团头鲂9个配套组合在2龄至3龄阶段的生长性能比较

为了解团头鲂(Megalobrama amblycephala)不同选育系的配套系育种潜力,筛选生长性能最佳的配套组合,对团头鲂3个配套选育系(A、B、C)进行了双列杂交,对其产生的9个配套组合在2龄至3龄阶段的生长性能进行了比较和分析。结果显示:9个配套组合在2龄阶段试验终末的体质量为512.14~751.01 g,特定增重率为0.391%/d~0.530%/d,相对增重率为280.89%~511.78%;9个配套组合在3龄阶段试验终末的体质量为1 150.9~1 307.1 g,特定增重率为0.158%/d~0.178%/d,相对增重率为81.32%~95.26%;组合7在2、3龄阶段的特定增重率和相对增重率均显著高于其它各组合(P0.05)。9个配套组合在整个试验期间(2龄至3龄阶段)的相对增重率为181.11%~301.44%,组合7的相对增重率比对照组(组合1)提高了9.69%,其它组合比对照组(组合1)下降了6.74%~34.10%。结果表明,组合7为生长性能最佳的配套组合,组合7的父本(团头鲂"浦江1号"选育系F10)和母本(雌核发育选育系F3♀)可分别作为配套系育种的最佳候选父系和母系。该结果为团头鲂配套选育系优良性状的聚合利用提供了科学依据。     

团头鲂优势配套系1龄至2龄阶段生长性能分析

为进一步验证团头鲂配套系间杂交子代的生长优势,以团头鲂"浦江1号"选育系F_(12)为对照组,通过开展同池养殖试验,分析了优势配套组合(优势组)——雌核发育群体F_4(♀)×"浦江1号"F_(11)(♂)子代在1龄至2龄阶段的生长性能。结果显示:在1龄阶段,优势组试验鱼的终末体质量比对照组高12.54%(P0.05),特定增重率比对照组高2.47%(P0.05);在2龄阶段,优势组试验鱼的相对增重率比对照组高8.97%(P0.05),特定增重率比对照组高4.62%(P0.05)。优势组在1龄和2龄阶段的体质量变异系数均低于对照组(P0.05)。与对照组相比,优势组在1龄阶段成活率略低,而在2龄阶段成活率略高(P0.05)。结果表明,团头鲂优势配套系在1龄和2龄阶段有明显的生长优势,但在整齐度和成活率方面与团头鲂"浦江1号"选育系F_(12)没有显著差异。     

团头鲂3个选育群体的选择压力分析

团头鲂选育群体基因组选择信号的检测有助于研究人工选育条件下基因组的进化机制,为团头鲂的进一步遗传改良提供依据。以团头鲂"浦江1号"选育奠基群体(F0)为对照组,以3个团头鲂选育群体为试验组,采用经典的Ewens-Watterson中性检验和基于3种算法(岛屿模型、分级岛屿模型、贝叶斯似然法)的FST-离群值点检验,在14个转录组微卫星位点上进行选择信号检测。结果显示,F0群体中所有位点均为中性位点,选育群体A在Mac927位点和Mac158位点受到了正向选择压力,选育群体B在Mac158位点受到了正向选择压力,选育群体C在Mac927位点和Mac158位点受到了正向选择压力。由此可见,自1985年起的连续世代人工选育已在团头鲂基因组中留下了可检测到的选择信号。3个选育群体均在Mac158位点检测到选择信号,表明3个选育群体受到的人工选择方向比较接近。     

团头鲂三个选育群体二龄阶段生长性状的配合力和遗传相关分析

为了解团头鲂不同选育群体的育种潜力和用途,对团头鲂3个选育群体(A、B、C)进行了双列杂交,对其产生的9个配套组合在二龄阶段体质量、体长和体高的配合力和遗传相关进行了分析。结果发现:(1)3个选育群体均存在极显著的一般配合力(P0.01),选育群体A的一般配合力(GCA)最高,选育群体C次之,选育群体B则为负向一般配合力,表明选育群体A和C有进一步直接选育潜力。(2)3个选育群体相互间均存在极显著的特殊配合力(P0.01),表明通过杂交可获得较好的利用效果。(3)体质量、体长和体高两两之间均存在极显著的表型相关和遗传相关(r0.899,P0.01),并存在极显著的加性和显性相关(r0.966,P0.01),有利于这些性状的共同选育。     

