Method for continuously producing complete male pelteobagrus fulvidraco by sex reversion and embryonic development

Method for continuously producing complete male pelteobagrus fulvidraco by sex reversion and embryonic development

  • CN 1,994,072 B
  • Filed: 12/28/2006
  • Issued: 08/03/2011
  • Est. Priority Date: 12/28/2006
  • Status: Active Grant
First Claim
Patent Images

1. sex reversal and gynogenesis continue to produce complete male yellow cartfish method, and it comprises sex reversal and gynogenesis and K.PearsonShi X 2Method of inspection is characterized in that being undertaken by following operation:

  • (1) artificial induced spawning;

    select body surface not have wound, belly and expand softness, raun and sexually matured milter that gonad development is good and carry out artificial induced spawning with the ovulation induction element with human chorionic gonadtropin and fish, in the disposable injection of every fish pectoral fin base portion, the ocyodinic using dosage;

    raun is human chorionic gonadtropin 800IU, fish is used the plain 10 μ

    g of ovulation, milter is gonadotrophin chorionic 400IU, and fish is used the plain 5 μ

    g of ovulation;

    Place 26 ℃

    of water temperatures, from the milter body, extrude seminal fluid after 24 hours or cut open the belly and get essence, extrude ripe ovum from raun and carry out artificial insemination;

    Long-snout catfish milter dosage is doubled;

    (2) hormonal reverses;

    as the hormone carrier, the ethinyl estradiol at 50~

    300 μ

    g/L soaked one hour in advance with artemia larvae, and suction filtration takes out the halogen worm as the living bait normal fry XX of 4 days yellow cartfish of the initial feeding ♀

    that throws something and feeds then;

    XY ♂

    respectively throws something and feeds once on every day is early, middle and late, and feeding volume is as the criterion to eat up in 30 minutes, feeds continuously 40 days;

    (3) gynogenesis of the XY ♀

    physiology raun of sex reversal(3.1) sperm deactivation, select long-snout catfish milter activation of spermatozoa yellow cartfish ovum gynogenesis for use, long-snout catfish milter after promoting the sexual maturity from hormone is cut open the belly and takes out spermary, shred with scissors, add 6-8 Hank '"'"' s liquid dilute filtration doubly to the culture dish of Φ

    9cm, seminal fluid thickness 2-3mm places then on the ice bath shaking table of 115 rev/mins of rotating speeds and shakes;

    With a wavelength 2537 The irradiation of 30W ultraviolet lamp tube, range 35cm, exposure intensity is 1 * 764 μ

    w/cm 2And the seminal fluid that takes a morsel at any time observes the sperm motility situation under anatomical lens, generally shines to get final product in 30 minutes;

    When sperm is activated the back major part slowly during swimming by water, promptly stop to shine;

    (3.2) the thelykaryon dliploid obtains, ripe fish-egg with the acquisition of XY ♀

    yellow cartfish physiology raun artificial induced spawning, carry out artificial insemination with the sperm of long-snout catfish deactivation, in insemination back 1-10 minute, handled 5~

    20 minutes with 0-5 ℃

    of frozen water, blocking-up reduction division second polar body is discharged, and recovers the thelykaryon dliploid, taking-up places 26 ℃

    constant incubator to hatch, and just can obtain candidate'"'"'s YY ♂

    supermale yellow cartfish and XX ♀

    raun;

    (4) offspring'"'"'s test cross of candidate YY ♂

    supermale yellow cartfish, the genotype of inferring the candidate milter from offspring'"'"'s female and male ratio is YY or XX, get sexual maturity candidate YY ♂

    supermale yellow cartfish and the normal raun test cross of XX ♀

    , press the bolti standard of formulations such as Mair, promptly use K.PearsonShi X 2Method of inspection is measured the observed value of its offspring'"'"'s property ratio and the degree of conformity of theoretical value;

    When meeting probability P>

    0.05, then be the XY type;

    When P<

    0.001, then be YY or XX type;

    With infer through offspring'"'"'s property than test cross be XY or YY or XX for examination fish genotype, thereby pick out the YY ♂

    supermale yellow cartfish of affirmation;

    (5) complete male yellow cartfish continues to produce, and will promptly get the complete male yellow cartfish of XY ♂

    through the YY ♂

    supermale yellow cartfish and the mating of normal X X ♀

    raun of offspring'"'"'s test cross affirmation;

    To promptly get YY ♂

    supermale yellow cartfish through the YY ♂

    supermale yellow cartfish and the mating of sex reversal supermale physiology raun of offspring'"'"'s test cross affirmation.

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