李梦君 1谭善芳 1吴长军 2唐华 2李艳 2陈海霞1
作者信息
- 1. 湖南农业大学 园艺学院,湖南 长沙 410128
- 2. 怀化林泉药业有限公司,湖南 怀化 418199
- 折叠
摘要
Abstract
[Objective]To screen the appropriate sterilization method for the primary rhizome explants of Polygonatum kingianum,and to provide a scientific reference for the optimization of tissue culture technology of P.kingianum as well as the sterilization technology for tissue culture of plants in the Polyg-onatum genus and other medicinal rhizome plants.[Method]Primary rhizomes of Polygonatum kingianum were used as the research object,and the exogenous contamination was eliminated through aseptic opera-tion.Seven different sterilization treatments were set up—after surface sterilization with 75％ethanol for 30 seconds:(A)soaking in 0.1％HgCl2 for 30 minutes;(B)soaking in 0.2％HgCl2 for 15 minutes;(C)soaking in 0.15％HgCl2 for 20 minutes;(D)soaking in 5％effective chlorine content NaClO for 10 minutes,rinsing with sterile water,then soaking in 0.1％HgCl2 for 10 minutes;(E)soaking in 5％effective chlorine content NaClO for 10 minutes,rinsing with sterile water,then soaking in 0.2％HgCl2 for 5 minutes;(F)soaking in 2.5％effective chlorine content NaClO for 5 minutes,rinsing with sterile water,then soaking in 0.15％HgCl2 for 10 minutes;(G)soaking in 2％effective chlorine content Na-ClO for 15 minutes,rinsing with sterile water,then soaking in 0.1％HgCl2 for 14 minutes.The explants were inoculated onto the adventitious bud induction medium,cultured for 45 days,and the contamination rate during the culture process,survival rate,and adventitious bud proliferation coefficient were deter-mined.[Result]The disinfection effect of single HgCl2 treatment was significantly better than that of the combined treatment of NaClO and HgCl2:1)Contamination rate:Group A had the lowest contamination rate(11.7％),which was significantly lower than all combined treatment groups(D:21.7％,E:35.0％,F:28.3％,G:26.7％)and showed no significant difference from Groups B and C;2)Surviv-al rate:Group A had the highest survival rate(88.3％),which was significantly higher than Groups D,E,F,and G and showed no significant difference from Groups B and C;3)Adventitious bud prolifera-tion coefficient:Group A had the maximum proliferation coefficient(1.4),which was significantly higher than Groups C,D,and G and showed no significant difference from Groups B,E,and F.The combined treatment groups generally exhibited high contamination rates,low survival rates,and some ex-plants showed browning and delayed germination of bud points.[Conclusion]The optimal sterilization protocol for the tissue culture of Polygonatum kingianum primary rhizomes is as follows:first soak the ex-plants in a 3％effective content carbendazim solution for 48 hours,then perform surface sterilization with 75％ethanol for 30 seconds,and finally soak them in 0.1％HgCl2 for 30 minutes.This protocol can provide a reference for the sterilization technology in the tissue culture of Polygonatum genus and other medicinal rhizome plants.关键词
滇黄精/初生根茎/灭菌剂/不定芽诱导Key words
Polygonatum kingianum/primary rhizome/sterilizing agent/induction of adventitious buds分类
农业科技
