Comparative transcriptomics revealed the ecological trap effect of linearly polarized light on Oratosquilla oratoria.

Although polarized light can assist many animals in performing special visual tasks, current polarized light pollution (PLP) caused by urban construction has been shown to induce maladaptive behaviors of PL-sensitive animals and change ecological interactions. However, the underlying mechanisms remain unclear. Our previous work hypothesized that linearly polarized light (LPL) is an ecological trap for Oratosquilla oratoria, a common Stomatopoda species in the China Sea. Here we explored the underlying negative effects of artificially LPL on O. oratoria based on comparative transcriptomics. We identified 3616 differentially expressed genes (DEGs) in O. oratoria compound eyes continuous exposed to natural light (NL) and LPL scenarios. In comparison with the NL scenario, a total of 1972 up- and 1644 down- regulated genes were obtained from the O. oratoria compound eyes under LPL scenario, respectively. Furthermore, we performed functional annotation of those DEGs described above and identified 65 DEGs related to phototransduction, reproduction, immunity, and synapse. Based on the functional information, we suspected that continuous LPL exposure could block the light transmission, disrupt the reproductive process, and lead to the progressive failure of the immune response of O. oratoria. In conclusion, this study is the first to systematically describe the negative effects of artificial LPL exposure on O. oratoria at the genetic level, and it can improve the biological conservation theory behind PLP.

Full-length transcriptome reveals the circularly polarized light response-related molecular genetic characteristics of Oratosquilla oratoria.

The mantis shrimp is the only animal that can recognize circularly polarized light (CPL), but its molecular genetic characteristics are unclear. Multi-tissue level full-length (FL) transcriptome sequencing of Oratosquilla oratoria, a representative widely distributed mantis shrimp, was performed in the present study. We used comparative transcriptomics to explore the critical genes of O. oratoria selected by CPL and the GNß gene associated with CPL signal transduction was hypothesized to be positively selected. Furthermore, the FL transcriptomes of O. oratoria compound eyes under five light conditions were sequenced and used to detect alternative splicing (AS). The ASs associated with CPL recognition mainly occurred in the LWS, ARR and TRPC regions. The number of FL transcripts with AS events and annotation information also provided evidence that O. oratoria could recognize LCPL. Additionally, 51 sequences belonging to the LWS, UV and Peropsin gene families were identified based on conserved 7tm domains. The LWS, UV and Peropsin opsins have similar 3D structures with seven domains across the cell membrane and conserved KSLRTPSN, DRY, and QAKK motifs. In conclusion, these results are undoubtedly valuable for perfecting the vision theory of O. oratoria and other mantis shrimp.

Transcriptional responses of liver and spleen in Lota lota to polyriboinosinic polyribocytidylic acid.

Introduction: The cultured Lota lota can meet the market demand in the context of the decline of wild resources, but the disease in the high-density culture process also deserves attention. Therefore, understanding the immune regulation mechanisms of L. lota will be the basis for obtaining high benefits in artificial culture. Methods: To explore the viral response mechanism of L. lota, RNA-seq was applied to identify the transcriptomic changes of the liver and spleen in L. lota by poly (I:C) stress. Results: The DEGs (liver: 2186 to 3123; spleen 1542 to 2622) and up-regulated genes (liver: 1231 to 1776; spleen 769 to 1502) in the liver and spleen increased with the prolongation (12h to 48h) of poly (I:C)-stimulation time. This means L. lota needs to mobilize more functional genes in response to longer periods of poly (I:C)-stimulation. Despite the responses of L. lota to poly (I:C) showed tissue-specificity, we hypothesized that both liver and spleen of L. lota can respond to poly (I:C) challenge may be through promoting apoptosis of DNA-damaged cells, increasing the activity of immune-enhancing enzymes, and increasing energy supply based on DEGs annotation information. Conclusions: Our results demonstrate the transcriptional responses of L. lota to poly (I:C)-stimulation, and these data provide the first resource on the genetic regulation mechanisms of L. lota against viruses. Furthermore, the present study can provide basic information for the prevention of viral diseases in L. lota artificial culture process.

Intestinal microbiota analyses of five economic fishery resources in the South China Sea.

