蝴蝶兰NaN_3离体化学诱变及再生植株RAPD检测

In vitro Mutation Induction by NaN_3 in Phalaenopsis and RAPD Detection for the Regenerated Plants

采用NaN3对离体培养的蝴蝶兰类原球茎薄切片进行诱变处理,研究不同浓度和不同处理时间对类原球茎薄切片生长的影响,并对再生苗DNA进行了RAPD检测。结果表明,NaN3对类原球茎薄切片生长产生重要影响,部分薄切片白化或褐化死亡,再生类原球茎生长受到抑制,再生苗数量减少,部分再生苗畸变,叶片皱缩、变厚。10条RAPD引物的PCR检测发现,再生苗DNA序列发生了一定程度的变异,产生RAPD图谱带型多态性。根据以上结果认为,本体系合适的NaN3诱变剂量为6mmol/L,处理2d。

The thin sections （TCLs） of Phalaenopsis protocorm-like bodies （PLB） were used as explants for in vitro mutation induction by NAN3. The effects of different concentration and time of treatment on the growth of TCLs were investigated and the DNA of the regenerated plants was detected with RAPD. The results showed that NaN3 had great effects on TCL growth. Some TCL after treatment got white or browning and died and grew slowly, and the number of regenerated plantlets was reduced, and the regenerated plantlets became abnormal with leaves erimpled and thicker. The RAPD reaction with 10 primers indicated that the molecule maps of RAPD had some polymorphisms which meant the sequence of DNA was changed. The concentration of 6 mmol/L NaN3 treatment for 2 days was a suitable dosage for Phalaenopsis mutation induction in this test system.