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International audience; The importance of poly-unsaturated fatty acids (PUFAs) in food is crucial for the animal and human development and health. As a complementary strategy to nutrition approaches, genetic selection has been suggested to improve fatty acids (FAs) composition in farmed fish. Gas chromatography (GC) is used as a reference method for the quantification of FAs; nevertheless, the high cost prevents large scale phenotyping as needed in breeding programs. Therefore, a calibration by means of Raman scattering spectrometry has been established in order to predict FA composition in rainbow trout Onchorhynchus mykiss adipose tissue. FA composition of visceral adipose tissue was analysed by both GC and Raman micro-spectrometry techniques on 268 individuals fed with three different feeds, which have different FA compositions. Among the possible regression methods, the ridge regression method, was found to be efficient to establish calibration models from the GC and spectral data. The best cross-validated R2 values were obtained for total PUFAs, omega-6 (Ω-6) and omega-3 (Ω-3) PUFA (0.79, 0.83 and 0.66, respectively). For individual Ω-3 PUFAs, α-linolenic acid (ALA, C18:3), eicosapentaenoic acid (EPA, C20:5) and docosahexenoic acid (DHA, C22:6) were found to have the best R2 values (0.82, 0.76 and 0.81, respectively). This study demonstrates that Raman spectroscopy could be used to obtain good correlation coefficients on adipocytes allowing to predict PUFAs, and calibration models can be used to predict PUFAs contents for large scale and high throughput phenotyping in rainbow trout

Background In response to major challenges regarding the supply and sustainability of marine ingredients in aquafeeds, the aquaculture industry has made a large-scale shift toward plant-based substitutions for fish oil and fish meal. But, this also led to lower levels of healthful n−3 long-chain polyunsaturated fatty acids (PUFAs)—especially eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids—in flesh. One potential solution is to select fish with better abilities to retain or synthesise PUFAs, to increase the efficiency of aquaculture and promote the production of healthier fish products. To this end, we aimed i) to estimate the genetic variability in fatty acid (FA) composition in visceral fat quantified by Raman spectroscopy, with respect to both individual FAs and groups under a feeding regime with limited n-3 PUFAs; ii) to study the genetic and phenotypic correlations between FAs and processing yields- and fat-related traits; iii) to detect QTLs associated with FA composition and identify candidate genes; and iv) to assess the efficiency of genomic selection compared to pedigree-based BLUP selection. Results Proportions of the various FAs in fish were indirectly estimated using Raman scattering spectroscopy. Fish were genotyped using the 57 K SNP Axiom™ Trout Genotyping Array. Following quality control, the final analysis contained 29,652 SNPs from 1382 fish. Heritability estimates for traits ranged from 0.03 ± 0.03 (n-3 PUFAs) to 0.24 ± 0.05 (n-6 PUFAs), confirming the potential for genomic selection. n-3 PUFAs are positively correlated to a decrease in fat deposition in the fillet and in the viscera but negatively correlated to body weight. This highlights the potential interest to combine selection on FA and against fat deposition to improve nutritional merit of aquaculture products. Several QTLs were identified for FA composition, containing multiple candidate genes with indirect links to FA metabolism. In particular, one region on Omy1 was associated with n-6 PUFAs, monounsaturated FAs, linoleic acid, and EPA, while a region on Omy7 had effects on n-6 PUFAs, EPA, and linoleic acid. When we compared the effectiveness of breeding programmes based on genomic selection (using a reference population of 1000 individuals related to selection candidates) or on pedigree-based selection, we found that the former yielded increases in selection accuracy of 12 to 120% depending on the FA trait. Conclusion This study reveals the polygenic genetic architecture for FA composition in rainbow trout and confirms that genomic selection has potential to improve EPA and DHA proportions in aquaculture species.

One of the top priorities of the aquaculture industry is the genetic improvement of economically important traits in fish, such as those related to processing and quality. However, the accuracy of genetic evaluations has been hindered by a lack of data on such traits from a sufficiently large population of animals. The objectives of this study were thus threefold: (i) to estimate genetic parameters of growth-, yield-, and quality-related traits in rainbow trout (Oncorhynchus mykiss) using three different phenotyping technologies [invasive and non-invasive: microwave-based, digital image analysis, and magnetic resonance imaging (MRI)], (ii) to detect quantitative trait loci (QTLs) associated with these traits, and (iii) to identify candidate genes present within these QTL regions. Our study collected data from 1,379 fish on growth, yield-related traits (body weight, condition coefficient, head yield, carcass yield, headless gutted carcass yield), and quality-related traits (total fat, percentage of fat in subcutaneous adipose tissue, percentage of fat in flesh, flesh colour); genotypic data were then obtained for all fish using the 57K SNP Axiom® Trout Genotyping array. Heritability estimates for most of the 14 traits examined were moderate to strong, varying from 0.12 to 0.67. Most traits were clearly polygenic, but our genome-wide association studies (GWASs) identified two genomic regions on chromosome 8 that explained up to 10% of the genetic variance (cumulative effects of two QTLs) for several traits (weight, condition coefficient, subcutaneous and total fat content, carcass and headless gutted carcass yields). For flesh colour traits, six QTLs explained 1–4% of the genetic variance. Within these regions, we identified several genes (htr1, gnpat, ephx1, bcmo1, and cyp2x) that have been implicated in adipogenesis or carotenoid metabolism, and thus represent good candidates for further functional validation. Finally, of the three techniques used for phenotyping, MRI demonstrated particular promise for measurements of fat content and distribution, while the digital image analysis-based approach was very useful in quantifying colour-related traits. This work provides new insights that may aid the development of commercial breeding programmes in rainbow trout, specifically with regard to the genetic improvement of yield and flesh-quality traits as well as the use of invasive and/or non-invasive technologies to predict such traits.