The investigation of intestinal microbiota can provide evidence for revealing the growth and development regulation, feeding habits, environmental adaptability and pollutant indication of marine organisms. To data, the intestinal microbiota of marine organisms in the South China Sea is relatively lacking. To supplement these information, we sequenced intestinal microbiota from five fishery resources (including Auxis rochei, A. thazard, Symplectoteuthis oualaniensis, Thunnus albacores, and Coryphaena equiselis) in the South China Sea using high-throughput Illumina sequencing technology. After filtering, a total of 18,706,729 reads were finally produced and then clustered into OTUs. The mean number of OTUs detected in A. rochei, A. thazard, C. equiselis, S. oualaniensis, and T. albacores was 127, 137, 52, 136, and 142, respectively. Although the Actinobacteria, Bacteroidetes, Cyanobacteria, Deferribacteres, Firmicutes, Proteobacteria, Spirochaetes, Tenericutes, [Thermi], and unclassified_Bacteria were the most abundant in the five species, Photobacterium is the most abundant microbiota. Meanwhile, intestinal microbiota showed species- and sampling sites- specificity, thus only 84 microbiota species were common to all species. Additionally, the potential functions of OTUs in the five species is mainly involved in the synthesis and metabolism of carbohydrate, amino acid, fatty acid and vitamin. This study can provide basic data for clarifying the diversity and species- specificity of intestinal microbiota of five species in the South China Sea, and help to improve the intestinal microbiota database of marine organisms.

Single-molecule Real-time (SMRT) Sequencing Facilitates Transcriptome Research and Genome Annotation of the Fish Sillago sinica.

As a newly described Sillaginidae species, Chinese sillago (Sillago sinica) needs a better understanding of gene annotation information. In this study, we reported the first full-length transcriptome data of S. sinica using the PacBio isoform sequencing Iso-seq and a description of transcriptome structure analysis. A total of 454,979 high-quality full-length transcripts were obtained by single-molecule real-time (SMRT) sequencing, which was corrected by Illumina sequencing data. After that, 66,948 non-redundant full-length transcripts were generated after mapping to the reference genome of S. sinica, including 49 fusion isoforms and 9,250 novel isoforms. 63,459 isoforms were successfully annotated by one of the Nr, Nt, SwissProt, Pfam, KOG, GO, and KEGG databases. Additionally, 30,987 alternative polyadenylation (APA) sites, 451,867 alternative splicing (AS) events, 21,928 long non-coding RNAs (lncRNAs) and 12,911 transcription factors (TFs) were identified. The full-length transcripts of S. sinica would provide a precious resource for characterizing the transcriptome of S. sinica and for the further study of gene function and regulatory mechanism of this species.

Whole-genome survey and phylogenetic analysis of Gadus macrocephalus.

Gadus macrocephalus (Pacific cod) is an economically important species on the northern coast of the Pacific. Although numerous studies on G. macrocephalus exist, there are few reports on its genomic data. Here, we used whole-genome sequencing data to elucidate the genomic characteristics and phylogenetic relationship of G. macrocephalus. From the 19-mer frequency distribution, the genome size was estimated to be 658.22 Mb. The heterozygosity, repetitive sequence content and GC content were approximately 0.62%, 27.50% and 44.73%, respectively. The draft genome sequences were initially assembled, yielding a total of 500,760 scaffolds (N50 = 3565 bp). A total of 789,860 microsatellite motifs were identified from the genomic data, and dinucleotide repeat was the most dominant simple sequence repeat motif. As a byproduct of whole-genome sequencing, the mitochondrial genome was assembled to investigate the evolutionary relationships between G. macrocephalus and its relatives. On the basis of 13 protein-coding gene sequences of the mitochondrial genome of Gadidae species, the maximum likelihood phylogenetic tree showed that complicated relationships and divergence times among Gadidae species. Demographic history analysis revealed changes in the G. macrocephalus population during the Pleistocene by using the pairwise sequentially Markovian coalescent model. These findings supplement the genomic data of G. macrocephalus, and make a valuable contribution to the whole-genome studies on G. macrocephalus.

Comparative transcriptome reveals the thermal stress response differences between Heilongjiang population and Xinjiang population of Lota lota.

