Replacement of Dietary Fish Oil with Vegetable Oils Improves the Growth and Flesh Quality of Large Yellow Croaker(Larmichthys crocea)

We investigated the effect of the replacement of dietary fish oil with vegetable oils on the growth and flesh quality of large yellow croaker(Larmichthys crocea). The basal diet(FO) was formulated to contain 66.5% fish meal and 6.4% menhaden fish oil; whereas the other 3 experimental diets were formulated by replacing the fish oil with 50% soybean oil(SO50), 100% soybean oil(SO100) and 100% palm oil(PO100), respectively. The 4 diets were randomly assigned to 4 floating sea cages(3.0 m × 3.0 m × 3.0 m), and each was stocked with 250 fish individuals with an initial average weight of 245.29 g ± 7.45 g. The fish were fed to apparent satiation twice a day at 5:00 and 17:00, respectively, for 12 weeks. Experimental analysis showed that the specific growth rate of fish fed SO50 or PO100 were significantly higher than that of fish fed FO or SO100(P<0.05), and crude lipid contents of ventral muscle and viscera were significantly lower in fish fed FO than in those fed the other 3 diets(P<0.05). No significant differences in condition factor, viscerosomatic index, hepatosomatic index, gutted yield and colorimetric values of fish among the dietary treatments were observed(P>0.05). Compared to FO diet, SO50, SO100 and PO100 diets led to substantial decreases in the liquid loss and water loss from fresh fillets(1 d, 4℃)(P<0.05). Similarly, thiobarbituric acid reactive substance(TBARS) values of fillets under different storage conditions(1 d, 4℃; 7 d, 4℃; 4 weeks,-20℃; 8 weeks,-20℃) decreased significantly after partial or complete replacement of fish oil with vegetable oils. These findings indicated that the growth performance and selected flesh quality properties(liquid holding capacity and TBARS value) of large yellow croaker were substantially improved by replacing dietary fish oil with vegetable oils.

Establishment of a Spleen Cell Line from Large Yellow Croaker Pseudosciaena crocea and its Primitive Application in Foreign Gene Transfection

A large yellow croaker,Pseudosciaena crocea,spleen(LYCS)cell line was established and the feasibility of using it for foreign gene transfection was evaluaed in this study.Primary culture of LYCS cells was initiated from spleen tissue pieces,which were cultured at 25℃ in Dulbecco's modiced Eagle medium/F12 medium(DMEM/F12,1:1)(pH7.2),supplemented with 20% fetal bovine serum,carboxymethyl chitosan,chondroitin sulfate,basic fibroblast growth factor(bFGF)and insulin-like growth factor-I(IGF-I).The cultured LYCS cells,in fibroblast shape,proliferated to 100% confluency 20 days later.Chromosome analyses indicated that the LYCS cells exhibited chromosomal aneuploidy with a modal chromosome number of 48 which displayed the normal diploid karyotype of P.crocea(6m+6sm+36t,NF=60).A LYCS cell line,with a population doubling time of 48.7 h at passage 60,has been established and subcultured to passage 70.Transgenic feasibility test demonstrated that positive green fluorescence protein(GFP)expression was observed in LYCS cells after pcDNA3.1-GFP plasmid transfection.In conclusion,a continuous foreign gene trans-fection feasible LYCS cell line has been established successfully.The cell line might serve as a valuable tool for studies of transgenic breeding and has potential applications for different kinds of cytotechnological studies.

Effect of Dietary Olaquindox on the Growth of Large Yellow Croaker(Pseudosciaena crocea R.) and the Distribution of Its Residues in Fish Tissues

