In the present study, we focused on whether Intracellular free Ca^2＋ （[Ca^2＋],） regulates the formation of mltochondrlal permeability transition pore （MPTP） In H2O2-induced apoptosis In tobacco protoplasts. It w...In the present study, we focused on whether Intracellular free Ca^2＋ （[Ca^2＋],） regulates the formation of mltochondrlal permeability transition pore （MPTP） In H2O2-induced apoptosis In tobacco protoplasts. It was shown that the decrease In mltochondrlal membrane potential （△ψm） preceded the appearance of H2O2-Induced apoptosls; pretreatment with the specific MPTP Inhibitor cyclosporine A, which also Inhibits Ca^2＋ cycling by the mitochondria, effectively retarded apoptosls and the decrease In △ψm. Apoptosls and decreased △ψm were exacerbated by CaCl2, whereas the plasma membrane voltage-dependent Ca^2＋ channel blocker lanthanum chloride （LaCl3） attentuated these responses. Chelation of extracellular Ca^2＋ with EGTA almost totally Inhibited apoptosls and the decrease In △ψmInduced by H2O2. The time-course of changes In [Ca^2＋]l In apoptosls was detected using the Ca^2＋ probe Fiuo-3 AM. These studies showed that [Ca^2＋]1 was Increased at the very early stage of H2O2-Induced apoptosls. The EGTA evidently Inhibited the Increase In [Ca^2＋]1 Induced by H=O=, whereas It was only partially Inhibited by LaCl3. The results suggest that H2O2 may elevate cytoplasmic free Ca^2＋ concentrations In tobacco protoplasts, which mainly results from the entry of extracellular Ca^2＋, to regulate mltochondrlal permeability transition. The signaling pathway of [Ca^2＋]1-medlated mltochondrlal permeability transition was associated with H2O2-Induced apoptosis In tobacco protoplaete.展开更多
Electrofusion of evacuolated (N. Tabacum L. cvs. Gexin no1) and vacuolated (N. Rustic) tobacco mesophyll protoplasts was performed on the Chinese spacecraft (Shenzhou No. 4, from 30th Dec. 2002 to 6th Jan. 2003). The ...Electrofusion of evacuolated (N. Tabacum L. cvs. Gexin no1) and vacuolated (N. Rustic) tobacco mesophyll protoplasts was performed on the Chinese spacecraft (Shenzhou No. 4, from 30th Dec. 2002 to 6th Jan. 2003). The results showed that the frequency of bi-nucleated and mul-tinucleated protoplasts was significantly increased under microgravity. Compared with the control samples incubated on the ground, the viability of protoplasts incubated in space was much higher. In addition, the influence of altered gravity on carbohydrates was also observed. These results confirmed the effect of microgravitation on electrofusion of plant cell protoplasts.展开更多
Nuclear migration and positioning are crucial for the morphogenesis of plant cells. We addressed the potential role of nuclear positioning for polarity induction using an experimental system based on regenerating prot...Nuclear migration and positioning are crucial for the morphogenesis of plant cells. We addressed the potential role of nuclear positioning for polarity induction using an experimental system based on regenerating protoplasts, where the induction of a cell axis de novo can be followed by quantification of specific regeneration stages. Using overexpression of fluorescently tagged extranuclear （perinu- clear actin basket, kinesins with a calponin homology domain （KCH）） as well as intranuclear （histone H2B） factors of nuclear positioning and time-lapse series of the early stages of regeneration, we found that nuclear position is no prerequi- site for polarity formation. However, polarity formation and nuclear migration were both modulated in the transgenic lines, indicating that both phenomena depend on factors affecting cytoskeletal tensegrity and chromatin structure. We integrated these findings into a model where retrograde signals are required for polarity induction. These signals travel via the cytoskeleton from the nucleus toward targets at the plasma membrane.展开更多
Laser tweezers can manipulate small particles, such as cells and organdies. When coupling them with laser microbeam selective fusion of two tobacco protoplasts containing some chloroplast was achieved. Physical and bi...Laser tweezers can manipulate small particles, such as cells and organdies. When coupling them with laser microbeam selective fusion of two tobacco protoplasts containing some chloroplast was achieved. Physical and biological variables that affect laser trapping and laser-induced fusion were also discussed. The results show that the effect of chloroplast content and distribution on the yield of cell fusion is remarkable.展开更多
The relationship among transfer and expression of auxin binding protein gene ( abp ), auxin (NAA) induced plasmalemma hyperpolarity and sensibility to auxin during protoplast culture was studied by measuring transmemb...The relationship among transfer and expression of auxin binding protein gene ( abp ), auxin (NAA) induced plasmalemma hyperpolarity and sensibility to auxin during protoplast culture was studied by measuring transmembrane potential difference (Em) and culturing the protoplasts of sense and antisense abp transgenic tobacco. The concentration of NAA inducing the highest degree of hyperpolarity of sense abp transgenic tobacco protoplasts was lower than the control, and in protoplast culture, their sensibility to auxin increased. The concentration of antisense abp transgenic tobacco protoplasts was higher than the control, and in protoplast culture, their sensibility to auxin decreased. These results demonstrated that ABP synthesized in endoplasmic reticulum needed to transport to cell membrane and functioned there.展开更多
基金National Natural Science Foundation of China （90102015,30170161） and Cooperation Project of Intemational in China and Greece （05-46）.
