Studies On The Culture In Vitro And Agrobacterium-Mediated Transformation Of Acacia Crsaaicarpa

In this paper, we studied culture in vitro of Acacia crassicarpa from three aspects: subculture and propagate of aseptic; callus induction and differentiation; root induction and sound plantlets and plantlets transplantation. Studied the agrobacterium-mediated transformation of Acacia crassicarpa from as the follow: effects of infection time and pre-culture and co-culture time on genetic transfrormation. The system of in vitro culture regeneration system and agrobacterium-mediated transformation of Acacia crassicarpa was established. The establishment of in vitro culture and agrobacterium-mediated transformation of Acacia crassicarpa can provide the important references for further research. The results were included in the following:1,The results of using 6-BA or KT single for aseptic seedling regeneration were not excellent. The suitable regeneration medium was amend MS+6-BA0.5 mg·L^-1+KT1.0 mg·L^0.1. In this medium the aseptic seedling could regeneration in high multiplication times as well as keep the strong growth potential, and the multiplication achieved to 3.4 times.2,Different explants of Acacia crassicarpa could induct callus effective by using 2,4-D, the highest rate of calus induction was reached 100%; but the callus were hard to differentiate a buds.3,Different explants of Acacia crassicarpa could induct callus effective by using 6-BA, and the callus could differentiate into buds well. The suitable differentiation medium was amend MS+6-BA2.5 mg·L^-1, in this medium the differentiation was highest. The rate of differentiation was depended on the explants. Explants -compound leaf was cultured in this media gained the highest differentiate rate of 55. 86%.4,NAA or IAA, IBA, ABT could promote the aseptic seedling of Acacia crassicarpa's root induction. MET didn't suit for the aseptic seedling of Acacia crassicarpa's root induction.5,The survival rate of plantlets transplantation was different from using different hormone categories or its combination. Plantlets transplant in yellow mud showed the highest survival rate, which roots was inducted in medium of ABT0.5-2.0 mg ? L^-1+MET0.5 mg ? L^-1, and the survival rate achieved to 86. 39%.6,There were different effects of different transplanting matrix on the survival rate of plantlets transplantation. When the yellow mud and nursery stock was in the ratio of one to one, the mixture matrix for transplanting gained the suitable rate of survival, the rate of survival was 84.36%.7,The explants of Acacia crassicarpa had high Km resistance background. The Km resistance level was 100-125 mg ? L^-1.8,Agrobacterium of At05 or At06, or A208SE could not grow when the medium contained Tb 300 mg ? L^-1, but explants could keep high differentiate rate in this medium.9,Different kinds of Agrobacterium, different concentration of the Agrobacterium, different infection time, different pre-culture time and co-culture time affected on the rate of GUS gene instantaneous expression.