高效降解秸秆的深绿木霉菌ATMT突变株的筛选  

Identification and Characterization of Trichoderma atroviride Mutants Carrying Capacities of Crop Straw Degradation

作　　者：马逸飞[1] 沈新迁[1] 刘雨晴[2] 曹珊珊[2] 黄博[3] 陈雪吟[4] 陈云鹏[1] 

机构地区：[1]上海交通大学农业与生物学院,上海200240 [2]上海中医药大学基础医学院,上海201203 [3]上海交通大学化学与化工学院,上海200240 [4]上海交通大学药学院,上海200240

出　　处：《上海交通大学学报（农业科学版）》2012年第1期15-22,共8页Journal of Shanghai Jiaotong University(Agricultural Science)

基　　金：上海市白玉兰科技人才基金项目(2010B046);农业部都市农业(南方)重点实验室开放课题(09UA005);上海交通大学第一期大学生创新实践计划项目(IPP1078)

摘　　要：以羧甲基纤维素粉或滤纸为唯一碳源,对深绿木霉菌T23的农杆菌介导的突变株进行了生长速度、产孢率、色素形成等形态学观测,同时,测定了滤纸酶酶活及羧甲基纤维素酶酶活,从中筛选出降解纤维素能力较出发菌T23显著变化的突变株7株。其中,多拷贝T-DNA插入突变株较单拷贝的突变株生长速度快,产孢率高。并且,这些突变株的发酵液呈明显的色素差异。突变株插入的拷贝数越多,酶活越强,在降解过程中这2种酶都观察到了产物的反馈抑制现象。The morphological differences including growth rate,sporulation ability and pigment production were determined in mutants derived from the wild-type strain T23 of Trichoderma atroviride via Agrobacterium-mediated transformation（ATMT） using Carboxymethyl cellulose（CMC） or filter paper as sole carbon source.Furthermore,the activities of filter paper enzyme（FPase） and CMCase were evaluated and seven mutants were identified to have obvious changes in cellulose degradation abilities compared with the original strain T23.Among of them,the mutants that harbouring double copies or triple copies of T-DNA integration in the genome of T.atroviride displayed apparent improvement in growth rate and sporulation ability.Moreover,color of zymotic fluid varied in these mutants,the result showed that they had produced different pigments in culture.The enzymatic activity of cellulose degradation in multiple-copied insertional mutants was higher than that of mutants with single-copied integration of exogenous DNA.Feedback inhibition caused by the degradation production of cellulose was observed by monitoring the changes in FPase and CMCase activities.

关 键 词：深绿木霉 ATMT 滤纸酶 纤维素酶 突变株 纤维素降解 

分 类 号：S182[农业科学—农业基础科学]