Protoplasts prepared from tobacco (Nicotiana tabacum L., cultivar BY-2) suspension cells have similar morphological characteristics to those in animal cells. The hallmarks of apoptosis such as condensation and periphe...Protoplasts prepared from tobacco (Nicotiana tabacum L., cultivar BY-2) suspension cells have similar morphological characteristics to those in animal cells. The hallmarks of apoptosis such as condensation and peripheral distribution of nuclei, TUNEL positive reaction, and DNA ladders were observed when tobacco protoplasts were treated with the hydroxyl radical generating system (1.0 mmol/L FeSO4/0.5 mmol/L H2O2). In animals, the loss of transmembrane potential (DeltaPsi(m)) and the exposure of phospholipid phosphatidylserine (PS) are believed to be the main apoptosis events. To test whether these significant processes take place in plants, flow cytometry was used to detect annexin V binding and changes in DeltaPsi(m). Results showed that the PS turned out from inner membrane and DeltaPsi(m) gradually decreased during the apoptosis. All these apoptotic characteristics proved that hydroxyl radicals can cause typical programmed cell death (PCD) in tobacco protoplasts and this design can be served as an effective experiment system to explore the mechanism of plant apoptosis.展开更多
Protoplasts of embryogenic suspension cells of loblolly pine (Pinus taeda L).were isolated at exponential growth stage.Influences of various concentrations of basal medium,levels of BA,and concentrations of inositol ...Protoplasts of embryogenic suspension cells of loblolly pine (Pinus taeda L).were isolated at exponential growth stage.Influences of various concentrations of basal medium,levels of BA,and concentrations of inositol on the differentiation of embryonal suspensor mass (ESM),early stage somatic embryos (ESE) ,and lae stage somatic embryos (LSE) were investigated .A study of the effect of various concentrations of LP basal medium sowed that the optimal basal medium concentration of ESM,ESE,and LSE differentiation was 1.25 LP medium.The effects of various levels of BA and inositol showed that the optimal concentrations of BA for the formation of ESM,ESE and LSE were 4 mg/L ,2mg/L and 1mg/L,respectively ,and the optimal concentrations of inositol for the ESM ,ESE and LSM formation were 400mg/L,800mg/L and 1,200mg/L,respectively.展开更多
Since nuclear extrusion was rediscovered in young Allium scale by S.H.WU in the years of 1950's systematic investigations on this phenomenon were carried out with various kinds of microscopic techniques a...Since nuclear extrusion was rediscovered in young Allium scale by S.H.WU in the years of 1950's systematic investigations on this phenomenon were carried out with various kinds of microscopic techniques and plant materials to collect more effective evidence to clarify the debate about whether the nuclear extrusion is an artifact or normal event. In the cooperative research of S. H. WU and C. H. LOU the normality of the occurrence of nuclear extrusion either in growing part of plant or in senescent tissue has been confirmed. This event is intimately associated with the physiological state of the tissues/cells and may play an important role in redistribution and reutilization of cell contents. Based on the results obtained a hypothesis of intercellular movement of protoplasm as a means of translocation of organic material in plants was suggested. Chromatin extrusion was also discovered in the pollen mother cells (cytomixis) of certain angiosperms by G. C. ZHENG and his team. Intercellular migration of chromatin appears most frequently at the stage of synizesis. Cytomixis has been studied in relation to variation and evolution. Chromosome aberration has been considered to be closely associated with chromatin extrusion. By vital microscopic observations of the live tissues of garlic (Allium sativum L.) bud and wheat (Triticum aestivum L.) ovule combined with cinemicroscopy and video recording it has been uncovered that, not only the nuclear material but also the cytoplasm could traverse the intercellular channels by vigorous contraction and expansion, and they may simultaneously extrude out of a cell but often asynchronously migrate from one cell to another. The involvement of cytoplasmic constituents in intercellular migration was also detected in pollen mother cells with electron microscopy. Regarding the mechanism of intercellular movement a series of experiments provide convincing evidence showing that this kind of movement is an active metabolic process closely coupled with energy metabolism, and the motive power for driving the extrusion may be supplied by the contractile proteins in protoplasm.展开更多
文摘Protoplasts prepared from tobacco (Nicotiana tabacum L., cultivar BY-2) suspension cells have similar morphological characteristics to those in animal cells. The hallmarks of apoptosis such as condensation and peripheral distribution of nuclei, TUNEL positive reaction, and DNA ladders were observed when tobacco protoplasts were treated with the hydroxyl radical generating system (1.0 mmol/L FeSO4/0.5 mmol/L H2O2). In animals, the loss of transmembrane potential (DeltaPsi(m)) and the exposure of phospholipid phosphatidylserine (PS) are believed to be the main apoptosis events. To test whether these significant processes take place in plants, flow cytometry was used to detect annexin V binding and changes in DeltaPsi(m). Results showed that the PS turned out from inner membrane and DeltaPsi(m) gradually decreased during the apoptosis. All these apoptotic characteristics proved that hydroxyl radicals can cause typical programmed cell death (PCD) in tobacco protoplasts and this design can be served as an effective experiment system to explore the mechanism of plant apoptosis.
文摘Protoplasts of embryogenic suspension cells of loblolly pine (Pinus taeda L).were isolated at exponential growth stage.Influences of various concentrations of basal medium,levels of BA,and concentrations of inositol on the differentiation of embryonal suspensor mass (ESM),early stage somatic embryos (ESE) ,and lae stage somatic embryos (LSE) were investigated .A study of the effect of various concentrations of LP basal medium sowed that the optimal basal medium concentration of ESM,ESE,and LSE differentiation was 1.25 LP medium.The effects of various levels of BA and inositol showed that the optimal concentrations of BA for the formation of ESM,ESE and LSE were 4 mg/L ,2mg/L and 1mg/L,respectively ,and the optimal concentrations of inositol for the ESM ,ESE and LSM formation were 400mg/L,800mg/L and 1,200mg/L,respectively.
文摘Since nuclear extrusion was rediscovered in young Allium scale by S.H.WU in the years of 1950's systematic investigations on this phenomenon were carried out with various kinds of microscopic techniques and plant materials to collect more effective evidence to clarify the debate about whether the nuclear extrusion is an artifact or normal event. In the cooperative research of S. H. WU and C. H. LOU the normality of the occurrence of nuclear extrusion either in growing part of plant or in senescent tissue has been confirmed. This event is intimately associated with the physiological state of the tissues/cells and may play an important role in redistribution and reutilization of cell contents. Based on the results obtained a hypothesis of intercellular movement of protoplasm as a means of translocation of organic material in plants was suggested. Chromatin extrusion was also discovered in the pollen mother cells (cytomixis) of certain angiosperms by G. C. ZHENG and his team. Intercellular migration of chromatin appears most frequently at the stage of synizesis. Cytomixis has been studied in relation to variation and evolution. Chromosome aberration has been considered to be closely associated with chromatin extrusion. By vital microscopic observations of the live tissues of garlic (Allium sativum L.) bud and wheat (Triticum aestivum L.) ovule combined with cinemicroscopy and video recording it has been uncovered that, not only the nuclear material but also the cytoplasm could traverse the intercellular channels by vigorous contraction and expansion, and they may simultaneously extrude out of a cell but often asynchronously migrate from one cell to another. The involvement of cytoplasmic constituents in intercellular migration was also detected in pollen mother cells with electron microscopy. Regarding the mechanism of intercellular movement a series of experiments provide convincing evidence showing that this kind of movement is an active metabolic process closely coupled with energy metabolism, and the motive power for driving the extrusion may be supplied by the contractile proteins in protoplasm.
