| This experimentation uses rieger begonia's (begonia×elatior) leaves, leafstalks and stems as explanted issue, inducing the formation of adventitious buds, culture and proliferate, and in the end induce the rhizogenesis, transplant reborn young plant, constitute the fast propagation system of rieger begonia. MS as the basic culture medium, this article analyses the affection of different explanted issue type, and different ratio of auxins and cytokines on the adventitious buds's differentiation, to explore the condign hormone ratio of rieger begonia multiplication culturing, and compare the different effect of white granulated sugar and sucrose. In the end, selects the optimal culture medium for rieger begonia's tube rhizogenesis, Ascertaines a series of technique parameter combination, effectively improves the breeding efficiency.Explore the genetic transformation of rieger begonia on the basis of fast propagation system, use agrobacterium tumefacien-mediated transformation of rieger begonia's leaf discs. The result shows that:1) The rieger begonia's explanted issues undergo different degree of differentiation in 6-BA0.2-2mg/L and NAA0.5-1mg/L, which illuminates that within certain range the hormone for rieger begonia's differentiation is not strict, several compound portions can make it happen, MS+BA0.2mg/L+NAA0.1mg/L as the condign differentiation culture medium;2) The condign explanted issue type for rieger begonia tissue culture is middle-typed mature lamina, which has low death rate, fast shooting and high differentiation ratio, being the good explanted issue provider. Compared with leaves, leafstalks and stems only has little differentiation of buds. When material is not abundance, leafstalks and stems as explanted issue also are appropriate.3) The time for subculture culturing can not exceed 60d, or the young plant would brown and die. Between 30-40s there are a great amount of effective young plants and the multiplication ratio is high.4) The difference between advanced white granulated sugar and pure sucrose in rieger begonia's multiplication growth and rhizogenesis effect is not remarkable, so the white granulated sugar can replace pure sucrose for reducing the culturing cost greatly.5) Rieger begonia's reborn young plant can take root easily, without any hormone the 1/2MS culture medium can induce the rooting, but the roots are very thin and weak. NAA can has great effect on the rooting , the condign culture medium formula for rieger begonia's reborn young plant rooting is 1/2MS+NAA0.05mg/L6) With or without pure sucrose in the rooting culture medium ,cultured rieger begonia roots are not quite different, but in the latten the plants are thinner and the laminas are flimsier and the color is weaker.7) Rieger begonia is quite sensitive to kan, the concentration of with should be relatively low in selective culture medium and rooting culture medium; rieger begonia is not sensitive to cef, so wash bacterium can be ignored in constructing the genetic transformation system, the cultured laminas can be directly inoculate in the MS+cef600mg/L+kan40mg/L culture medium to undergo the selective culturing;8) The condign time of begonia elatior infection is about 10min.9) The explanted issues undergoing 3d beforehand-culture or not before infection are not quite different in differentiation time, and re-rooting frequency, so in order to shorten the transformation time, rieger begonia transformation can ignore the beforehand-culture.10) Using rieger begonia's leaves as material to undergo genetic transformation, no blue pigmentation stains appear under GUS pigmentation, and the targeted genes are not transformed into received body, which is consistent with the experiential results of Sanae Kishmoto. |
