Studies On Tissue Culture Of Pinus Nigra Var. Austriaca

The study was used mature zygotic of Pinus nigra var. austriaca that bought from Germany as the initiation explants of tissue culture, with organogenesis, to investigate the technology of induction, differentiation, elongation, rooting of adventitious buds and transplanting of regeneration plantlet.The result showed that the seeds of Pinus nigra var. austriaca were pretreatment with 4℃for 15 days and were soaked in water for 24 hours . The well-marinated seeds were sterilized by using 0.1%KMnO4 for 30min, after flushing by sterile distilled water, seedcoat was removed.Then the embryo was soaked in 70%alcohol for 45s, transferred them into 0.1%HgCl2 for 8min,and washed by sterile distilled water.Under disinfective condition ,the endosperm was striped to get mature embryo into adventitious buds medium.The rate of pollution and browning were low. And we could get more adventitious buds when the mature embryos were aclinic on mediums. The inducement of adventitious buds generally initiate on the cotyledon or the upper hypocotyls.In the study, the best mediun of induction adventitious buds was 3/4WPM supplemented with 2.0mg.L^-16-BA and 30g.L^-1 sucrose, which could produce a great deal of adventitious buds. The rate of induction was more than 55 per cent. And GD+6-BA0.8mg.L^-1+ NAA0.01mg.L^-1+white sugar50g.L^-1 was the best combination for propagation. The rate of propagation was about 900 per cent.Then transferred them on the elongation medium of MS+0.02mg.L^-1NAA+AC1.0g.L^-1.GA3 went against the growth of elongation.The study also used the plantlets of sterile seedlings and seedlings in soil to induce tufted buds. The result was that the seedlings in soil would have more influence on induction of tufted buds because of the more rate of pollution. The highest propagation index of tufted buds of sterile seedlings was 6.23 on the 1/2GD+3.0mg.L^-16-BA+0.05mg.L^-1NAA+ 0.2mg.L^-1IBA+20g.L^-1sucrose, and sterile seedlings cultured on medium for 20 to 30 days were better.In the end, experiment had studied the rooting of adventitious buds and the transplanting of regeneration plantlets. The study showed that rooting in tube was much better than out tube of adventitious buds of Pinus nigra var. austriaca.WPM+NAA0.02mg.L^-1+ IBA0.5mg.L^-1+sucrose2.0% was the best combination for rooting in-tube than others; The...