Tissue Culture And Construction Of Genetic Transformation Elementary Study In Rose

The Rose(Rose hybrid L.)originated in China,is one of the four fresh cut flowers in the world.With the development of Rose industry,the demands of consumers have become increasingly diversified.However,due to the limitation of traditional cross breeding technology and narrow genetic background of original rose varieties.It's difficult to achieve a breakthrough in the improvement of the intrinsic quality.Modern molecular breeding technology can be targeted to the rapid transfer of good target genes into the recipient plant,so as to achieve the purpose of improving its intrinsic quality.Molecular breeding technology will play an irreplaceable role in the Rose breeding for its pertinence and high efficiency.In this thesis,Stem segments with axillary buds and leaves of Rose.‘Peach Avalanche',Rosa.‘Avalanche' and Rosa.‘Nirvana' were used as explants.The effects of multiple combinations of plant growth regulators on the germination of axillary buds and the induction of leaf callus were studied.The effects of different infestation time and co-cultivation time of agrobacterium on the genetic transformation efficiency of rose callus were analyzed Resistant callus of rose were gotten after transformation.All the findings of this study were as follows:1.From the study for induction of Rose.‘Peach Avalanche',Rosa.‘Avalanche' and Rosa.‘Nirvana' axillary buds.germination,We can get the conclusion that optimum medium for axillary bud germination of Rose.‘Peach Avalanche',Rosa.‘Avalanche' and Rosa.‘Nirvana' are MS+6-BA 1.5 mg/L+2,4-D 0.1 mg/L,MS+6-BA 1.5 mg/L+2,4-D 0.5 mg/L and MS+ 6-BA 1.5 mg/L+2,4-D 0.5 mg/L,respectively.2.Callus induction was performed using aseptic leaves and isolated leaves of Rose.‘Peach Avalanche',Rosa.‘Avalanche' and Rosa.‘Nirvana' Callus induction efficiency is the highest on the MS medium with 4.0 mg/L 2,4-D.The callus induction efficiency of the aseptic leaf is significantly higher than the isolated leaves.The callus induction efficiency of Rose.‘Peach Avalanche',Rosa.‘Avalanche' and Rosa.‘Nirvana' aseptic leaves reached 94.64%,97.78% and 89.58%,respectively.3.Effects of different infection time and co-cultivation time on the transformation efficiency of the callus genetic transformation system mediated by Agrobacterium tumefaciens were studied.The results showed that 25 minutes infection time with Agrobacterium tumefaciens and 3 days co-culture time treatment on the callus of Rose.‘Peach Avalanche',Rosa.‘Avalanche' and Rosa.‘Nirvana' all had the highest GUS expression rate of 79.26%,69.33% and 65.27%,respectively.4.The physiologically active resistant callus was obtained by reducing the concentration of Hyg and Cef in the selection medium.The resistant callus of Rose.‘Peach Avalanche'and Rosa.‘Avalanche',which with HygB,grew normally in the medium of MS+2,4-D 2.0 mg/L+Hyg 10 mg/L+Cef 250 mg/L,and the physiological activity was restored.