Fusarium moniliforme Sheld.is a rice pathogenic fungus and causes the disease called Bakanae,which has increasingly damaged rice production in the recent years. Trichoderma spp. has been one of the most widely used bi...Fusarium moniliforme Sheld.is a rice pathogenic fungus and causes the disease called Bakanae,which has increasingly damaged rice production in the recent years. Trichoderma spp. has been one of the most widely used biological control agent of plant disease. By geneticaly labelling F. moniliforme with the GFP reporter gene, we have studied the antagonistic action of Trichoderma viride against this pathogenic fungus. The binary GFP reporter vector pCHF3-35S∷GFP was constructed, which carries the gfp gene driven by the CaMv35S promoter. The vector was transformed into F. moniliforme via Agrobacterium.The mycoparasitism of T.viride against F.moniliforme was tested by dual culture and examined with fluorescence microscope. The result of the dual culture showed that the T.viride maintained a strong competitive ability against F. moniliforme , by growing on the top of the pathogen colony. Fluorescence microscope observation indicated that attacked hyphae of F. moniliform were distorted, swollen or broken. This indicate an enzymatic by T.viride to degrade the host cell walls and used the cell contents as a source of nutrients (Fig 1) .展开更多
This study describes a novel dark-green spore producing strain of Trichoderma harzianum exhibiting higher activities of cellulase, hemicellulase and ligninase on specific plate assays. To assess the cellulase producti...This study describes a novel dark-green spore producing strain of Trichoderma harzianum exhibiting higher activities of cellulase, hemicellulase and ligninase on specific plate assays. To assess the cellulase production in detail, basal salt medium (BSM) was supplemented with synthetic [carboxymethyl cellulose (CMC), glucose, sucrose, dextrose, lactose or maltose] and natural (flours of banana, banana peel, jack seed, potato or tapioca) carbon as well as nitrogen (yeast extract, beef extract, peptone, NaNO3 or NH4NO3) sources. Temperature and pH optima were 28?C and 4, respectively for the growth of the fungus in CMC- BSM with 146 U/ml cellulase activity. Flours of potato and banana supported comparable yields of cellulase to that of CMC (147 U/ml and 168 U/ml, respectively), while sodium nitrate was the preferred nitrogen source (150 U/ml). The water soluble yellowish-green pigment (a probable siderophore) extracted from the spores showed an absorption maximum at 414 nm. To comprise, this fungus shows the complete lignocellulolytic potential which offers great industrial significance, especially for the ethanol production from the lignocellulosic waste coupled with the production of a new pigment.展开更多
In this study we describe a novel dark-green strain of Trichoderma viride exhibiting complete ensemble of cellulase, hemicellulase and ligninase activities on specific plate assays. To assess the cellulase production ...In this study we describe a novel dark-green strain of Trichoderma viride exhibiting complete ensemble of cellulase, hemicellulase and ligninase activities on specific plate assays. To assess the cellulase production in detail, basal salt medium (BSM) was fortified with synthetic (carboxymethyl cellulose (CMC), glucose, sucrose, dextrose, lactose or maltose) and natural (flours of banana, banana peel, jack seed, potato or tapioca) carbon as well as nitrogen (yeast extract, beef extract, peptone, NaNO3 or NH4NO3) sources. Temperature and pH optima were 28°C and 4, respectively for the growth of the fungus in CMC-BSM with 137 U/mL cellulase activity, which was enhanced to 173 U/mL at 1.25% CMC concentration. Flours of potato and banana peel supported comparable yields of cellulase to that of CMC, while sodium nitrate was the preferred nitrogen source. The water soluble bluish-green pigment (a probable siderophore) extracted from the spores showed an absorption maximum at 292 nm. To sum up, the complete lignocellulolytic potential of this fungus offers great industrial significance, coupled with the production of a new pigment.展开更多
目的针对植物生物反应器中外源基因表达水平普遍较低的问题,探索和评价丝状真菌瑞氏木霉(Trichode rma re e s e i)编码的小分子疏水蛋白(Hydrophobin,HFBI)作为融合标签在植物生物反应器中提高系统表达量的应用潜力;分析其提高外源基...目的针对植物生物反应器中外源基因表达水平普遍较低的问题,探索和评价丝状真菌瑞氏木霉(Trichode rma re e s e i)编码的小分子疏水蛋白(Hydrophobin,HFBI)作为融合标签在植物生物反应器中提高系统表达量的应用潜力;分析其提高外源基因表达水平的可能机制。方法以绿色荧光蛋白(GFP)为报道基因,采用基因体外合成技术和亚克隆技术构建GFP和GFPHFBI融合植物表达载体。利用农杆菌渗滤技术接种植物本明烟(Nicotiana be nthamiana)。通过GFP荧光观察,荧光显微镜镜检,Western blot,ELISA和RT-PCR等实验手段测定报告基因GFP在植物中的表达情况,探明HFBI融合标签在植物中表达外源基因的作用效果和特点并分析其可能的作用机制。结果 HFBI融合标签对植物细胞无明显的细胞毒性;GFP-HFBI融合蛋白在植物中的积累水平显著高于对照;GFP-HFBI融合蛋白在细胞内形成致密的蛋白质颗粒。结论 HFBI融合标签能够显著提高外源基因在植物中的积累水平。推测形成的蛋白质颗粒隔绝了细胞内源性蛋白酶对目的表达产物的降解,进而提高了外源基因产物在细胞中的积累。展开更多
文摘Fusarium moniliforme Sheld.is a rice pathogenic fungus and causes the disease called Bakanae,which has increasingly damaged rice production in the recent years. Trichoderma spp. has been one of the most widely used biological control agent of plant disease. By geneticaly labelling F. moniliforme with the GFP reporter gene, we have studied the antagonistic action of Trichoderma viride against this pathogenic fungus. The binary GFP reporter vector pCHF3-35S∷GFP was constructed, which carries the gfp gene driven by the CaMv35S promoter. The vector was transformed into F. moniliforme via Agrobacterium.The mycoparasitism of T.viride against F.moniliforme was tested by dual culture and examined with fluorescence microscope. The result of the dual culture showed that the T.viride maintained a strong competitive ability against F. moniliforme , by growing on the top of the pathogen colony. Fluorescence microscope observation indicated that attacked hyphae of F. moniliform were distorted, swollen or broken. This indicate an enzymatic by T.viride to degrade the host cell walls and used the cell contents as a source of nutrients (Fig 1) .