Olaquindox(OLA), one of quinoxaline-N, N-dioxides, has been put under ban. However it was used as a medicinal feed additive early; it promotes the growth of livestock and prevents them from dysentery and bacterial enteritis. In this study, we evaluated the effect of dietary OLA on the growth of large yellow croaker(Pseudosciaena crocea R.) and the histological distribution of OLA and its metabolite 3-methyl-quinoxaline-2-carboxylic acid(MQCA) in fish tissues. Four diets containing 0(control), 42.5, 89.5 and 277.2 mg kg-1 OLA, respectively, were formulated and tested, 3 cages(1.0 m × 1.0 m × 1.5 m) each diet and 100 juveniles(9.75 ± 0.35 g) each cage. The fish were fed to satiation twice a day at 05:00 am and 17:00 pm for 8 weeks. The survival rate of fish fed the diet containing 42.5 and 89.5 mg kg-1 OLA was significantly higher than that of fish fed the diet containing 0 and 277.2 mg kg-1 OLA(P < 0.05), while the weight gain rate of fish fed the diet containing 42.5 and 89.5 mg kg-1 OLA was significantly higher than that of fish fed the diet without OLA(control)(P<0.05), but similar to that of fish fed the diet with 277.2 mg kg-1 OLA. Fish fed the diet with 277.2 mg kg-1 OLA had the highest content of OLA and MQCA in liver(3.44 and 0.39 mg kg-1, respectively), skin(0.46 and 0.09 mg kg-1, respectively) and muscle(0.24 and 0.06 mg kg-1, respectively). In average, fish fed the diet containing OLA had the highest content of OLA and MQCA in liver which was followed by skin and muscle(P < 0.05), whereas OLA and MQCA were not detectable in control. Our findings demonstrated that OLA and MQCA accumulated in large yellow croaker when it was fed with the diet containing OLA, thus imposing a potential safety risk to human health.

Temperature Tolerance of Large Yellow Croaker,Pseudosciaena Crocea(Richardson)Associated with Summer Season

L ethal temperature tolerance was determined for about 8 cm, age 0 Pseudosciaena crocea using both slow heating and rapid transfer protocol. The acclimatization temperature was 28 ℃ with summer season, lethal temperature ( LT50 value ) of slow heating protocol ( CTMax ) was 35.0 ℃, and the upper and lower incipient lethal temperatures of rapid transfer protocol were 34.2 ℃ and 17.5 ℃ respectively.

Artificial induction of mito-gynogenetic diploids in large yellow croaker(Pseudosciaena crocea) by hydrostatic pressure

The present study investigated conditions for inducing mito-gynogenetic(endomitosis) diploids by hydrostatic pressure in the large yellow croaker Pseudosciaena crocea.In haploid control groups,the development of eggs was activated with ultraviolet radiated semen.All fry presented typical haploid syndrome in the haploid control groups,and were verified as haploids using cytometry.After hydrostatic pressure treatment,morphologically normal fry reappeared at different frequencies according to the intensity and time of pressure shock.Fry with normal appearance in the pressure treated groups were verified as gynogenetic double haploids(GDHs),containing only one allele from the female parent at all four diagnostic microsatellite loci.For a fixed duration of 3 min,the optimal intensity of blocking the first mitosis was determined to be 40 Mpa,which was similar to that of blocking the second meiosis.There was a "window" of starting time,from 36.1 min to 38.1 min post-insemination at 25.0±1.0°C,within which the production of GDHs was not significantly different.Maximum production of morphologically normal fries,9.36%±2.97% of developed eggs,was found when the eggs were shocked with hydrostatic pressure at 40 Mpa for 3 min,starting from 38.1 min post insemination at 25.0±1.0°C.

Diagnosis of iridovirus in large yellow croaker by PCR

A rapid and sensitive PCR-based method for the detection of the large yellow croaker iridovirus (LYCIV) is described, which involves the amplification of a 295 bp fragment of the LYCIV ATPase gene from DNA isolated from naturally infected fish spleen. Sequencing of LYCIV ATPase gene fragment showed it shared 100% nucleotide sequence homology with the corresponding region of the ATPase gene of red sea bream iridovirus (RSIV) and sea bass iridovirus (SBIV), suggesting that LYCIV was homologous with RSIV and SBIV at least in part of the gemone. The specificity and sensitivity of the PCR procedure were tested on the iridovirus-infected fishes, the expected fragment was detected from spleen DNA samples of infected fishes, whereas no fragments were amplified from healthy fish spleen DNA, white spot syndrome baculoviruses (WSBV) DNA and pseudorabies virus (PRV) DNA. Detection limit of this method was 10(-7) ng positive plasmid DNA containing target sequence, equal to about 100 virions. In the infected experiment, first positive detection (1/4) appeared at Day 3 post-infection, all fish (4/4) tested positive at Day 7, however obvious symptoms were observed at Day 8, so LYCIV infection could be detected prior to the appearance of obvious symptoms. These results indicate that this PCR method could be used for early, rapid and specific detection of LYCIV infection.

