Improvement On Industrial Production Technique In Vitro Culture Of Anthurium Andraeanum

Because of high ornamental and economic value, Anthurium andraeanum, as a tropical flower, shows a wide market prospect. With the development of modern biological technology, tissue culture had been applied to in vitro propagation of A. andraeanum, which could greatly enhance the propagation coefficient. However, the protocol of micropropagation should be simplified to reduce the cost of industrial production. In this study, 14 cultivars were employed to investigate the effect of factors which affected in vitro culture of A. andraeanum. A system for callus induction and regeneration at very high frequency was established. The transplanting technique for test-tube plantlet was optimized. Furthermore, in order to improve the genetic resources of A. andraeanum, colchicine was employed to induce polyploidy and 12 tetraploid plants were obtainedThe main results are as followings:The effects of 6-BA (1.0-2.0 mg.L^-1),2, 4-D (0-0.5 mg.L^-1),NAA (0-0.1 mg.L^-1)and IAA(0-0.1 mg.L^-1) in different combinations and at different concentrations on proliferation of Anthurium andraenum were studied by orthogonal design L₉ (3)⁴.The results shows that the hormone concentration had no significant effect on the regeneration ability of the calli.The visual analysis, variance analysis and multiple comparison proved that stem was the optimum explant. 1/2MS+BA 2.0 mg.L^-1 +2,4-D 0.5mg.L^-1 +IAA 0.1 mg.L^-1,with a callus induction rate of up to 97.25%,was the optimal callus induction medium, from which the calli were yellowish and loose in texture; the optimal medium for the adventitious buds induction was1/2MS+BA 2.0 mg.L^-1+2,4-D 0.5mg.L^-1,with a rate of 157.3 % and many buds; the optimal medium for callus induction from the roots was 1/2MS+BA 2.0mg.L^-1 +2,4-D 0.5mg.L^-1,with a rate of 18.3%.Taking the sterile fragments as explants,the rapid propagation technique was established.calli,adventitious buds and roots could be induced on three kinds of media simultaneously, but the media of l/2MS+6-BA2.0mg.L^-1+NAA0.05mg.L^-1 was the optimal media for calli induction, the media of 1/2MS + 6-BA 1.5mg.L^-1+NAA 0.05 mg.L^-1+ IAA0.05mg.L^-1 is for to adventitious buds induction, and the media of 1/2MS+6-BA1.0mg.L^-1+NAA0.05mg.L^-1 is for root induction. The propagation duration could be reduced to todays. The explants alternately cultured on above three media and a some or even better effect was observed. However, the genotype had a significant effect on callus induction and bud differentiation, which resulted in a cullum induction rate varied from 100% to 42.9%; and a buds regeneration rate varied from 305%to38.9 %, ,in 14 cultivars tested,13 cultivars could regenerate roots directly and be transplantedTransplanting technique of tube seedlings was studied on A. andraeanum and the results indicated that the optimal procedure was to soaking the seedling with 0.1% of KMnO₄ for 15 mins before transplantion; in the media of cocobran: turves:perlite with a rate of 2:2:1 or 2:1:1,the management after transplanted for two months was also critical to the product of tissue culture seedling on a large scale. Based on the above method, the survival rate of transplanting was up to 96% and the survival plants was very vigorous.The hydroponics technique and medium transplantation technique of A.andraeanum tube-seedling was compared. The results showed that the hydroponics in vitro is superior to another on the survival rate and the growth plantlet. The cultivars of "Toscane", "Clarinervium" and "Rapido" were superior to the other varieties.There are many other advantages of hydroponics, such as high survival rate,low infection rate and growing rapid.The test-tube stem explants were immersed into the solution of 0.1% colchcine for 72 hours to induce its autotetrapliody and induction rate was up to 40% . The length of stomata on the lower epidermis of leaves from tetraploid plant was notably longer than those of the diploidy, while the density of stomata was obviously decreased.In addition,, Autotetraploid plants of Anthurium andraeanum were tall and large with thicker and bottle green leaves. It is a new resource of good quality and ornamental of Anthurium andraeanum.The cost of propagating 1000000 tube-seedlings of Anthurium andraeanum per year was analyzed.Using former tissue culture methed, the cost of per container tube-seedling isï¿¥0.329,and the cost of per container tube-seedlings reduced toï¿¥0.177.When using the optimited methed. For one million test-tube plantlets output, the total input can be savedï¿¥152000. In this thesis, based on the combination of industrial production and laboratory research, a regeneration system of A. andraeanum for its industrial production has been set up, which will direct the industrialization of other flowers.