Potassium antimonite was used to localize Ca2+ in the apical bud cells of spruce from July 1999 to May 2000. During the period of active growth (July 14), Calcium precipitates, an indication of Ca2+ localization, were...Potassium antimonite was used to localize Ca2+ in the apical bud cells of spruce from July 1999 to May 2000. During the period of active growth (July 14), Calcium precipitates, an indication of Ca2+ localization, were mainly distributed in vacuoles, intercellular spaces and cell walls. Few Ca2+ deposits localized in the cytosol and nucleus, showing a low level of the cytosolic and nuclear Ca2+ concentration in the warm summer. In August, some Ca2+ deposits appeared in the cytosol and nuclei, indicating that Ca2+ influx occurred in the cytosol and nucleus as the day length became shorter. From September to November, high levels of the cytosolic and nuclear Ca2+ remained. During the mid-winter (December and January), the distribution of Ca2+ deposits and the ultrastructures in the cells were altered dramatically. Plasmolysis occurred in many cells due to the protoplasmic dehydration. In addition plasmalemma invagination and nuclear chromatin aggregation also occurred. A large number of Ca2+ deposits appeared in the space between the plasmalemma and the cell wall. And also some Ca2+ deposits were distributed in the plastids. However, few Ca2+ deposits were observed in the cytosol and nuclei. By spring of the next year (May), when plants were de-acclimated and resumed active growth, Ca2+ subcellular localization essentially restored to that observed in July of the last year, i.e., the cells contained low cytosolic and nuclear Ca2+ concentrations; Ca2+ deposits were mainly distributed in the vacuoles, cell walls and intercellular spaces. The relationships between the seasonal changes of intracellular Ca2+ concentration and the development of dormancy/cold acclimation, as well as plasmolysis associated with dormancy and cold hardiness were discussed.展开更多
Three genes encoding for fungal cell wall degrading enzymes (CWDEs), ech42, nag70 and gluc78 from the biocontrol fungus Trichoderma atroviride were inserted into the binary vector pCAMBIA1305.2 singly and in all possi...Three genes encoding for fungal cell wall degrading enzymes (CWDEs), ech42, nag70 and gluc78 from the biocontrol fungus Trichoderma atroviride were inserted into the binary vector pCAMBIA1305.2 singly and in all possible combinations and transformed to rice plants. More than 1800 independently regenerated plantlets in seven different populations (for each of the three genes and each of the four gene combinations) were obtained. The ech42 gene encoding for an endochitinase increased resistance to sheath blight caused by Rhizoctonia solani, while the exochitinase-encoding gene, nag70, had lesser effect. The expression level of endochitinase but exochitinase was correlated with disease resistance. Nevertheless, exochitinase enhanced the effect of endochitinase on disease resistance when the two genes co-expressed in transgenics. Resistance to Magnaporthe grisea was found in all kinds of regenerated plants including that with single gluc78. A few lines expressing either ech42 or nag70 gene were immune to the disease. Transgenic plants are being tested to further evaluate disease resistance at field level. This is the first report of multiple of expression of genes encoding CWDEs from Trichoderma atroviride that result in resistance to blast and sheath blight in rice.展开更多
Trichoderma harzianum strain T22 controls various diseases of maize and other crops, including seedling and root rots caused by Pythium ultimum. Seedlings of inbred line Mo17 were grown from T22-treated or untreated s...Trichoderma harzianum strain T22 controls various diseases of maize and other crops, including seedling and root rots caused by Pythium ultimum. Seedlings of inbred line Mo17 were grown from T22-treated or untreated seeds in field soil or in field soil intested with the pathogen. Five days after planting, seedlings of Mo17 (5-days-old) were smaller in the presence of P. ultimum and larger in the presence of T22 relative to the control. The combination of T22 with P. ultimum (T22+ P. ultimum ) resulted in plants as large as T22 alone. Methods for protein extraction and 2-D gel electrophoresis were developed. Proteins in seedlings roots from the various treatments were separated on 2-D gels and analyzed using PDQuest TM. 2-D software. With seedlings produced from T22-treated seeds, there were 104 unmatched proteins and 164 matched proteins relative to the control, and 97 and 150 from the treatment with T22+ P. ultimum, respectively, however, with P. ultimum alone the numbers were much lower than above two treatments. Comparatively, there was very lower similarity of proteome patterns of seedling roots with T22 or P. ultimum or both to control seedlings, the correlative coefficient values were 0.72, 0.51 and 0.49 for the comparisons among control with T22, P. ultimum and T22+ P. ultimum, respectively. Moreover, correlative coefficient of proteome patterns between T22 with P. ultimum was only 0.65, and T22 fungal proteome were also not same as any one of seedling roots with various treatments. Taken together, the components in seedling root proteome seemed to be mostly coming from Mo17 plants themselves and affected strongly by either microbes, but the effects appeared to be stronger by P. ultimum than by T22. 