All-male production by marker-assisted selection for sex determining loci of admixed Oreochromis niloticus and Oreochromis aureus stocks.

Crossing Oreochromis niloticus (On) females with Oreochromis aureus (Oa) males results in all-male progeny that are essential for effective tilapia farming. However, a reproductive barrier between these species limits mating and mass-fry production. One approach to overcoming this barrier is to select parental stocks of mixed genetic backgrounds, which allow interspecific reproductive recognition, while closely maintaining the genetic profiles for sex-determination (SD) of the respective purebred species. Here, we test this approach in a data set of 160 On × Oa spawns of 109 male and 100 female parents randomly collected from admixed stocks, and genotyped for microsatellite markers representing the known SD loci on linkage groups (LGs) 1, 3, and 23. Following crossbreeding, the most significant paternal effects on male proportions in progeny were found for LG1-BYL018 (P < 2 × 10-32 ) and for LG3-UNH168 × LG23-UNH898 interaction (P < 1 × 10-17 ; R2  = 0.98). Furthermore, a maternal effect for LG3-UNH168 (P < 9 × 10-7 ) was associated with low female proportions in progeny (<7%), indicating a non-Mendelian effect on SD. Eighty-four males (77%) and 30 females (30%) were selected as parents, based on their genetic profiles for the SD loci that were associated with male production. Of these, 51 of 53 crosses produced all-male progeny, while two crosses had low female proportions in their progeny (<4%). This suggests that selection could be improved using the causative sequence variation underlying SD on LG3, since the large non-recombining block of the SD region in purebred Oa readily breaks down in hybrids. Nevertheless, marker-assisted selection for sex determining loci of admixed parental stocks may be used for all-male production.

A duplication of the Anti-Müllerian hormone gene is associated with genetic sex determination of different Oreochromis niloticus strains.

Sex determination (SD) mechanisms are ancient and conserved, yet much diversity is exhibited in primary sex-determining signals that trigger male or female development. In O. niloticus, SD is associated with a male-specific locus on linkage group (LG) 23 which harbors the Y-linked Anti-Müllerian hormone (amh) gene, and a truncated duplication, denoted amhΔy. We have evaluated the possible role of identified indels and SNPs in the amh gene on SD, based on conservation in different O. niloticus strains. A fluorescent assay for the detection of a 5 bp insertion in amhΔy exon VI, efficiently discriminated between XX, XY, and YY genotypes. Concordance rate between amhΔy and sex varied in six Oreochromis strains, from 100% (Ghana) through 90% (Swansea) to 85% (Thai-Chitralada). The association of amhΔy with sex was found to be conserved in all tested O. niloticus strains, and thus supports its key role in SD. However, the previously identified missense SNP (C/T) in amh exon II was found only in the Swansea strain, thus excluding its candidacy for the causal variation of SD across all strains. Effects of markers on LGs 1, 3, and 23 (amhΔy) fully explained sex distribution in one Thai-Chitralada family (R2 = 1.0), whereas in another family only the major effect of LG23 (amhΔy) was significant (R2 = 0.37). Thus, amhΔy on LG23 is associated with genetic SD, either as a single causal gene in different O. niloticus strains, or in combination with segregating genes on LGs 1 and 3 in the Thai-Chitralada hybrid strain.

Distinguishing between Bread Wheat and Spelt Grains Using Molecular Markers and Spectroscopy.

The increasing demand for spelt products requires the baking industry to develop accurate and efficient tools to differentiate between spelt and bread wheat grains. We subjected a 272-sample spelt-bread wheat set to several potential diagnostic methods. DNA markers for γ-gliadin-D ( GAG56D), γ-gliadin-B ( GAG56B), and the Q-gene were used, alongside phenotypic assessment of ease-of-threshing and near-infrared spectroscopy (NIRS). The GAG56B and GAG56D markers demonstrated low diagnostic power in comparison to the Q-gene genotyping, which showed full accordance with the threshing phenotype, providing a highly accurate distinction between bread wheat and spelt kernels. A highly reliable Q classification was based on a three-waveband NIR model [Kappa (0.97), R-square (0.93)], which suggested that this gene influences grain characteristics. Our data ruled out a protein concentration bias of the NIRS-based diagnosis. These findings highlight the Q gene and NIRS as important, valuable, but simple tools for distinguishing between bread wheat and spelt.