不同增重性能大黄鱼(Larimichthys crocea)幼鱼间体尺性状对体质量影响效果的差异

分析影响大黄鱼幼鱼同生群内速生子群和普通子群体质量的体尺形态性状组合差异,进而解构导致两者间增重机制差异的内在逻辑,对于指导大黄鱼幼鱼速生种质发掘与选育具重要价值。于象山西沪港海域,随机取样经板式网箱养殖3个月的3000尾大黄鱼同生群幼鱼并全数称量后,按体质量大小筛得速生子群[体质量取值居于前5%,范围(3.01~4.81 g),记为FG]和普通子群[体质量取值居中,出现率50%,范围(1.61~2.33)g,记为CG]。随机取FG和CG子群各30尾,逐尾测得体质量(BW)、体长(X_(1))、肛长(X_(2))、侧线长(X_(3))、尾柄高(X_(4))、腹鳍间距(X_(5))、头长(X6)和头宽(X7)后,采用多元分析方法定量研究了体尺性状对FG和CG子群体质量影响效果的差异。结果表明:(1)所涉各项生物学性状的测定值均呈FG>CG(P<0.05),且两者变异系数最大的性状均为X_(4),分别达30.07%和26.70%;(2)经主成分分析,提取到2个特征根值大于1的主成分,其中PC1主要影响变量为X_(1)、X_(2)和X_(3),PC_(2)主要影响变量仅为X_(4),两者的方差累计贡献率为83.285%;经判别分析筛留的X_(1),对两者的综合判别准确率为91.67%;(3)经通径分析,FG、CG子群被保留的体尺性状组合对BW的直接作用排序分别为X_(5)>X_(2)>X_(3)和X_(1)>X6>X_(4)>X7,间接作用分别为X_(2)>X_(3)>X_(5)和X7>X6>X_(1)>X_(4),它们的决定系数加和值和复相关指数均相同,分别为0.848和0.888;(4)经偏回归分析,获得了用以估算FG、CG子群BW的多元线性回归方程组。研究结果可为大黄鱼速生种质甄别与发掘提供参考。

Large yellow croaker(Larimichthys crocea)mitofusin 2 inhibits type I IFN responses by degrading MAVS via enhanced K48‑linked ubiquitination

In mammals,mitofusin 2(MFN2)is involved in mitochondrial fusion,and suppresses the virus-induced RIG-I-like receptor(RLR)signaling pathway.However,little is known about the function of MFN2 in non-mammalian species.In the present study,we cloned an MFN2 ortholog(LcMFN2)in large yellow croaker(Larimichthys crocea).Phylogenetic analysis showed that MFN2 emerged after the divergence of amphioxus and vertebrates.The protein sequences of MFN2 were well conserved from fsh to mammals.LcMFN2 was expressed in all the tissues/organs examined at diferent levels,and its expression was upregulated in response to poly(I:C)stimulation.Overexpression of LcMFN2 inhibited MAVS-induced type I interferon(IFN)promoter activation and antiviral gene expression.In contrast,knockdown of endogenous LcMFN2 enhanced poly(I:C)induced production of type I IFNs.Additionally,LcMFN2 enhanced K48-linked polyubiquitination of MAVS,promoting its degradation.Also,overexpression of LcMFN2 impaired the cellular antiviral response,as evidenced by the increased expression of viral genes and more severe cytopathic efects(CPE)in cells infected with spring viremia of carp virus(SVCV).These results indicated that LcMFN2 inhibited type I IFN response by degrading MAVS,suggesting its negative regulatory role in cellular antiviral response.Therefore,our study sheds a new light on the regulatory mechanisms of the cellular antiviral response in teleosts.

