| Vernicia fordii and Camellia oleifera, both are unique woody oil tree species in southern China, has an important economic value. Vernicia fordii is an important industrial oil tree species, tung oil is a good raw material for the manufacture of paint, ink and biodiesel. Camellia oleifera is a high quality woody edible oil tree, the unsaturated fatty acid content of camellia oil is more than 90%, has the ability of antioxidant, scavenging free radicals and prevention of cardiovascular and cerebrovascular diseases and so on, is a recognized high quality healthy cooking oil. Vernicia fordii, Camellia oleifera, walnut and tallow known as the four woody oil tree species in China. In this study, mature embryos of Vernicia fordii and Camellia oleifera as materials, studied embryo disinfection treatment, sterile seedlings of Vernicia fordii and Camellia oleifera were obtained. Induction of adventitious buds by petiole,cotyledon and hypocotyl of sterile seedling, then the adventitious bud propagation, elongating, rooting, seedling training and transplanting, established the tissue culture technology system, played the foundation for genetic transformation of Vernicia fordii and Camellia oleifera.The main research results are as follows:1.The results show that the optimal disinfedtion time of embryos is 8 minutes by 0.1% mercuric chloride, the appropriate medium of embryo is 1/2MS+1.0 mg/L GA3, and seeding rate is up to 85.42%. Using germ-free petioles of Vernicia fordii as explants, the fittest medium for shoot differentiation of petioles was 1/2MS+3.0 mg/L 6-BA+0.05 mg/L IAA, with a 91.67% shoot induction rate; the best medium for shoot multiplication was 1/2MS+3.0 mg/L 6-BA+0.05 mg/L IBA+1.0 mg/L GA3, with the multiplication coefficient reaching 4.89; the best rooting medium was 1/2MS+0.05 mg/L IBA, with a 96.18% rooting rate; after the culture-bottle seedlings were transplanted into the matrix composed of peat soil, perlite and loess (2:1:1), the survival rate was up to 93.55%.2.Using germ-free hypocotyls of tung tree as explants, the fittest medium for callus induction of hypocotyls were WPM+5.0 mg/L6-BA+1.0 mg/LKT+0.1 mg/L NAA, with a 100% callus induction rate; the fittest medium for shoot differentiation were WPM+1.0 mg/L 6-BA+0.05 mg/L NAA+2.0 mg/L GA3, with a 82.46% differentiation rate; the best medium for shoot multiplication were WPM+2.0 mg/L KT+0.05 mg/L IAA+2.0 mg/L GA3, with the multiplication coefficient reaching 6.8; the best medium for elongation culture were WPM+1.0 mg/LNAA+2.0 mg/L GA3; the best rooting medium was 1/2MS+0.1 mg/L IBA, with a 97.12% rooting rate; after the culture-bottle seedlings were transplanted into the matrix composed of peat soil, perlite and loess (1:1:1), the survival rate was up to 91.67%.3.The optimum disinfection method for Camellia oleifera embryos was using 75% alcohol immersion 30 seconds, washed with sterile water for 5 times, then using 0.1% HgCl2 immersion 6 mintues, the appropriate medium of embryo is 1/2MS basic medium, and seeding rate is up to 81.11%. Hypocotyls and cotyledons of sterile seedling were used as explants, the fittest medium for shoot differentiation were 1/2MS+3.0 mg/L 6-BA+0.05 mg/L IAA and 1/2MS+3.0 mg/L 6-BA+0.05 mg/L IAA+0.05 mg/L NAA+2.0 mg/L GA3, with a 87.28% and 83.35% shoot induction rate; the best medium for shoot multiplication was 1/2MS+3.0 mg/L 6-BA+0.05 mg/L IBA+1.0 mg/L GA3, with the multiplication coefficient reaching 7.0 and 7.4; the best elongation medium was WPM+0.5 mg/L NAA+5.0 mg/L GA3; the best rooting medium was 1/2MS+1.0 mg/L IBA+2.0 mg/L NAA+35 g/L perlite, with a 91.02% rooting rate; after the culture-bottle seedlings were transplanted into the matrix composed of peat soil, perlite and loess (2:1:1), the survival rate was up to 86.67%. |
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