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The shell color of Pacific oyster (Crassostrea gigas) is a desirable trait; but the nutritional studies on C. gigas with different shell colors have not been conducted. Through successive selective breeding, five shell color strains of black (B), purple (P), orange (O), golden (G) and white (W) C. gigas have been developed. The aim of this study was to evaluate the chemical composition and nutritional value of five shell color strains and one commercial population with a common color. The biochemical composition including moisture, total protein, glycogen, ash, total fat, fatty acids (FA), amino acids and minerals was detected. The results indicated that the protein (50.76%-56.57%) was the major component. The content of glycogen showed a significant difference between orange shell and golden shell strains, as well as between commercial population and golden shell strain. In addition, all shell color strains contained a large amount of essential amino acids (12.20-14.15 g (100 g)−1), of them leucine (2.81-3.29 g (100 g)−1) and lysine (2.79-3.28 g (100 g)−1) were predominant. The oysters were rich in polyunsaturated fatty acids (42.26%-45.24% of total fatty acid) with high levels of DHA (18.53%-21.16% of total fatty acid) and EPA (17.23%-18.68% of total fatty acid). Significant differences of mineral contents (Mg, Zn, Fe and Cu) were identified among the six populations. These results indicated that C. gigas with different shell colors presented rich nutritional value with high protein, glycogen, essential amino acids and polyunsaturated fatty acids. The biochemical composition obtained in this study is useful for selective breeding of C. gigas with different shell colors. [ABSTRACT FROM AUTHOR]

Porphyrins are a widespread group of pigments in nature which are believed to contribute to shell colors in mollusks. Previous studies have provided candidate genes for porphyrin shell coloration, however, the linkage analysis between functional genes and porphyrin pigmentation remains unclear in mollusks. RNA interference is a powerful molecular tool for analyzing the loss of functions of genes in vivo and alter gene expression. In this study, we used unicellular alga Platymonas subcordiformis and Nitzschia closterium f. minutissima as vectors to feed oysters with Escherichia coli strain HT115 engineered to express double-stranded RNAs targeting specific genes involved in porphyrin synthesis. A strain of Crassostrea gigas with orange shell was used to target key haem pathway genes expression using the aforementioned approach. We show here that feeding the oysters with E. coli, containing dsRNA targeting pigmentation genes, can cause changes in the color of the newly deposited shell. For example, the RNAi knockdown of CgALAS and CgPBGD resulted in the loss of uroporphyrin pigmentation from the shell due to the accumulation of the pigment in the oyster's mantle. The study probed the crucial role of ALAS and PBGD genes potential functions of uroporphyrin production and shell color pigmentation in C. gigas. [ABSTRACT FROM AUTHOR]

• CgTyp-1 and CgTyp-3 were expressed at higher levels in the black shell than that in the white shell color oysters. • Two tyrosinase-like genes were located in the outer epithelium of the outer fold either in the black or white shell color oysters. • Knockdown of each Typ resulted in obvious decreases of the tyrosinase activity and melanosome count in mantles. • CgTyp-1 and CgTyp-3 were involved in the biosynthesisof melanin. Color polymorphism in Mollusca is of great interest for consumer preference. Although the heritability of shell color variation has been conducted by experimental crossing, little is known about molecular basis involved in these patterns. Tyrosinase-like proteins are important enzymes which are members of the type-3 copper protein superfamily. In this research, two tyrosinase-like protein genes including CgTyp-1 and CgTyp-3 were identified in the Pacific oyster Crassostrea gigas. Tissue expression analysis showed that CgTyp-1 and CgTyp-3 were dominantly expressed in the mantle. Particularly, they were expressed significantly higher in the edge mantle than that in the central mantle whether on the left or right mantles. Additionally, expressions of CgTyp-1 and CgTyp-3 were mainly found in the black shell color oysters, with relative lower levels in the white shell color oysters. In situ hybridization showed that positive signals for CgTyp-1 and CgTyp-3 were both detected within the outer epithelium of the outer fold either in the black or white shell color oysters. After interference, the expression levels of CgTyp-1 and CgTyp-3 mRNA were significantly attenuated, and the efficiency of RNAi reached 84.72% and 71.58%, respectively. Besides, knockdown CgTyp-1 or CgTyp-3 , obviously decreased the tyrosinase activity of mantles. Furthermore, the number of the melanosomes within epithelium of the outer fold was sharply reduced by silencing of each Typ. These findings argue that CgTyp-1 and CgTyp-3 may be involved in the melanin synthesis, which lends insight into regulation mechanism of shell pigmentation in C. gigas. [ABSTRACT FROM AUTHOR]

