摘要：
Seeds of Zanthoxylum dissitium Hemsl are rich in secondary metabolites with complex composition,such as polysaccharide and phenol,which seriously harden the isolation of its total RNA.As a consequence,general extraction method would make the total RNA seriously degrade,or could not obtain total RNA at all.Therefore,this paper through four methods including modified CTAB method,Trizol method,Plant RNA Kit and traditional CTAB method,were used to extract the total RNA with the extraction results being compared and analyzed by agarose gel electrophoresis,BioPhotometer plus and RT-PCR verification.The result showed that Trizol method,Plant RNA Kit and traditional CTAB method failed to extract the Total RNA from Zanthoxylum dissitium Hemsl seeds,whereas the modified CTAB method successfully extracted the total RNA with relatively good integrity and high purity.The 28S rRNA and 18S rRNA bands of the extracted RNA were clear and tidy,with the ratio of OD260/OD280 being 2.07,which makes the extracted total RNA very suitable for RT-PCR experiments.The isolation of high quality RNA from Zanthoxylum dissitium Hemsl seeds by modified CTAB method indicated that polysaccharide and phenol compounds could be effectively eliminated through this approach,and could satisfy the requirement of following molecular biology study.%蚬壳花椒种子中富含油脂、多糖和多酚类等成分复杂的次生代谢物质,影响其总RNA的提取,一般的提取方法无法获得总RNA或者总RNA降解严重.本实验采用改良CTAB法、TRNzol法、试剂盒法、传统CTAB法提取总RNA,通过琼脂糖凝胶电泳,核酸蛋白检测仪检测以及RT-PCR验证等对提取结果进行比较与分析.结果表明:TRNzol法、试剂盒法、传统CTAB法未能从蚬壳花椒种子中提取到总RNA,改良CTAB法能提取完整性较好、纯度较高的RNA,28S及18S条带清晰整齐,D260/D280值为2.07,提取的总RNA适用于RT-PCR实验,可以为后续的分子生物学研究奠定了基础.
