The characters and ultrastructure of the intercellular connection were revealed in the outer epidermis of the garlic clove sheath by means of fluorescent probe TRITC_Phalloidin (TRITC_Ph) labeling combined with confoc...The characters and ultrastructure of the intercellular connection were revealed in the outer epidermis of the garlic clove sheath by means of fluorescent probe TRITC_Phalloidin (TRITC_Ph) labeling combined with confocal laser scanning microscopy (CLSM), immuno_gold labeling and transmission electron microscopy. These results show that transcellular channel is a complex of rod_like cytoplasm channel and grouped plasmodesmata (PDs) in pit. The former remains a portion of the cell protoplast. The diameter of PD is normally 60-70 nm. The PDs are the real intercellular symplasmic connections of the cells. The transcellular fibers labeled with the TRITC_Ph obviously become narrow in the primary pit fields, which is the same as the characters observed under the electron microscope. The bright fluorescent spot in the primary wall reflects the grouped PDs in pit, and hence the presence of F_actin in the PDs can be confirmed. In immuno_gold labeling experiment, a lot of gold particles were massively distributed in the rod_like cytoplasm channel and grouped PDs. The result provides effective support that these fluorescent filaments possibly are the existing form of F_actin.展开更多
With light and electron microscopy the substructural change and the ATPase activity of corn (Zea mays L.) root cap cells after short-term osmotic stress were studied. Some spoke-like fine strands originating from the ...With light and electron microscopy the substructural change and the ATPase activity of corn (Zea mays L.) root cap cells after short-term osmotic stress were studied. Some spoke-like fine strands originating from the departed periplasm and stretching towards cell wall could be observed even after plasmolysis. By observing the precipitation of ATPase activity product (lead phosphate) at plasma membrane and plasmodesmata, it was found that the fine strands were plasma membrane-lined channels surrounding the cytoplasm and that they still firmly connected to the plasmodesmata during plasmolysis. Compared with the control (unstressed), a sharp decrease of ATPase activity in the plasmodesmata of the stressed cells was observed. Inhibition of energy metabolism in these limited locales would affect the physiological activity, maybe including the regulation of permeability and the change of size exclusion limit (SEL) of plasmodesmata.展开更多
Plasmodesmata (PDs) are cytoplasmic structures that link adjacent cells to form the symplast of a plant. PDs are involved extensively in a plant's life by mediating symplastic transport of a wide range of ions and...Plasmodesmata (PDs) are cytoplasmic structures that link adjacent cells to form the symplast of a plant. PDs are involved extensively in a plant's life by mediating symplastic transport of a wide range of ions and molecules. Major components of a plasmodesma (PD) include a plasma membrane, a desmotubule, and a cytoplasmic annulus, all of which are readily detectable by electron microscopy. Both the plasma membrane and the desmotubule contain proteinaceous particles, thought to be involved in altering the size of the cytoplasmic annulus. Cytoskeleton elements (actin and myosin) are essential for maintaining the integrity of PDs. Together with these elements, calcium_binding proteins probably play a significant role in regulating PD function. Symplastic transport occurs through the cytoplasmic annulus for the great majority of solutes, while other substances may traverse through the desmotubule internal compartment, the desmotubule shell, or the plasma membrane. The symplast is subdivided into several domains with varying molecular size exclusion limits (ranging from <1 kD to >10 kD). Plasmodesmata can be either primary or secondary; the former are developed during new wall formation and the latter are made in existing walls. The dynamic nature of plasmodesmata is also reflected by their changing frequencies, which, in turn, depend on the developmental and physiological status of the tissue or the entire plant. While diffusion is the major mechanism of symplastic transport, plasmodesmata are selective for certain ions and molecules. Upon viral infection, viral movement proteins interact with PD receptor proteins and, as a result of yet unknown mechanisms, the plasmodesmata are remarkably dilated to allow viral movement proteins and the bound viral genome to enter healthy cells. Some proteins of plant origin are also able to traverse plasmodesmata, presumably in ways similar to viral movement proteins. Some of these plant proteins are probably signal molecules contributing to cell differentiation and other activities. Other proteins move cell_to_cell in a non_specific manner.展开更多
