黄海中南部小黄鱼生物学特征的变化

国家重点基础研究规划项目（2006CB400608）;国家自然科学基金项目（30371104,40906086）;山东省泰山学者专项基金;中央级公益性科研院所基本科研业务费专项基金资助     

A panel of polymorphic EST-derived microsatellite loci for the small yellow croaker (Larimichthys polyactis)

Small yellow croaker (Larimichthys polyactis) is an important economic species of marine fishery. We developed and evaluated simple sequence repeat (SSR) markers from expressed sequence tags (ESTs) of Pseudosciaena crocea, Paralichthys olivaceus and Psetta maxima. Characteristics of nine EST–SSR loci were investigated using 46 L. polyactis individuals. The number of alleles per locus ranged from 2 to 6. The observed heterozygosity ranged from 0.0652 to 0.7391, while the expected heterozygosity ranged from 0.0638 to 0.7754. Seven loci departed from Hardy–Weinberg equilibrium (P < 0.01) significantly. These loci and markers will be useful for population genetics and systemic evolution among species of small yellow croaker.     

Development of microsatellite markers for the small yellow croaker Larimichthys polyactis (Sciaenidae) by cross-species amplification

The small yellow croaker (Larimichthys polyactis) is a highly valued fish for human consumption found in the Western Pacific that was considered endangered until recently because of overfishing. We selected microsatellite markers for this species from markers developed for Miichthys miiuy, also of the family Sciaenidae. Among 43 markers polymorphic for M. miiuy, 11 were found to be polymorphic for L. polyactis. Characterization of these 11 loci was made based on 30 L. polyactis individuals collected by trawling in the Zhoushan Fishing Ground, Zhejiang Province, China. Total genomic DNA was isolated from fin clips. The number of alleles per locus ranged from 4 to 10, with a mean of 5.82, while the effective number of alleles ranged from 1.64 to 10.00, with a mean of 3.22. Observed and expected heterozygosities ranged from 0.17 to 0.72 and from 0.39 to 0.81, respectively. Significant deviation from Hardy-Weinberg equilibrium was found at four loci, after applying Bonferroni's correction. There was no significant association between any of the pairs of microsatellite loci, hence allelic variation at these loci was considered independent. These 11 polymorphic microsatellite loci will be useful for genetic diversity analysis and molecular-assisted breeding for L. polyactis.     

Factors affecting morphological development of the sagittal otolith in juvenile and adult small yellow croaker (Larimichthys polyactis Bleeker, 1877)

The morphology and morphometrics of the sagittal otoliths of small yellow croaker (Larimichthys polyactis) from the southern Yellow Sea were investigated. Study objectives were to evaluate the shape variability and morphometric variables of sagittae of juveniles and adults as related to developmental changes and habit shift. A total of 152 fish were sampled from April to June of 2012 and 2013, along the coastal waters of the Lüsi spawning ground in the southern Yellow Sea. Changes in otolith shapes from the juvenile to the adult were presented with the rim development through the entire‐lobed‐entire transition and with the curvature of the cauda toward the ventral margin. The otolith elongation in the juvenile stage occurred at 10–20 mm standard length (SL) and was likely associated with the formation of otolith accessory growth centers from larvae to juvenile. The L. polyactis sagittal otoliths acquired their definitive shape at 130 mm SL maturity. Ontogeny on otolith shape might be related, for example, to the factors of metamorphic development, feeding habitat, and ambient water salinity, which varied throughout the growth of L. polyactis.     

黄海和东海小黄鱼遗传多样性的RAPD分析

小黄鱼（Pseudosciaena polyactis）是我国近海重要经济鱼类之一。本文分析了采自黄海和东海5个海区共计48个个体小黄鱼的随机扩增DNA多态性（RAPD）。从40个10 bp引物中选取20个用于群体遗传多样性分析,共检测出145个位点,其中132个（91.03%） 显多态性。用Shannon多样性指数量化的平均遗传多态度为1.93（1.50～2.44）,群体内和群体间的遗传变异比例分别为69%和31%;群体间的平均遗传相似度和遗传距离分别为0.9139和0.0861。用非加权配对算数平均法（UPGMA）聚类分析的结果表明,所分析的5个群体可分为3个地理群系,从分子水平上支持了过去有关学者把黄海和东海的小黄鱼划分为北、中、南3个地理群系的观点。     

A morphometric investigation of the small yellow croaker (Larimichthys polyactis Bleeker, 1877): evidence for subpopulations on the Chinese coast

In this study the morphometric characteristics based on the truss network system were constructed from the body shapes and used to investigate the population structure of the small yellow croaker (Larimichthys polyactis Bleeker, 1877). From April to June 2013, a total of 431 fish samples were collected from commercial catches acquired with bottom trawlers (20 × 20 mm) in seven main spawning grounds along the coast of China. The results of principal component and discriminant function analyses indicated a clear distinction among the regional stocks and within the Haizhou Bay (HZB) stock. Observed differences primarily involved head measurements, body length and tail size, which were associated with environmental factors. Latitudinal gradient of the stock distribution and the relationship among the stocks provided further evidence for the existence of subpopulations and the re‐division of the East China Sea group. Moreover, the temporal inter‐structure of the HZB stock is a potential reason for the contrast in results from previous studies. Additional thorough studies of migration trajectories are required for a conclusive and improved understanding of the population structure.     

