镰刀菌诱导白木香结香部位总RNA提取方法的研究

A Research of different method for isolating total RNA from artificial agarwood induced by Fusarium sp.A2

目的白木香树在自然条件下不产生树脂,以自然、微生物或人工方式可使之形成沉香树脂。＂小孔滴注法＂接种镰刀菌人工诱导白木香结香,8个月后野外采集白木香结香部位（分泌黑色树脂的树干组织）,提取其总RNA,为阐明沉香特征产物的代谢途径、揭示人工诱导白木香结香的分子机制奠定基础。方法 ＂小孔滴注法＂人工造香,手工割取白木香结香部位。分别采用CTAB-Li Cl法、Trizol试剂法、RN09试剂盒法、改良异硫氰酸胍-CTAB法和Qiagen试剂盒法提取总RNA并检测完整性。结果与其它四种提取方法相比,Qiagen试剂盒法提取的RNA条带清晰,OD260/OD280比值在1.8-2.0的范围内。结论 Qiagen试剂盒法适用于镰刀菌人工诱导白木香结香部位总RNA提取,对其它次生代谢物质含量较高的植物中提取总RNA具有一定的借鉴意义。

Objective To establish an effective method for extracting total RNA from the resin - deposited parts of the trunk of Aquilaria sinensis （ Lour. ） Gilg induced by Fusarium sp. A2. Methods By respectively employing CTAB - LiCl method, Trizol reagent method, EASYspin RN09 RNA isolation kit, modified guanidinium thiocyanate - CTAB method or Qiagen RNeasy Plant Mini Kit to extract total RNA from artificial inducement of agarwood （ the black resinous portions of A. sinensis ）. Results Compared to another four methods, the total RNA extracted by Qiagen RNeasy Plant Mini Kit was high - quality. The results showed the clear bands with little RNA decomposed, and the ratio of OD260/OD280 were 1.8 - 2.0. Conclusion Total RNA from artificial inducement of agarwood that induced by the biological method was isolated by Qiagen RNeasy Plant Mini Kit efficiently, which could provide some advise to the total RNA extract from other plant with complicated secondary metabolites.