High-efficiency In Vitro Regeneration In Hippeastrum Vittatum ‘Blossom Peacock’ And ‘Royal Velvet’ Based On Different Explants

Hippeastrum vittatum are internationally famous bulb flowers with bright colors,large flowers and wide application range.They have low natural reproduction rate,usually using ball splitting,cutting and sowing methods,but the reproductive efficiency are low,which are difficult to meet the growing market demand.Tissue culture technology for in vitro rapid propagation is one of the most effective methods of industrial breeding in horticultural plants.However,the tissue culture has not achieved industrial production,and the technical system is still not perfect of Hippeastrum vittatum.In order to further improve the high-efficiency in vitro regeneration technology system of Hippeastrum vittatum.Effects about different concentrations as well as ratios of plant growth regulators on adventitious bud induction and callus induction were studied with the scales,flower organs,and plantlet leaves of ’Blossom Peacock’ and ’Royal Velvet’.In order to lay a technical foundation for the industrialization of of Hippeastrum vittatum bulbs.The main test results are as follows:1.Explants with ’Blossom Peacock’ and ’Royal Velvet’ scales of Hippeastrum vittatum.The concentration of TDZ significantly affects the induction.The best explants are the base scales(0.5 cm).The optimal callus induction medium was MS+2.0 mg/L 6-BA+1.0 mg/L NAA+2.0 mg/L TDZ,in which ’Blossom Peacock’ formed callus(embryonic callus was 100%)after 50 days of culture,the induction rate reached 80.00%,continue to culture for 30 days to form somatic embryos,and the developed somatic embryos were subcultured in MS medium for 35 days to form plantlets;’Royal Velvet’ formed callus after 60 days of culture,the induction rate reached 64.29%,continue to culture for 60 d to form adventitious buds.The optimal small bulb induction medium was MS+0.5 mg/L NAA+3.0 mg/L TDZ,the induction rates of ’Blossom Peacock’ and ’Royal Velvet’ reached 71.43% and 70.00%,respectively.2.Explants with ’Blossom Peacock’ and ’Royal Velvet’ petal and stalk of Hippeastrum vittatum.When the petals are used as explants(no coloration in young flower buds),the optimal culture condition is under dark culture for three weeks to light culture(lighting16 h/d,light intensity 36 μmol/m2/s).The best adventitious buds induction medium was MS+0.5mg/L NAA+1.0 mg/L TDZ,and the induction rates of ’Blossom Peacock’ and ’Royal Velvet’reached 83.33% and 77.78%,respectively.The stalk was used as the explant,and the optimal culture conditions is to under light culture;the optimal induction of adventitious bud medium was MS+0.5 mg/L NAA+2.0 mg/L TDZ,the induction rates of ’Blossom Peacock’ and ’Royal Velvet’ reached 37.50% and 29.17%,respectively.3.Explants with ’Blossom Peacock’ and ’Royal Velvet’ plantlet leaves of Hippeastrum vittatum.The results showed that the best explants were the base of young leaves(0.5 cm) formed by culturing in MS medium for 10 days.The photoperoid is 16 h/d(light intensity 36μmol/m2/s)and cultured the leaf convex surface in contact with the medium.The best medium for adventitious buds induction of Hippeastrum vittatum was MS+2.0 mg/L6-BA+1.0 mg/L NAA+2.0 mg/L TDZ.The adventitious buds of both varieties occurred in an indirect way,and the ’Blossom Peacock’ formed callus after 40 days of culture,adventitious buds formed on 55 days,the induction rate reached 69.44%;’Royal Velvet’ formed callus after45 days of culture,and adventitious buds formed on 65 days,the induction rate reached66.67%.The optimal adventitious bud proliferation medium was MS+2.0 mg/L 6-BA+1.0mg/L NAA+1.0 mg/L TDZ,and the proliferation coefficients of ’Blossom Peacock’ and ’Royal Velvet’ reached 4.67 and 3.46,respectively.The optimal somatic embryo induction medium was MS+2.0 mg/L 6-BA+1.0 mg/L PIC,and the induction rate of ’Blossom Peacock’ and’Royal Velvet’ reached 66.67% and 63.89%,respectively.4.Comprehensively visible,the best explants were the base of young leaves of’Blossom Peacock’ and ’Royal Velvet’.The photoperoid is 16 h/d(light intensity 36 μmol/m~2/s).The best medium for adventitious buds induction was MS+2.0 mg/L 6-BA+1.0 mg/L NAA+2.0 mg/L TDZ.The optimal adventitious bud proliferation medium was MS+2.0 mg/L6-BA+1.0 mg/L NAA+1.0 mg/L TDZ.Obtaining a large number of plantlet of Hippeastrum vittatum.Rooting culture was carried out in MS medium without added hormones,and the rooting rate of both varieties reached 100% after 30 days.The Hippeastrum vittatum plantlets after 30 days of rooting culture were moved to room temperature for 3 days.Then remove the sealing film domestication for 3 days.They were transplanted to the high temperature sterilized charcoal: vermiculite(volume ratio)1:1 matrix,and then the survival rate reached 100%.