Development and preliminary evaluation of a genomewide single nucleotide polymorphisms resource generated by RAD‐seq for the small yellow croaker (Larimichthys polyactis)

Recent advances in high‐throughput sequencing technologies have offered the possibility to generate genomewide sequence data to delineate previously unidentified genetic structure, obtain more accurate estimates of demographic parameters and to evaluate potential adaptive divergence. Here, we identified 27 556 single nucleotide polymorphisms for the small yellow croaker (Larimichthys polyactis) using restriction‐site‐associated DNA (RAD) sequencing of 24 individuals from two populations. Significant sources of genetic variation were identified, with an average nucleotide diversity (π) of 0.00105 ± 0.000425 across individuals, and long‐term effective population size was thus estimated to range between 26 172 and 261 716. According to the results, no differentiation between the two populations was detected based on the SNP data set of top quality score per contig or neutral loci. However, the two analysed populations were highly differentiated based on SNP data set of both top F_ST value per contig and the outlier SNPs. Moreover, local adaptation was highlighted by an F_ST‐based outlier tests implemented in LOSITAN and a total of 538 potentially locally selected SNPs were identified. blast2go annotation of contigs containing the outlier SNPs yielded hits for 37 (66%) of 56 significant blastx matches. Candidate genes for local adaptation constituted a wide array of biological functions, including cellular response to oxidative stress, actin filament binding, ion transmembrane transport and synapse assembly. The generated SNP resources in this study provided a valuable tool for future population genetics and genomics studies of L. polyactis.     

Isolation and characterization of 10 polymorphic microsatellite loci from small yellow croaker (Pseudosciaena polyactis)

Small yellow croaker (Pseudosciaena polyactis) is an economically important marine fish species. About 43 microsatellite loci were isolated from two enriched genomic library of Pseudosciaena polyactis. Ten of these loci were polymorphic in a test population with alleles per locus ranging from two to six, and observed and expected heterozygosities per locus from 0.3750 to 0.8750 and from 0.3112 to 0.8121, respectively. No loci significantly deviated from Hardy–Weinberg equilibrium after Bonferroni correction and no significant linkage disequilibrium was found between pairs of loci. These polymorphic microsatellite loci would be useful for investigating genetic diversity of Pseudosciaena polyactis.     

Climate‐induced habitat suitability changes intensify fishing impacts on the life history of large yellow croaker (Larimichthys crocea)

Intense fishing pressure and climate change are major threats to the fish population and coastal fisheries. Larimichthys crocea (large yellow croaker) is a long‐lived fish, which performs seasonal migrations from its spawning and nursery grounds along the coast of the East China Sea (ECS) to overwintering grounds offshore. This study used length‐based analysis and habitat suitability index (HSI) model to evaluate the current life‐history parameters and overwintering habitat suitability of L. crocea, respectively. We compared recent (2019) and historical (1971–1982) life‐history parameters and overwintering HSI to analyze the fishing pressure and climate change effects on the overall population and overwintering phase of L. crocea. The length‐based analysis indicated serious overfishing of L. crocea, characterized by reduced catch, size truncation, constrained distribution, and advanced maturation causing a recruitment bottleneck. The overwintering HSI modeling results indicated that climate change has led to decreased sea surface temperature during L. crocea overwintering phase over the last half‐century, which in turn led to area decrease and an offshore‐oriented shifting of optimal overwintering habitat of L. crocea. The fishing‐caused size truncation may have constrained the migratory ability, and distribution of L. crocea subsequently led to the mismatch of the optimal overwintering habitat against climate change background, namely habitat bottleneck. Hence, while heavy fishing was the major cause of L. crocea collapse, climate‐induced overwintering habitat suitability may have intensified the fishery collapse of L. crocea population. It is important for management to consider both overfishing and climate change issues when developing stock enhancement activities and policy regulations, particularly for migratory long‐lived fish that share a similar life history to L. crocea. Combined with China''s current restocking and stock enhancement initiatives, we propose recommendations for the future restocking of L. crocea in China.     

