魔芋(Amorphophallus spp.)四种组织器官RNA提取方法的比较

Comparative Study on RNA Extraction Methods from Konjac(Amorphophallus spp.) of Four Organs

由于魔芋植株各部位富含多糖,较难获得高质量的RNA。为了筛选出魔芋各组织器官总RNA提取最适合的方法,本研究采用Trizol法、异硫氰酸胍法、改良CTAB法、RNAprep Pure Plant Kit和Plant RNA Kit五种方法分别对花魔芋叶片、茎杆、根和球茎四种组织部位的总RNA进行了提取,并用琼脂糖凝胶电泳和紫外分光光度计对RNA质量进行综合评价。结果表明:用Trizol法和异硫氰酸胍法提取魔芋各器官的RNA效果较差,杂质含量较高,降解严重;采用RNAprep Pure Plant Kit和Plant RNA Kit两种商业试剂盒仅提取出魔芋叶片的RNA,茎秆、根、球茎三种组织器官未能提取质量完整的RNA,所提取的RNA效果不稳定且浓度较低。改良CTAB法提取的魔芋四种组织器官RNA均能得到完整、清晰的RNA条带,且四个器官的OD260/OD280均在1.8~2.0之间,采用改良CTAB法提取魔芋各组织器官总RNA较为理想。本研究为不同器官来源魔芋植株样品的RNA提取方法提供了科学依据。

Due to the polysaccharide content in different parts of konjac plants, it is difficult to obtain high quality RNA. For various screening konjac tissue and organs for total RNA extraction is the most suitable method, this research adopts the Trizol method, different guanidine thiocyanate method and modified CTAB method, RNAprep Pure Plant Kit, Plant RNA Kit five methods respectively to konjac leaf, stem, root, bulb four tissue of total RNA extraction, using agarose gel electrophoresis and ultraviolet spectrophotometer to evaluate the quality of RNA. The results showed that the RNA extracted from the organs of konjac by Trizol method and guanidine isothiocyanate method had poor effect, high impurity content and serious degradation. RNAprep Pure Plant Kit and Plant RNA Kit were used as commercial kits to extract only RNA from konjac leaves. The three tissues and organs of konjac,namely stem, root and bulb, failed to extract RNA with complete quality. The extracted RNA had unstable effect and low concentration. The RNA bands of the four tissues and organs of konjac extracted by the improved CTAB method were complete and clear, and the OD260/OD280 of the four organs were between 1.8 and 2.0. The total RNA of the tissues and organs of konjac extracted by the improved CTAB method was ideal. This study provides a scientific basis for RNA extraction of konjac plant samples from different organs.