Institute of Marine Research, 5984, Matredal, Norway.
Institute of Aquaculture, School of Natural Sciences, University of Stirling, Stirling, FK9 4LA, Scotland, UK.
BMC Genet. 2020 Jul 8;21(1):71. doi: 10.1186/s12863-020-00878-8.
In several fish species homozygous and heterozygous clonal lines have been produced using gynogenetic and androgenetic techniques. These lines are standardized and can be reproduced over generations. In rainbow trout such lines have existed for decades and has become important research tools in genome studies as well as in studies of commercially important traits. The Atlantic salmon is one of the best studied fish species globally, but all experiments are done on fish of wild or domesticated origin and access to standardized immortal fish lines would be of great benefit. Here, we describe the protocols developed to produce mitotic gynogenes, and from these the first clonal lines in Atlantic salmon.
Atlantic salmon eggs fertilized with UV irradiated sperm combined with a pressure shock applied at 4700-4800 minC at 8 °C gave all homozygous (doubled haploid) gynogenetic progeny with high survival. From the six first maturing females, five all homozygous clonal lines were produced by meiotic gynogenesis and were verified as clonal and identical to their mother with microsatellite markers.
We have now produced the first documented cloned Atlantic salmon lines. This work demonstrates the potential for production of further Atlantic salmon clonal lines, potentially with distinct characteristics. Such lines will provide an important resource for further elucidation of phenotypic and genetic traits in this globally important species.
在一些鱼类物种中,通过雌核发育和雄核发育技术已经产生了纯合和杂合的克隆系。这些系已经标准化,可以在几代中繁殖。在虹鳟鱼中,这些系已经存在了几十年,并且已经成为基因组研究以及商业上重要性状研究的重要研究工具。大西洋鲑是全球研究最多的鱼类之一,但所有实验都是在野生或驯养的鱼类上进行的,获得标准化的永生鱼类系将非常有益。在这里,我们描述了生产有丝分裂雌核发育鱼的方案,并从这些鱼中产生了第一批大西洋鲑克隆系。
用紫外线照射的精子受精的大西洋鲑卵,结合在 8°C 下施加的 4700-4800 minC 的压力冲击,产生了具有高存活率的所有纯合(双倍单倍体)雌核发育后代。从最初成熟的六只雌鱼中,通过减数分裂雌核发育产生了五只全部纯合的克隆系,并通过微卫星标记验证为与它们的母亲相同的克隆系。
我们现在已经生产了第一批有记录的克隆大西洋鲑鱼系。这项工作表明,有可能生产出更多具有不同特征的大西洋鲑鱼克隆系。这些系将为进一步阐明这个全球重要物种的表型和遗传特征提供重要资源。