41 spots were selected for protein mass fingerprinting identification, and most detected-spots were intensified in abundance by T22 or T22+ P.ultimum treatments such as pathogenesis-related protein and endochitnase etc. SOD (Mn) was found to be involved in the defensive reaction of host against P. ultimum because the protein only appeared in the treatment with T22 or T22+ P.ultimum. Besides, some proteins associated with host respiration, nutrition synthesis and transport appeared to be in coordination with defensive-related proteins against the damping off.展开更多
Three genes encoding for fungal cell wall degrading enzymes (CWDE), ech42, nag70 and gluc78 from the biocontrol fungus Trichoderma atroviride were inserted into the binary vector pCAMBIA1305.2 singly and in all possib...Three genes encoding for fungal cell wall degrading enzymes (CWDE), ech42, nag70 and gluc78 from the biocontrol fungus Trichoderma atroviride were inserted into the binary vector pCAMBIA1305.2 singly and in all possible combinations. The coding sequences were placed downstream of the rice actin promoter and all vectors were used to transform rice plants. A total of more than 1,800 independently regenerated plantlets in seven different populations (for each of the three genes and each of the four gene combinations) were obtained. Expression in plant was obtained for all the fungal genes used singly or in combinations. The ech42 gene encoding for an endochitinase increased resistance to sheath blight caused by Rhizoctonia solani, while the exochitinase-encoding gene, nag70, had a lesser effect. The expression level of endochitinase but not of the exochitinase was correlated with disease resistance. Nevertheless, exochitinase enhanced the positive effect of endochitinase on disease resistance when two genes were co-expressed in transgenic rice. Improved resistance to Magnaporthe grisea was found in all types of regenerated plants, including those with the gluc78 gene alone, while a few lines expressing either ech42 or nag70 appeared to be immune to this pathogen. Transgenic plants expressing the gluc78 gene alone were stunted and only few of them survived, even though they showed resistance to M. grisea. However, combination with either one of the two other genes (ech42, nag70) as included in the same T-DNA region, reduced the negative effect of gluc78 on plant growth. This is the first report of single or multiple of expression of transgens encoding CWDEs that results in resistance to blast and sheath blight in rice.展开更多
基金Partly supported by the National Natural Science Foundation of China (Grant No.60050003)
文摘Potassium antimonite was used to localize Ca2+ in the apical bud cells of spruce from July 1999 to May 2000. During the period of active growth (July 14), Calcium precipitates, an indication of Ca2+ localization, were mainly distributed in vacuoles, intercellular spaces and cell walls. Few Ca2+ deposits localized in the cytosol and nucleus, showing a low level of the cytosolic and nuclear Ca2+ concentration in the warm summer. In August, some Ca2+ deposits appeared in the cytosol and nuclei, indicating that Ca2+ influx occurred in the cytosol and nucleus as the day length became shorter. From September to November, high levels of the cytosolic and nuclear Ca2+ remained. During the mid-winter (December and January), the distribution of Ca2+ deposits and the ultrastructures in the cells were altered dramatically. Plasmolysis occurred in many cells due to the protoplasmic dehydration. In addition plasmalemma invagination and nuclear chromatin aggregation also occurred. A large number of Ca2+ deposits appeared in the space between the plasmalemma and the cell wall. And also some Ca2+ deposits were distributed in the plastids. However, few Ca2+ deposits were observed in the cytosol and nuclei. By spring of the next year (May), when plants were de-acclimated and resumed active growth, Ca2+ subcellular localization essentially restored to that observed in July of the last year, i.e., the cells contained low cytosolic and nuclear Ca2+ concentrations; Ca2+ deposits were mainly distributed in the vacuoles, cell walls and intercellular spaces. The relationships between the seasonal changes of intracellular Ca2+ concentration and the development of dormancy/cold acclimation, as well as plasmolysis associated with dormancy and cold hardiness were discussed.
