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For hundreds of years, the color diversity of Mollusca shells has been a topic of interest for humanity. However, the genetic control underlying color expression is still poorly understood in mollusks. The pearl oyster Pinctada margaritifera is increasingly becoming a biological model to study this process due to its ability to produce a large range of colors. Previous breeding experiments demonstrated that color phenotypes were partly under genetic control, and while a few genes were found in comparative transcriptomics and epigenetic experiments, genetic variants associated with the phenotypes have not yet been investigated. Here, we used a pooled‐sequencing approach on 172 individuals to investigate color‐associated variants on three color phenotypes of economic interest for pearl farming, in three wild and one hatchery populations. While our results uncovered SNPs targeting pigment‐related genes already identified in previous studies, such as PBGD, tyrosinases, GST, or FECH, we also identified new color‐related genes occurring in the same pathways, like CYP4F8, CYP3A4, and CYP2R1. Moreover, we identified new genes involved in novel pathways unknown to be involved in shell coloration for P. margaritifera, like the carotenoid pathway, BCO1. These findings are essential to possibly implement future breeding programs focused on individual selection for specific color production in pearl oysters and improve the footprint of perliculture on the Polynesian lagoon by producing less but with a better quality. [ABSTRACT FROM AUTHOR]

Abstract Aquaculture of nacreous gems, such as cultured pearls, mabé or keshi, is done mostly using different mollusc species grown in countries of the Indo-Pacific region. To date, no single species has been exploited for the simultaneous generation of more than one of these bioproducts, but all require animals with colourful shells. Historically, Pinctada species have mainly been used for nucleated pearl production, selecting the rarer colourful individuals to be used as graft donors. By contrast, colourful Pteria species have mostly been used for mabé production, as the grafting operation for pearl production is associated with low yield. In this study, we report the potential for cumulating cultured pearl and mabé (MP), or keshi and mabé (MK) production, using a colourful hatchery-produced G2 family of P. margaritifera. For these trials, MP and MK combinations were compared with the operations to produce pearls (P), mabé (M) or keshi (K) alone in an experimental design using groups of small and large recipients from the G2 family. Results showed no significant impact of combining operation types on subsequent pearl weight, keshi weight, or mabé thickness within recipient oyster size group. By contrast, significant differences were observed between the large and small recipients. The small group produced the thickest mabé, while the large group produced the heaviest pearls and keshi. These contrasting results revealed: 1) the relative independence between the two tissues capable of biomineralisation activities, the mantle (shell and mabé growth) and the pearl sac (pearl or keshi growth); 2) the potential compensatory growth of the small recipient oyster group, which had the highest shell growth performance; and 3) the regulation capacity of the larger oyster group of pearl sac activity. With the same growing area and number of cultured oysters, it would be possible for the P. margaritifera pearl industry to benefit from hatchery propagation of selected colourful shell and produce valuable keshi and mabé together with cultured pearls. Highlights • Simultaneous pearls and mabé or keshi and mabé production is possible. • Colourful hatchery-produced P. margaritifera could serve as recipients. • Recipient compensatory growth could affect both mabé and shell growth. • Pearls and keshi are heaviest when grown in large recipient oysters. • Product diversification is possible for the P. margaritifera pearl industry. [ABSTRACT FROM AUTHOR]

Cultured pearl production from Pinctada margaritifera uses the biomineralization capacities of the mantle graft, the saibo , which is usually obtained from only the middle mantle section of the donor oyster. To evaluate the potential for using other parts of the mantle, this study explores and describes the cultured pearl quality traits, pearl size, shape, surface defects and colour parameters obtained with saibo from the entire length of the mantle, comprising the four following sections: 1) posterior, 2) connection with the gills, 3) middle, (the section usually used commercially), and 4) anterior. Rates of nucleus retention and oyster mortality were also recorded and compared between sections. For this, two experimental grafts were designed and conducted in two contrasting culture sites, using 10 selected wild donor oysters in each to perform a total of 1536 grafts. Mantle section comparison revealed that the anterior section was different from the three other sections, showing: 1) the lowest nacre deposition rate in terms of weight and thickness, 2) the palest pearls, with lowest rate of the attractive overtone colour and the 3) a lower rate of pearls with lustre. For pearl circles and shape, no difference was recorded among the different mantle sections. Posterior, connection and middle sections showed similar pearl quality traits, revealing how the number of high quality saibo obtainable from the same batch of donors can easily be increased, thus benefitting the P. margaritifera pearl industry. This finding could provide significant benefits to pearl farmers and the further development of current pearl grafting practices. [ABSTRACT FROM AUTHOR]

