深绿木霉T2发酵液蛋白分离物诱导抗病作用研究

Studyon induced resistant of protein isolate from fermentation broth of Trichoderma atroviride T2

采用生长速率法和产孢量测定法,研究了不同培养基和温度、pH、接菌量、培养基量对深绿木霉T2生长和产孢量的影响。结果表明,PDA培养基有利于深绿木霉T2的生长;深绿木霉T2发酵产孢最优条件为温度25℃、pH 7、接菌量为0.05mL/L、培养基量为100mL/250mL,此条件下发酵72h时产孢量最大,为1.5×109个/mL;76h时产孢量下降;发酵液通过盐析及凝胶层析获得具有3个洗脱峰的蛋白质液,分别为Ⅰ,Ⅱ和Ⅲ;采用生长速率法和诱导活体接种法分别测定不同浓度的3个洗脱液对百合灰霉菌的抑菌率及其诱导抗病效果,结果表明洗脱峰Ⅰ蛋白质的100倍液对百合灰霉菌的抑菌率最小,仅为1.16%,而其诱导抗病效果最好,为84.77%。

The effects of medium,temperature,pH value,inoculation amount on the growth and sporulation of Trichoderma atroviride T2 were studied and the result indicated that PDA medium performed better and its sporulation optimum fermentation conditions were 25℃,pH 7,inoculation amount of 0.5 mL/L,medium content of 100mL/250 mL,sporulation reached the maximum at 72 hunder these conditions,it was 1.5×109/mL sporulation began to decline at 76 hby growth rate method and spore production measurement method.Three proteins eluted peaks from fermentation broth Trichoderma atroviride T2 were obtained by（NH4）2SO4precipitation and gel filtration chromatography,they wereⅠ,Ⅱ and Ⅲ,respectively and their inhibition rates and induced resistance effects on their different concentrations to lily Botrytis cinerea were measured by growth rate method and the induced in vivo inoculation method.The inhibition rate of protein elution peakⅠ100times was the minimum（1.16%）,while its induced resistance was the best（84.77%）.