Studies On Hardwood S Lignin Synthesis Enzymes Critical Genes Genetic Transformation Of Chinese Fir

Chinese fir ,Cunninghamia lanceolata (Lamb.) Hook., is one of the evergreen coniferous species, native species, and native to the People's Republic of China. Because of its rapid growth, high wood quality, Chinese fir has been widely cultivated wood for a long time in Southern of China. Been widely cultivated, it accounts for the top of the Southern coniferous plantations, timber plantations forefront of the accumulation ranks at present accounts for the top of the South conifer plantations, timber plantations forefront of the accumulation ranks. However, restrictions on the chemical structure of lignin, the woody biomass energy conversion in the restricted. This test attempts to use hardwood lignin synthase key enzyme gene-CAld5H to transformed Chinese fir. The main contents and results were as follows:High-efficient tissue culture system was established with stem section as explants. And studied the effect of different factors for callus induction, subculture and differentiation of adventitious buds. It showed that the optimal medium for callus induction was P6 + BA 1.5mg/L+ 2,4-D 0.2mg/L + NAA 0.1 mg/L,the rate was 100%; The best callus subculture medium was DCR + 2,4-D 0.5mg/L + BA 0.5mg/L + KT 0.5mg/L + TDZ 0.008 mg/L, the proliferative multiple was 5.4 after cultured 15d. Adventitious buds can be induced on DCR + BA 1.5 mg/L + NAA 0.1 mg/L + KT 1.5 mg/L, the differentiation rate reached 18.0±2.5%, and there were 5.5±0.43 buds. All media was added 30g/L sugar and 6g/L Agar. This test also studied callus browning, adding inhibitors and changing the culture conditions had not significant effect.Research on gene transformation system has found that the selective pressure should be controlled 10～20mg/L in induction phase and 5～8mg/L in differentiation phase. Through the transient expression of GFP gene,we tried to explore the optimum parameters,it showed as follows:target distance at 9cm, 300μg gold particle per shoot, vacuum pressure at 28 inches Hg, target DNA of 1.5μg per bombardment,2 times bombardment(s6cm﹠9cm).In the test of transformation of CAld5H gene, there wasn't resistant callus obtained, indicating that the foreign gene was not integrated into the plant genome. So the new methods need to be further researched.