Studies On The Fast Propagation In Vitro Of High-yeild Resin Slash Pine

Mature zygotic of Pinnus elliottii that bought from America was used as theinitiation explants of tissue culture, with organogenesis, to investigate the technology ofinduction, differentiation, elongation, rooting of adventitious buds and the study of opentissue culture of Pinnus elliottii. In this experiment, through the study of tissue culture ofPinnus elliottii, to establish a stable, efficient and low-cost system of plantletregeneration for slash pine.The contents of the study including: the technology of induction，differentiation, elongation,rooting of adventitious buds. With the methods of Plant morphology and anatomy were used tounderstand organogenesis and the tentative exploration of open tissue culture of Pinnus elliottii. Theresults showed:To sterilize the explant, with the orthogonal test of L9（32） to reassure the time ofsterilization. The time of2%NaClO:5min、10min、15min;0.1%HgCl₂:30s、60s、90s.The results showed that the best treatment are2%NaClO soak it for5min, thentransformed to0.1%HgCl₂90s to sterilize. Contamination rate was to8%, and thebrowning rate was about18%.The entire embryo is the optimal explant, the effect of embryo which get rid of rootis similar to it, but it is easy to transform to callus tissue, which is suitable for theinduction of indirect organogenesis. With the orthogonal test of L₉（3⁴） to induction. Thebest medium to induce the adventitious buds is: MS+2.0mg·L^-16-BA+0.05mg·L^-1NAA,the rate of induction is up to67.3%.With the orthogonal test of L₉（3⁴） to refirm the best condition of bud proliferation.The DCR medium supplemented with6-BA3.0mg·L^-1and NAA0.05mg·L^-1, the averageof the number of bud proliferation is up to6.90, and the bud is healthy. The best mediumfor increasing the height of cluster buds is DCR medium of0.05%AC, which is nosupplementation on hormonal.With the orthogonal test of L₉（3⁴） to select the best condition of rooting. PDAmedium is the best basal medium to the induction of roots, with0.1mg·L^-1IBA in it.Coconut shell powder was the best substrate, the rooting rate reached to73%and suitableto transplant.Through observation, differentiations of organ were understood. The callus has twostyles: the callus with/without the capacity of inducing organ. The callus which compacthas the capacity to form meristematic tissue and had the capacity of differentiation.Initially callus cells formed in the cortex, trails of tracheids were present but not yetobviously connected from primordia to the main tracheids of the stem. Then tracheidsdeveloped and formed primordia leading to external roots. On the basis of traditional tissue culture, the open tissue culture of Pinus elliottiiwas explored. Through comparing the sterilization effect of different concentrationssodium hypochlorite soaked vaccination appliance, chose0.8%sodium hypochlorite isthe optimal concentration of sterilization, which can controle the infect effectively.Adding0.01%NaClO to substitute for high-temperature autoclave, during the opentissue culture, chose DCR as the basal medium with5g·L^-1suger can decrease the costsand control the infect.