Actin and myosin were found to be associated with the cytoplasmic sleeve of plasmodesmata. As cytoskeletal proteins, actin and myosin are believed to regulate the conductivity of plasmodesmata （PDs） in higher plants...Actin and myosin were found to be associated with the cytoplasmic sleeve of plasmodesmata. As cytoskeletal proteins, actin and myosin are believed to regulate the conductivity of plasmodesmata （PDs） in higher plants. Using immunocytochemical methods, we found the two proteins to be co-localized - and closely linked to each other - in plasmodesmata and ectodesmata-like structure in ageing parenchymatous cells of Allium sativum L. We suggest that intercellular communication is affected by the interaction between actin and myosin.展开更多
Callose,aβ-1,3-glucan plant cell wall polymer,regulates symplasmic channel size at plasmodesmata(PD)and plays a crucial role in a variety of plant processes.However,elucidating the molecular mechanism of PD callose h...Callose,aβ-1,3-glucan plant cell wall polymer,regulates symplasmic channel size at plasmodesmata(PD)and plays a crucial role in a variety of plant processes.However,elucidating the molecular mechanism of PD callose homeostasis is limited.We screened and identified an Arabidopsis mutant plant with excessive callose deposition at PD and found that the mutated gene wasα1-COP,a member of the coat protein I(COPI)coatomer complex.We report that loss of function ofα1-COP elevates the callose accumulation at PD by affecting subcellular protein localization of callose degradation enzyme Pd BG2.This process is linked to the functions of ERH1,an inositol phosphoryl ceramide synthase,and glucosylceramide synthase through physical interactions with theα1-COP protein.Additionally,the loss of function ofα1-COP alters the subcellular localization of ERH1 and GCS proteins,resulting in a reduction of Glc Cers and Glc HCers molecules,which are key sphingolipid(SL)species for lipid raft formation.Our findings suggest thatα1-COP protein,together with SL modifiers controlling lipid raft compositions,regulates the subcellular localization of GPI-anchored PDBG2 proteins,and hence the callose turnover at PD and symplasmic movement of biomolecules.Our findings provide the first key clue to link the COPI-mediated intracellular trafficking pathway to the callose-mediated intercellular signaling pathway through PD.展开更多
Pathogens use effector proteins to manipulate their hosts. During infection of tomato, the fungus Fusarium oxysporum secretes the effectors Avr2 and Six5. Whereas Avr2 suffices to trigger I-2-mediated cell death in he...Pathogens use effector proteins to manipulate their hosts. During infection of tomato, the fungus Fusarium oxysporum secretes the effectors Avr2 and Six5. Whereas Avr2 suffices to trigger I-2-mediated cell death in heterologous systems, both effectors are required for I-2-mediated disease resistance in tomato. How Six5 participates in triggering resistance is unknown. Using bimolecular fluorescence complementation assays we found that Avr2 and Six5 interact at plasmodesmata. Single-cell transformation revealed that a 2xRFP marker protein and Avr2-GFP only move to neighboring cells in the presence of Six5. Six5 alone does not alter plasmodesmatal transduction as 2xRFP was only translocated in the presence of both effectors. In SIX5-expressing transgenic plants, the distribution of virally expressed Avr2-GFP, and subsequent onset of I-2-mediated cell death, differed from that in wild-type tomato. Taken together, our data show that in the presence of Six5, Avr2 moves from cell to cell, which in susceptible plants contributes to virulence, but in I-2 containing plants induces resistance.展开更多
Plant plasmodesmata (PDs) are specialized channels that enable communication between neighboring cells. The intercellular permeability of PDs, which affects plant development, defense, and responses to stimuli, must b...Plant plasmodesmata (PDs) are specialized channels that enable communication between neighboring cells. The intercellular permeability of PDs, which affects plant development, defense, and responses to stimuli, must be tightly regulated. However, the lipid compositions of PD membrane and their impact on PD permeability remain elusive. Here, we report that the Arabidopsis sld1 sld2 double mutant, lacking sphingolipid long-chain base 8 desaturases 1 and 2, displayed decreased PD permeability due to a significant increase in callose accumulation. PD-located protein 5 (PDLP5) was significantly enriched in the leaf epidermal cells of sld1 sld2 and showed specific binding affinity to phytosphinganine (t18:0), suggesting that the enrichment of t18:0-based sphingolipids in sld1 sld2 PDs might facilitate the recruitment of PDLP5 proteins to PDs. The sld1 sld2 double mutant seedlings showed enhanced resistance to the fungal-wilt pathogen Verticillium dahlia and the bacterium Pseudomonas syringae pv. tomato DC3000, which could be fully rescued in sld1 sld2 pdlp5 triple mutant . Taken together, these results indicate that phytosphinganine might regulate PD functions and cell-to-cell communication by modifying the level of PDLP5 in PD membranes.展开更多
