Studies On The Tissue Culture Of Abies Concolor

The techniques research about the tissue culture of Abies concolor will lay a foundation for its vegetative propagation and application of other biotechnology. In this thesis, the technology about tissue culture of Abies concolor was studied in detail. The whole technique system included sterilization, optimal explants selection and the optimal media selection for initiation, proliferation, rooting.The results were as follows:(1) Different disinfectant times were tested with the 70% alcohol and 0.1% HgCl2. For seeds, disinfested in 0.1% HgCl2 for 8 minutes was the best and its pollution rate was the lowest (1.11%). For shoot tips, disinfested in 0.1% HgCl2 for 2 minutes was the best and its pollution rate and browning rate were the lowest.(2) Mature embryos, cotyledons, cotyledon-hypocotyls, hypocotyls, shoot tips, terminal buds from mature embryos of different provenances of Abies concolor were taken as explants. Mature embryos, shoot tips and terminal buds were the suitable explant materials.(3) The optimal induction medium of adventitious buds consisted of one-half strength MS medium and 2.7mg/L 6-BA. The results indicated that there were differences among various provenances. Adventitious buds had achieved from mature embryos of the provenance 4 of which induction frequency reached 23.33%. The suitable initiation media of shoot tips were MS medium or MS medium containing 0.1mg/L 6-BA.Significant differences were detected among provenances, the initiation frequency ranged from 80.0% to 93.3%. The best germination medium of mature embryos was DCR medium. The germination frequency ranged from 83.33% to 98.33%. The callus induction was successfully taken place in darkness on the SH medium containing 1.8mg/L 2,4-Dand 1.2mg/L6-BA.(4) Adventitious buds elongate difficultly and browned to death with the culture time gone by. It is a difficult problem that callus would browned to death, after it wastransferred 2 to 3 times. The selected induction medium of axillary buds was MS medium containing 0.3mg/L 6-BA and 0.1mg/L IBA. The shoot tips of provenance 8 produced more axillary buds than provenance 12. The height of shoot tips of provenance 12 was higher than that of provenance 8. The differentiation medium of terminal buds was SH medium containing 0.4~0.5mg/L 6-BA and 0.3mg/L IBA. The differentiation frequency ranged from 40.00% to 76.67%. The axillary buds and dormant buds could elongate 1-2 cm height on MS medium.(5) The shoots of Abies concolor were difficult to root. At present, the roots were only achieved from shoots of provenance 513. The rooting medium was SH medium containing 0.5mg/L 6-BA, 0.5mg/L IBA and 0.5mg/L NAA.All media contained 30g/L sucrose (rooting medium contained 20g/L) and 6.5g/L agar. The pH of all media was adjusted to 5.8 prior to autoclaving. The temperature in the tissue culture room was adjusted at 24C +2C, with an illumination time of 12-14 hours and illumination intensity 1200~1600Lx.Shoots were achieved from mature embryos, shoot tips, terminal buds, but which were difficult to root and further work was needed.