Effective CRISPR/Cas9-based genome editing in large yellow croaker (Larimichthys crocea)

The large yellow croaker (Larimichthys crocea) is an economically important marine species with the highest annual production among the farmed marine fishes in China. However, the aquaculture industry of this species is suffering from severe problems that include weakened disease resistance, decreased growth rate, and reduced meat quality due to frequent inbreeding. Genome editing, which has a huge potential for solving those problems by introducing favorable genetic changes, is not yet available for the large yellow croaker. Here, we pioneered the techniques of embryo microinjection and genome editing using the CRISPR/Cas9 system in this species. Recombinant plasmids encoding green fluorescent protein (GFP) were introduced into the fertilized eggs of L. crocea by microinjection before the chorion had hardened. A high survival rate (40%) and GFP-positive larvae rate (81.8%) were achieved, indicating that the microinjection technique in L. crocea was successfully established. On this basis, Cas9 mRNA and sgRNA targeting the tyrosinase a gene in L. crocea (Lc-tyra) were co-injected into fertilized eggs of L. crocea. Mutant individuals with insertion and deletion mutations of Lc-tyra were detected. These results indicated that the CRISPR/Cas9-based genome editing technology established herein could efficiently introduce mutations at a specific site in the L. crocea genome. This method provides the potential for genetic improvement and functional genomic study in this species. This is the first report on effective CRISPR/Cas9-based genome editing in L. crocea.

Synthesized acoustic signal characteristics of net-cage-cultured Large yellow croaker,Pseudosciaena crocea

This study performs the quantitative analysis and comparison to acoustic signal characteristics of Large yellow croaker （Pseudosciaena crocea） at two different ages. The sounds were recorded from the fishes in a net-cage. Two exponential oscillation functions are built to fit the acoustic signal of the fishes. The signal characteristic of the oscillation frequency and attenuation coefficient was described quantitatively. Simulation curves of the function could fit well acoustic signals. Both the average oscillation frequency and attenuation coefficient of the fitted signals from the 13-15-month-old fishes are lower than those from the 7-8-month-old fishes. The results suggest that the oscillation frdquency and attenuation coefficient of the acoustic signal flmction may be relevant to the physical process of sound production and age characteristics of Large yellow croaker. This study may be valuable for the acoustic application to the artificial culture of the species.

闽粤群和岱衢群养殖大黄鱼(Pseudosciaena crocea)及其杂交子代遗传差异的SSR分析

采用SSR方法进行了闽粤群(MY)和岱衢群(DQ)及其正交和反交大黄鱼的遗传差异的研究。结果表明,Shannon’s多样性指数为DQ群高于MY群,分别为2.14、2.04;反交群高于正交群分别为2.13、2.03;杂交群体中为2.24与其亲体群的相似(2.22)。亲体群间基因分化系数Fst值为0.020,低于杂交后代的0.026。AMOVA显示,亲体群和后代群基因差异均很低,且亲体群基因分化度低于和杂交后代群,Fst值分别为0.011、0.032。Nei遗传距离显示子代间遗传距离大与亲代代间的遗传距离分别为0.39、0.22。UPGMA聚类将其分为3组,其中DQ群与MY群遗传距离最近。可以认为养殖的DQ群与MY群大黄鱼间遗传背景差异较低,其中DQ族遗传背景较好,杂交后代群的基因多样性和基因差异性稍有提高,其中反交效果较好。

