Abstract:

In order to select a time-saving, simple and efficient total DNA extraction method for Huperzia crispata’s endophyte fungi mycelia which were isolated from Huperzia crispata. In this study, using benzyl chloride method to isolate total DNA of endophyte fungi and comparing quartz sand grind method with ultrasonic break method two break treatments for endophyte fungi mycelia DNA extraction efficiency. Results showed that the DNA extraction efficiency and quality obtained by ultrasonic break method was superior to quartz sand grind method. Using quartz sand grind this method, the extracted DNA had more impurity and appeared serious tailing. But using ultrasonic break this method, the extracted DNA were concentrated in 23.1 kb band, little polysaccharides and proteins were polluted near loading well. Ultrasonic break treatment was a fast, efficient and time-saving method for endophyte fungi mycelia DNA extraction, especially suitable for large sacle DNA extraction. The genomic DNA extracted by this two different break treatments were digested by RNase A and amplified with ITS primers, agarose gel electrophoresis checking results showed that this two different break treatments could obtain good quality PCR effects. But, the ultrasonic break method was a faster and more efficient method for DNA extraction from Huperzia crispata’s endophyte fungi mecylia than quartz sand grind method, especially for large scale samples and firm texture fungi colony.