团头鲂3个选育群体遗传纯度的微卫星分析

为从基因层面了解团头鲂3个选育群体的遗传纯度,以团头鲂"浦江1号"选育奠基群体(F0)为对照组,采用14个多态性转录组微卫星标记评估了团头鲂3个选育群体的遗传纯度和遗传多样性。结果显示,3个选育群体在14个微卫星位点的平均观察纯合度为0.425 6~0.508 9,平均期望纯合度为0.2482~0.258 7,平均Shannon信息指数为1.597 1~1.682 5,群体内个体间的平均遗传距离为1.138 2~1.285 0,个体间的平均遗传一致度为0.2767~0.3204。3个选育群体的平均Shannon信息指数均高于F0群体,但差异不显著(P0.05)。遗传一致度和期望纯合度在3个选育群体间的变化趋势一致,即:A群体C群体B群体。群体间Nei氏标准遗传距离(DS)为0.180 8~0.798 9,A群体与B群体间遗传距离最小(DS=0.180 8),F0群体与C群体间遗传距离最大(DS=0.798 9)。总体而言,C群体遗传多样性水平最高,F0群体遗传多样性水平最低。对于3个选育群体而言,A群体的遗传纯度最高,B群体的遗传纯度最低。研究结果为选育群体的进一步选育纯化及种质资源聚合利用提供了科学依据。     

团头鲂3个选育群体遗传潜力的微卫星分析

为从遗传多样性的角度了解团头鲂(Megalobrama amblycephala)3个选育群体的遗传潜力,该研究以团头鲂"浦江1号"选育奠基群体(F_0)为对照组,采用14个多态性转录组微卫星标记评估了团头鲂3个选育群体的遗传多样性,分析其遗传潜力。结果显示,3个选育群体平均每个位点的等位基因数(A)为7.928 6~8.785 7,有效等位基因数(A_E)为4.409 4~4.878 4,观察杂合度(H_O)为0.491 1~0.574 4,期望杂合度(HE)为0.741 3~0.751 8,多态信息含量(PIC)为0.691 2~0.705 2,近交系数(FIS)为0.229~0.352。3个选育群体的遗传多样性水平(AE、HE)均高于F0群体,但不存在显著差异(P0.05)。3个选育群体的有效群体大小(N_e)为11.0~29.3,在近期可能经历过遗传瓶颈。3个选育群体间D_A、D_(SW)遗传距离分别为0.175 4~0.358 8、0.804 7~1.054 4。该结果表明,3个选育群体的遗传多样性较高,遗传潜力较大,但因有效群体数量较少和瓶颈效应的影响,存在杂合度下降和近交衰退的风险,今后需采取科学措施来保护选育群体的遗传潜力。     

采用标记方法比较不同群体团头鲂的生长情况

以电子标记和剪鳍标记方法分别对4个相同数量、不同群体的团头鲂鱼种进行标记,放入同一池塘进行养殖,并测定其初始及试验结束时的生长指标,以研究不同群体团头鲂生长性能差异.试验结果表明:团头鲂"浦江1号"3个群体间生长速度的差异不显著(P＞0.05),但均极显著大于普通鲂(P＜0.01);生长速度的顺序为:09z2鲂＞09z1鲂＞自产鲂＞普通鲂."浦江1号"3个群体间体长/体高的比值差异不显著(P＞0.05),但普通鲂体长/体高的比值极显著大于09z2鲂(P＜0.01),显著大于自产鲂和09z1鲂(P＜0.05).体型性状对团头鲂鱼种的生长有一定影响,体型优良的团头鲂生长优势明显.采用电子标记法和剪鳍标记法的试验结果一致,且这两种标记方法均对团头鲂鱼种的生长无显著影响.     

