Institute of Marine Research, Bergen, Norway.
University of Stirling, Stirling, Scotland, UK.
Mol Ecol Resour. 2024 Oct;24(7):e14004. doi: 10.1111/1755-0998.14004. Epub 2024 Aug 6.
Polyploidy occurs naturally across eukaryotic lineages and has been harnessed in the domestication of many crops and vertebrates. In aquaculture, triploidy can be induced as a biocontainment strategy, as it creates a reproductive barrier preventing farm-to-wild introgression, which is currently a major conservation issue for the industry. However, recent work suggests that triploidisation protocols may, on occasion, produce 'failed triploids' displaying diploidy, aneuploidy and aberrant inheritance. The potentially negative consequences for conservation and animal welfare motivate the need for methods to evaluate the success of ploidy-manipulation protocols early in the production process. We developed a semi-automated version of the MAC-PR (microsatellite DNA allele counting - peak ratios) method to resolve the allelic configuration of large numbers of individuals across a panel of microsatellite markers that can be used to infer ploidy, pedigree and inheritance aberrations. We demonstrate an application of the approach using material from a series of Atlantic salmon (Salmo salar) breeding experiments where ploidy was manipulated using a hydrostatic pressure treatment. We validated the approach to infer ploidy against blood smears, finding a > 99% agreement between these methods, and demonstrate its potential utility to infer ploidy as early as the embryonic stage. Furthermore, we present tools to assign diploid and triploid progeny to families and to detect aberrant inheritance, which may be useful for breeding programmes that utilise ploidy manipulation techniques. The approach adds to the ploidy verification toolbox. The increased precision in detecting ploidy and inheritance aberrations will facilitate the ability of triploidisation programmes to prevent farm-to-wild introgression.
多倍体在真核生物谱系中自然发生,并已被用于许多作物和脊椎动物的驯化。在水产养殖中,三倍体可以作为一种生物控制策略来诱导,因为它会产生一种生殖障碍,防止从养殖场到野外的基因渗入,这是该行业目前的一个主要保护问题。然而,最近的研究表明,三倍体化方案有时可能会产生“失败的三倍体”,表现出二倍体、非整倍体和异常遗传。这对保护和动物福利的潜在负面影响促使人们需要在生产过程早期就评估倍性操作方案的成功。我们开发了 MAC-PR(微卫星 DNA 等位基因计数 - 峰比)方法的半自动版本,以解决大量个体在微卫星标记面板上的等位基因配置问题,该方法可用于推断倍性、系谱和遗传异常。我们使用一系列大西洋鲑(Salmo salar)繁殖实验中的材料展示了该方法的应用,这些实验使用静水压力处理来操纵倍性。我们验证了该方法推断倍性与血涂片的一致性,发现这两种方法的一致性超过 99%,并证明其在胚胎阶段推断倍性的潜在用途。此外,我们还提出了将二倍体和三倍体后代分配给家庭并检测异常遗传的工具,这对于利用倍性操作技术的繁殖计划可能很有用。该方法增加了倍性验证工具包。在检测倍性和遗传异常方面的精度提高,将有助于三倍体化计划防止从养殖场到野外的基因渗入。