蝴蝶兰EMS离体化学诱变及再生植株RAPD检测

用EMS对离体培养的蝴蝶兰类原球茎薄切片进行化学诱变,研究了不同浓度、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EMS对类原球茎薄切片生长产生重要影响,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。诱变剂EMS使部分再生苗畸变,叶片皱缩、变厚。并产生2株叶形、叶色突变体。部分再生苗表现出RAPD图谱带型的多态性,暗示DNA序列发生了一定的变异。EMS适合诱变剂量为浓度0．4％处理2d～4d。

EMS Chemical Induction for Phalaenopsis in in vitro Culture and RAPD Detecting for Regenerated Plants

Ethyl Methansulfonate （EMS） was used as mutagen for chemical induction with thin cell layers（TCLs） of Phalaenopsis protoeorm-like bodies（PLBs）. 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 planflets were tested by RAPD reaction. The results showed that mutagen EMS had great effects on TCLs growth. Some TCLs got browning and died, the PLBs grew slowly and the number of regenerated planflets were reduced . There were obvious injured effects on the explants, PLBs and planflets regenerated from the TCLs. The injured effects got more and more serious with higher concentration of mutagens and longer time treatment. The mutagen EMS made some plantlets aberrance, leaves erimpled and got thicker. Two mutants of leaf shape and color were produced. The RAPD maps of some regenerated plants showed polymorphisms which means the sequence variation of DNA had happened. The concentrations of 0.4 % EMS treatment for 2 - 4 days are suitable dosage for Phalaenopsis chemical induction in vitro.