ones. The proportion of night feed demands increased in cold periods independent of fish group. The relationship between temperature and feed intake was species-dependent. Finally, this study indicated that feeding expression in hybrids was related to the combination of the parental genome, supporting our hypothesis for a hereditary influence on feeding activity in fish.     

Ontogeny of tolerance to and avoidance of ultraviolet radiation in red sea bream Pagrus major and black sea bream Acanthopagrus schlegeli

ABSTRACT:   Ontogenetic changes of tolerance to, and avoidance of, ultraviolet-B radiation (UV-B) were examined in red sea bream Pagrus major and black sea bream Acanthopagrus schlegeli . In the tolerance experiment, larvae and juveniles (age 13–46 days) were put in beakers, and were exposed to one of five different levels of UV-B radiation (1.8, 1.1, 0.2, 0.1, and 0 W/m²) for one hour. Their survival rates were calculated either 12 or 24 h later. In the avoidance experiment, fish (age 3–49 days) were put in a long experimental tank, half of which was covered with UV-blocking film and placed under two levels of UV-B radiation (1.1 and 0.2 W/m²), and their avoidance indices were calculated. Black sea bream had significantly better survival compared to red sea bream for most ages. Only black sea bream of ages 37 and 49 days showed significant avoidance of UV radiation under the higher level of UV-B, whereas both species did not show avoidance on any days at the lower level. The present results suggest that black sea bream are significantly better adapted to habitats with high UV-B radiation, than red sea bream, reflecting that back sea bream live in shallower waters through their early life stages.     

Ploidy level,morphological characteristics and performance of interploid hybrids by Megalobrama amblycephala♀× allo‐tetraploids ♂

Interploid hybrids were produced by mating allo‐tetraploid males, an interspecific tetraploidy induced by blunt snout bream (Megalobrama amblycephala) ♀× black bream (Megalobrama terminalis) ♂, with blunt snout bream females. The fertilization rate was high in the interploid hybrids. Although significant (P<0.05) mortalities were observed at hatching, swim‐up and 30‐day stages, respectively, the survival percentage of the interploid hybrids became stabilized after the 60‐day stage. In different interploid hybridization batches, 64–97% of juveniles were identified as 3n and 1–36% as 2n. Less than 2% of tetraploids were obtained in these interploid hybridization batches. The daily growth rate of the interploid 3n increased 7.5%, compared with that of diploid blunt snout breams during the 180‐day culture. Moreover, a method using seven morphological parameter ratios was set up with a relatively high classification accuracy of 93.8% to harmlessly discriminate interploid 3n from allo‐tetraploids, black bream and blunt snout bream.     

延迟投饵对真鲷,牙鲆仔鱼早期阶段摄食,存活及生长的影响

试验结果表明：（１）２日龄真鲷,牙鲆仔鱼初次摄食,在６￣７日龄和５￣６日龄,不能建立外源性营养的仔鱼分别进入ＰＮＲ期,而卵黄囊均在此前１￣２天已耗尽。混合营养期分别为３￣４天和１￣２天。（２）真鲷和牙鲆仔鱼在不同饥饰饿阶段初次摄食率的变化式型是：开始较低,此后逐步上升,高峰期出现在卵黄囊接近耗尽时,此后开始下降。记录到的最高初次摄食率分别为８６．７％和３５％。（３）牙鲆仔鱼体长的增长率随延迟投饵天     

Discard composition and release fate in the snapper and grouper commercial hook-and-line fishery in North Carolina, USA

Abstract  Minimum size limits may be ineffective for reef fishes because they often sustain barotrauma when caught from deep (>20 m) waters. A study was undertaken in conjunction with hook-and-line commercial fishermen to calculate discard percentages and evaluate potential release mortality of eight economically important species: black sea bass, Centropristis striata (Linnaeus), red grouper, Epinephelus morio (Valenciennes), snowy grouper, Epinephelus niveatus (Valenciennes), gag, Mycteroperca microlepis (Goode and Bean), scamp, Mycteroperca phenax (Jordan and Swain), vermilion snapper, Rhomboplites aurorubens (Cuvier), white grunt, Haemulon plumieri (Lacepède) and red porgy, Pagrus pagrus (Linnaeus). Fishing with baited hook and line occurred in 2004 and 2005 in Onslow Bay, NC, in waters 19–150 m deep. Sub-legal discard rates were 15% for vermilion snapper, 25% for red porgy, 7% for red grouper, 33% for gag, 35% for scamp and 12% for black sea bass. Although mortality based on post-release behaviour was relatively low, higher mortalities estimated from models incorporating hooking location and depth of capture suggest that minimum size limits may not provide the population benefits intended by management in the North Carolina reef fishery.     

Spawning season,spawning grounds,and egg production of red sea bream in Hiuchi-nada,Seto Inland Sea

The spawning season and grounds of red sea bream in Hiuchi-nada, the central part of the Seto Inland Sea, were described using a new method based on monoclonal antibodies for identifying Pagrus major eggs, and the daily egg production (standardized by the incubation time and survival rate) was estimated. At the peak of spawning (May), the ranges of sea temperature, salinity, and chlorophyll a where red sea bream eggs occurred were 14.8–17.4°C, 32.0–33.0, 0.5–4.4 μg/L, respectively. The main spawning grounds of the red sea bream were confirmed as being the areas near the Geiyo Islands, Misaki Peninsula, Saijyo, Niihama. The spatial spread of red sea bream eggs increased with the egg developmental stage. The ranges of daily egg production in 2005 and 2006 were 0.3–19.3 and 0.2–6.7 × 10⁹ eggs/day, respectively. In Hiuchi-nada, aquaculture farms are located close to the spawning grounds, and the potential spawning population from the aquaculture farms equaled or exceeded that of the estimated spawning population obtained by the egg production method. Red sea bream eggs in Hiuchi-nada might be produced by both wild and aquaculture-based spawning populations.     

活饵料中n-3高度不饱和脂肪酸对黑鲷仔稚鱼生长和存活的影响

采用乳化油直接添加法,用n－3高度不饱和脂肪酸（n－3HUFA）含量不等的4种乳化油分别强化轮虫、卤虫活饵料,培育4组黑鲷仔鱼和稚鱼,各自历时15d,结果表明,n－3HUFA对黑鲷仔鱼和稚鱼的生长和存活均有重要影响。在该条件下,轮虫体内n－3HUFA含量为0．233％（湿重计）,卤虫体内n－3HUFA含量为4．273％（湿重计）时,仔鱼和稚鱼达到最佳生长和成活率。