Long-chain polyunsaturated fatty acids (LC-PUFAs) are essential in important physiological processes. However, the endogenous PUFA biosynthesis pathway is poorly understood in marine bivalves. Previously, a fatty acyl desaturase (Fad) with Δ5 activity was functionally characterized and an elongase termed Elovl2/5 was reported to efficiently elongate 18:2n–6 and 18:3n–3 to 20:2n–6 and 20:3n–3 respectively in Chlamys nobilis. In this study, another elongase and another Fad were identified. Functional characterization in recombinant yeast showed that the newly cloned elongase can elongate 20:4n–6 and 20:5n–3 to C22 and C24, while the newly cloned scallop Fad exhibited a Δ8-desaturation activity, and could desaturate exogenously added PUFA 20:3n–3 and 20:2n–6 to 20:4n–3 and 20:3n–6 respectively, providing the first compelling evidence that noble scallop could de novo biosynthesize 20:5n–3 and 20:4n–6 from PUFA precursors though the “Δ8 pathway”. No Δ6 or Δ4 activity was detected for this Fad. Searching against our scallop transcriptome database failed to find any other Fad-like genes, indicating that noble scallop might have limited ability to biosynthesize 22:6n–3. Interestingly, like previously characterized Elovl2/5, the two newly cloned genes showed less efficient activity toward n–3 PUFA substrates than their homologous n–6 substrates, resulting in a relatively low efficiency to biosynthesize n–3 PUFA, implying an adaption to marine environment.

The aim of this study was to investigate whether total carotenoid content (TCC) in noble scallop Chlamys nobilis is related to body tissue, shell colour, and gender. TCC was determined by a UV–vis recording spectrophotometer in tissue of gonad, adductor, mantle, and gill separately sampled from male and female individuals with orange and brown shell colours from the same cultured population. TCC was significantly different among body tissues, depending on shell colour and gender, ranging from 0.73 to 59.85 μg g−1. In general, TCC was greater in the order of gonad > mantle > adductor > gill. In the same gender, orange shell colour individuals contained significantly higher TCC than brown ones in all four tissues (P < 0.05). In the same shell colour, female contained significantly higher TCC than male in gonad and adductor tissues (P < 0.05).

•A novel Toll gene called CnTLR-1 was cloned in the Chlamys nobilis.•Scallops with different carotenoids content were acutely challenged by different pathogens.•CnTLR-1 transcripts were all up-regulated after challenge of Vibrio parahaemolyticus, LPS and Poly I:C groups.•Transcripts were significantly higher in orange scallops than that of brown ones with and without pathogenic challenges.•CnTLR-1 is an important immune factor against pathogens in the noble scallop.To investigate whether toll like receptors (TLRs) genes do have an immune influence on noble scallop Chlamys nobilis under pathogen stress, acute challenges lasting 48 h to Vibrio parahaemolyticus, lipopolysaccharide (LPS), polyinosinic polycytidylic acid (Poly I:C), and PBS were conducted in two scallop stains of orange and brown with different carotenoids content. A novel toll-like receptor gene called CnTLR-1 was cloned and its transcripts under different challenges were determined. Meantime, total carotenoids content (TCC) of different immune responses were determined to investigate whether there was a relationship between gene expression and carotenoids content. The full length cDNA of CnTLR-1 is 2982 bp with an open reading frame (ORF) of 1920 bp encoding 639-deduced amino acids, which contains five leucine-rich repeats (LRR), two LRR-C-terminal (LRRCT) motifs and a LRR-N-terminal (LRRNT) motif in the extracellular domain, a transmembrane domain and a Toll/Interleukin-1 Receptor (TIR) of 138-amino acids in the cytoplasmic region. Phylogenetic tree analysis showed that CnTLR-1 could be clustered with mollusk TLRs into one group and especially was related closely to Crassostrea gigas and Mytilus galloprovincialis TLRs. CnTLR-1 transcripts were detected in decreasing levels in the mantle, hemocytes, gill, kidney, gonad, hepatopancreas, intestines and adductor. Compared with PBS control group, CnTLR-1 transcripts were up-regulated in V. parahaemolyticus, LPS and Poly I:C groups. Further, CnTLR-1 transcripts were significantly higher in orange scallops than that of brown ones with and without pathogenic challenges. TCC, which is higher in orange scallops, was initially increased and then decreased during a 48 h immune challenge in the hemocytes. The present results indicate that CnTLR-1 is an important factor involved in the immune defense against pathogens in the noble scallop.