大黄鱼(Larimichthys crocea)补体调节因子CFH和CFHR2基因的分子特征及表达分析

补体是鱼类免疫系统重要的组分,具有识别和消除外来病原体,激活免疫细胞,调控获得性免疫等功能。在免疫组织中,补体调节因子大约占据了补体成分的二分之一,当受到外界病原刺激时会迅速活化补体成分,并且进一步聚合形成酶复合物发挥一系列免疫效应。CFH和CFHR2是补体替代途径重要的调节因子,对于补体系统正常运转必不可少。为此,本文测定了大黄鱼补体调节因子CFH和CFHR2基因的c DNA全序列,并对基因组织特异性表达和溶藻弧菌刺激后基因m RNA表达量的变化等方面进行了研究。CFH和CFHR2序列全长分别为1332 bp和1170 bp,分别编码443和389个氨基酸,N端信号肽序列分别为24和32个氨基酸。推导的氨基酸序列结构分析表明大黄鱼CFH和CFHR2基因具有RCA蛋白家族的典型特征,即含有多个保守的CCP结构(补体控制蛋白)。实时荧光定量PCR结果显示,CFH和CFHR2在健康大黄鱼的肝、脾、肾、肠、脑、胃、心和肌肉这8种组织中都有表达,其中肝脏的表达量显著高于其它几种组织。溶藻弧菌(Vibrio alginolyticus)侵染了健康的大黄鱼之后,CFH和CFHR2的m RNA表达量均明显上调,并且随着感染时间的变化呈现不同的上升趋势。结果表明补体调节因子m RNA表达量的变化与溶藻弧菌的侵染密切相关,表明了CFH和CFHR2可能在大黄鱼自身免疫机制中发挥重要的作用。

诱导性成熟雌性大黄鱼性逆转的初步研究

【目的】大黄鱼雌鱼生长显著快于雄鱼,全雌苗种受到养殖业者期待。用雄激素处理性腺未分化的遗传雌性鱼苗,使其发育成为生理性雄鱼(XX♂),进而与原系雌鱼(XX♀)杂交,是制备全雌鱼苗的常规方法,但存在周期长、大黄鱼苗经雄激素甲基睾酮(MT)处理后生长受到严重抑制等问题。若能直接诱导雌性成鱼性逆转成为生理性雄鱼则可避免上述问题。本研究旨在探索通过诱导性成熟雌性大黄鱼性逆转培育为生理性雄鱼的可能性和诱导方法,为高效制备生理性雄鱼开辟一条新途径。【方法】在MT中加入一定比例的PHTPP作为诱导药物,将混合药物用壳聚糖制成胶条或溶解于大豆油中制成缓释液,注射到大黄鱼成熟雌鱼的腹腔进行诱导性逆转实验,观察性腺的组织学变化,并检测血清中性激素浓度和性腺中雌激素受体基因表达量的变化。【结果】MT-PHTPPA-大豆油混合液的诱导效果优于壳聚糖-MT-PHTPP药条;用(40μg MT+4μg PHTPP)/kg鱼体质量的剂量腹腔注射MT-PHTPPA-大豆油混合液,持续处理80 d后,实验组大黄鱼的卵巢退化,形态变成与精巢相似,其中形成大量精原细胞,血清中雌二醇(E2)浓度显著下降、11-酮基睾酮(11-KT)浓度显著上升(P<0.05),性腺中E2受体Esr2基因的表达受到严重抑制。【结论】通过诱导性腺分化完成乃至发育成熟的雌性大黄鱼性逆转培育生理性雄鱼是可能的。

中国大黄鱼产业:生产、市场及展望

大黄鱼具有很高的食用价值、营养价值、药用价值和经济价值。进入21世纪,中国常年生产大黄鱼的捕捞省份为沿海9省、养殖为沿海4省,主产捕捞省份为粤辽琼、养殖省份为闽浙粤。全国大黄鱼产量的13%来自捕捞、87%来自养殖。中国出口大黄鱼商品有3个品种,闽浙一直是大黄鱼主要出口省份。常年出口对象国家和地区为21个,其中中国香港、韩国和美国一直是主要出口市场,而中国香港和美国的市场拓展较明显,韩国市场显著缩小。同时,国内大黄鱼消费潜力较大,共同为大黄鱼产业发展奠定了市场根基。继而,从生产层面、内需层面和出口层面进行了大黄鱼产业的前景展望。

