| To achieve Sapindus(Sapindus mukorossi Gaertn.)the clone plantation model of efficient cultivation of raw material,this study conducted on selection set up rather county of fujian province with superior individual tender stem section,young stem section of greenhouse seedling and young leaves of solid seedlings as explant of no preliminary exploration of sub system of tissue culture,the main results were as follows:(1)The factors affecting the disinfection effect of the explants were as follows:explant>0.1%HgCl2>2%NaClO>75%ethanol.The best way to sterilize the young stem segments of seedlings in greenhouse is:firstly,75%ethanol is used to sterilize 20 s,then 0.1%HgCl2 is used to sterilize 6 min,and finally 2%NaClO is used to sterilize 2 min.The rate of sterile explants obtained by this method is 87.75%.The best way to sterilize the tender stem segment of the optimal plant is:first soak in 75%ethanol for 10 s,then sterilize with 0.1%HgCl2 for 8 min,and finally soak in 2%NaClO for 2 min.The survival rate of the optimal plant reaches 76.37%,which is 50.34%higher than other disinfection methods.The best way to disinfect the young leaves of wannanseeds is:first,soak them in 75%ethanol for 10 s,then treat them with 0.1%HgCl2 for 6 min,and finally soak them in 2%NaClO for 2 min.The survival rate is the highest(89.37%).(2)In march,the contamination rate and Browning rate of the young stem segment of the excellent plant were significantly lower than that of the explants collected in other months(P<0.05),which were 28.80%and 17.78%,respectively.The contamination rate and Browning rate of overwintering bud explants collected in February were the highest,reaching 86.67%and 84.44%respectively.(3)The best medium to induce callus from young leaves was MS+NAA O.lmg/L+2,4-d 2.0mg/L+IBA 0.15mg/L,with the highest induction rate(97.5%)and the fastest average start-up time(6.96d).(4)The optimal culture medium for the germination of axillary buds in the young stem segment of excellent plant was MS+NAA 0.2mg/L+6-ba 3.0mg/L+IBA 0.05mg/L,with the highest induction rate(57.53%)and the shortest average start-up time of axillary buds(8.68d).The optimal medium combination to induce the germination of axillary buds in the tender stem segment of the seedless seedlings in greenhouse was MS+NAA 1.0mg/L+IBA 0.05mg/L+6-ba 1.0mg/L,with the highest germination rate(89.67%)and the shortest average start-up time of axillary buds(7.00d).(5)MS+NAA 0.2mg/L+2mg/L6-ba+0.4mg/L KT was the most suitable medium to induce the proliferation of axillary buds in the tender stem segment of seedless seeds,and the proliferation coefficient was 2.24 after 30 days.(6)VC has an obvious effect on controlling the Browning of the explants without seeds.When 1 g/L VC is added into the culture medium,the probability of Browning of the explants is low(41.73%).The effect of PVP was the second.When the concentration was 0.1gg/L,the Browning rate was the lowest,43.39%.The effect of AC on controlling Browning was the least satisfactory,and the Browning rate of the explants at the three concentrations was up to 50%.In the experiment of setting different illumination duration to explore the influence on the Browning of tissue culture seedlings without diseased seeds,the control effect of completely dark culture was the best,with the Browning rate of 26.55%.The Browning rate of the explants cultured under full light was the highest,which was 83.68%.Therefore,the Browning rate could be reduced by placing the explants in the condition of no light.Conclusion:this experiment was to not have children disinfection method of tissue culture,cultivation,transgenerational in early generation and Browning control aspects has carried on the different levels of inquiry,suggested that still need to improve in the future of successive transfer culture multiplication and rooting culture more in-depth exploration,in order to promote without risk of seed propagation system set up and improve. |
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