Using pollen as a vector of foreign genes for plant transformation is an active research area. An important link in this transformation system is the delivery of foreign genes into pollen. However, DNA transfer into p...Using pollen as a vector of foreign genes for plant transformation is an active research area. An important link in this transformation system is the delivery of foreign genes into pollen. However, DNA transfer into pollen is difficult because of the existence of a thick pollen wall, whereas pollen protoplast deprived of pollen wall should be advantageous展开更多
The formation of the polarity of pollen proto-plast and the dynamics of actin cytoskeleton were observed by non-fixation, Alexa-Phalloidin probing and confocal laser scanning microscopy. Our results showed that the pr...The formation of the polarity of pollen proto-plast and the dynamics of actin cytoskeleton were observed by non-fixation, Alexa-Phalloidin probing and confocal laser scanning microscopy. Our results showed that the protoplast obtained from stored pollen contained numerous crystalline fusiform bodies to constitute a storage form of actin. When dormant pollen was hydrated, the actin cytoskeleton forms a fine network spreading uniformly in the protoplast. In the process of polarity formation and germination of pollen pro-toplast, actin filaments marshaled slowly to the brim, and then formed multilayer continuous actin filament bundles surrounding the cortical of the protoplast. When the proto-plast was exposed to actin filament-disrupting drugs, such as Latrunculin A and Cytochalasin D, continuously arranged actin bundles were disturbed and in this condition, the pro-toplast could not germinate. But when exposed to actin fila-ment stabiling drug-phalliodin, the dynamics of actin fila-ments in the protoplasts behaved normally and the proto-plasts could germinate normally. These results were also confirmed by the pharmacology experiments on pollen grains. And when Latrunculin A or Cytochalasin D was washed off, the ratio of pollen germination was resumed partly. All the results above show that the dynamic organiza-tion of the actin cytoskeleton are critical in the cell polarity formation and germination of pollen protoplast, and that the reorganization of actin cytoskeleton is mainly due to the re-arrangement of actin filament arrays.展开更多
文摘Using pollen as a vector of foreign genes for plant transformation is an active research area. An important link in this transformation system is the delivery of foreign genes into pollen. However, DNA transfer into pollen is difficult because of the existence of a thick pollen wall, whereas pollen protoplast deprived of pollen wall should be advantageous
文摘The formation of the polarity of pollen proto-plast and the dynamics of actin cytoskeleton were observed by non-fixation, Alexa-Phalloidin probing and confocal laser scanning microscopy. Our results showed that the protoplast obtained from stored pollen contained numerous crystalline fusiform bodies to constitute a storage form of actin. When dormant pollen was hydrated, the actin cytoskeleton forms a fine network spreading uniformly in the protoplast. In the process of polarity formation and germination of pollen pro-toplast, actin filaments marshaled slowly to the brim, and then formed multilayer continuous actin filament bundles surrounding the cortical of the protoplast. When the proto-plast was exposed to actin filament-disrupting drugs, such as Latrunculin A and Cytochalasin D, continuously arranged actin bundles were disturbed and in this condition, the pro-toplast could not germinate. But when exposed to actin fila-ment stabiling drug-phalliodin, the dynamics of actin fila-ments in the protoplasts behaved normally and the proto-plasts could germinate normally. These results were also confirmed by the pharmacology experiments on pollen grains. And when Latrunculin A or Cytochalasin D was washed off, the ratio of pollen germination was resumed partly. All the results above show that the dynamic organiza-tion of the actin cytoskeleton are critical in the cell polarity formation and germination of pollen protoplast, and that the reorganization of actin cytoskeleton is mainly due to the re-arrangement of actin filament arrays.