Shell coloration in many molluscs is highly variable. A shell color segregation study with progenies from a full factorial cross generated among Pacific oysters exhibiting distinct shell colors (golden, white and black) was conducted to investigate the inheritance of the golden shell color and its correlation with dark pigmentation. Random samples from twenty-three full-sib families were obtained and the shell coloration of offspring within each family was recorded. Results revealed that golden coloration was inherited in a different pattern from dark pigmentation, indicating its different genetic basis. Dark pigmentation was identified as a foreground color while golden or white color were background ones. The locus controlling background colors has two alleles with the allele for golden background being dominant to the allele for white background. In addition, the overlying foreground pigmentation of shells with a golden background was significantly lighter than that of shells with a white background, which suggested an epistatic effect of background color on shell foreground pigmentation. All these findings will facilitate the selection of elite oyster lines with desired shell coloration for aquaculture. [ABSTRACT FROM AUTHOR]

Molluscs exhibit diverse shell colors. The molecular regulation of shell coloration is however not well understood. To investigate the connection of shell coloration with pigment synthesis, we analyzed the distribution of porphyrins, a widespread group of pigments in nature, in four Pacific oyster strains of different shell colors including black, orange, golden, and white. The porphyrin distribution was analyzed in oyster mantles and shells by fluorescence imaging and UV spectrophotometer. The results showed that red fluorescence emitted by porphyrins under the UV light was detected only on the nacre of the orange-shell strain and mantles of orange, black and white-shell strains. Extracts from newly deposit shell, nacre and mantle tissue from orange-shell specimens showed peaks in UV–vis spectra that are characteristic of porphyrins, but these were not observed for the other shell-color strains. In addition, genes of the haem synthetic pathway were isolated and characterized. Phylogenetic analysis of CgALAS , CgALAD , CgPBGD , CgUROS , and CgUROD provide further evidence for a conserved genetic pathway of haem synthesis during evolution. Differential expression of the haem genes expressed in mantle tissues support these findings and are consistent with porphyrins being produced by the orange strain only. Tissue in situ hybridization demonstrated the expression of these candidate genes at the outer fold of C. gigas mantles where shell is deposited. Our studies provide a better understanding of shell pigmentation in C. gigas and candidate genes for future mechanistic analysis of shell color formation in molluscs. • Distribution of porphyrin pigments in the outer mantle edge and the shell of Crassostrea gigas was observed by spectral analysis. • The coding sequences of key haem pathway genes were identified and characterized in C. gigas. • The shell color of C. gigas may be affected by the accumulation and degradation of porphyrin caused by the expression of key haem pathway genes. [ABSTRACT FROM AUTHOR]

The Pacific oyster (Crassostrea gigas) is one of the most important species which has the largest production among all cultured shellfish. Recently, several shell color strains of C. gigas have been developed to improve the commercial values, however the inheritance of shell pigmentation is still unclear. A total of 29 families cross-mated between different pigmented oysters were produced to investigate the inheritance of the shell pigmentation and purple pigmentation. Random samples from each family were obtained and their shell pigmentation were recorded. Results revealed that shell pigmentation was controlled by two genetic locus, among which one is responsible for secretion of pigmentation and the other is responsible for distribution mode of pigmentation. The locus controlling the secretion of pigmentation has two alleles with the allele for the presence of pigmentation being dominant to the allele for the absence of pigmentation. Similarly, another locus controlling the distribution mode of pigmentation has two alleles with the allele for striped distribution being dominant to the allele for solid distribution. In addition, one independent locus with two alleles was suggested to control the purple-striped pigmentation, which one allele for shell devoid of purple pigmentation is dominant to another allele for shell with purple pigmentation. The findings will provide valuable information for the efficient selective breeding of shell color strains in the Pacific oyster. • A two-locus model for inheritance of shell pigmentation were demonstrated. • The allele for secretion of pigmentation is dominant to the other allele for absence of pigmentation. • The allele for striped distribution of pigmentation is dominant to the allele for solid distribution of pigmentation. • An indenpendent loci was found to control the inheritance of purple striped pigmentation. [ABSTRACT FROM AUTHOR]