文摘The characters and ultrastructure of the intercellular connection were revealed in the outer epidermis of the garlic clove sheath by means of fluorescent probe TRITC_Phalloidin (TRITC_Ph) labeling combined with confocal laser scanning microscopy (CLSM), immuno_gold labeling and transmission electron microscopy. These results show that transcellular channel is a complex of rod_like cytoplasm channel and grouped plasmodesmata (PDs) in pit. The former remains a portion of the cell protoplast. The diameter of PD is normally 60-70 nm. The PDs are the real intercellular symplasmic connections of the cells. The transcellular fibers labeled with the TRITC_Ph obviously become narrow in the primary pit fields, which is the same as the characters observed under the electron microscope. The bright fluorescent spot in the primary wall reflects the grouped PDs in pit, and hence the presence of F_actin in the PDs can be confirmed. In immuno_gold labeling experiment, a lot of gold particles were massively distributed in the rod_like cytoplasm channel and grouped PDs. The result provides effective support that these fluorescent filaments possibly are the existing form of F_actin.
基金Supported by the grants from the National Natural Science Foundation of China.
文摘With light and electron microscopy the substructural change and the ATPase activity of corn (Zea mays L.) root cap cells after short-term osmotic stress were studied. Some spoke-like fine strands originating from the departed periplasm and stretching towards cell wall could be observed even after plasmolysis. By observing the precipitation of ATPase activity product (lead phosphate) at plasma membrane and plasmodesmata, it was found that the fine strands were plasma membrane-lined channels surrounding the cytoplasm and that they still firmly connected to the plasmodesmata during plasmolysis. Compared with the control (unstressed), a sharp decrease of ATPase activity in the plasmodesmata of the stressed cells was observed. Inhibition of energy metabolism in these limited locales would affect the physiological activity, maybe including the regulation of permeability and the change of size exclusion limit (SEL) of plasmodesmata.
文摘Plasmodesmata (PDs) are cytoplasmic structures that link adjacent cells to form the symplast of a plant. PDs are involved extensively in a plant's life by mediating symplastic transport of a wide range of ions and molecules. Major components of a plasmodesma (PD) include a plasma membrane, a desmotubule, and a cytoplasmic annulus, all of which are readily detectable by electron microscopy. Both the plasma membrane and the desmotubule contain proteinaceous particles, thought to be involved in altering the size of the cytoplasmic annulus. Cytoskeleton elements (actin and myosin) are essential for maintaining the integrity of PDs. Together with these elements, calcium_binding proteins probably play a significant role in regulating PD function. Symplastic transport occurs through the cytoplasmic annulus for the great majority of solutes, while other substances may traverse through the desmotubule internal compartment, the desmotubule shell, or the plasma membrane. The symplast is subdivided into several domains with varying molecular size exclusion limits (ranging from <1 kD to >10 kD). Plasmodesmata can be either primary or secondary; the former are developed during new wall formation and the latter are made in existing walls. The dynamic nature of plasmodesmata is also reflected by their changing frequencies, which, in turn, depend on the developmental and physiological status of the tissue or the entire plant. While diffusion is the major mechanism of symplastic transport, plasmodesmata are selective for certain ions and molecules. Upon viral infection, viral movement proteins interact with PD receptor proteins and, as a result of yet unknown mechanisms, the plasmodesmata are remarkably dilated to allow viral movement proteins and the bound viral genome to enter healthy cells. Some proteins of plant origin are also able to traverse plasmodesmata, presumably in ways similar to viral movement proteins. Some of these plant proteins are probably signal molecules contributing to cell differentiation and other activities. Other proteins move cell_to_cell in a non_specific manner.
基金supported by the National Natural Science Foundation of China (30070365, 30470861, 30971706)the Natural Science Foundation of Hebei Province, China (C2008000321)the Specialized Research Fund for the Doctoral Program of Higher Education, China (20060086003)
文摘Actin and myosin were found to be associated with the cytoplasmic sleeve of plasmodesmata. As cytoskeletal proteins, actin and myosin are believed to regulate the conductivity of plasmodesmata （PDs） in higher plants. Using immunocytochemical methods, we found the two proteins to be co-localized - and closely linked to each other - in plasmodesmata and ectodesmata-like structure in ageing parenchymatous cells of Allium sativum L. We suggest that intercellular communication is affected by the interaction between actin and myosin.
