Method for constructing allotetraploid of bluntsnout bream
A technique of allotetraploidy and method establishment, applied in the field of fish breeding
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Embodiment 1
[0059] The establishment of embodiment 1 optimal induction condition
[0060] Orthogonal is diploid bream ♀ × diploid trigonum bream ♂; reciprocal is diploid bream ♀ × diploid bream ♂.
[0061] Set from 1.0τ after fertilization 0 ~2.6τ 0 (That is, when the water temperature is 22°C, from 27min to 60min; at 24°C, from 17min to 50min, τ 0 Proportional to water temperature and time), at different stages of the first mitotic stage of fertilized eggs of bream, heat shock treatment was performed at three different temperatures of 41.0°C, 40.0°C and 39.0°C every 3 minutes, and the duration of treatment 2min or 3min. The fertilized eggs were stuck on the petri dish, and the heat shock was carried out in a constant temperature water bath. Each experimental group was set up with 3-4 parallels, and the embryonic chromosome analysis was carried out in order to establish the appropriate conditions for tetraploid induction.
[0062] The results show that the orthogonal heat shock induct...
Embodiment 2
[0063] The establishment of embodiment 2 head bream allotetraploid
[0064] In the present embodiment, divide into three groups to carry out test altogether:
[0065] Test group 1: adopt the following allotetraploid establishment steps (1)-(7);
[0066] Test group 2: adopt the following allotetraploid establishment steps (1)-(7);
[0067] Experimental group 3: Only interspecies hybridization of bream ♀×triangular bream ♂ was carried out without the heat shock induction step.
[0068] (1) Choose different genetic distance combinations of distant hybridization: Orthogonal bream ♀ × trigonal bream ♂; reciprocal crossing triangular bream ♀ × trigonal bream ♂.
[0069] (2) Select well-mature female and male broodstock, use ovulation-stimulating hormone No. 2 (LHRHa) and human chorionic gonadotropin (HhCG) for mixed induced labor and dry insemination.
[0070] (3) Carry out forward and reverse hybridization.
[0071] (4) Heat shock induction was performed on the hybrid fertilize...
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