The black-lipped pearl oyster, Pinctada margaritifera, is the most important farmed species in French Polynesia and the basis of the most valuable export industry. Mass production of black pearls relies on a surgical operation requiring tissue from a donor pearl oyster to be grafted, together with a nucleus made of shell, into the gonad of a recipient oyster. Improving pearl size through family selection remains one of the main challenges for future aquaculture development. This study analyses the relative contribution of donor and recipient oysters to pearl size. To this end, hatchery-produced donor oysters of two batches, large and small (based on shell height), were used to supply grafts for recipients, which were then monitored individually for their growth performance by recording shell height, width, and thickness, and total live weight (flesh + shells) every 6 months (four biometric measurement times) over 20 months of culture. Pearls issued from the two batches of donors showed no significant differences in nacre weight or thickness. In contrast, recipient oyster shell height and total weight were increasingly positively correlated with these pearl size parameters over the culture period, becoming significant at 8 months post-grafting. Potential therefore exists to use shell height and oyster weight as phenotypic indicators for selective breeding of recipient oysters with high growth performance to increase pearl size in P. margaritifera. [ABSTRACT FROM AUTHOR]

The black-lipped pearl oyster, Pinctada margaritifera, is the most important farmed mollusc species in French Polynesia. Donor oyster selection among wild P. margaritifera individuals, chosen according to their inner shell colour, makes it possible to obtain the broadest range of cultured pearl colours of any species. This study demonstrates the relative influence of using black [B] or red [R] outer shell phenotypes, combined with green [G] or yellow [Y] inner shell phenotypes, on pearl darkness level, colour categories and lustre. A large scale grafting experiment was designed and carried out over five grow-out locations, covering three archipelagos: Tuamotus, Society and Gambier. Results revealed that the [B + G] phenotypes may be used as donors to produce dark green pearl, which suit the demands of the Asian market; whereas, phenotypes incorporating [R] and/or [Y] phenotypes may be used to obtain multicolour pearls of medium/light darkness, which suit the demands of the European market. From an environmental point of view, the 1) [B] phenotype showed no significant variation for light and other pearl colour production, and 2) [Y] phenotype produced both the same rate of pearl darkness level and green colour pearls whatever the grow-out location. A classification tree model was built to predict, according to shell phenotype and culture location, the colour and darkness level of harvested pearls. Lustre was shown to be more influenced by the environment than by phenotype. These results should be taken into account in pearl farm production management and in selective breeding programmes. [ABSTRACT FROM AUTHOR]

Pinctada margaritifera is an economically important marine bivalve species for cultured pearl production in French Polynesian aquaculture. In order to evaluate the influence of donor oyster age on pearl quality traits, experiments were conducted over 6 years using both grafts and surgreffe operations. At harvest, six pearl quality traits were recorded and compared: surface defects, lustre, grade, darkness level and visual colour. Analysing the quality traits of pearls harvested in the initial graft process and those of pearls obtained from surgreffe experiments allowed a comparison of the influence of pearl sac cells originating from the initial mantle graft, which aged together with their recipient oysters. The results demonstrated a significant decrease between these successive grafts in lustre, grade (A-B-C), darkness level, and green colour - traits that are of major importance in the pearl market. The duplicated graft experiment allowed the comparison of donor oyster families at 2 and 5 years old, where a mantle graft was inserted into recipient oysters aged 2.5 years. The results showed the same tendencies to a lesser extent, with (i) an improved pearl grade, predominantly through a most important rate of 0 surface defect category, and (ii) a green/grey ratio in favour of the younger donor. A comparison between the graft-surgreffe and the duplicated graft experiments also highlighted: (i) the indirect role played by the younger recipient oysters, which must be optimized for optimal pearl quality realization, and (ii) the complex interplay between the donor and recipient oysters. [ABSTRACT FROM AUTHOR]

