Study On In Vitro Culture Of Unfertilized Ovary Of Garlic

Took six different genotypes garlic varieties as experimental materials, the garlic floralorgan could get well development by means of garlic stem culture in vitro. Throughunfertilized ovary in vitro culture respectively, garlic haploid plantlet regeneration system wasestablished preliminary. The suitable culture condition and the influence factors of callus ofgarlic unfertilized ovary culture were experimentally studied. And through the multiplicationof the callus of ovary, regeneration plants were obtained. The results showed as follows,1. Garlic callus induction and differentiation culture need suitable culture mediumconditions. Took ‘Caosuan’ unfertilized ovary as experimental materials for in vitro culture,callus could be formed on B5medium and MS medium, the result of B5medium was betterthan MS medium; N6medium was unfavorable because no ovary garlic callus formed on it.The suitable garlic ovary callus induction hormone combinations was2mg·L^-16-BA+1mg·L^-1NAA, and callus induction frequency reached11.11%. B5medium with3mg·L^-16-BA+1mg·L^-1NAA had the highest adventitious bud differentiation frequency46.15%. The optimalculture medium for root induction was B5+0.05mg·L^-1NAA.2. Low temperature pretreatment and short-term heat shock treatment both improved theovary garlic callus induction rate. Took unfertilized ovary before flowering1²d of‘Caosuan’ and ‘Zhengyuezao’ as experimental materials, giving4℃low temperaturepretreatment for0（CK）,1,2,4,6d and35℃heat shock treatment for0（CK）,12,24,36and48h. The result showed that4℃for4d treatment could significantly improve the callusinduction rate,35℃for48h treatment could significantly increase the induction rate. But withheat shock treatment of ovary whatever time length, browning was relatively serious andbrowning rate was over60%, so the method should not be adopted.3. Different genotypes of ovary had different callus induction rate. Take the unfertilizedovary before flowering1²d of ‘Caosuan’,‘Ershuizao’,‘Puke’,‘Zhengyuezao’,‘Hongqixing’as experimental materials, the genotype of garlic which had the highest ovary callus induction11.11%is ‘Caosuan’, the followings were ‘Zhengyuezao’,‘Puke’,‘Hongqixing’,‘Erjizao’,‘Ershuizao’ had the lowest ovary callus induction0.4. The development degree of ovary has a significant influence on induction rate ofregenerative plants. Took ‘Caosuan’,‘Puke’,‘Zhengyuezao’ as experimental materials, theovaries on the flowering day has higher ovary callus induction rate than the ovaries at thestage2³d and6⁷d before flowering day, and those three induction rate were13.82%, 8.00%,13.25%respectively.5. Taking normal diploid garlic leaves for comparison, flow cytometry was used toidentify of regenerated plants. The result showed that14of16regenerated plants from calluswere haploid plants and the rest2regenerated plants were diploid plants, the2regeneratedplants from gynogenesis embryos were haploid plants.