Establishing of an economic and efficient genetic transformation system of protoplast for Fusarium graminearum

• 1、College of Plant Protection, Nanjing Agricultural University, Nanjing 210095

Abstract：The genetic transformation of protoplast for Fusarium graminearum is a important technique of establishing mutation library and studying functional genome. In this research, JS399-19HR strain Y2021A of Fusarium graminearum was used for studying the effect of the different digesting materials, enzyme combination, enzyme concentration, digesting time, digesting temperature and weight of the best digesting material on protoplast preparation. Then the DNA segment（1.7kb）of hygromycin resistant gene B（hph） was transformed into the protoplasts of JS399-19HR strain Y2021A. The efficiency of transformation was calculated and the tranformants were detected by PCR. The optimum condition for preparing protoplasts was: one milliliter enzyme mixture of 2% Driselase and 2% Snailase digested the 0.12 g germLings at 30 ℃ for 1.5 hours. The efficiency of transformation was up to 40-50 transformants per microgram of DNA and the fragment hph had inserted into the genome of transformants by the identification of PCR. An economic and efficient genetic transformation system of protoplast for Fusarium graminearum was successfully established, which was an essential technology for establishing mutation library and studying functional genome Fusarium graminearum.

Keywords： Genetics Fusarium graminearum Protoplast preparation Genetic transformation