Some cold-water fishes are particularly sensitive to the water temperature increasing caused by current global warming. However, the alterations in the physiology and behavior of infraspecific populations living in heterogeneous landscapes in response to water temperature increasing were significantly different. Consequently, understanding the impact of temperature increasing on different populations may be crucial for the conservation of cold-water fishes in the context of global warming. The burbot is the only freshwater specie in Gadiformes. To better understand the differences of different populations of burbot under similar thermal stress, Lota lota was selected as the research objects. Firstly, RNA-seq was applied to identify the transcriptomic responses of Heilongjiang population exposed to three temperature gradients (0 °C, 18 °C and 28 °C). Compared with 0 °C, 4216 and 12,657 genes were significantly differentially expressed at 18 °C and 28 °C, respectively. Meanwhile, 49 genes were significantly differentially expressed in three temperature pairs and these genes were presumed to involve in stress response process, immunologic process, reproductive process, development process, material metabolism process, signal transduction process, spermatogenesis process and cell apoptosis process. The response differences of two L. lota populations to similar thermal stress were compared and the results showed that they have different gene expression responses (the number of differentially expression genes and biological processes). The lower annual temperature of the Heilongjiang River might make it more sensitive to temperature increasing. Based on the comparative transcriptome analyses, 12 orthologous genes were considered as the potential regulators of L. lota preference for cold-water environment and these genes are potentially related to the immunologic process, reproductive process, development process, signal transduction process, and cell apoptosis process. Those results can provide basic information for the rational development of conservation strategies of different L. lota populations under the background of global warming.

Characterization and analysis of transcriptome complexity using SMRT-Seq combined with RNA-Seq for a better understanding of Acanthogobius ommaturus in response to temperature stress.

Acanthogobius ommaturus is a eurythermic fish, which is widely distributed in coastal, estuarine and bay waters of China, Japan and Korea. Due to the lack of whole genomic information, full-length transcriptome of A. ommaturus was firstly generated by single molecule real-time sequencing (SMRT-seq) in this study. A total of 49,833 full-length non-redundant transcripts (FLNRTs), 2255 alternative splices, 46,856 simple sequence repeats, 5094 long non-coding RNAs and 2708 transcription factors were obtained. Additionally, FLNRTs were used as reference sequences for the following transcriptome analysis of the temperature stress (7 °C, 14 °C, 21 °C (control), 28 °C and 35 °C). GO and KEGG enrichment analysis using GSEA were performed on all genes in 10 response modules which were screened out by WGCNA. Enrichment analysis showed that protein degradation, immune response and energy metabolism play an active role in the temperature stress of A. ommaturus. The differentially expressed hub genes (DEHGs) in response modules were closely related to adhesion, vascular remodeling and disease. The results of this study provided the first systematical full-length transcriptome profile of A. ommaturus and characterized its temperature stress responses, which will serve as the foundation for further exploring the molecular mechanism of the temperature stress in fish.

CsIL-11, a teleost interleukin-11, is involved in promoting phagocytosis and antibacterial immune defense.

Interleukin (IL)-11 is a multifunctional cytokine belonging to the IL-6 family, which plays essential roles in immune response. However, much less is known about the immunological functions of IL-11 in teleost. In this study, we investigated the immune properties of a teleost IL-11 homologue (CsIL-11) from tongue sole Cynoglossus semilaevis. CsIL-11 possesses four conserved α-helices and conserved CsIL-11 receptor binding residues L86 and R187, and shares 23.3%-80.1% identities with other IL-11 homologues. CsIL-11 expression was constitutive in tissues, with most abundant in blood and least abundant in spleen, and upregulated by bacterial challenge in blood, spleen, and head kidney. Recombinant CsIL-11 (rCsIL-11) in the native form of monomer, could bind to peripheral blood leukocytes (PBLs) membrane and enhance the activation and phagocytosis of PBLs. When administered in vivo, rCsIL-11 could markedly promote the host to defend against microbial infection. Overall, our findings show that CsIL-11 plays a pivotal role in regulating PBLs phagocytosis and antibacterial immunity.

New perspective on the genetic structure and habitat adaptation of Pampus minor off the coast of China based on RAD-seq.

Understanding the genetic diversity and habitat adaptation patterns of different geographic populations of marine organisms is essential for the rational maintenance, development, and utilization of their resources. Pampus minor Liu and Li 1998 is an economically valuable marine fish species. To determine the population genetic structure and elucidate the genetic mechanism underlying the habitat adaptation of this species, restriction site-associated DNA sequencing (RAD-seq) was used to scan the whole genomes of three P. minor putative populations along the coast of China for single-nucleotide polymorphism (SNPs) and outlier SNPs. Our population genetic structure analysis based on 2388 SNPs and 731 outlier SNPs throughout the genome revealed no significant genetic differentiation among the three populations. Results suggested that the life-cycle characteristics of P. minor, its relatively large population sizes, and ocean current transport might have shaped its current genetic pattern. The annotation information of both assembled sequences (which included outlier SNPs) and candidate loci associated with adaptations indicated that genes involved in many processes, including ion exchange, osmotic pressure regulation, metabolism, and the immune response, have been very important in the adaptations of P. minor to its heterogeneous habitats. Against the background of increased human activities, increased fishing intensity, and destruction of marine habitats, the results of this study provide basic genetic information for the accurate division of protection units of P. minor.