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.

Molluscs display a vast range of shell colours both between and within species. However, only a few species show colour variation in their soft tissues. In French Polynesia, the pearl oyster Pinctada margaritifera has three tissue morphotypes: the black wild-type and two rare mutations: white albino and orange mantle. Phenotypic transmission is known to occur from these phenotypes when they are used as graft donors for pearl production, leading to multicoloured and white pearls from black and albino mantle grafts, respectively. The present study furthers this knowledge by examining the phenotypic association between the orange mantle tissue morphotype and hard tissues: shells and cultured pearls. Based on a large experimental graft, shell colour quantification and pearl qualification showed that the orange morphotype is associated with light-coloured shells and pearls. Expression analysis of some candidate genes previously identified in the white mantle mutant, tested here on both graft and pearl sac tissues from orange mantle donors, confirmed the involvement of genes associated with shell matrix protein (shem4) and the melanin biosynthesis pathway (zinc). This study provides fundamental information on the mechanism behind mantle tissue colour in P. margaritifera and its association with biomineralisation and pigmentation processes that will be potentially valuable in future selection programs.

Nucleated pearls are produced by molluscs of the Pinctada genus through the biomineralisation activity of the pearl sac tissue within the recipient oyster. The pearl sac originates from graft tissue taken from the donor oyster mantle and its functioning is crucial in determining key factors that impact pearl quality surface characteristics. The specific role of related gene regulation during gem biogenesis was unknown, so we analysed the expression profiles of eight genes encoding nacreous (PIF, MSI60, PERL1) or prismatic (SHEM5, PRISM, ASP, SHEM9) shell matrix proteins or both (CALC1) in the pearl sac (N = 211) of Pinctada margaritifera during pearl biogenesis. The pearls and pearl sacs analysed were from a uniform experimental graft with sequential harvests at 3, 6 and 9 months post-grafting. Quality traits of the corresponding pearls were recorded: surface defects, surface deposits and overall quality grade. Results showed that (1) the first 3 months of culture seem crucial for pearl quality surface determination and (2) all the genes (SHEM5, PRISM, ASP, SHEM9) encoding proteins related to calcite layer formation were over-expressed in the pearl sacs that produced low pearl surface quality. Multivariate regression tree building clearly identified three genes implicated in pearl surface quality, SHEM9, ASP and PIF. SHEM9 and ASP were clearly implicated in low pearl quality, whereas PIF was implicated in high quality. Results could be used as biomarkers for genetic improvement of P. margaritifera pearl quality and constitute a novel perspective to understanding the molecular mechanism of pearl formation.

Grafting mantle tissue of a donor pearl oyster into the gonad of a recipient oyster results in the formation of a chimera, the pearl sac. The phenotypic variations of this chimera are hypothesized to be the result of interactions between the donor and recipient genomes. In this study, the heritability of phenotypic variation and its association with gene expression were investigated for the first time during Pinctada margaritifera pearl production. Genetic variance was evaluated at different levels, 1) before the graft operation (expression in graft tissue), 2) after grafting (pearl sac tissue expression in chimera), and 3) on the product of the graft (pearl phenotype traits) based on controlled biparental crosses and the F1 generation. Donor-related genetic parameter estimates clearly demonstrate heritability for nacre weight and thickness, darkness and color, and surface defects and grade, which signifies a genetic basis in the donor oyster. In graft relative gene expression, the value of heritability was superior to 0.20 in for almost all genes; whereas in pearl sac, heritability estimates were low (h2 < 0.10; except for CALC1 and Aspein). Pearl sac expression seems to be more influenced by residual variance than the graft, which can be explained by environmental effects that influence pearls sac gene expression and act as a recipient additive genetic component. The interactions between donor and recipient are very complex, and further research is required to understand the role of the recipient oysters on pearl phenotypic and gene expression variances.

