黑核桃组培中消毒及防止褐化最适浓度分析

Study on disinfection and prevention of browning in tissue culture of black walnut

【目的】筛选出消毒的最佳时间和抗褐化剂的最适浓度,研究外植体细胞结构在褐化过程中的变化,为黑核桃的组培研究和工厂化育苗提供理论基础。【方法】采用不同时长对黑核桃叶片和茎段进行消毒处理,观察其对污染率、褐化率的影响;以AC、PVP、VC 3种抗褐化剂进行正交试验,研究其对黑核桃茎段褐化率和萌发率的影响;观察褐化外植体徒手切片。【结果】(1)叶片以0.1%HgCl_(2)消毒5 min,污染率和褐化率最低;(2)茎段以二次消毒法,0.1%HgCl_(2)加吐温80消毒5 min,污染率和褐化率最低;(3)以AC 2 g/L、PVP 110 mg/L、VC 100 mg/L处理的褐化率最低;以AC 1 g/L、PVP 70 mg/L、VC 50 mg/L处理的萌发率最高。【结论】叶片以0.1%HgCl_(2)消毒5min为最佳消毒时间;茎段以二次消毒法,0.1%HgCl_(2)加吐温80消毒5 min为最佳消毒时间;最适的抗褐化剂浓度配比为AC1g/L、PVP 110 mg/L、VC100 mg/L。

【Objective】The optimal time for disinfection and the optimal concentration of anti-browning agent were screened out,and the changes in the cell structure of the explants during the browning process will be discussed in the hope of providing theoretical basis for tissue culture research and industrial seedling of black walnut.【Methods】The leaves and stems of black walnut were disinfected with different lengths of time to observe their effects on the pollution rate and browning rate.Three anti-browning agents,including AC,PVP and VC,were used in orthogonal experiments to study their effects on the browning rate and germination rate of black walnut stems.After that,freehand section of the browned explants was observed.【Results】(1)When leaves were disinfected with 0.1%HgCl_(2) for 5 min,the contamination rate and browning rate were the lowest.(2)When the stem segment was disinfected with 0.1%HgCl_(2) and tween 80 for 5 min,the contamination rate and browning rate were the lowest by the two-step sterilization.(3)Treatment with AC 2 g/L,PVP 110 mg/L,VC 100 mg/L had the lowest browning rate and the germination rate of AC 1 g/L,PVP 70 mg/L and VC 50 mg/L was the highest.【Conclusion】The best disinfection time is to disinfect the leaves with 0.1%HgCl_(2) for 5 minutes.The best disinfection time is to disinfect the stem segment with 0.1%HgCl_(2) plus tween 80 for 5 min by the two-step sterilization.The most suitable anti-browning agent concentration ratio is AC 1 g/L,PVP 110 mg/L,VC 100 mg/L.