The Establishment Of Regeneration System Of Five Species Of Primula

Primula is rich in species resources.Primula having showy brightly colored flowers and high ornamental value.Its main breeding method is seed reproduction.But its seed life is short.Its ermination rate,seedling proliferation rate and proliferation rate is low,so it is difficult to satisfy the planting requirements.The use of tissue culture technology can accelerate the breeding speed and better maintain fine varieties.Primula vulgaris,Primula obconica,Primula denticulata‘Cashmeriana Ruby',Primula bulleyana and Primula×pubescens were used as material in experiment.The epicotyls and hypocotyls,leaves of sterile seedlings,petioles of sterile seedlings and axillary buds of sterile seedlings were used as explants.Through the exploration of seed germination conditions,callus and adventitious bud induction,multiplication of adventitious buds,rooting culture and transplanting,the plant regeneration system of these five species of Primula were established.The results show that:?1?Different kinds of sterilizing agents,sterilization time,illumination and temperature affected the effect of seed germination of Primula vulgaris and Primula obconica.The optimum seed sterilization of Primula obconica was 30 seconds of0.1%HgCl₂.The best method of sterilizing Primula vulgaris seeds was 10 minutes of6%NaClO.The seeds of Primula vulgaris and Primula obconica germinated under the condition of 12 hours of illumination and temperature of 20²5?,which had relatively high germination rate.?2?The optimal medium of Primula vulgaris for inducing adventitious shoots from epicotyls explants was MS+NAA 0.4 mg/L+6-BA 0.6 mg/L,the induction rate was21.67%.The optimal medium for callus induction from petioles explants was1/2MS+NAA 1 mg/L+6-BA 0.1 mg/L,the induction rate was 77.78%;the optimal medium for differentiation was 1/2MS+NAA 0.5 mg/L+6-BA 0.1 mg/L,the differentiation rate was 88.89%.The optimal medium for callus induction from leaves explants was 1/2MS+NAA 1 mg/L+6-BA 0.5 mg/L,the induction rate was 84.44%;the optimal medium for differentiation was 1/2MS+NAA 0.5 mg/L+6-BA 0.5 mg/L,the differentiation rate was 82.22%.The optimal medium for inducing adventitious buds from axillary bud explants was MS+NAA 0.1 mg/L+6-BA 1 mg/L,and the induction rate was 97.78%.The hypocotyls explants could not induce adventitious buds.The best medium for induction of multiple buds was MS+NAA 0.6 mg/L+6-BA 0.5 mg/L,the proliferation rate was 62.96%.The optimal medium for rooting was MS,the rooting rate was 95.59%.?3?The optimal medium of Primula obconica for inducing adventitious buds from epicotyls explants was MS+NAA 0.1 mg/L+6-BA 0.6 mg/L,the induction rate was26.67%.The optimal medium for callus induction from petioles explants was MS+NAA1 mg/L+6-BA 0.1 mg/L,the induction rate was 82.22%;the optimal medium for differentiation was MS+NAA 0.1 mg/L+6-BA 0.5 mg/L,the differentiation rate was82.22%.The optimal medium for callus induction and differentiation from leaves explants was MS+NAA 0.5 mg/L+6-BA 0.1 mg/L,the induction rate was 84.44%and the differentiation rate was 88.89%.The optimal medium for inducing adventitious buds from axillary bud explants was MS+NAA 0.5 mg/L+6-BA 0.1 mg/L,and the induction rate was 68.89%.The hypocotyls explants could not induce adventitious bud.The best medium for induction of multiple buds was MS+NAA 0.6 mg/L+6-BA 0.2 mg/L,the proliferation rate was 29.63%.The optimal medium for rooting was MS+NAA 0.1 mg/L,the rooting rate was 96.30%.?4?The optimal medium of Primula denticulata‘Cashmeriana Ruby'for callus induction and differentiation from leaves explants was MS+NAA 1 mg/L+6-BA 0.1mg/L,the induction rate was 77.78%and the differentiation rate was 82.22%.The optimal medium for inducing adventitious buds from axillary bud explants was MS+NAA1mg/L+6-BA 0.1 mg/L,and the induction rate was 73.33%.The petiole explants could not induce callus and adventitious buds.The best medium for induction of multiple buds was MS+NAA 1 mg/L+6-BA 0.5 mg/L,the proliferation rate was 85.19%.The optimal medium for rooting was 1/2MS+NAA 0.2 mg/L,the rooting rate was 95.59%.?5?The optimal medium of Primula bulleyana for callus induction and differentiation from petioles explants was MS+NAA 0.1 mg/L+6-BA 0.5 mg/L,the induction rate was 75.56%and the differentiation rate was 80%.The optimal medium for inducing adventitious buds from axillary bud explants was MS+NAA 0.1 mg/L+6-BA0.5 mg/L,and the induction rate was 100%.The leaf explants could not induce callus and adventitious buds.The best medium for induction of multiple buds was MS+NAA0.6mg/L+6-BA 2 mg/L,the proliferation rate was 92.59%.The optimal medium for rooting was 1/2MS+IBA 0.1 mg/L,the rooting rate was 88.89%.?6?The optimal medium of Primula×pubescens for inducing adventitious buds from axillary bud explants was 1/2MS+NAA 0.5 mg/L+6-BA 2 mg/L,and the induction rate was 73.33%.The leaf explants and petiole explants were browning during induction,and callus and adventitious buds could not be induced.The best medium for induction of multiple shoots was MS+NAA 0.2 mg/L+6-BA 0.5 mg/L,the proliferation rate was96.30%.The optimal medium for rooting was MS+NAA 0.2 mg/L+IBA 0.3 mg/L,the rooting rate was 92.60%.?7?Tissue culture of these five species of Primula seedlings were transplanted into the mixture of peat and perlite,and the volume ratio was 3:1,with a maximum survival rate of 96.67%.