Induced diploid gynogenesis and polyploidy in the ornamental (koi) carp Cyprinus carpio L.
The results of a series of experiments conducted in our laboratory on the ornamental common carp (koi), aimed at optimizing heat-shock chromosome-set manipulation procedures, are described. The timing of heat-shock initiation was expressed in the relative unit of embryological age (τ0) in order to standardize this parameter, the absolute time for heat-shock initiation being calculated from duration of one τ0 at two different pre-treatment water temperatures. Heat shocks were applied within the periods of 0.05–0.60 τ0 and 1.20–2.20 τ0 which, respectively, cover the successive phases of the 2nd meiotic division and the 1st cleavage. The highest production of diploid gynogenetic offspring was observed when heat shocks were initiated at 0.15–0.25 τ0 and at 1.5 τ0, after insemination, corresponding to anaphase of meiosis-II, and metaphase of the 1st cleavage, respectively. Similar results were obtained irrespective of the different pre-treatment water temperatures, thus confirming the possibility of standardizing heat-shock timing by τ0.
Gynogenesis and sex determination in large-scale loach Paramisgurnus dabryanus (Sauvage)
Gynogenesis was induced using heterologous sperms in large-scale loach, Paramisgurnus dabryanus (Sauvage), in which a ZW/ZZ sex determination was previously proposed. Three microsatellite loci were used to monitor exclusive maternal inheritance of gynogenetic progenies. The results showed that high percentages of meiogynogens were produced at 4 min post-fertilization and mitogynogens were produced at 18 min post-fertilization by heat shocks, while meiotic gynogenesis was induced by cold shocks within a wide period and high heterozygosity was even observed in gynogens produced at 24 min post-fertilization. The sex ratios of the F1 progenies in three gynogenetic families were significantly deviated from 1:1 expectation with a female bias in two families and a male bias in one family (P < 0.05), and the other four gynogenetic families showed approximate 1:1 sex ratios. Moreover, the self-mating between gynogenetic F1 progenies and mating between gynogenetic F1 progenies and normal individuals produced all-female progenies or identical proportions of females and males. The data of sex ratios generally confirmed that the sex determination in large-scale loach was determined by the putative ZW/ZZ system, and the possible reasons causing the biased sex ratios are discussed.
Histocompatibility analyses in tetraploids induced from clonal triploid crucian carp and in gynogenetic diploid goldfish
Histocompatibility analyses in goldfish were performed using the tetraploid goldfish-crucian carp hybrid and the first generation of gynogenetic diploid (GD1) goldfish. Tetraploids were obtained by crossing clonal triploid crucian carp with goldfish. GD1 goldfish were produced by the suppression of the second meiotic division. Tetraploid scale grafts on triploid clone members evoked an acute rejection in 4–6 days, whereas the reverse transplants were accepted or rejected chronically. Reciprocal grafting between tetraploids showed subacute rejection in 10–12 days, although some fish showed chronic rejection in 20–30 days. On the other hand, scale grafts reciprocally exchanged among triploids were intact even 3 months after grafting, although some of them showed a unidirectional rejection pattern. Furthermore, allograft rejection among gynogens occurred between 5 and 20 days, whereas all the scale allografts between members of control siblings were rejected within 9 days. In addition, neither accelerated acute rejection nor acceptance of allografts was observed in grafts exchanged among GD1 goldfish. These results suggest that single doses of histocompatibility alleles are effective in eliciting acute rejection, and each of the fourth haploid set of chromosomes originating from paternal goldfish might share the same histocompatibility antigens to a large extent. This experiment also indicates that the genecentromere recombination rate is quite high with respect to the histocompatibility loci in this species.