AFLP技术鉴别罂粟、虞美人和大麻种属差异的初步研究

目的探讨采用AFLP技术检测植物DNA,鉴别罂粟、虞美人和大麻植物种属间差异的方法。方法收集罂粟根、茎、叶、花、果以及虞美人和大麻叶检材,用AxyPrep DNA试剂盒提DNA,经EcoRI/MseI酶切,人工接头及PCR预扩增,用E-ACA/M-CAG、E-ACT/M-CTC、E-ACC/M-CTA、E-ACC/M-CTG、E-AGC/M-CTT、E-AGG/M-CTA6对标记了荧光的选择性引物进行PCR扩增,其产物在的CEQ8000遗传分析仪上检测。结果6对引物分别在罂粟、大麻和虞美人样本中检出27~46、5~20、4~31条扩增片段,种属间存在明显差异;同一罂粟根、茎、叶、花和果的DNA检测结果相同。结论罂粟、虞美人和大麻3种植物的AFLP分析结果显示出的差异性,同一植株不同部位DNA AFLP结果的同一性,有可能用于检测未知植物检材的种属来源。

A preliminary study on species differences among Papaver somniferum L, Papaver rhoeas L and Cannabis sativa L by AFLP technique

Objective To disclose the inter-species differences among Papaver somniferum L, Papaver rhoeas L and Cannabis sativa L using AFLP analysis. MethodsThe root, stem, foliage, flower, seed of Papaver somniferum L, foliage of Papaver rhoeas L and Cannabis sativa L were collected respectively. DNA was isolated with AxyPrep Kit, and double-digested by EcoR I and Mse I, then oligonucleotide adapters were ligated. After Pre-amplification, selective amplification was performed using 6 pairs of fluorescent primer. The DNA fragments were analyzed using a CEQ 8000 DNA Fragment Analyzer. Results27 to 46, 5 to 20 and 4 to 31 pieces of amplified products were detected from Papaver somniferum L, Cannabis sativa L and Papaver rhoeas L respectively, and significant difference was observed among these three groups of products. The identical species-specific peaks were identified from root, stem, leaf, flower and seed DNA from the same Papaver somniferum L. ConclusionThe diacritical results from Papaver somniferum L, Papaver rhoeas L and Cannabis sativa L, as well as the identical results of different parts from the same plant demonstrated that AFLP analysis could be used possibly to determine the species origin of unknown plants samples.