Marine Biotechnology

, Volume 12, Issue 4, pp 380–385

Production of Viable Homozygous, Doubled Haploid Channel Catfish (Ictalurus punctatus)

  • Geoffrey C. Waldbieser
  • Brian G. Bosworth
  • Sylvie M. A. Quiniou
Original Article

DOI: 10.1007/s10126-009-9221-2

Cite this article as:
Waldbieser, G.C., Bosworth, B.G. & Quiniou, S.M.A. Mar Biotechnol (2010) 12: 380. doi:10.1007/s10126-009-9221-2

Abstract

Production of doubled haploids via mitotic gynogenesis is a useful tool for the creation of completely inbred fish. In order to produce viable doubled haploid channel catfish, we utilized hydrostatic pressure or thermal treatments on eggs fertilized with sperm that had been exposed to ultraviolet light. At 1.5 h post-fertilization, the embryos were exposed to either 590 kg/cm2 hydrostatic pressure for 3 min, 37°C for 5 min, or 41°C for 3 min. In the pressure-treated group, only 21 offspring hatched from five spawns with family sizes of one, two, two, four, and 12 offspring each. Eight embryos from the 37°C treatment and 32 embryos from the 41°C treatment survived to hatch. Genotype analysis using microsatellite loci demonstrated all 21 offspring resulting from pressure treatment were homozygous at the 64 loci tested, and none contained alleles unique to the donor male. Eleven of 32 offspring from the 41°C treatment were homozygous at the 18 loci tested, while 21 offspring were heterozygous at six to 12 of these loci. Again, no offspring contained alleles unique to the donor male. However, all eight offspring from the 37°C treatment were heterozygous at multiple loci, and one contained unambiguous paternal alleles. These experiments demonstrated our ability to produce viable homozygous, doubled haploid channel catfish. Doubled haploid catfish can be used to create completely inbred populations for genetic analyses, and homozygous genomic templates will be useful in gene identification and genome characterization.

Keywords

Channel Catfish Homozygous Doubled haploid Gynogenesis 

Supplementary material

10126_2009_9221_MOESM1_ESM.doc (34 kb)
ESM Table 1Microsatellite loci used for genotype analysis of doubled haploid catfish. Fluorophore is denoted at beginning of forward primer (DOC 33 kb)
10126_2009_9221_MOESM2_ESM.doc (358 kb)
ESM Table 2Genotypes of gynogenetic fish produced by pressure treatment of activated catfish eggs. Only one allele is displayed in homozygous fish (DOC 358 kb)
10126_2009_9221_MOESM3_ESM.doc (49 kb)
ESM Table 3Genotypes of gynogenetic fish produced by 37°C—5 min treatment of activated catfish eggs. Only one allele is displayed in homozygous fish (DOC 49 kb)
10126_2009_9221_MOESM4_ESM.doc (135 kb)
ESM Table 4Genotypes of gynogenetic fish produced by 41°C—3 min treatment of activated catfish eggs. Only one allele is displayed in homozygous fish (DOC 135 kb)

Copyright information

© US Government 2009

Authors and Affiliations

  • Geoffrey C. Waldbieser
    • 1
  • Brian G. Bosworth
    • 1
  • Sylvie M. A. Quiniou
    • 1
  1. 1.Catfish Genetics Research Unit, Agricultural Research ServiceUS Dept of AgricultureStonevilleUSA

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