The pearl oyster Pinctada margaritifera has the specific ability to produce pearls with the widest range of colours among all pearl oyster species. This pearl colour diversity originates from the mantle biomineralising tissue (graft) of the donor oyster, which is originally responsible for the variety of colours of the inner shell surface. This study aimed to: 1) assess the geographic distribution and establish a first stocklist of the colourful oyster phenotypes used as donors in French Polynesia, and 2) investigate the phenotypic relation between inner shell colouration and the corresponding colour of harvested pearls. With the support of a pearl farmers' network, we investigated the different donor phenotype frequencies among five collection sites (Ahe, Apataki, Takaroa, Takume and Mangareva). This donor evaluation was made during grafting of pearl oysters (N = 49,938) obtained from collector stations. Results showed that pearl production is mainly based on six common colourful donor phenotypes classified as monochromatic and polychromatic profiles, which shown different frequencies among the collection sites. Experimental grafts (N = 4640) were then realised and subsequent culture conducted at a single site in order to avoid pearl colour variation due to environmental influences. Traceability between donors (N = 232) and pearls (N = 2776), revealed that: 1) yellow (gold) and aubergine (reddish) pearls could be mostly obtained by using the monochromatic yellow and red donor phenotypes, respectively, and 2) one third to one quarter of grey pearls was inevitably harvested, whatever the polychromatic phenotype chosen as the donor, which leaves at least half the harvest composed of the attractive green and peacock colours. This preliminary stocklist of colour range together with analysis of the colour phenotype transmission between inner shell and pearl provide the basis for producing multiple pearl oyster “colour lines” through hatchery propagation and would be helpful for future selective breeding programs. Statement of relevance Donor shell colour selection predict colours of pearls [ABSTRACT FROM AUTHOR]

In French Polynesia, the aquaculture of Pinctada margaritifera is carried out in numerous grow-out sites, located over three archipelagos (Gambier, Society and Tuamotu). To evaluate the impact of macro-geographical effects of these growing sites on pearl quality traits, five hatcheries produced families were used as homogeneous donor oysters in an experimental graft. The molluscs were then reared in two commercial locations: Tahaa Island (Society) and Rangiroa atoll (Tuamotu). At harvest, eight pearl quality traits were recorded and compared: surface defects, lustre, grade, circles, shape categories, darkness level, body and secondary colour and visual colour categories. Overall inter-site comparison revealed that: (1) all traits were affected by grow-out location except for lustre and round shape, and (2) a higher mean rate of valuable pearls was produced in Rangiroa. Indeed, for pearl grade, Rangiroa showed twice as many A-B and less reject samples than Tahaa. This was related to the number of surface defects (grade component): in Rangiroa, twice as many pearls had no defects and less pearls had up to 10 defects. Concerning pearl shape, more circled and baroque pearls were found in Tahaa (+10%). For colour variation, 10% more pearls have an attractive green overtone in Rangiroa than in Tahaa, where more grey body colour were harvested. Lustre does not seem to be affected by these two culture site (except at a family scale). This is the first time P. margaritifera donor family have been shown to vary in the quality of pearls they produce depending on their grow-out location. [ABSTRACT FROM AUTHOR]

Cultured pearls produced with Pinctada margaritifera, using the surgreffe method (implantation of a second nucleus following pearl harvest) were studied for the first time to: (1) examine family effect on nacre thickness, nacre weight and nacre deposition speed and (2) compare variation in these three traits with that obtained from the cultured pearls previously harvested after the corresponding initial grafts. A surgreffe experiment using 783 recipient oysters was realized in Rangiroa atoll (French Polynesia). After 24 months of culture, 389 cultured pearls were harvested. Significant donor family effect was found for the harvested pearl rate from surgreffe ( P = 0.046). Highly significant donor family effect was recorded for nacre thickness ( P = 0.004). Very highly significant donor family effects were recorded for nacre weight and nacre deposition speed ( P < 0.0001). Comparison between surgreffe and initial graft showed: (1) no significant effect for the average cultured pearl rate harvested ( P = 0.052) and average cultured pearl nacre deposition speed ( P = 0.622) and (2) very highly significant differences ( P < 0.0001) for the average cultured pearl nacre thickness and nacre weight. This study highlighted three major implications for pearl industry management: (1) donor family effect was maintained from initial graft to surgreffe, for nacre thickness, weight and deposition speed, (2) the persistence of the pearl sac metabolic activity over three years of culture and (3) the relation between harvested pearl rate and the size of the nucleus inserted in the pearl sac. [ABSTRACT FROM AUTHOR]

