Improvement Of In Vitro Rapid Multiplication In Butterfly Orchids

The improvement of in vitro rapid multiplication in butterfly orchids was systematically studied. The main results were as follows:(1) The experiments using the pedicle axillary bud, as explant showed that the sterilization for 15min was optimal. The highest efficiency of shoot formation was more than 90% in MS medium with 3mg/L BA.(2) Shoot tips produced the higher percentage of PLB with the highest percentage up to 100% on MS medium supplemented with 5.0 mg/L 6-BA and 0.5mg/L NAA. The percentage of PLB was increased by the cutting of shoot tips.(3) The optimal medium for the proliferation of PLB was MS medium with 3mg/L BA , 0.2 mg/L NAA and 1.5%. activated carbon and the highest proliferation rate reached 2.3. The volume of PLB affected the proliferation rate of PLB and PLB of 3.0 mm in diameter gave a highest proliferation rate up to 2.3.(4) PLB turned green and differentiated through the long culture. The 1/2MS medium with 6-BA and NAA was beneficial for the rooting with a average rooting rate of 3.2.(5) The survival rate of plantlets was increased by 12.5% by transition cuLture in comparison with the control. Before thansplanting to pots with fog,the plantlets treated for 3d in room and then 3d in the greenhouse gave the highestsurvival rate up to 90%.Twq somaclones, round leaf of butterfly orchids and split leaf of butterfly orchids, were obtained in this study.(6) A. tumefaciens GV3101 harboring the plasmid pBI121 was used for transformation of GFP gene driven by the CaMV 35S promoter. The 129 putative transgenic plants were regenerated on the selection medium containing 20mg/L kanamycin. PCR analysis indicated that 59 of the 129 putative transgenic plants were transgenic plants of butterfly orchids.