关键词: 蛹虫草, 基因枪, 遗传转化, 绿色荧光蛋白

Abstract: An effective and convenient transformation method of Cordyceps militaris were established by means of particle bombardment. Coating microcarriers with plasmid pDHt-gpdA-GFP-bar，we transferred C. militaris CM01 by particle bombardment. After being selected by glufosinate-ammonium，two transformants were obtained under three conditions differing in the two parameters of target distance （6 cm or 9 cm） and helium pressure（7.58 × 106 Pa or 8.96 × 106 Pa）. The transformation frequency was 0.4 cfu? μg-1. PCR and southern blot results indicated that the selective marker gene Bar was integrated into the genomic DNA，a single copy or multiple copies integration，and genetically stable. Green fluorescence of the C. militaris transformant mycelia could be observed under fluorescent microscope，which indicated that the expression of green fluorescent protein was successful in the C. militaris transformants. Transformation by particle bombardment enriched the transformation methods of C. militaris. However，sample preparation and parameter setting need to be optimized，in order to improve the efficiency of transformation.

Key words: Cordyceps militaris, particle bombardment, genetic transformation, green fluorescent protein