Cultured pearls quality produced by the mollusk Pinctada margaritifera var. cumingi is determined by a panel of criteria, of which shape is one of the most relevant. Cultured pearls with round and semi-round (RS) shapes are the most valuable. Decreasing the proportion of other pearl shapes (OT: baroque, button, drop, oval), and especially the circled pearls (CL) and rejects ( rebuts : RT), is an important challenge for the pearl industry. The present study examined the effect of grafter skill and season of graft on the variation in proportions of CL, RS, OT and RT cultured pearl shape categories in a mono-site culture (Mangareva island lagoon). Six large-scale grafting experiments, carried out in the same way as commercial grafting sessions, were made by seventeen experienced grafters over three seasons. After two years of culture, a total of 42,575 cultured pearls were harvested and sorted into shape categories. This study is the first to apply large-scale traceability to effects of individual grafter skill on cultured pearl shape in P. margaritifera . Grafter effects were clearly demonstrated on the proportions 1) of CL and RS, 2), to a in a lesser extent, of OT and RT, but 3) were low, affecting only a few percent of the overall means. For a high quality cultured pearl production, grafter effect could reduce CL rate by 7% and increase RS rate by 4%. In addition, the results indicate a significantly indirect influence of grafting season on shape class proportions, as these differ in autumn grafts, CL (− 12%), RS (+ 13%) and OT (+ 24%), in comparison to winter and spring. Possible implications of “oyster” physiological reproductive status are discussed, as the difference in grafting season corresponds to differences in maturity of the recipient “oysters”. Rates of RT were only increased (+ 14%) during the seasons when the lagoon water temperature was the coldest (winter) and the warmest (transition to summer). [ABSTRACT FROM AUTHOR]

The top aquaculture species in French Polynesia is Pinctada margaritifera , a mollusc grown for the production of a unique gem: the black pearl. One of the challenges facing the pearl farming industry is to “produce less but better pearls” through genetic improvement. An experimental hatchery system was used to generate full-sib families to be tested for their potential as donor “oysters”. A large-scale grafting experiment was done and seven cultured pearl quality traits: grade, surface defects, lustre, darkness level, visual colour categories, circles and shape categories were recorded. Our results revealed, for the first time, significant phenotypic relationships between these quality traits. The grade A cultured pearl class had the largest proportion of pearls with a green overtone (65%), the lowest number of circled pearls (15%) and the maximum of round-shaped pearls (45%). In contrast, the “reject” cultured pearl class had the largest proportion of pearls with grey body color (65%), the greatest number of circled pearls (35%) and the maximum with a baroque shape (nearly 60%). When grade components were studied separately, cultured pearls in the zero surface defect class exhibited the same tendencies as grade A pearls, contrasting with the class where there were more than ten defects on the surface of each pearl. When cultured pearls were classified according to the presence or absence of lustre, pearls with lustre mostly had a green overtone colour, whilst pearls without lustre did not. These findings have major implications for cultured pearl quality improvement, as modern genetic breeding methods can increase the proportion of high quality cultured pearls though selected lines of donor oysters capable of producing pearls with a green overtone. Selection of appropriate donor phenotypes, incorporation of pigmentation traits into a pearl oyster breeding programme and production of lines with desirable colours will be developed for oyster aquaculture in French Polynesia. [ABSTRACT FROM AUTHOR]