Identification of putative key genes for thermal adaptation in the Japanese mantis shrimp (Oratosquilla oratoria) through population genomic analysis.

Little is known about the mechanisms underlying the relationship between genetic variation and the adaptation of Oratosquilla oratoria populations to different habitat temperature. Here, the genome-wide genetic information of three O. oratoria populations were obtained by IIB restriction site-associated DNA (2b-RAD) sequencing and 2403 single-nucleotide polymorphisms (SNPs) were identified. Based on the 2403 SNPs, we found a remarkable genetic differentiation between the Yellow Sea and the East China Sea groups of O. oratoria. Furthermore, 63 SNPs are thought to be associated with different sea temperatures. Based on the 63 SNPs, it is hypothesised that the long-term temperature differences may contribute to the variation of genes associated with multiple biological functions, such as material metabolism, cytoskeleton, cellular processes, inflammatory response and hormonal regulation. This study provides new information for elucidating the molecular mechanisms underlying the relationship between genetic variation and the adaptation of Oratosquilla oratoria populations to different temperature.

Comprehensive phylogeny of Konosirus punctatus (Clupeiformes: Clupeidae) based on transcriptomic data.

Konosirus punctatus is an important species for the structure of marine ecosystems. Meanwhile, it is a native species in the northwest Pacific Ocean and supports important commercial fishery. In the present study, we generated the whole transcriptome of K. punctatus from combined tissues (muscle, liver, gill, heart, kidney, swim bladder and sexual gland) using Illumina RNA-seq technology and a total of 46087110 clean reads were obtained, corresponding to 6531521430 nucleotides. Meanwhile, 10000 clean reads were randomly selected and compared with NT database to examine the possible contamination. Results showed that 6754 clean reads were distributed among some species closely related with K. punctatus, indicating no-pollution. De novo assembly was performed and all clean reads were assembled to produce 71610 longest unigenes with an N50 of 906 bp. Among all the unigenes, 43974 unigenes were annotated in at least one database and 3172 unigenes were annotated in all databases. All unigenes were further analyzed to predict the gene structure and we have obtained a total of 54864 coding sequences (CDSs) and 17326 simple sequence repeats (SSRs). Saturation analyses were applied to evaluate the accuracy of gene expression and we hypothesized that the detection of gene expression might be effective. Finally, single-copy orthologous genes were applied to construct the phylogenetic relationship of K. punctatus. Results showed that K. punctatus diverged from the common ancestor with Alosa alosa, Alosa pseudoharengus and Sardina pilchardus at approx. 61.16-92.52 MYA. The present study will provide a foundational molecular information for the biological research of K. punctatus.

Genomic characteristics and profile of microsatellite primers for Acanthogobius ommaturus by genome survey sequencing.

Acanthogobius ommaturus is one of the suitable species to study the genetic mechanism of adaptive evolution, but there are few reports on its genetics. In the present study, the genomic survey sequencing method was used to analyze the genome characters of A. ommaturus. A total of 50.50 G high-quality sequence data were obtained in the present study. From the 19-mer distribution frequency, the estimated genome size was 928.01 Mb. The calculated sequence repeat rate was about 38.31%, the heterozygosity was approximately 0.17%, and the GC% content was approximately 40.88%. Moreover, 475,724 simple sequence repeats (SSRs) were identified. Among them, dinucleotide repeats were the most (53.70% of the total SSRs), followed by tri- (35.36%), hexa- (4.59%), tetra- (4.57%) and penta- (1.77%) nucleotide repeats type. This is the first genome-wide feature of this species to be reported.

Gill Transcriptome Sequencing and De Novo Annotation of Acanthogobius ommaturus in Response to Salinity Stress.

