Tissue Culture And Rapid Propagation Of Dianthus Superbus L.

This experiment got sterile seedlings by mature seeds of Dianthus superbus L., using its stem segment as explant,and then studied the technology of tissue culture under those works such as explant initiation culture,proliferation and subculture,root growing culture and transplantation.In order to achieve factory cultivating work,summarized some factors that influenced the test-tube seedling proliferation,such as inoculate seedling status and number,subculture period and frequency.Firstly,tried to use non-tube rootage work to tissue culture of D. superbus,and explored a new method of rapid propagation,and then provided data foundation for factory cultivating work.The results were as follows:1.The condition of initiation culture and proliferation should be with agar concentration8g/L,sucrose concentration30g/L,25℃and30001x light with12h-14h in chamber.In the rootage procedure,used the same condition except sucrose concentration15g/L.Kept lighting all the time and22℃-24℃when transplanting at early stage and dropped temperature after15d.2.The optimal method for seed sterilization:75%alcohol soaked30s,then cleaned it5times using sterile water,then used0.1%HgCl+Tween to sterilize for5min,and in this condition pollution rate was0,death rate was10.00%.3.Explant initiation culture:the best explant was sterile seedlings stem,whose initiantion rate was the highest with93.33%.The best initiation medium was MS,the best cell mitogen was6-BA,the best auxin was NAA,and the optimal proportion was6-BA3.0mg/L+NAA0.3mg/L.4.Proliferation and subculture:the best proliferation medium was MS with proliferation coefficient3.07.The best cell mitogen was KT,the best auxin was NAA,and the optimal proportion was KT4.0mg/L+NAA0.05mg/L.In this condition the leaves were dark green and plant height was2.5cm-3.5cm and had no callus.5. The optimal inoculant seedling amount was10per bottle in subculture,and then test-tube seedling grew well with4or5buds splitted each seedling,leaf area were also nearly uniform.6.The best subculture period was30d with proliferation coefficient achieved to4.0.Cluster buds differentiated clearly,and leaves’ color and area were uniform.Subcultured4or5times were the optimal choose,and the average proliferation coefficient was4.1without glass seedling phenomenon.7.1n rootage work,the optimal medium was1/4MS+sucrosel5g/L and auxin was NAA2.0mg/L,and average root account was15.33,and average root length was3.27cm,and root developed and grew well,and leaf area were also nearly uniform.Because of less influence adding active carbon,suggested adding no active carbon in rootage work.8.Non-tube rootage:after4-5times proliferation,it should be taken to transplant as standard that leaves area were uniform,the color of leaves were dark green and plant height were higher than2.5cm.9.Transplant and naturalization:the optimal medium for transplanting was garden soil:turf:sandy soil=1:1:1,with23℃and lighting all time.The best training seedlings method was kept the cap opening.