Individual Pinctada margaritifera molluscs were collected from the Takapoto atoll (Tuamotu Archipelago, French Polynesia) and used to produce ten first generation full-sib families in a hatchery system, following artificial breeding protocols. After three years of culture, these progenies were transferred to Rangiroa atoll (Tuamotu Archipelago, French Polynesia) and tested for their potential as graft donors. A large-scale grafting experiment of 1500 grafts was conducted, in which a single professional grafter used ten individual donor oysters from each of the ten families, grafting 15 recipient oysters from each donor. The recipient oysters were all obtained from wild spat collection in Ahe (Tuamotu Archipelago, French Polynesia). After 18 months of culture, 874 pearls were harvested. Highly significant donor family effects were found for nucleus retention, nacre thickness, nacre weight, pearl colour darkness and visually-perceived colour (bodycolor and overtone), pearl shape categories, surface defects and lustre, the last two of which are components of the Tahitian classification grade. No significant difference was recorded between the ten G1 families for the absence or presence of rings. The progenies could be ranked from “best” (i.e., the donor whose grafts produced the greatest number of grade A pearls) to the “worst”. Some progenies had extreme characteristics: family B presented the greatest number of pearls with lustre (98%) and a high proportion of dark gray to black with green overtone pearls (70%). These results have important implications for the selective breeding of donor pearl oysters: it may be possible to reach a point where specific donor lines whose grafts produce pearls with specific quality traits could be identified and maintained as specific breeding lines. [ABSTRACT FROM AUTHOR]

Producing high quality cultured black pearls from Pinctada margaritifera is one of the major challenges for the “pearl oyster” industry in French Polynesia. In order to assess donor effect on cultured pearl quality, wild Pinctada margaritifera originating from the Tuamotu Archipelago were used in a duplicated grafting experiment. After 12 months of culture, nucleus retention was assessed and seven pearl quality traits recorded on the 454 cultured pearls harvested from the experiment. The traits scored were nacre thickness and pearl weight, surface defects, lustre, grade, and the colour components: 1) darkness of cultured pearl colour, and 2) visual perception of colour class (bodycolor and/or overtone). Our results demonstrate for the first time that individual wild donors of implanted mantle grafts significantly affect these seven quality traits in P. margaritifera cultured pearls. This finding was repeated in two series of grafts made by different professional grafters. The wild donors could be ranked from “best” (e.g., the donor whose grafts produced the cultured pearl with the maximum lustre) to the “worst”. Moreover, we showed strong correlations between: 1) cultured pearl nacre thickness and grade, with grade A showing the greatest nacre thickness on average compared with grade D and rejects; and 2) nacre thickness/cultured pearl weight and colour components (darkness and visual “colour categories”), with the palest cultured pearls (i.e. white cultured pearls) being the smallest (lowest nacre thickness and weight). Thus, one way of enhancing P. margaritifera foundation stocks for a selective breeding program could be to select the “best” donors, using appropriate molecular tools. Generation of selected donor lines from these stocks through hatchery production would be one way to increase the quality of cultured pearl farming of P. margaritifera in French Polynesia. [ABSTRACT FROM AUTHOR]

The shell color of the Mollusca has attracted naturalists and collectors for hundreds of years, while the molecular pathways regulating pigment production and the pigments themselves remain poorly described. In this study, our aim was to identify the main pigments and their molecular pathways in the pearl oyster Pinctada margaritifera—the species displaying the broadest range of colors. Three inner shell colors were investigated—red, yellow, and green. To maximize phenotypic homogeneity, a controlled population approach combined with common garden conditioning was used. Comparative analysis of transcriptomes (RNA-seq) of P. margaritifera with different shell colors revealed the central role of the heme pathway, which is involved in the production of red (uroporphyrin and derivates), yellow (bilirubin), and green (biliverdin and cobalamin forms) pigments. In addition, the Raper–Mason, and purine metabolism pathways were shown to produce yellow pigments (pheomelanin and xanthine) and the black pigment eumelanin. The presence of these pigments in pigmented shell was validated by Raman spectroscopy. This method also highlighted that all the identified pathways and pigments are expressed ubiquitously and that the dominant color of the shell is due to the preferential expression of one pathway compared with another. These pathways could likely be extrapolated to many other organisms presenting broad chromatic variation. [ABSTRACT FROM AUTHOR]