基于线粒体DNA COII基因序列团头鲂3个选育群体的遗传变异分析

为了从遗传多样性的角度评估团头鲂不同选育群体的选育潜力,以团头鲂"浦江1号"选育奠基群体(F_0)为对照组,采用线粒体DNA COII基因标记评估了团头鲂3个选育群体的遗传多样性,分析了它们的选育潜力。结果显示,在3个选育群体的72条序列中,共确定了15种单倍型,群体间存在6种共享单倍型,单倍型多样性(H)的范围为0.731 9～0.818 8,核苷酸多样性(π)的范围为0.001 108～0.001 765,平均核苷酸差异数(K)的范围为0.649～1.034,群体内核苷酸序列间平均遗传距离的范围为0.000 930～0.001 432,3个选育群体的遗传多样性水平(H、π、K)均高于F_0群体。3个选育群体间Kimura双参数遗传距离和遗传分化指数(F_(ST))的范围分别为0.001 249～0.001 450和0.012 34～0.124 50,A群体与C群体间成对F_(ST)值差异显著(P0.05),F_0群体与A群体间成对F_(ST)值差异极显著(P0.01)。结果表明,3个选育群体的遗传多样性较高,选育潜力较大;同时,选育群体间(A群体与C群体间)、选育群体与奠基群体间(A群体与F_0群体间)均存在显著的遗传分化,表明不同方向上的累代人工选育已在一定程度上改变了选育群体的遗传结构。     

中华鳖黄河群体选育F1-3 世代生长性能比较试验

为了解中华鳖黄河群体选育世代的养殖性能,对其3个选育世代F1、F2、F3从温室苗种饲养阶段到室外成鳖饲养阶段的生长性能进行了连续观测比较试验.结果显示:(1)个体日均增重:苗种阶段的顺序为F3≈F2＞F1,F2和F3极显著地优于F1(P＜0.01);成鳖阶段的顺序为F3＞F2＞F1,F3极显著地优于F1和F2(P＜0.01);(2)每平方米日均产量:苗种阶段的顺序为F3＞F2＞F1,F3和F2极显著地优于F1(P＜0.01);成鳖阶段的顺序为F3＞F2＞F1,F3极显著地优于F1和F2(P＜0.01);(3)体重变异系数:苗种阶段的顺序为F1＞F2＞F3;成鳖阶段的顺序为F2＞F1＞F3;F3生长最整齐;(4)裙边比:在成鳖阶段,裙边比的大小顺序为F3(18.1%)＞F2(17.79%)＞F1(17.47%).对以上4项指标的综合分析表明,黄河鳖选育群体的养殖性能逐代提高.     

团头鲂双向选育效应研究

报道１９８６￣１９９９年团头鲂正向选育和逆向选育的两种截然不同的效果。正向选育采用混合选育法,每代从鱼苗到亲鱼的总选择率约为万分之四,选留健壮的较大个体,第五代成鱼阶段的平均日增重率比对照系（原种）提高２９．１％,即选育效应每代平均为＋５．８％。逆向选育为全兄妹近交,后代随机留养,近交系第三代成鱼阶段的平均日增重率比对照系（原种）降低１６．６％,即选育效应每代平均为－５．５％;近交系每第四代当年苗种阶段畸形率高达２．８％,越冬成活率比对照系低２８．４％,比选育系低４８．３％;选育系的越冬成活率比对照系提高３８．４％。本研究证明,科学的良性选育可大大提高养殖性能;近亲交配可导致养殖性能的迅速降低,必须严加防范。     

Dietary histidine requirement of juvenile blunt snout bream (Megalobrama amblycephala)

An eight‐week feeding trial was conducted to determine the dietary histidine requirement of juvenile blunt snout bream. The results showed that final body weight, weight gain rate and specific growth rate significantly increased with increasing dietary histidine levels up to 9.9 g/kg (p < .05) and decreased gradually thereafter, while feed conversion ratio showed a converse trend. The survival rate, condition factor, viscerosomatic and hepatosomatic index were not significantly affected by the graded dietary histidine levels (p > .05). About 9.9 g/kg dietary histidine level significantly improved whole‐body protein and lowered moisture content. The contents of plasma total protein, cholesterol, urea and triglyceride levels were not significantly affected by dietary histidine levels. About 9.9 g/kg dietary histidine level significantly upregulated target of rapamycin and insulin‐like growth factor mRNA levels (p < .05), and 12.1 g/kg and 14.2 g/kg dietary histidine levels significantly upregulated eukaryotic translation initiation factor 4E‐binding protein 2 and ribosomal protein S6 kinase‐polypeptide 1 mRNA levels (p < .05). Based on second‐degree polynomial regression analysis of weight gain rate, and specific growth rate against dietary histidine levels, the dietary histidine requirement for juvenile blunt snout bream was estimated to be 11.2 g/kg of diet, corresponding to 36.1 g/kg of dietary protein.     