饲料中添加甘氨酸可提高大黄鱼(Larimichthys crocea)的抗氧化和抗应激能力

为研究饲料甘氨酸对大黄鱼(Larimichthys crocea)抗氧化和抗应激反应的影响,将初始体重为(130.35±8.37)g的大黄鱼随机分成6组,每组3个重复,每个重复(网箱)50尾鱼。在基础饲料中分别添加不同梯度的甘氨酸(0、0.6%、1.2%、2.4%、4.8%和6.0%),配制出甘氨酸实测含量分别为(1.58%、2.15%、2.75%、3.96%、6.33%和7.51%)的6种实验饲料。经过30 d养殖后,对大黄鱼进行拖网应激实验。结果表明,养殖实验结束后,饲料处理未对大黄鱼的存活和体成分产生显著影响。肝脏总抗氧化能力在饲料甘氨酸含量为2.75%时达到最大值,而丙二醛的含量在甘氨酸含量为3.96%时达到最小值,但与2.75%组无显著差异(P>0.05)。以肝脏总抗氧化能力为评价指标,根据二次曲线回归模型得出大黄鱼饲料中甘氨酸的适宜含量为3.57%。血清谷草转氨酶和谷丙转氨酶的活性均在饲料甘氨酸含量为2.75%时呈现最小值(P<0.05)。在拖网应激前,饲料甘氨酸含量为2.75%时,大黄鱼血清皮质醇的含量最高。在拖网应激后,该处理的大黄鱼血清皮质醇的变化幅度最小,而此时的血糖维持在较高水平。综合考虑大黄鱼的抗氧化和抗应激能力,本研究推荐大黄鱼饲料中甘氨酸的适宜含量为2.75%–3.57%。

大黄鱼(Larimichthys crocea)补体C3和C4基因的分子特征及表达分析

本文测定了大黄鱼C3(L.c-C3)和C4(L.c-C4)基因的c DNA全序列。结果表明,L.c-C3和L.cC4序列全长分别为4962bp和5088bp,分别编码1653和1695个氨基酸,N端信号肽序列分别为23和19个氨基酸。推导的氨基酸序列结构分析表明大黄鱼C3和C4与已报道的补体C3、C4同样都具有在功能上比较重要的残基以及保守的硫酯区。分子进化分析表明,L.c-C3和L.c-C4分别与鮸鱼C3、C4的氨基酸同源性最高。实时荧光定量PCR结果显示,L.c-C3和L.c-C4在健康大黄鱼的肝脏、脾脏、肠、鳃、心脏、脑、肌肉和胃这8种组织中都有表达,其中肝脏的表达量最高。在大黄鱼胚胎不同发育时期(从2细胞期到初生仔鱼)中,L.c-C3在各个阶段没有明显的变化,而L.c-C4的表达量有明显升高。溶藻弧菌(Vibrio alginolyticus)侵染的大黄鱼肝脏和脾脏中,L.c-C3和L.c-C4的m RNA表达量均明显上调。该结果表明,大黄鱼肝组织C3和C4基因表达变化与溶藻弧菌的侵染密切相关,揭示了C3和C4在大黄鱼抗细菌免疫反应中具有重要的作用。

不同饵料及养殖环境对大黄鱼(Larimichthys crocea)形态差异的影响

本研究通过采集不同环境和不同饲料养殖的岱衢洋大黄鱼(Larimichthys crocea)形态数据,利用聚类分析、主成分分析以及判别分析对3组大黄鱼形态差异进行分析。结果显示,不同环境对大黄鱼体高、尾长存在显著影响,而不同饲料对大黄鱼体高、尾高、尾长、体厚存在显著影响,同时对D3–4、D3–6、D4–6、D5–6框架形态存在显著影响,差异主要集中于躯干前部。表明不同环境下大黄鱼形态变化可能与流速适应相关,而饵料对体型的影响可能是因为营养组成及物质积累的不同。聚类分析显示,配合饲料组与深水网箱组形态更接近,而与冰鲜饲料相距较远。通过主成分分析提取3个主成分,其中配合饲料组与深水网箱组在主成分1上相近,但在主成分3上存在差异,而冰鲜饲料组在各主成分上均离散,表明小网箱冰鲜饲料养殖大黄鱼形态分离较大,体型一致性低。利用判别分析,构建3种条件下大黄鱼体型判别函数,经交互验证,判别函数与预期无显著差异,可用于不同养殖模式大黄鱼的形态判别。本研究初步表明,通过改变养殖环境水流、养殖空间大小、饲料可在一定程度实现养殖大黄鱼形态改良。