Regulation of adiponectin on lipid metabolism in large yellow croaker (Larimichthys crocea)

Adiponectin (APN), an adipose tissue-derived hormone, plays a key role in regulating energy metabolism in mammals. However, its physiological roles in teleosts remain poorly understood. In the present study, the apn gene was cloned from large yellow croaker, which was mainly expressed in the adipose, muscle and liver. Further studies showed that adaptor protein phosphotyrosine interaction PH domain and leucine zipper 1 (APPL1) was localized in the cytoplasm near the cell membrane and was directly bounded to adiponectin receptors (AdipoRs). Meanwhile, APN played a crucial role in lipid metabolism of primary muscle cells by promoting the synthesis, oxidation and transport of fatty acids, and the promoting effects were blocked by knockdown of appl1 and AdipoRs. Furthermore, the activation/inhibition of peroxisome proliferators activated receptor γ (PPARγ) enhanced/suppressed the APN-mediated lipid metabolism. Overall, results showed that APN mediated lipid metabolism through AdipoRs-APPL1 activated PPARγ and further regulated the synthesis, oxidation and transport of FA. This study will facilitate the investigation of APN functions in lipid metabolism and energy homeostasis and reveal the evolution of lipids utilization and energy homeostasis in vertebrates.     

Seasonal, diel and ontogenetic variation in feeding patterns of small yellow croaker in the central Yellow Sea

Stomach contents of 1603 small yellow croaker Pseudosciaena polyactis , sampled from seasonal bottom trawl surveys in the central Yellow Sea between March 2001 and January 2002, were examined. The results showed that small yellow croaker was a carnivorous predator and >30 prey species were identified from stomach contents analysis. Crustaceans (mainly euphausiids and decapods) were the most important prey, occurring in 93·1% of the stomachs containing food, and accounting for 77·6% of the total food by mass. Feeding activity was highest in autumn and lowest in spring and winter. Decapods were more important in summer, whereas euphausiids were more important during other seasons. Ontogenetic differences were found in the diet composition and feeding activity within the range of size (standard length, L _S) studied. The importance of fishes and decapods increased with L _S, whereas euphausiids, copepods and amphipods decreased in importance with L _S. Dietary breadth increased markedly for adults. A positive relationship was found between L _S and prey size. In each season the maximum diel feeding activity occurred at 0800 and 2400 hours, indicating that there was crepuscular and nocturnal feeding by small yellow croaker.     

Mitochondrial DNA variation and population genetic structure in the small yellow croaker at the coast of Yellow Sea and East China Sea

Small yellow croaker is one of the most important fishery species in China. The mass–scale artificial propagation of this fish species was initially developed in 2015 with the aim of facilitating the fish production stock enhancement and aquaculture programs in the future. In the present study, the wild broodfish and its corresponding progeny along with three other wild populations were sampled and subjected to sequence analysis of the mitochondrial cytochrome c oxidase subunit I gene. The genetic diversity and population genetic structure were evaluated with a total sample size of 141 individuals representing the populations of the Yellow Sea (Qingdao and Lvsi populations) and the East China Sea (Xiangshan and Ningde populations). The wild populations were characterized by high haplotype diversity (0.925–0.976) and low nucleotide diversity (0.376%–0.560%). The hierarchical analysis of molecular variance (AMOVA) analysis and the values of pairwise Ф-statistics (Ф_ST) indicated non-significant genetic differentiation among the four wild populations. The hatchery stock XSH exhibited lower indices of genetic diversity compared with the wild populations that could be attributed to the small effective population size. The findings of the present study have valuable insight to the sustainable management and utilization of this resource.     

黄海南部和东海小黄鱼(Larimichthys polyactis)产卵场分布及其环境特征

根据2003～2005年和2007年4月份在黄海南部和东海海域进行的底拖网调查数据,分析了小黄鱼产卵场分布和产卵场环境特征等.结果表明,目前小黄鱼产卵场范围较过去有扩大,范围已经扩展到外海海域,产卵场可分为黄海南部产卵场和东海产卵场;其中黄海南部产卵场主要集中在33°00′～34°00′N,122°30′～124°00′E,产卵场最适水温范围为9.65～12.17℃,最适盐度范围为32.25～34.54,最适水深范围为29.74～76.49m;东海产卵场主要集中在30°30′～31°00′N,124°00′～125°00′E海域和30°30′～32°30′N,125°00′～126°00′E海域,产卵场最适水温范围为10.13～16.64℃,最适盐度范围为32.50～34.37,最适水深范围为40.23～85.61m;黄海南部和东海产卵场水温分布差异极显著(P<0.01);小黄鱼产卵场较过去发生较大变化的主要原因可能是在过度捕捞等扰动因素的影响下,小黄鱼对环境适应性明显提高.     