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, 6 pearl quality traits were recorded and compared: surface defects, luster, grade, darkness level, and visual color. Analyzing 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 luster, grade (A-B-C,) darkness level, and green color – 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 old. The results showed the same tendencies to a lesser extent, with 1) an improved pearl grade, predominantly through a most important rate of 0 surface defect category, and 2) a green / grey ratio in favor of the younger donor. A comparison between the graft-surgreffe and the duplicated graft experiments also highlighted: 1) the indirect role played by the younger recipient oysters, which must be optimized for optimal pearl quality realization, and 2) the complex interplay between donor and recipient oysters.

Cultured pearl nacre deposition rate in Pinctada margaritifera oysters was studied to examine its influence on Tahitian classification grade and colour and to further our understanding of using selected donor oysters produced in a hatchery system for the improvement of these traits. A large-scale grafting experiment using 150 farmed donor oysters was realised in Rangiroa atoll (French Polynesia). After 18 months of culture, 874 pearls were harvested and 7 pearls quality traits were recorded: (1) nacre deposition rate assessed by nacre thickness and weight, (2) classification grade, lustre and surface defects, and (3) colour assessed by pearl colour darkness level and visual colour categories. Nacre deposition rate significantly influence pearl grade, surface defects, darkness level and colour categories, but not the lustre. Grade A pearls had the thickest and heaviest nacre on average, compared with grade D pearls and rejects. In addition, pearls without any surface defect had thicker and heavier nacre. Pearls with low darkness were the thinnest and the lightest. Pearls of aubergine–peacock secondary colours were the thickest and the heaviest. These results highlight the importance of rapid nacre deposition rate for increasing the production of grade A pearls, with less surface defects and dominance of a black tone colour. Fast nacre deposition may, however, have a negative impact on other pearl quality traits, such as pearl lustre. This study demonstrates the importance of examining relationships between pearl quality traits in a way that provides solid information that can be directly used for breeding and genetic selection programmes.

International audience; n 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.

Ageing is defined as the progressive decline in tissue and organ functions over time. This study aims to evaluate the ageing effect on cultured pearl quality phenotypes (including size and quality traits) in the graft-recipient animal model: Pinctada margaritifera. For this, eight uniform grafting experiments were designed using two hatchery-produced pearl oyster families as donors, which were followed through time, between 7 and 30 months in age. For each age category, 20 donors were studied for each culture site giving a total of 2400 grafted oysters. Several phenotypic measurements were made: 1) donor family growth performance from shell size records, 2) pearl size and corresponding quality traits, and 3) expression of some genes related to biomineralization processes on both the mantle graft and on pearl sac tissues. Results showed that: 1) donor age has an impact on pearl size, with grafts coming from the youngest donors yielding the biggest pearls; and 2) grafts from donors between 12 and 18 months in age produced pearls of the highest quality (grade and surface quality), a result supported by an analysis where the level of expression for a panel of genes associated with biomineralization was greatest in donors within the 12 to 18 months age group. These results indicate that donors aged between 12 and 18 months have high potential for biomineralisation and nacre deposition, and likely produce larger and higher quality cultured pearls than older donors.

International audience; Breeding strategies and parental effects play a major role in genetic selection programmes for quality trait improvement. The black-lip pearl oyster, Pinctada margaritifera, is a protandrous hermaphrodite species in which the number of mature females is low, and these only appear after 4 years of culture. This difference underlines the importance of studying the contributions of dams and sires in the production and selection of donor pearl oyster families to choose the most appropriate breeding strategies for cultured pearl quality trait improvement. Laboratory crosses were made using wild dams and sires to produce half-sib families from polyandry and polygyny mating designs. The resulting half-sib families were used as donor oysters in a large grafting experiment of 900 grafts with random recipient oysters, using nuclei of the same size and brand, the same operating technician and the same grow-out site. After 18 months of culture, 497 cultured pearls were harvested. Comparison between half-sib pairs revealed that for nacre thickness, weight and shape, a polygyny effect exists, whereas no significant effect was observed for the polyandry mating. By contrast, no evidence of dam- or sire-related benefits was found for the presence/absence of circles, darkness level and colour categories or grade and its components: lustre and surface defects. These results should have important implications for the selective breeding of donor pearl oysters for hatchery management.

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.

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 artifi- cial 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 collec- tion 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.

Premise of the study: We developed nuclear microsatellite primers to explore the genetic diversity, population genetic structure, and evolutionary history of the fonio (Digitaria exilis), an understudied cereal cultivated in West Africa. o Methods and Results: We used a microsatellite-enriched library approach to isolate and characterize 38 nuclear primer pairs (31 di-, five tri-, and two tetranucleotide repeats), of which 21 were polymorphic and exhibited a clear pattern in 36 accessions from West Africa. The number of alleles per locus ranged from two to 22, with a mean of 4.71, and expected heterozygosity ranged from 0.03 to 0.93. o Conclusions: The developed set of 21 polymorphic SSR markers will provide tools for population and evolutionary genetics studies of the cultivated fonio. (Resume d'auteur)