不同倍性团头鲂群体的线粒体DNA 分析

测定了来自5个不同倍性团头鲂（Megalobrama amblycephala）群体（4n-F1、正交3n、反交3n、异源3n、二倍体2n）57个个体的线粒体DNA控制区、细胞色素b基因、16S rRNA基因和COⅡ基因序列,通过对这4段基因序列的联合分析,研究了5个不同倍性团头鲂群体的遗传变异。这4个基因片段的长度分别为：控制区937bp,细胞色素b基因1140bp,16S rRNA基因片段564bp,COⅡ基因片段642bp。将4个基因片段合并为1条总长度为3283bp的片段：在这条3283bp片段中,T、C、A、G4种核苷酸的平均含量分别为27．18％、25．45％、30．71％、16．66％。57个个体中确定了55种单倍型,未发现群体间有共享的单倍型,五群体的单倍型多样度在0．9848-1．0000之间,显示不同倍性团头鲂群体内单倍型类型丰富。5个群体内各序列平均核苷酸差异数（目在8．000-24．273之间,核苷酸多样性指数（π）在0．2483％～0．7866％之间：群体间遗传距离范围为0．0039～0．0106,显示不同倍性团头鲂群体遗传多态性丰富。核苷酸多样性指数（π）、平均核苷酸差异数（K）和核苷酸序列间平均遗传距离在5个群体间的变化趋势一致,由大到小依次为反交3n、异源3n、正交3n、同源4n-F1、二倍体2n。而且,反交3n、异源3n、正交3n和同源4n-F1群体的核苷酸多样性指数（π）、平均核苷酸差异数（K）以及核苷酸序列间平均遗传距离均显著大于2n群体（P〈0．05）。结果表明,反交3n、异源3n、正交3n和同源4n-F1群体的遗传多样性水平显著高于2n群体。[中国水产科学,2008,15（2）：222-229]     

Dietary biotin requirement of juvenile blunt snout bream,Megalobrama amblycephala

This study was conducted to determine the optimal dietary biotin requirement of juvenile Megalobrama amblycephala. Quadruple groups of fish (initial average weight 2.01 ± 0.01 g) were fed thrice daily with six isonitrogenous and isoenergetic purified diets containing 0 (basal diet), 0.015, 0.049, 0.158, 0.624 and 2.49 mg kg^?1 biotin, respectively, for 8 weeks. Results showed that survival rate, final weight, feed intake, specific growth rate, protein efficiency ratio and nitrogen retention efficiency all increased significantly (P < 0.01) as dietary biotin levels increased from 0 to 0.049 mg kg^?1, whereas the opposite was true for feed conversion ratio. Dressout percentage, condition factor, hepatosomatic index, viscera/body ratio all showed no significant difference (P > 0.05) within the biotin range tested. Contrary to moisture content, whole‐body protein and lipid contents showed a positive correlation with dietary biotin levels. In addition, liver biotin content increased significantly (P < 0.05) with increasing dietary biotin levels up to 0.624 mg kg^?1. Hepatic pyruvate carboxylase (PC) and acetyl‐CoA carboxylase (ACC) activities both showed an increasing trend as dietary biotin levels increased. Based on the regression analysis of weight gain, hepatic PC and ACC activities, the optimal dietary biotin requirement of juvenile Megalobrama amblycephala is estimated to be 0.063, 0.071 and 0.075 mg kg^?1, respectively.     

