| Efficiency and stable of plant regeneration provides critical basis for genetic transformation, while Prunus persica (L.) Batsch which belongs to Prunus spp. is believed a genus hard to regeneration, thus, factors effecting plant regeneration should be optimized. Adventitious buds were successfully regenerated in the present study from hypocotyls of five genotypes'Qinmi','Shahong','Tiantaowang','Jintong5'and'97-8-4'of Prunus persica (L.) Batsch. The effect of genotypes, composition of basal medium and combination of plant growth regulators on adventitious bud formation was conducted. Adventitious buds derived from leaves of in vitro grown of Prunus persica (L.) Batsch were also obtained, four genotypes'Bumuzaosheng','Tiantaowang','Ruijiang'and'97-8-4'were applied and several factors in this system were studied.The main results are as the follows:1. The regeneration rate from hypocotyls is significantly genotype dependent. In the same culture condition, the regeneration percentages of'Tiantaowang','Jintong5'and'97-8-4'are significant higher than'Qinmi'and'Shahong'.2. Considering regeneration percentage and numbers of adventitious buds per hypocotyl, The most superior medium for genotypes of'Tiantaowang','Jintong5'and'97-8-4'is LP medium when comparing with MS and G medium. Regeneration percentages of 65.79%, 68.44% and 68.98% can be obtained from hypocotyls in the presence of 2.0 mg/ L BA and 0.02 mg/ L NAA.3. Callus induction and regeneration of adventitious buds from leaves are founded genotype-dependent. Up to 100% callus induction were obtained for genotypes of'97-8-4'and'Tiantaowang', a higher percentage of adventitious buds regeneration in this two genotypes than'Bumuzaosheng'(10%), while no adventitious bud was obtained in the genotype of'Ruijiang'.4. The optimal medium for callus induction is MS medium, the optimal medium for regeneration of adventitious buds is LP medium. In the genotype'97-8-4', callus could be inducted in the all tested medium: MS, LP and G, the most effective medium for callus induction is MS, while LP medium show the best reaction ( up to 18.33% ) for regeneration of adventitious buds, which is significant higher than MS and G medium.5. The regeneration ability can be affected by the development degree of leaves, the fully expanded younger leaves show best reaction for callus induction and adventitious buds regeneration, up to 96.67% and 18.33% was obtained, respectively. Callus can be also efficiently induced (83.33%) from the non-fully expanded leaves and subsequently regeneration of adventitious buds, while non-expanded and mature leaves have shown lower callus induction and no adventitious buds were obtained.6. Different parts of a single leave have been identified showing different regeneration ability. Petiole and the basal part are better than the medium and upper part of a leave, the upper part could not regenerate adventitious buds.7. At a certain concentration of cytokine, the higher concentration of NAA, the higher percentage of callus induction and callus formation per leave were obtained as well as more compact of callus8. When the leaves cultured in the dark, the optimal combination and concentration of plant growth regulators are 2.0 mg/ L BA + 0.2 mg/ L NAA. While in the light condition, 4.0 mg/ L BA+ 0.1 mg/ L NAA or 6.0 mg/ L BA+ 0.2 mg/ L NAA have shown the best results. In the optimized condition, the regeneration percentages of adventitious buds derived form the fully expanded younger leaves are 18.33%, 16.67% and 13.33% for'97-8-4','Tiantaowang'and'Bumuzaosheng'respectively. |
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