•A big defensin gene from Chlamys nobilis (CnBD) was cloned.•Healthy adult scallops were acutely challenged by Vibrio parahaemolyticus.•CnBD transcripts were up-regulated after challenge of V. parahaemolyticus.•CnBD plays an immune role against bacterial infection in noble scallop.The noble scallop Chlamys nobilis has been an important marine cultured bivalve in the Southern Sea of China for decades. However, large-scale mortality events often occurred during the scallop’ cultivation. As one of AMPs (antimicrobial peptides), big defensin is an important component of the innate immunity against pathogenic microorganisms in invertebrates. In order to investigate whether the big defensin can play a role in the immune defense against pathogenic microorganisms in noble scallop, a big defensin gene from the hemocytes of Chlamys nobilis (CnBD) was cloned, and the mRNA level was measured after an acute Vibrio parahaemolyticus challenge of 36 h. The CnBD cDNA contains an open reading frame (ORF) of 381 bp encoding a peptide of 126 amino acids residues. The deduce amino acid sequence of CnBD shows a high similarity with that from Argopecten irradians and displays common features of big defensin, indicating that CnBD is a new member of the big defensin family. Compared with the control group, the relative mRNA level of CnBD was significantly up-regulated at 3, 24 and 36 h. The present result indicated that CnBD played an immune role against bacterial infection in noble scallop.

•Scallops with different carotenoids content were treated by an acute low temperature stress.•The complete CuZnSOD gene was cloned and sequenced.•Transcript levels were significantly higher in the orange scallops than in the brown ones.•Gene expression levels have a significantly positive correlation with carotenoids content.•Carotenoids upregulate gene expression under low temperature stress in the noble scallop.The noble scallop Chlamys nobilis belongs to a warm-water mollusk and has been cultured in the sea of southern China since 1980s'. However, accidents of massive mortality have often occurred during the winter, and one of the reasons could be accumulation of harmful reactive oxygen species caused by lower temperature. Carotenoids are well known for their anti-oxidant function. To investigate whether carotenoids do play a role in mollusks' antioxidant defense system under lower temperature stress, an acute lower temperature experiment was conducted by using two types of scallops: the orange with higher carotenoids content and the brown with lower carotenoids content. Their CuZnSOD gene was cloned, mRNA expression levels were determined, and SOD activity and carotenoids content were measured. The complete CuZnSOD cDNA consists of 1078 nucleotides with an open reading frame encoding 154 amino acid residues, which has high identity with that of its sister species Chlamys farreri. The mRNA expression levels in both the mantle and gill from the orange scallops were significantly higher (P < 0.05) than that of the brown ones, but the result was the opposite in the blood. SOD activity in the mantle and gill from the orange scallops was significantly higher than (P < 0.05) that from the brown ones. Further, significantly positive correlations were found among CuZnSOD gene transcript levels, SOD activity and total carotenoids content in the orange scallops. The present results suggested that carotenoids could play roles in antioxidant defense system by upregulating gene expression under lower temperature stress in the noble scallop.

•Scallops with different carotenoids content were treated by an acute low temperature stress.•The complete CuZnSOD gene was cloned and sequenced.•Transcript levels were significantly higher in the orange scallops than in the brown ones.•Gene expression levels have a significantly positive correlation with carotenoids content.•Carotenoids upregulate gene expression under low temperature stress in the noble scallop.The noble scallop Chlamys nobilis belongs to a warm-water mollusk and has been cultured in the sea of southern China since 1980s'. However, accidents of massive mortality have often occurred during the winter, and one of the reasons could be accumulation of harmful reactive oxygen species caused by lower temperature. Carotenoids are well known for their anti-oxidant function. To investigate whether carotenoids do play a role in mollusks' antioxidant defense system under lower temperature stress, an acute lower temperature experiment was conducted by using two types of scallops: the orange with higher carotenoids content and the brown with lower carotenoids content. Their CuZnSOD gene was cloned, mRNA expression levels were determined, and SOD activity and carotenoids content were measured. The complete CuZnSOD cDNA consists of 1078 nucleotides with an open reading frame encoding 154 amino acid residues, which has high identity with that of its sister species Chlamys farreri. The mRNA expression levels in both the mantle and gill from the orange scallops were significantly higher (P < 0.05) than that of the brown ones, but the result was the opposite in the blood. SOD activity in the mantle and gill from the orange scallops was significantly higher than (P < 0.05) that from the brown ones. Further, significantly positive correlations were found among CuZnSOD gene transcript levels, SOD activity and total carotenoids content in the orange scallops. The present results suggested that carotenoids could play roles in antioxidant defense system by upregulating gene expression under lower temperature stress in the noble scallop.