Dietary isoleucine requirement of juvenile blunt snout bream,Megalobrama amblycephala

A 9‐week feeding trial was conducted to estimate the dietary isoleucine requirement of juvenile blunt snout bream. Six isonitrogenous and isoenergetic experimental diets were formulated to contain graded isoleucine levels ranging from 5.3 to 20.1 g kg^?1 dry diet. At the end of the experiment, weight gain (WG), specific growth rate (SGR), feed efficiency ratio (FER) and protein efficiency ratio (PER) increased with increasing dietary isoleucine level up to 11.1 g kg^?1 dry diet, and dietary isoleucine level above 14.2 g kg^?1 dry diet declined these performances. Dietary isoleucine levels (14.2 and 17.3 g kg^?1 dry diet) significantly improved whole‐body protein content, but decreased whole‐body lipid, plasma triglyceride and cholesterol contents. Significantly lower visceral fat index (VFI) in fish fed with 14.2 g kg^?1 dietary isoleucine was observed compared to those fed with deficient or excessive isoleucine. Dietary isoleucine supplementation significantly increased plasma isoleucine concentration, while plasma valine and leucine concentrations showed a reversed trend. Dietary isoleucine levels regulated the target of rapamycin (TOR) gene expression and improved plasma superoxide dismutase (SOD) activity in juvenile blunt snout bream. Based on second‐order polynomial regression model analysis of SGR and FER, the optimum dietary isoleucine requirement was estimated to be 13.8 g kg^?1 dry diet (40.6 g kg^?1 dietary protein) and 14.0 g kg^?1 dry diet (41.2 g kg^?1 dietary protein), respectively.     

湖北淤泥湖团头舫mtDNA限制性片段长度多态性的研究

用ＢａｍＨＩ、ＢｇｌＩ、ＣｌａＩ、ＥｃｏＲＩ、ＨｉｎｄⅢ、ＫｐｎＩ、ＰｓｔＩ，ＰｖｕⅡＳａｃⅡ、ＳａｃＩ、ＳｃａＩ、ＸｂａＩ，ＸｂａＩ和ＸｈｏＩ十四种限制性内切酶对来源于湖北淤泥湖的团头舫进行线粒体ＤＮＡ限制性片估长度多态性研究。初步表明ＤＮＡ上切点呈现不同程度的多态性。共检测到六种母集集团。发现一尾团头舫ｍｔＤＮＡ分子大小比普通型鱼ｍｔＤＮＡ小约０．７０ｋｂ。呈现出ｍｔＤＮＡ发子大小多态现象。     

Effects of dietary docosahexaenoic acid on growth performance,fatty acid profile and lipogenesis of blunt snout bream (Megalobrama amblycephala)

Six diets were designed to investigate the effects of dietary docosahexaenoic acid (22:6n‐3; DHA) levels (0.5, 1.3, 2.3, 4.2, 8.1 and 15.9 g/kg diets) on growth performance, fatty acid profile and expression of some lipogenesis‐related genes of blunt snout bream (Megalobrama amblycephala). Fish (average weight: 26.40 ± 0.11 g) were randomly fed one of six diets for 8 weeks. Results indicated that the final body weight (FBW) and specific growth rate (SGR) of fish fed 1.3 g/kg DHA were significantly higher than other groups except for the 2.3 g/kg DHA (p < .05). Compared with other groups, the number of lipid droplet clusters of the liver stained with oil red O in the 2.3 g/kg DHA group was the highest, which was consistent with the lipid contents of whole body and liver. The DHA proportion in liver and muscle significantly increased with the increasing dietary DHA levels (p < .05), which reflected fatty acid profiles of diets. The highest mRNA expressions of acetyl‐CoA carboxylase α (ACCα), fatty acid synthase (FAS) and sterol regulatory element‐binding protein‐1 (SREBP‐1) occurred in the 1.3 g/kg DHA group, followed by 2.3 g/kg DHA. In summary, the supplementation of 1.3–2.3 g/kg DHA could improve growth performance and lipogenesis, and the dietary DHA could improve DHA and PUFA proportion in liver and muscle.     