基金Project (No.3997002) supported by the National Natural Science Foundation of China
文摘Three genes encoding for fungal cell wall degrading enzymes (CWDEs), ech42, nag70 and gluc78 from the biocontrol fungus Trichoderma atroviride were inserted into the binary vector pCAMBIA1305.2 singly and in all possible combinations and transformed to rice plants. More than 1800 independently regenerated plantlets in seven different populations (for each of the three genes and each of the four gene combinations) were obtained. The ech42 gene encoding for an endochitinase increased resistance to sheath blight caused by Rhizoctonia solani, while the exochitinase-encoding gene, nag70, had lesser effect. The expression level of endochitinase but exochitinase was correlated with disease resistance. Nevertheless, exochitinase enhanced the effect of endochitinase on disease resistance when the two genes co-expressed in transgenics. Resistance to Magnaporthe grisea was found in all kinds of regenerated plants including that with single gluc78. A few lines expressing either ech42 or nag70 gene were immune to the disease. Transgenic plants are being tested to further evaluate disease resistance at field level. This is the first report of multiple of expression of genes encoding CWDEs from Trichoderma atroviride that result in resistance to blast and sheath blight in rice.
文摘Trichoderma harzianum strain T22 controls various diseases of maize and other crops, including seedling and root rots caused by Pythium ultimum. Seedlings of inbred line Mo17 were grown from T22-treated or untreated seeds in field soil or in field soil intested with the pathogen. Five days after planting, seedlings of Mo17 (5-days-old) were smaller in the presence of P. ultimum and larger in the presence of T22 relative to the control. The combination of T22 with P. ultimum (T22+ P. ultimum ) resulted in plants as large as T22 alone. Methods for protein extraction and 2-D gel electrophoresis were developed. Proteins in seedlings roots from the various treatments were separated on 2-D gels and analyzed using PDQuest TM. 2-D software. With seedlings produced from T22-treated seeds, there were 104 unmatched proteins and 164 matched proteins relative to the control, and 97 and 150 from the treatment with T22+ P. ultimum, respectively, however, with P. ultimum alone the numbers were much lower than above two treatments. Comparatively, there was very lower similarity of proteome patterns of seedling roots with T22 or P. ultimum or both to control seedlings, the correlative coefficient values were 0.72, 0.51 and 0.49 for the comparisons among control with T22, P. ultimum and T22+ P. ultimum, respectively. Moreover, correlative coefficient of proteome patterns between T22 with P. ultimum was only 0.65, and T22 fungal proteome were also not same as any one of seedling roots with various treatments. Taken together, the components in seedling root proteome seemed to be mostly coming from Mo17 plants themselves and affected strongly by either microbes, but the effects appeared to be stronger by P. ultimum than by T22. 41 spots were selected for protein mass fingerprinting identification, and most detected-spots were intensified in abundance by T22 or T22+ P.ultimum treatments such as pathogenesis-related protein and endochitnase etc. SOD (Mn) was found to be involved in the defensive reaction of host against P. ultimum because the protein only appeared in the treatment with T22 or T22+ P.ultimum. Besides, some proteins associated with host respiration, nutrition synthesis and transport appeared to be in coordination with defensive-related proteins against the damping off.
文摘Three genes encoding for fungal cell wall degrading enzymes (CWDE), ech42, nag70 and gluc78 from the biocontrol fungus Trichoderma atroviride were inserted into the binary vector pCAMBIA1305.2 singly and in all possible combinations. The coding sequences were placed downstream of the rice actin promoter and all vectors were used to transform rice plants. A total of more than 1,800 independently regenerated plantlets in seven different populations (for each of the three genes and each of the four gene combinations) were obtained. Expression in plant was obtained for all the fungal genes used singly or in combinations. The ech42 gene encoding for an endochitinase increased resistance to sheath blight caused by Rhizoctonia solani, while the exochitinase-encoding gene, nag70, had a lesser effect. The expression level of endochitinase but not of the exochitinase was correlated with disease resistance. Nevertheless, exochitinase enhanced the positive effect of endochitinase on disease resistance when two genes were co-expressed in transgenic rice. Improved resistance to Magnaporthe grisea was found in all types of regenerated plants, including those with the gluc78 gene alone, while a few lines expressing either ech42 or nag70 appeared to be immune to this pathogen. Transgenic plants expressing the gluc78 gene alone were stunted and only few of them survived, even though they showed resistance to M. grisea. However, combination with either one of the two other genes (ech42, nag70) as included in the same T-DNA region, reduced the negative effect of gluc78 on plant growth. This is the first report of single or multiple of expression of transgens encoding CWDEs that results in resistance to blast and sheath blight in rice.
