文心兰NaN3离体化学诱变及RAPD检测

Study on NaN3 chemical induction for Oncidium in in vitro culture and RAPD screening

关键词:  文心兰  叠氮化钠  离体培养  化学诱变  RAPD

Key words:  Oncdium  NaN3  in vitro culture  chemical induction  RAPD

摘要:

采用3种浓度的NaN3分别对离体培养的文心兰类原球茎薄切片进行不同时间诱变处理，考察了不同浓度、不同时间诱变处理对类原球茎薄切片生长、类原球茎再生及再生苗生长的影响，并对再生苗DNA进行了RAPD检测。结果表明：诱变剂对类原球茎薄切片生长产生严重影响，部分薄切片褐化死亡，再生类原球茎生长受到抑制，再生苗数量减少，表现出明显的伤害作用，这种伤害作用随诱变剂浓度的加大和处理时间的延长而加重；NaN3抑制再生苗生长，试管苗普遍矮小。10条RAPD引物PCR检测表明，再生苗DNA发生了一定的变异，表现在RAPD图谱带型的多态性，随着诱变剂浓度的加大和处理时间延长，多态性比例增高。NaN3适宜诱变浓度为6 mmol/L，适宜处理时间为2～4 d。

Abstract:

Three kinds of concentrations of NaN3 were used as mutagens for chemical induction with thin cell layers(TCLs)of Oncidium protocorm like bodies(PLBs)in different time. The effects of different concentration and time treatments on the growth of TCLs，PLBs and plantlets which regenerated from the TCLs were investigated. Meanwhile，the DNA of the plantlets were tested by RAPD reaction. The results showed that mutagen NaN3 had great effects on TCLs growth. Some TCLs got browning and died，the PLBs grew slowly and the number of regenerated plantlets were reduced. There were obvious injured effects on the explants，PLBs and plantlets regenerated from the TCLs. The injured effects got more and more serious with higher concentration of mutagen and longer time treatment. The mutagen NaN3 inhibited plantlets growing. RAPD reaction indicated that the molecule maps had some polymorphisms which mean the sequence of DNA had changed. The ratio of polymorphisms got higher with higher concentration of mutagen and longer time treatment. The concentrations of 6 mmol/L NaN3 treatment for 2-4 days are suitable dosage for Oncdium chemical induction in vitro.