Dietary methionine requirement of juvenile blunt snout bream (Megalobrama amblycephala) at a constant dietary cystine level

A 9‐week feeding trial was conducted to investigate the dietary methionine requirement of juvenile Megalobrama amblycephala at a constant dietary cystine level. Six semipurified diets were formulated to contain graded dietary methionine levels from 3.9 to 15.4 g kg^?1 in about 2.5 g kg^?1 increments in the presence of 2.2 g kg^?1 cystine. Results showed that specific growth rate (SGR) and protein efficiency ratio (PER) significantly increased with increasing dietary methionine levels from 3.9 to 12.4 g kg^?1 and thereafter kept stable. Maximum protein productive value (PPV), nitrogen retention efficiency (NRE) and liver weight were observed in 8.5 g methionine kg^?1 diet. Protein contents in whole fish body were positively correlated with dietary methionine level, while lipid contents were negatively correlated with it. Morphological index and hepatic glutamate‐pyruvate transaminase (GPT) activities were independent of dietary methionine levels. However, dietary methionine supplementation significantly improved haematological parameters, plasma methionine and total essential amino acid contents and hepatic glutamate‐oxaloacetate transaminase (GOT) activities. Analysis of dose response using broken‐line regression on the basis of SGR and PPV versus dietary methionine level estimated the optimum dietary methionine requirements of juvenile M. amblycephala to be between 8.5 and 8.4 g kg^?1 of diet (25.0 and 24.7 g kg^?1 of protein) in the presence of 2.2 g kg^?1 cystine, respectively. Hence, the corresponding total sulphur amino acids requirements of this species were calculated to be 10.7 and 10.6 g kg^?1 of diet (31.5 and 31.2 g kg^?1 of dietary protein).     

果寡糖对急性氨氮应激下团头鲂非特异性免疫指标的影响

为研究氨氮应激条件下果寡糖对团头鲂(Megalobrama amblycephala)非特异免疫指标的影响,试验选择360尾初重为13.5 g的团头鲂随机分为3组,每组4个重复,每缸30尾鱼,对照组投喂基础日粮,试验组分别投喂果寡糖(FOS)添加量为0.4%和0.8%的日粮,饲养8周,养殖试验结束后,用10 mg/L的NH4Cl对24尾鱼进行氨氮应激试验。试验结果显示,氨氮应激前,0.4%FOS组的血液谷丙转氨酶(GPT)活性显著低于对照组和0.8%FOS组,在氨氮应激后,0.4%FOS组的GOT活性有降低的趋势,但仅在24 h时显著低于0.8%组,0.8%组和对照组之间并无显著差异;血清酸性磷酸酶、补体3(C3)、补体4(C4)、酚氧化酶都出现先升高后降低的趋势,分别在应激后的3、24、12、12 h达到最大值。此外,应激前0.4%FOS组补体C3、C4、酚氧化酶的活性都显著高于对照组,应激后0.4%FOS组C3在3 h处显著高于对照组,但是在其它时间点各组差异并不显著,果寡糖添加水平和采样时间对这些免疫指标都无显著的交互作用。结果表明饲料中添加0.4%的果寡糖能够提高团头鲂的免疫力指标活性,提高了团头鲂抗氨氮应激的能力。     

Establishment and characterization of a fin cell line from blunt snout bream, Megalobrama amblycephala

This study established and characterized a new cell line (MAF) from the fin of blunt snout bream (Megalobrama amblycephala), a freshwater fish cultivated in China. MAF cells proliferated well in medium 199 supplemented with 10 % fetal bovine serum at 28 °C and have been subcultured more than 95 times in almost a year. MAF cells were revived at 90–95 % viability after 3–6 months of storage in liquid nitrogen. Karyotyping indicated that the modal chromosome number of MAF cells was 48. The MAF cell line consisted predominantly of fibroblastic and epithelial-like cells from M. amblycephala, which was confirmed by immunofluorescence and mitochondrial 12s rRNA sequencing. Viral susceptibility tests showed that MAF cells were susceptible to infection by snakehead rhabdovirus, spring viremia carp virus, and channel catfish virus, which was demonstrated by the presence of cytopathic effect, high viral titers, and PCR products. Bacterial cytotoxicity studies showed that extracellular products from Aeromonas hydrophila were toxic to MAF cells. Cu²⁺ was also cytotoxic to MAF cells, and the 24-h IC₅₀ value was 144.48 μmol/l. When MAF cells were transfected with pEGFP-N1 plasmid, bright fluorescent signals were observed, and the transfection efficiency reached up to 5 %. These results suggest that the MAF cell line may provide a valuable tool for studying virus pathogenesis, as well as cytotoxicity testing and genetic manipulation studies.