大黄鱼(Larimichthys crocea)CRFB13基因的克隆与表达分析

IFN-γ是哺乳动物的一个关键免疫调节因子,通过特异性地结合IFN-γ受体1(IFNGR1)和IFN-γ受体2(IFNGR2)组成的异源四聚受体复合物,来发挥其生物学功能.目前,在鱼类中已经克隆出IFNGR1基因的2个亚型:细胞因子受体家族B 13(CRFB13)和细胞因子受体家族B17(CRFB17).本研究从大黄鱼(Larimichthys crocea)中克隆得到了一个CRFB13基因(Lyc CRFB13),其开放阅读框全长1 158个核苷酸,编码由385个氨基酸组成的1个蛋白.Lyc CRFB13蛋白具有典型的II型细胞因子受体特征,包括1个信号肽、1个胞外的FN III-like结构域,1个单次跨膜结构域,以及胞内的JAK1和STAT1结合位点.氨基酸序列分析表明鱼类的CRFB13都具有JAK1和STAT1结合位点.系统进化分析表明鱼类的2种IFNGR1,CRFB13和CRFB17形成2个独立的分支,Lyc CRFB13和鱼类的CRFB13聚在一起,与雀鲷(Stegastes partitus)CRFB13的亲缘关系最近.组织分布显示Lyc CRFB13为组成型表达,在大黄鱼鳃中的表达量最高,而在小肠中的表达量最低.Poly(I:C)刺激后,大黄鱼鳃和头肾中CRFB13的mRNA转录水平都显著升高.此外,poly(I:C)和LPS还能够诱导大黄鱼头肾白细胞中Lyc CRFB13的表达,这些结果表明Lyc CRFB13可能在大黄鱼抗病毒、抗细菌免疫反应中发挥重要作用.

大黄鱼(Pseudosciaena crocea)leptin和cholecystokinin基因的克隆和表达特性研究

本文克隆了两种大黄鱼(Pseudosciaena crocea)摄食调控因子胆囊收缩素(CCK)、瘦素(LEP)基因的全长序列,并对其表达特性进行了研究。结果表明,克隆得到CCK基因属硬骨鱼类的CCK1亚家族,全长900nt,编码137个氨基酸,与其他鱼类的CCK1相比序列组成非常保守,特别是在20氨基酸的信号肽和8个氨基酸的CCK-8活性结构区域;而克隆得到LEP基因属硬骨鱼类的LEPA亚家族,全长1290nt,编码161个氨基酸,与其他鱼类LEP相比基因同源性较差,但在3D结构上却保留了LEP经典的4个α螺旋特征。两种因子在所检测的所有组织中均有表达,但尤以脑、胃、肠消化道、肝脏等摄食及能量平衡相关组织中表达活性最高。8天的饥饿能使大黄鱼脑、消化道组织的CCK和肝脏、脂肪组织中的LEP表达显著降低(P<0.05)。该结果说明CCK和LEP可能在大黄鱼的摄食生理和能量平衡中起着重要的调控作用;本研究将为深入了解鱼类食欲调控的神经内分泌机理提供基础。

Single nucleotide polymorphisms in intron 1 and intron 2 of Larimichthys crocea growth hormone gene are correlated with growth traits

The growth hormone gene (GH) affects animal growth and is a potential target for genetic studies of variation related to growth traits. In this study, we analyzed single nucleotide polymorphisms (SNPs) in GH intron regions and their associations with growth traits in large yellow croaker, Larimichthys crocea, from Zhejiang and Fujian stocks. The results of PCR-single strand conformation polymorphism showed two haplotypes of intron 1, named AA and AB genotypes, in Zhejiang stock. AB exhibited an SNP at position 196 (G A) that was negatively correlated with body height and positively correlated with standard length/body height (P 0.05). Two different genotypes, CC and CD, were identified in intron 2 in Fujian stock, with CD showing an SNP at position 692 (T C). The CD genotype had a significantly positive correlation with both weight and total length (P 0.01). These basic data highlight the potential for using GH as a genetic marker of fish growth in marker assisted selection.