Growth and mortality of small yellow croaker (Larimichthys polyactis) inhabiting Haizhou bay of China

Length frequency data of small yellow croaker(Larimichthys polyactis) were acquired from the survey vessel in May,July,September and December,2011 in Haizhou Bay of China.In this study,921 fish individuals were analyzed for the estimation of growth and mortality parameters.Between length and weight,the power coefficient b was 2.7321,2.9703,3.0418 and 2.7252 for the 4 surveying months,respectively.The estimated von Bertalanffy growth function parameters were 230mm(L ∞) and 0.26yr-1(K) as were calculated with ELEFAN method equipped in FiSAT computer package.With length-converted catch curve analysis,the total mortality rate(Z) and its 95% confidence interval were 2.16(1.69-2.64) yr-1,0.59(0.15-1.04) yr-1,1.16(0.80-1.52) yr-1 and 0.96(0.70-1.23) yr-1 for the 4 surveying months,respectively,with the pooled data the value was 1.15(0.81-1.48) yr-1.The natural mortality rate(M) was 0.516 yr-1 as was calculated with Pauly’s equation(the annual average sea water temperature was 11℃).Therefore,fish mortality rate was 0.634 yr-1.The yield-per-recruit analysis indicated that when t c was 1,F max was 0.7 and F 0.1 was 0.55.Currently,the age at first capture is about 1 year and F current was 0.634.Therefore,F current was larger than F 0.1 and less than F max.This indicates that current fish mortality is at a dangerously high level.With Gulland method,the biological reference point for fishery(F opt) was estimated as 0.516 yr-1,lower than current fish mortality.Accordingly,reducing catch in the region was strongly recommended.     

Isolation and characterization of polymorphic microsatellite loci in large yellow croaker, Larimichthys crocea

An (AC)n-microsatellite-enriched library for Larimichthys crocea was constructed in this study. Primers for fifty simple sequence repeat (SSR) loci were synthesized and genotyped on 30 L. crocea individuals from Guanjingyang wild population (WP) in Fujian Province and 38 individuals from Ningbo cultured population (CP) in Zhejiang Province. Only 21 loci were successfully amplified and polymorphic in two populations. In WP, the observed heterozygosity (H_O ) ranged from 0.233 to 0.900 and the expected heterozygosity (H_E ) ranged from 0.326 to 0.893, with an average of 7.8 alleles/locus, the polymorphism information content (PIC) ranged from 0.283 to 0.866 (mean 0.731). In CP, the H O ranged from 0.189 to 0.892 and the H E ranged from 0.333 to 0.800, with an average of 4.4 alleles/locus. The probability test showed significant departures from HWE in 9 and 2 of the 21 loci in WP and in CP, respectively. Deficiency of heterozygotes at four loci showed the presence of null alleles (P <0.01). The PIC of 20 microsatellite loci in WP were greater than 0.50. Overall, these novel markers are potentially useful for the study of population genetics, construction of genetic linkage and quantitative trait loci maps in large yellow croaker.     

大黄鱼β-actin 抗体的制备与组织表达谱分析

为探讨大黄鱼(Larimichthys crocea)β-actin基因(Lc Actb)作为内参基因进行相关研究的可行性,从转录和翻译水平研究其组织表达谱,作者以大黄鱼肝脏为材料,扩增Lc Actb的开放阅读框序列,并将其亚克隆至原核表达载体p ET32a(+)。酶切、测序结果表明,p ET32a-Lc Actb构建成功。将p ET32a-Lc Actb转化到大肠杆菌BL21(DE3)中进行优化表达,表达蛋白经纯化后进行Western blotting验证;原核表达结果表明,IPTG可诱导一个分子量约45 ku的特异蛋白,且主要以包涵体的形式存在,优化表达条件为:28℃、0.4mmol/L IPTG、200 r/min诱导表达5 h。用表达蛋白作为抗原,免疫新西兰白兔制备抗Lc Actb的多克隆抗体;用纯化的多抗进行Western blotting的结果表明,获得了特异性的Lc Actb抗体;通过实时荧光定量RT-PCR检测大黄鱼多种组织m RNA的表达,其次,制备了多种组织匀浆蛋白,使用纯化的抗体进行Western blotting分析,结果显示,Lc Actb在大黄鱼成体各组织中转录和翻译水平的表达差异均不显著,可做为研究大黄鱼成体组织中其他基因表达和翻译水平的内参标准。     

大黄鱼的声刺激行为研究

为了解水下强噪声对大黄鱼的影响,结合行为学方法开展了3个年龄的大黄鱼声刺激实验．结果发现：3个年龄的大黄鱼在水中声压约10Pa时均能对声波发生条件反应,但是,它们的声波敏感频率和直接致死的声压阈值差异较大;1个月幼苗和8个月小鱼的声波敏感频率分别为800Hz和600Hz,直接致死的声压阈值约为40Pa和4kPa．13个月大鱼的声波敏感频率也在600Hz,但当声压达到4kPa时,鱼群受惊吓明显,且未能直接致死．另外,这些曾经暴露在强声波中的各年龄段的大黄鱼在随后48h里较多出现相继死亡的现象．表明这些长时间暴露在水下噪声中的大黄鱼可能会因累积效应引起行为模式改变和间接致死等慢性危害．     

大黄鱼Lcnkl3基因的分子结构、表达特征及多肽片段抗菌活性

作为重要的免疫基因, NK-lysin参与了鱼类非特异性免疫系统对抗病原感染的免疫应答过程。在之前的研究中, 我们已经发现重要经济鱼类大黄鱼(Larimichthys crocea)体内存在2种类型的NK-lysin基因。在本文中我们报道了1种新型大黄鱼NK-lysin基因Lcnkl3, 并对基因序列、表达特征及其多肽片段的抗菌活性进行了分析。Lcnkl3基因的开放阅读框长度为435 bp, 编码144个氨基酸残基, 其多肽序列Lcnkl3中包含Saposin B结构域及6个高度保守的半胱氨酸残基。系统进化分析表明Lcnkl3与大黄鱼Lcnkl2最为相似, 与其他鱼类NK-lysin多肽则差异较大。Lcnkl3基因在未受感染的大黄鱼各组织中具有不同的基础表达量, 表达水平最高的组织为心脏。在病原诱导的免疫应答过程中, Lcnkl3基因在各组织不同感染阶段的表达模式存在差异。源于Lcnkl3成熟肽序列的多肽片段对不同细菌的抑制和杀灭效果差异显著。以上结果表明Lcnkl3属于大黄鱼NK-lysin基因家族, 并参与了大黄鱼对抗病原感染的免疫过程。