橡胶树白粉菌收集及DNA和RNA提取方法比较

Comparison the collection and DNA and RNA extraction methods of the rubber tree powdery mildew

橡胶树白粉菌(Oidium heveae)是专性寄生的病原真菌,难以获取足量和纯度高的菌体,因而不易提取到足够多和质量高的核酸,阻碍了该菌的分子生物学研究,目前关于橡胶树白粉菌核酸提取方法少有报道。采用软毛笔刷取和醋酸纤维乙酯撕取两种方法收集白粉病菌。在尝试使用常规的菌物RNA提取方法基础上,通过优化操作步骤、方法等,研究出改良的Trizol法,用该法能高效率提取到纯度高的橡胶树白粉菌RNA。比较不同DNA提取方法的效率,推荐使用得率高、纯度好的OMEGA FungalDNA kit提取法,用于提取橡胶树白粉菌的DNA。

Obligate parasitic fungus Oidium heveae B. A. Steinmann caused the rubber tree powdery mildew, as it is difficult to obtain sufficient and high purity fungus, molecular biology research of O.heveae is hampered by the quality and quantity of powdery mildew＇s nucleic acid. At present, nucleic acid extraction methods of the rubber tree powdery mildew are rarely reported. The soften brush bushing method and cellulose acetate ethyl removed method are used to obtain the sufficient O.heveae. Based on the common fungal nucleic acid extraction methods, through optimized the operation steps, developed an improved Trizol method for the rubber tree powdery mildew RNA extraction, which showed that it has a high efficient and the isolated RNA is in high purity. Comparison of different DNA extraction methods to rubber tree powdery mildew, we recommended the OMEGA Fungal DNA kit as it has high efficiency and good purity.