文摘In the present study, we focused on whether Intracellular free Ca^2＋ （[Ca^2＋],） regulates the formation of mltochondrlal permeability transition pore （MPTP） In H2O2-induced apoptosis In tobacco protoplasts. It was shown that the decrease In mltochondrlal membrane potential （△ψm） preceded the appearance of H2O2-Induced apoptosls; pretreatment with the specific MPTP Inhibitor cyclosporine A, which also Inhibits Ca^2＋ cycling by the mitochondria, effectively retarded apoptosls and the decrease In △ψm. Apoptosls and decreased △ψm were exacerbated by CaCl2, whereas the plasma membrane voltage-dependent Ca^2＋ channel blocker lanthanum chloride （LaCl3） attentuated these responses. Chelation of extracellular Ca^2＋ with EGTA almost totally Inhibited apoptosls and the decrease In △ψmInduced by H2O2. The time-course of changes In [Ca^2＋]l In apoptosls was detected using the Ca^2＋ probe Fiuo-3 AM. These studies showed that [Ca^2＋]1 was Increased at the very early stage of H2O2-Induced apoptosls. The EGTA evidently Inhibited the Increase In [Ca^2＋]1 Induced by H=O=, whereas It was only partially Inhibited by LaCl3. The results suggest that H2O2 may elevate cytoplasmic free Ca^2＋ concentrations In tobacco protoplasts, which mainly results from the entry of extracellular Ca^2＋, to regulate mltochondrlal permeability transition. The signaling pathway of [Ca^2＋]1-medlated mltochondrlal permeability transition was associated with H2O2-Induced apoptosis In tobacco protoplaete.
文摘Electrofusion of evacuolated (N. Tabacum L. cvs. Gexin no1) and vacuolated (N. Rustic) tobacco mesophyll protoplasts was performed on the Chinese spacecraft (Shenzhou No. 4, from 30th Dec. 2002 to 6th Jan. 2003). The results showed that the frequency of bi-nucleated and mul-tinucleated protoplasts was significantly increased under microgravity. Compared with the control samples incubated on the ground, the viability of protoplasts incubated in space was much higher. In addition, the influence of altered gravity on carbohydrates was also observed. These results confirmed the effect of microgravitation on electrofusion of plant cell protoplasts.
基金supported by a Ph.D. fellowship of State of BadenWürttemberg (Landesgraduiertenfrderung) to L. Brochhausenfunds from the Deutsche Forschungsgemeinschaft (NI 324/19-1)
文摘Nuclear migration and positioning are crucial for the morphogenesis of plant cells. We addressed the potential role of nuclear positioning for polarity induction using an experimental system based on regenerating protoplasts, where the induction of a cell axis de novo can be followed by quantification of specific regeneration stages. Using overexpression of fluorescently tagged extranuclear （perinu- clear actin basket, kinesins with a calponin homology domain （KCH）） as well as intranuclear （histone H2B） factors of nuclear positioning and time-lapse series of the early stages of regeneration, we found that nuclear position is no prerequi- site for polarity formation. However, polarity formation and nuclear migration were both modulated in the transgenic lines, indicating that both phenomena depend on factors affecting cytoskeletal tensegrity and chromatin structure. We integrated these findings into a model where retrograde signals are required for polarity induction. These signals travel via the cytoskeleton from the nucleus toward targets at the plasma membrane.
基金Project supported by the National Natural Science Foundation of China (Grant No. 39470205)State Science and Technology Commision, and Science and Technolgy Commision of Anhui Province
文摘Laser tweezers can manipulate small particles, such as cells and organdies. When coupling them with laser microbeam selective fusion of two tobacco protoplasts containing some chloroplast was achieved. Physical and biological variables that affect laser trapping and laser-induced fusion were also discussed. The results show that the effect of chloroplast content and distribution on the yield of cell fusion is remarkable.
文摘The relationship among transfer and expression of auxin binding protein gene ( abp ), auxin (NAA) induced plasmalemma hyperpolarity and sensibility to auxin during protoplast culture was studied by measuring transmembrane potential difference (Em) and culturing the protoplasts of sense and antisense abp transgenic tobacco. The concentration of NAA inducing the highest degree of hyperpolarity of sense abp transgenic tobacco protoplasts was lower than the control, and in protoplast culture, their sensibility to auxin increased. The concentration of antisense abp transgenic tobacco protoplasts was higher than the control, and in protoplast culture, their sensibility to auxin decreased. These results demonstrated that ABP synthesized in endoplasmic reticulum needed to transport to cell membrane and functioned there.
