| In this study,we used different parts of Euonymus as explants and discussed the effect on stem disinfection of sampling time and sterilization method,the influence factors of seed germination.We studied the effects of sand reservoir a,mature and immature,with aril and without aril,different concentrations of GA3 on seed germination;the effects of NAA(0.5,1,2.0mg/L)and 6-BA(0,1 mg/L)on the induction of adventitious buds in mature embryos of sand and without sand;the effects of adventitious bud induction on stem segment with NAA(0.5,0.05mg/L),6-BA(1,2 mg/L)and agar(6.5,7.0g/L);the effects of callus induction on floral organs(calyx,petal,stamen and pistil)and leaf with basic medium,2,4-D,TDZ and dark culture time,and choose the best explants and the induction method.Then selected the optimal medium for adventitious bud proliferation from basic medium,6-BA,NAA and GA3 of three levels.Finally,the best method of in vitro rooting was selected from 28 treatments of adding NAA,IAA,IBA(0.2-4.0mg/L)and GA3(0.2-0.8mg/L),the best in vitro rooting methods were compared with out vitro rooting methods of substrate,rooting agent and concentration in four levels,selected the best rooting method.The results are as follows:(1)Mid-May is the best sampling time of Euonymus stem segments,the infection rate was 3%and the survival rate was 76.67%;mercuric chloride combine two drops of tween-20 for 8min is the best method for Euonymus stem segments sterilization,the infection rate was 3%and the survival rate was 76.67%;(2)Aril sand storage on seed germination rate is the highest(10.2%)in MS+GA32.0mg/L medium compared with the other treatment,and cotyledons germination after 20d.The callus induction rate was higher in leaves with leaf midrib in WPM+2,4-D 0.8 mg/l+ TDZ mg/1 medium(65%-67%),but pistil callus induction rate(mean of 86.5/%)is highest in B5+2,4-D 1.2 mg/l+ TDZ 2 mg/l+ 20 d dark culture.However,pistil callus was loose green,it is not the best callus morphology.Therefore callus induction did not succeed,remains to be studied.The best medium for the induction of adventitious buds on embryo was MS+6-BA1.0mg/L+.NAA0.5mg/L,the induction rate of adventitious buds was 12.8%.the adventitious induction rate of stem segments in 6-BA2.0mg/L+ NAA0.05mg/L medium was 70.67%,which was much higher than embryo.So,stem segments is the best explants for adventitious bud induction.(3)MS+ 6-BA1.0mg/L+NAA0.05mg/L is the best medium for adventitious bud proliferation of Euonymus,the proliferation coefficient was 23.51,that is,the number of buds is 23.51.After subculture 3 times,the number of buds could be increased to 48.5.(4)The best in vitro rooting method is l/2MS+GA30.4mg/L for Euonymus,rooting rate is 72.5%,the average rooting number is 4.1.The best condition for out vitro rooting is vermiculite +IAA2500mg/L,the rooting rate is 100%,the average rooting number is 5.3.Two kinds of methods can effectively promote the rooting of Euonymus plantlets rooting,but out vitro rooting is the best rooting method. |
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