稗草生防潜力菌Helminthosporium gramineum f. sp. echinochloae的原生质体制备和再生

禾长蠕孢菌（Helminthosporium gramineum f. sp. echinochloae，HGE)是稗草生防潜力菌,为建立HGE的遗传转化体系,研究了菌龄、胞壁降解酶、稳渗剂等对其原生质体制备和再生的影响。结果表明,HGE菌原生质体制备的适宜条件为：26℃下采用1.0%崩溃酶处理菌龄为6 h的HGE菌丝体2 h,0.7 mol/L NaCl为稳渗剂。原生质体再生的适宜条件为：32℃下采用05%崩溃酶处理菌龄为16 h的HGE菌丝体4 h,0.7 mol/L NaCl为稳渗剂,R1为高渗再生培养基。

Preparation and regeneration of protoplasts from a potential barnyard grass biocontrol fungus Helminthosporium gramineum f. sp. echinochloae

Helminthosporium gramineum f.sp.echinochloae(HGE)is a promising biological control barnyard grass agent.To establish the genetic transformation system of HGE,the optimum conditions including mycelium incubation time,various compounding of enzymes and osmotic stabilizers for protoplast isolation and regeneration from HGE protoplasts were examined.The results indicated that protoplasts were optimally isolated when mycelia of HGE had been cultured for 6 h.and digested with 1% Drislase at 26℃ for 2 h in osmotic stabilizer of 0.7mol·L~(-1) NaCl solution.The maximum regeneration ratio was obtained on the RI regeneration medium when the mycelia of HGE were inocuhted after 16 h.and digested with 0.5% Drislase at 32℃ for 4 h in 0.7mol·L~(-1) NaCl solution.