Shell color polymorphism is common in nature for the marine mollusks. The Pacific oyster Crassostrea gigas , is widely cultured and one of the most economically important bivalve species with various shell color variants. Here, a total of 1061 single nucleotide polymorphism markers, developed from expressed sequence tags (EST-SNPs), were used to construct two genetic linkage maps based on reciprocal-cross between black shell and white shell oysters (MF-A and MF-B) respectively. The two separate linkage maps were combined into an integrated map spanning 947.31 cM in total length. The integrated map was composed of 351 EST-SNP markers distributing on 10 linkage groups, with an average interval of 2.78 cM between adjacent markers. For the MF-A, 11 quantitative trait loci (QTLs) for shell color traits and five QTLs for growth traits were detected, explaining 8.1%–13.8% (mean = 11.76%) and 10.4%–11.0% (mean = 10.66%) phenotypic variance respectively. For the MF-B, nine QTLs for shell color traits and eight QTLs for growth traits were detected, explaining 8.8%–21.7% (mean = 12.84%) and 8.4%–9.4% (mean = 8.88%) phenotypic variance respectively. QTLs clustering was found on the linkage group 2 of the MF-A and the linkage group 8 of the MF-B, and the QTLs showed that pleiotropism could affect at most three traits. Three shared-QTLs associated with shell color traits were identified on the integrated map, and one of them (qA- a * 4) was significantly homologous to C. gigas calmodulin-like protein. The QTLs identified in the present study could be useful in finding candidate genes for the shell color and growth-related traits in future, and potentially applied to marker-assisted selection breeding programs for C. gigas . [ABSTRACT FROM AUTHOR]

Molluscan shellfish have a diverse form of shell coloration and sex determination, understanding the genetic mechanism under these two traits is crucial to their germplasm exploitation and conservation. In our breeding program, a novel orange-shell variant of C. gigas was obtained, and an F2 family was constructed by crossing an orange-shell male with a black-shell female. This F2 family provided an ideal material for QTL mapping of orange shell color and sex traits of C. gigas simultaneously. Here, a high-density genetic linkage map containing 1799 SNP markers was constructed using this F2 family by genotyping-by-sequencing. These markers were distributed on 10 linkage groups and the total map size was 828.31 cM, with an average marker interval of 0.58 cM. Based on this map, six chromosome-level significant QTLs associated with orange shell color were detected, with the phenotypic variance explained ranging from 9.10% to 12.30%. Within the QTL regions, four candidate genes, 3-ketoacyl-CoA thiolase A, fibroblast growth factor receptor 4, poly [ADP-ribose] polymerase tankyrase-1 and tripartite motif-containing protein 45-like, related to the organic matrix of mollusk shells or collagen were identified. Additionally, only one genome-wide significant sex-related QTL was detected, which explained 19.30% of phenotypic variance. Two sex-related genes, proteasome subunit beta type-3 and G-protein coupled estrogen receptor 1 were identified within the QTL region. The QTLs and candidate genes identified in this study provide valuable resources for further investigations on the molecular mechanisms of shell color and sex in C. gigas. • A high-density genetic linkage map containing 1799 SNP markers was constructed for C. gigas using an F2 family. • Six chromosome-level significant QTLs associate with orange shell color were detected in C. gigas. • Only one genome-wide significant sex-related QTL was detected in C. gigas. • Different description methods of phenotype have a great impact on the result of QTL mapping. [ABSTRACT FROM AUTHOR]