| Ormosia hosiei is an evergreen tree belonging to the ormosia genera of Leguminous family.It is a unique native tree species in China and a national class II key protected precious tree species.It is of great significance that to study the tissue culture technology of O.hosiei and improve its reproduction coefficient for the rapid cultivation of O.hosiei seedlings and the expansion cultivation.In this study,the stem segments,apical buds,leaves,petioles and mature embryos of O.hosiei were used as explants.On the basis of screening suitable explants,the start-up culture conditions were carried out.The tufted buds were induced,the proliferation of different explants was cultured,the rooting inside and outside the bottle were compared,and the transplanting technology of test tube seedlings were studied in order to construct a complete tissue culture and breeding system.The main findings as follows:1.The most suitable explant for the tissue culture of O.hosiei was stem segment,with 100% survival rate and 91.55% bud induction rate and to produce clump shoot;both the top bud survival rate and bud induction rate were 100%.but its apical dominance was more obvious,which inhibited the production of clustered shoots,with the low proliferation rate;neither leaves nor petiole could induce the buds.2.The best condition for the establishment of the aseptic system of O.hosiei was :0.1% KMn O410 min + 75% ethanol 30 s + 0.1% Hg Cl28 min with lower than6.98% pollution rate and 100% survival rate;The effects of start-cultivate for O.hosie were the best when in the basic medium 1/2MS,sucrose concentration 30g/L,agar 8g/L and p H6.0 conditions,the induction rate of buds was 85.19%.3.The stems segment had the best inducing effect in 1 / 2MS + TDZ0.02 mg / L+ NAA0.2mg / L medium,with 91.43% induction rate,the number of clusters of buds up to 6.12.TDZ and NAA had a significant effect on the clump shoot number and the browning induced by explants.By the multiplication culture experiment of three explant(stem segments,buds,original Stem segments),the results showed the stemsegments was the best explant for multiplication culture,the best culture medium was WPM+6-BA1mg/L+NAA0.5mg/L,with 96.15% proliferation rateand the 5.88 multiplication multiple.4.The optimum medium for the tube inside rooting was 1/2MS+NAA 0.5mg/L,with 87.37% rooting rate.The type of medium had a significant effect on the rooting rate.The best method for rooting outside the bottle was NAA 150mg/L and putting the bottom of rootless seedlings soaked quickly about 4~5s,and then cuutting into matrix of peat soil: perlite: vermiculite(3:1:3),with 97.78% rooting rate.Compared with the tube inside rooting,the rooting rate of outside the test tube was 10.14%higher than that inside the tube,and it could save time and cost,increase economic,so we suggest that best rooting method for O.hosiei is rooting outside the test tube.5.The optimum ventilation condition and nutrition condition are the key factors for the survival of domestication and transplanting of O.hosiei tissue culture seedlings.The best transplanting matrix are peat soil,Perlite,vermiculite at rate of3:1:3,with 93.33% survival rate,and the growth of seedlings is better. |
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