Population structure of large yellow croaker (Larimichthys crocea) revealed by single nucleotide polymorphisms

The large yellow croaker, Larimichthys crocea, is one of the most commercially important marine species native to China Seas. In this study, Sequenom MassARRAY was used to analyze 120 individuals from three geographic populations in the East and South China Seas and a cultured stock. Forty-four of 60 single nucleotide polymorphism (SNP) loci were ascertained. Eleven loci, scf117, scf708, scf247, scf595, scf100, scf166, scf305, scf664, scf117, scf187 and scf511 each showed population-specific genotypes. A total of 127 alleles were detected. Averagely, the effective number of alleles was between 1.328 and 1.341, expected heterozygosity and observed heterozygosity ranged from 0.273 to 0.320, and 0.190 to 0.235, respectively, Pic value ranged from 0.215 to 0.247, and F_IS ranged from 0.174 to 0.279. The neighbor-joining tree based on calculated genetic distances showed a concordant topology with the traditional morphological characterization. When the population size of wild large yellow croaker drastically has shrunk over the past 20 years, the genetic diversity in both wild and farmed stocks has declined as well.     

应用混合模型和分位数回归分析东黄海小黄鱼肥满度空间异质性

基于2008-2010年黄海南部近海(SYS)、东海北部外海(NECS)和东海中部近海(MECS)小黄鱼体长和体质量数据,采用均值回归和分位数回归模型,解析了小黄鱼幼鱼和成鱼群体体长-体质量关系的空间变异.结果表明: 协方差模型和线性混合模型的残差标准误基本一致,线性模型残差标准误最高.从线性混合模型对特定区域和总体区域平均体质量计算的相对比值来看,SYS和NECS幼鱼群体的平均体质量高于总体平均值,但MECS低于总体平均值;成鱼群体则为NECS平均体质量高于总体平均值,MECS和SYS低于总体平均值.分位回归估计的肥满度和异速生长指数结果显示,幼鱼群体在不同分位的估计参数呈显著变化,SYS异速生长指数均值为2.85,在0.1~0.95分位的估计值变化范围为2.63~2.96.MECS和NECS参数估计值和置信区间在各分位数呈异质性变化,低分位时,NECS估计值在3个调查区域中最低,MECS最高;高分位时,MECS和NECS均高于SYS.对低分位0.25、中分位0.5和高分位0.75分位数的异速体长体质量关系的方差分析结果显示,低分位和高分位数之间体长 体质量关系极为显著(0.25∶0.75,F=6.38,df=1737,P<0.01),低分位数和中分位数之间为显著(0.25∶0.5,F=2.35,df=1737,P=0.039),中分位数和高分位数之间接近显著(0.5∶0.75,F=2.21,df=1737,P=0.051).成鱼群体SYS异速生长指数均值为3.01,在0.1~0.95分位的估计值变化范围为2.77~3.10.低分位和高分位数之间体长 体质量关系差异达到显著水平(0.25∶0.75,F=3.31,df=2793,P=0.01),低分位和中分位之间差异不显著(0.25∶0.5,F=0.98,df=2793,P=0.43),而高分位和中分位之间则差异极显著(0.5∶0.75,F=3.56,df=2793,P<0.01).     

Population Genetic Studies Revealed Local Adaptation in a High Gene-Flow Marine Fish,the Small Yellow Croaker (Larimichthys polyactis)

The genetic differentiation of many marine fish species is low. Yet local adaptation may be common in marine fish species as the vast and changing marine environment provides more chances for natural selection. Here, we used anonymous as well as known protein gene linked microsatellites and mitochondrial DNA to detect the population structure of the small yellow croaker (Larimichthys polyactis) in the Northwest Pacific marginal seas. Among these loci, we detected at least two microsatellites, anonymous H16 and HSP27 to be clearly under diversifying selection in outlier tests. Sequence cloning and analysis revealed that H16 was located in the intron of BAHCC1 gene. Landscape genetic analysis showed that H16 mutations were significantly associated with temperature, which further supported the diversifying selection at this locus. These marker types presented different patterns of population structure: (i) mitochondrial DNA phylogeny showed no evidence of genetic divergence and demonstrated only one glacial linage; (ii) population differentiation using putatively neutral microsatellites presented a pattern of high gene flow in the L. polyactis. In addition, several genetic barriers were identified; (iii) the population differentiation pattern revealed by loci under diversifying selection was rather different from that revealed by putatively neutral loci. The results above suggest local adaptation in the small yellow croaker. In summary, population genetic studies based on different marker types disentangle the effects of demographic history, migration, genetic drift and local adaptation on population structure and also provide valuable new insights for the design of management strategies in L. polyactis.