Acanthogobius ommaturus is a euryhaline fish widely distributed in coastal, bay and estuarine areas, showing a strong tolerance to salinity. In order to understand the mechanism of adaptation to salinity stress, RNA-seq was used to compare the transcriptome responses of Acanthogobius ommaturus to the changes of salinity. Four salinity gradients, 0 psu, 15 psu (control), 30 psu and 45 psu were set to conduct the experiment. In total, 131,225 unigenes were obtained from the gill tissue of A. ommaturus using the Illumina HiSeq 2000 platform (San Diego, USA). Compared with the gene expression profile of the control group, 572 differentially expressed genes (DEGs) were screened, with 150 at 0 psu, 170 at 30 psu, and 252 at 45 psu. Additionally, among these DEGs, Gene Ontology (GO) analysis indicated that binding, metabolic processes and cellular processes were significantly enriched. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways analysis detected 3, 5 and 8 pathways related to signal transduction, metabolism, digestive and endocrine systems at 0 psu, 30 psu and 45 psu, respectively. Based on GO enrichment analysis and manual literature searches, the results of the present study indicated that A. ommaturus mainly responded to energy metabolism, ion transport and signal transduction to resist the damage caused by salinity stress. Eight DEGs were randomly selected for further validation by quantitative real-time PCR (qRT-PCR) and the results were consistent with the RNA-seq data.

Comprehensive Transcriptome Analysis Reveals Insights into Phylogeny and Positively Selected Genes of Sillago Species.

Sillago species lives in the demersal environments and face multiple stressors, such as localized oxygen depletion, sulfide accumulation, and high turbidity. In this study, we performed transcriptome analyses of seven Sillago species to provide insights into the phylogeny and positively selected genes of this species. After de novo assembly, 82,024, 58,102, 63,807, 85,990, 102,185, 69,748, and 102,903 unigenes were generated from S. japonica, S. aeolus, S. sp.1, S. sihama, S. sp.2, S. parvisquamis, and S. sinica, respectively. Furthermore, 140 shared orthologous exon markers were identified and then applied to reconstruct the phylogenetic relationships of the seven Sillago species. The reconstructed phylogenetic structure was significantly congruent with the prevailing morphological and molecular biological view of Sillago species relationships. In addition, a total of 44 genes were identified to be positively selected, and these genes were potential participants in the stress response, material (carbohydrate, amino acid and lipid) and energy metabolism, growth and differentiation, embryogenesis, visual sense, and other biological processes. We suspected that these genes possibly allowed Sillago species to increase their ecological adaptation to multiple environmental stressors.

Single-molecule real-time (SMRT) sequencing facilitates Tachypleus tridentatus genome annotation.

Tachypleus tridentatus is a keystone species in marine ecosystems. Its hemolymph also provides the limulus amebocyte lysate (LAL) for detection of bacterial endotoxin in human medical service. Here we combined SMRT sequencing and Illumina RNA-seq to characterize the novel isoforms, novel genetic loci, fusion isoforms formation and transcriptome structure and further to unveil the transcriptome complexity of T. tridentatus. We identified 26,705 non-redundancy isoforms form 10,919 genetic loci, including 25,713 novel isoforms, 2403 novel genes and 170 fusion isoforms. In addition, 1578 novel genes and 23,172 novel isoforms were annotated in the NR, Pfam, KOG, COG, eggNOG, Swiss-Prot, KEGG and GO databases. Meanwhile, we have obtained 4671 gene family clustering based on genetic loci. Furthermore, there are 17,296, 4887, 1054, and 1435 APAs, AS events, lncRNAs, and TFs were identified in the T. tridentatus long-read transcriptome and the target genes of 1054 lncRNA sequences were also predicted. Overall, our work firstly provided the long-read transcriptome and these data are very necessary to improve the annotation information of T. tridentatus genome and optimize the boundaries of 12,342 original reference annotated genes. Furthermore, these information are a potential resource to study LAL secretion mechanisms in T. tridentatus.

Effect of salinity fluctuation on the transcriptome of the Japanese mantis shrimp Oratosquilla oratoria.

Salinity fluctuation may detrimentally affect the composition and biological processes of crustaceans. As a euryhaline crustacean, Oratosquilla oratoria can survive at salinities ranging from 20 psu to 40 psu. Therefore, we designed five salinity gradients (20, 25, 30, 35, and 40 psu) and 66.39 Gb clean transcriptome data were obtained after O. oratorias were exposed to each gradient for 24 h. All clean data were spliced into 50,482 unigenes and 17,035 unigenes were annotated in at least one database. Compared with 30 psu, 1010, 851, 1733 and 2188 differentially expressed genes were obtained at 20, 25, 35 and 40 psu, respectively. Results also showed that the osmoregulation of O. oratoria is primarily regulated by lipid and amino acid metabolism, amongst others. No significant up-regulated pathways were enriched at 25 psu and 35 psu, although more significant down-regulated pathways were obtained at 35 psu. Therefore, we assumed that the optimum survival salinity of O. oratoria may range from 25 psu to 35 psu. However, 35 psu may be more suitable for O. oratoria. In addition, 55 unigenes that encode putative inorganic ion exchanges were identified. This study aims to provide fundamental information for understanding the osmoregulation mechanisms of crustaceans.