基金supported by the National Research Foundation of Korea(NRF)grant funded by the Korea Government(MSIT)(Grant Nos.NRF 2018R1A2A1A05077295,2020M3A9I4038352,2022R1A2C3010331,2020R1A6A1A03044344,and 2022R1A 5A1031361)a grant from the New Breeding Technologies Development Program(Grant No.PJ01653202),Rural Development Administration(RDA),Republic of Korea。
文摘Callose,aβ-1,3-glucan plant cell wall polymer,regulates symplasmic channel size at plasmodesmata(PD)and plays a crucial role in a variety of plant processes.However,elucidating the molecular mechanism of PD callose homeostasis is limited.We screened and identified an Arabidopsis mutant plant with excessive callose deposition at PD and found that the mutated gene wasα1-COP,a member of the coat protein I(COPI)coatomer complex.We report that loss of function ofα1-COP elevates the callose accumulation at PD by affecting subcellular protein localization of callose degradation enzyme Pd BG2.This process is linked to the functions of ERH1,an inositol phosphoryl ceramide synthase,and glucosylceramide synthase through physical interactions with theα1-COP protein.Additionally,the loss of function ofα1-COP alters the subcellular localization of ERH1 and GCS proteins,resulting in a reduction of Glc Cers and Glc HCers molecules,which are key sphingolipid(SL)species for lipid raft formation.Our findings suggest thatα1-COP protein,together with SL modifiers controlling lipid raft compositions,regulates the subcellular localization of GPI-anchored PDBG2 proteins,and hence the callose turnover at PD and symplasmic movement of biomolecules.Our findings provide the first key clue to link the COPI-mediated intracellular trafficking pathway to the callose-mediated intercellular signaling pathway through PD.
文摘Pathogens use effector proteins to manipulate their hosts. During infection of tomato, the fungus Fusarium oxysporum secretes the effectors Avr2 and Six5. Whereas Avr2 suffices to trigger I-2-mediated cell death in heterologous systems, both effectors are required for I-2-mediated disease resistance in tomato. How Six5 participates in triggering resistance is unknown. Using bimolecular fluorescence complementation assays we found that Avr2 and Six5 interact at plasmodesmata. Single-cell transformation revealed that a 2xRFP marker protein and Avr2-GFP only move to neighboring cells in the presence of Six5. Six5 alone does not alter plasmodesmatal transduction as 2xRFP was only translocated in the presence of both effectors. In SIX5-expressing transgenic plants, the distribution of virally expressed Avr2-GFP, and subsequent onset of I-2-mediated cell death, differed from that in wild-type tomato. Taken together, our data show that in the presence of Six5, Avr2 moves from cell to cell, which in susceptible plants contributes to virulence, but in I-2 containing plants induces resistance.
基金This research was supported by grants from the National Science and Technology Major Project(2016ZX08010-001)the National Natural Science Foundation of China(31570283).
文摘Plant plasmodesmata (PDs) are specialized channels that enable communication between neighboring cells. The intercellular permeability of PDs, which affects plant development, defense, and responses to stimuli, must be tightly regulated. However, the lipid compositions of PD membrane and their impact on PD permeability remain elusive. Here, we report that the Arabidopsis sld1 sld2 double mutant, lacking sphingolipid long-chain base 8 desaturases 1 and 2, displayed decreased PD permeability due to a significant increase in callose accumulation. PD-located protein 5 (PDLP5) was significantly enriched in the leaf epidermal cells of sld1 sld2 and showed specific binding affinity to phytosphinganine (t18:0), suggesting that the enrichment of t18:0-based sphingolipids in sld1 sld2 PDs might facilitate the recruitment of PDLP5 proteins to PDs. The sld1 sld2 double mutant seedlings showed enhanced resistance to the fungal-wilt pathogen Verticillium dahlia and the bacterium Pseudomonas syringae pv. tomato DC3000, which could be fully rescued in sld1 sld2 pdlp5 triple mutant . Taken together, these results indicate that phytosphinganine might regulate PD functions and cell-to-cell communication by modifying the level of PDLP5 in PD membranes.