文摘This study describes a novel dark-green spore producing strain of Trichoderma harzianum exhibiting higher activities of cellulase, hemicellulase and ligninase on specific plate assays. To assess the cellulase production in detail, basal salt medium (BSM) was supplemented with synthetic [carboxymethyl cellulose (CMC), glucose, sucrose, dextrose, lactose or maltose] and natural (flours of banana, banana peel, jack seed, potato or tapioca) carbon as well as nitrogen (yeast extract, beef extract, peptone, NaNO3 or NH4NO3) sources. Temperature and pH optima were 28?C and 4, respectively for the growth of the fungus in CMC- BSM with 146 U/ml cellulase activity. Flours of potato and banana supported comparable yields of cellulase to that of CMC (147 U/ml and 168 U/ml, respectively), while sodium nitrate was the preferred nitrogen source (150 U/ml). The water soluble yellowish-green pigment (a probable siderophore) extracted from the spores showed an absorption maximum at 414 nm. To comprise, this fungus shows the complete lignocellulolytic potential which offers great industrial significance, especially for the ethanol production from the lignocellulosic waste coupled with the production of a new pigment.
文摘In this study we describe a novel dark-green strain of Trichoderma viride exhibiting complete ensemble of cellulase, hemicellulase and ligninase activities on specific plate assays. To assess the cellulase production in detail, basal salt medium (BSM) was fortified with synthetic (carboxymethyl cellulose (CMC), glucose, sucrose, dextrose, lactose or maltose) and natural (flours of banana, banana peel, jack seed, potato or tapioca) carbon as well as nitrogen (yeast extract, beef extract, peptone, NaNO3 or NH4NO3) sources. Temperature and pH optima were 28°C and 4, respectively for the growth of the fungus in CMC-BSM with 137 U/mL cellulase activity, which was enhanced to 173 U/mL at 1.25% CMC concentration. Flours of potato and banana peel supported comparable yields of cellulase to that of CMC, while sodium nitrate was the preferred nitrogen source. The water soluble bluish-green pigment (a probable siderophore) extracted from the spores showed an absorption maximum at 292 nm. To sum up, the complete lignocellulolytic potential of this fungus offers great industrial significance, coupled with the production of a new pigment.
文摘目的针对植物生物反应器中外源基因表达水平普遍较低的问题,探索和评价丝状真菌瑞氏木霉(Trichode rma re e s e i)编码的小分子疏水蛋白(Hydrophobin,HFBI)作为融合标签在植物生物反应器中提高系统表达量的应用潜力;分析其提高外源基因表达水平的可能机制。方法以绿色荧光蛋白(GFP)为报道基因,采用基因体外合成技术和亚克隆技术构建GFP和GFPHFBI融合植物表达载体。利用农杆菌渗滤技术接种植物本明烟(Nicotiana be nthamiana)。通过GFP荧光观察,荧光显微镜镜检,Western blot,ELISA和RT-PCR等实验手段测定报告基因GFP在植物中的表达情况,探明HFBI融合标签在植物中表达外源基因的作用效果和特点并分析其可能的作用机制。结果 HFBI融合标签对植物细胞无明显的细胞毒性;GFP-HFBI融合蛋白在植物中的积累水平显著高于对照;GFP-HFBI融合蛋白在细胞内形成致密的蛋白质颗粒。结论 HFBI融合标签能够显著提高外源基因在植物中的积累水平。推测形成的蛋白质颗粒隔绝了细胞内源性蛋白酶对目的表达产物的降解,进而提高了外源基因产物在细胞中的积累。