Transcriptome analyses reveal alterations in muscle metabolism, immune responses and reproductive behavior of Japanese mantis shrimp (Oratosquilla oratoria) at different cold temperature.

Low temperature reduction is thought to cause widespread effects on the physical and behavioral traits of marine organisms, which include metabolic processes, immune responses, and reproductive behavior. Crustaceans are generally considered sensitive to temperature reduction due to the lack of efficient regulators. To better understand the molecular regulatory mechanisms of crustacean exposure to cold stress, Japanese mantis shrimp (Oratosquilla oratoria) was chosen as a representative crustacean. Transcriptomic responses in O. oratoria from five temperatures (25 °C, 22 °C, 19 °C, 16 °C, and 13 °C) were studied using RNA-seq. A total of 64.91 Gb of clean transcriptomic data were generated in 10 libraries and then spliced into 52,107 unigenes with an average length of 1089 bp and an N50 length of 1872 bp. A total of 14,841 unigenes was annotated in at least one database using Blastx alignment. Compared with the control temperature (25 °C), 7, 21, 58, and 236 unigenes were significantly differentially expressed at 22 °C, 19 °C, 16 °C, and 13 °C, respectively. GO analysis showed that 6, 20, 27, and 35 terms were significantly enriched at 22 °C, 19 °C, 16 °C, and 13 °C, respectively. In addition, 2, 5, 2, and 10 significant pathways were presented at 22 °C, 19 °C, 16 °C, and 13 °C, respectively. Combining NR, GO, and KEGG annotation information, many genes significantly differentially expressed at low temperatures may be associated with metabolic processes, immune response, and reproductive behavior. Additionally, we reconstructed the phylogenetic relationship based on 366 orthologous genes and the predicted differentiation time of O. oratoria and P. vannamei range from 212.82 to 365.30 Mya. Furthermore, 16 orthologous genes were identified as PSGs and 30 orthologous genes were identified as FEGs and these adaptive genes were associated with energy metabolism, stress response and immunity, and multiple cellular processing. These results provide fundamental information about molecular mechanisms regulating cold stress response of O. oratoria.

Transcriptomic Responses of Two Ecologically Divergent Populations of Japanese Mantis Shrimp (Oratosquilla oratoria) under Thermal Stress.

Crustaceans are generally considered more sensitive to ocean warming due to their lack of certain efficient regulators. However, the alterations in the physiology and behavior of crustaceans in response to thermal stress differ vastly even among the infraspecific populations of heterogeneous landscapes. Consequently, understanding the impact of temperature fluctuation on crustacean infraspecific populations might be essential for maintaining a sustainable persistence of populations at existing locations. In the present study, we chose the Japanese mantis shrimp (Oratosquilla oratoria) as the representative crustacean population, and conducted transcriptome analyses in two divergent O. oratoria populations (the Zhoushan and Qingdao populations) under same thermal stress (20-28 °C) to identify the population-specific expression response to thermal stress. The results showed significant differences in gene expressions, GO terms and metabolic pathways between the two populations. We hypothesized that intraspecific mutations in the same or different genes might lead to thermal adaptive divergences. Temperature increases from 20-28 °C produced significant enrichment in GO terms and altered the metabolic pathways in the Zhoushan population despite the lack of differentially expressed unigenes. Therefore, several functional genes with large pleiotropic effects may underlie the response to thermal stress in the Zhoushan population. Furthermore, the most significantly enriched biological processes of the Qingdao population were associated with the state or activity of cells and its significant enriched pathways with genetic information processing as well as immune and environmental information processing. In contrast, the differentially regulated unigenes of the Zhoushan population were primarily involved in the regulatory cellular and transcription processes and the most significant pathways found were metabolic and digestive. Consequently, the regulatory mechanisms of the Zhoushan population are probably more efficient than those of the Qingdao population under the same thermal stress.