The black-lipped pearl oyster, Pinctada margaritifera , produces the largest and most valuable coloured pearls in the world. Cultured pearl size remains one of the most important quality traits. Despite the great geographical area covered by pearl farms in the atolls of French Polynesia, little is known about the influence of grow-out site effects on pearl size attained. To explore the genetic and environmental impact on the size of pearls as well as the genetic × environment interaction, a uniform experimental graft was designed on two contrasting macro-geographical lagoons. Five biparental families of donor oysters were grafted and then reared at both sites. After 18 months of culture, phenotypic parameters corresponding to pearl size, i.e. nacre weight and thickness, were recorded among the harvested pearls. The expression of Pif-177 gene, a biomarker encoding protein in the aragonite nacreous layer, was analysed in the corresponding pearl sac. The results show a family effect for nacre weight and thickness on both sites, with family F058 producing the heaviest and thickest nacre, and F805 the lightest and thinnest. By contrast, inter-site comparison revealed no significant site effects for these two parameters. In addition, grow-out location did not modify the relative gene expression of Pif-177 in the pearl sac between donor families in either culture site. Both nacre weight and thickness were positively correlated with the level gene expression of Pif-177. These results suggest that pearl size parameters were not affected by the environment in the present study and this is supported by the relative gene expression of Pif-177 observed. This knowledge constitutes an initial step in the study of pearl size trait inheritance, which will be helpful in the near future for the diffusion of genetically selected donor oyster lines produced by hatchery systems throughout production sites. [ABSTRACT FROM AUTHOR]

Size is the most important and valuable quality of the cultured black-lip pearl, Pinctada margaritifera . As this pearl aquaculture is carried out at numerous grow-out sites, this study analyzes the environmental influence on pearl size parameters (nacre weight and thickness) in relation to the recipient oyster biometric parameters (shell thickness, height, width, and oyster weight) at harvest time. Toward this end, an experimental graft was designed by using a homogeneous donor oyster phenotype. The recipient oysters were randomly and equally transferred and reared in five commercial and contrasting grow-out locations. Overall inter-site comparisons revealed that the cultured pearl size (N = 2168) and the biometric parameters of the recipient oysters were highest for sites with warmer temperatures with low seasonal variation in comparison to the southern latitude sites. These results were supported by positive correlations between pearl nacre thickness and recipient oyster shell thickness, height, and width. In parallel, the biomineralization potential of the mantle graft was screened through four genes encoding aragonite ( Pif 177 , MSI60 ) and calcite ( shematrin 9 , aspein ). As the gene expression levels were the same among all the donor oysters, this finding demonstrates that: 1) the pearl sac that originated from the mantle graft was not isolated from environmental variations during the culture period and 2) the phenotypic expressions of the two biomineralizing tissues in the recipient oyster were consistent (shell and pearl). In the near future, this knowledge will be helpful at the production sites of genetically selected donor oyster lines for growth produced in hatchery systems. [ABSTRACT FROM AUTHOR]

The present study investigated the effect of electrolysis on the biomineralization capacities of juveniles of the mollusk Pinctada margaritifera for the first time. Size-selected individuals from two groups, “Medium” and “Large”, from a multi-parental family produced in a hatchery system were subjected to electrolysis under a low voltage current over a nine-week experimental period. The growth of the juveniles was individually monitored and assessed weekly by wet weight and shell height measurements. At the end of the experiment, mantle tissue was sampled for biomineralization-related gene expression analysis. Electrolysis significantly increased pearl oyster growth in terms of shell height and wet weight for Large juveniles from the 5th and the 2nd week, respectively, until the end of the experiment. However, differences were only significant for Medium individuals from the 7th week for shell height and from the 9th week for wet weight. Furthermore, transcriptional analysis of six known biomineralization genes coding for shell matrix proteins of calcitic prisms and/or nacreous shell structures revealed that five were significantly overexpressed in the mantle mineralizing tissue under electrolysis: three in common between the two size class groups and two that were expressed exclusively in one or the other group. Finally, we found no statistical difference of the shell thickness ratio between individuals undergoing electrolysis and control conditions. Taken together, our results indicate, for the first time in a calcifying marine organism, that electrolysis influences molecular mechanisms involved in biomineralization and may stimulate some parameters of pearl oyster growth rate. [ABSTRACT FROM AUTHOR]