关键词: 烤烟, 烤烟, 光合作用, 配方肥增效剂

Abstract: 【OBJECTIVE】In order to explore the method of extracting high quality total RNA from muscle larvae of T.spiralis【METHOD】Total RNA of muscle larvae of T.spiralis were extracted by three methods of using Trizol agentv, (After washing worms with DEPC-treated water, spin down worms at 10000g in microfuge for 1 minute and then remove excess water. Total RNA of muscle larvae of T.spiralis were extracted by three methods of using Trizol reagent:(1)method combining liquid nitrogen and Trizol : After putting liquid nitrogen into mortar up to T.spiralis prepared can be frozen ,put fresh samples into mortar and quickly grinding and add 1ml of Trizol Reagent per 50~100 mg of T.spiralis before thawing ,and transfer the liquid into the eppendorf tubev(2)liquid method: A: Put the fresh samples into mortar precooled at -20℃, In the course of of grinding, add liquid nitrogen unceasing to keep samples frozen and then transfer the powders into the eppendorf tube prepared. B: Put fresh T.spiralis into liquid nitrogen and store at -80℃ prior to grinding, the rest procedure is the same as A.(3)homogenization method: Homogenize T.spiralis samples in 1 ml of Trizol reagent per 50~100 mg of tissue for 20 minutes using a glass-homogenizer putting ice water mixture. The protocols are carried according to the methods of Trizol after grinding or homogenating. The total RNA were compared by gel electrophoresis and the ratio of OD260/OD280)【RESULT】The method of combining liquid nitrogen and Trizol was the suitable method in extracting total RNA from muscle larvae of T.spiralis and obtained the best purification and yield of total RNA; 【CONCLUSION】The high quailty total RNA can be obtained with fresh (samples) and combining liquid nitrogen and lysate.