The microstructure of plasmodesmata in internodal stem tissue of the Saccharum hybrid var. NCo376 : evidence for an apoplasmic loading pathway
- Botha, Christiaan E J, Cross, Robin H M, Gerber, Jacques
- Authors: Botha, Christiaan E J , Cross, Robin H M , Gerber, Jacques
- Date: 2004
- Language: English
- Type: Article
- Identifier: vital:6494 , http://hdl.handle.net/10962/d1004487
- Description: The distribution, structure and functional state of plasmodesmata were investigated to gain a clearer understanding of the sucrose transport pathway to the storage parenchyma cells in stem tissue in Saccharum officinarum var. NCo376. Evidence from structural studies on sugarcane stems by electron microscopy indicated that there are numerous plasmodesmata from the vascular bundles through to the storage parenchyma cells in mature stem tissue. Our studies, supported by fluorescence microscopy and iontophoresis, indicate that there are functional plasmodesmata in the phloemunloading pathway from transport phloem tissue to the bundle sheath in Saccharum, which could support symplasmic transport; plasmodesmata outside of the sheath cells in the storage parenchyma appear to be constricted by sphincter-like structures within their neck regions. Staining with Aniline Blue revealed evidence of large callose deposits, which co-localized with plasmodesmatal aggregates in the walls of the storage parenchyma cells. This suggests that the sucrose transport into, and accumulation by, storage parenchyma of mature stem tissue is under apoplasmic control.
- Full Text:
- Date Issued: 2004
- Authors: Botha, Christiaan E J , Cross, Robin H M , Gerber, Jacques
- Date: 2004
- Language: English
- Type: Article
- Identifier: vital:6494 , http://hdl.handle.net/10962/d1004487
- Description: The distribution, structure and functional state of plasmodesmata were investigated to gain a clearer understanding of the sucrose transport pathway to the storage parenchyma cells in stem tissue in Saccharum officinarum var. NCo376. Evidence from structural studies on sugarcane stems by electron microscopy indicated that there are numerous plasmodesmata from the vascular bundles through to the storage parenchyma cells in mature stem tissue. Our studies, supported by fluorescence microscopy and iontophoresis, indicate that there are functional plasmodesmata in the phloemunloading pathway from transport phloem tissue to the bundle sheath in Saccharum, which could support symplasmic transport; plasmodesmata outside of the sheath cells in the storage parenchyma appear to be constricted by sphincter-like structures within their neck regions. Staining with Aniline Blue revealed evidence of large callose deposits, which co-localized with plasmodesmatal aggregates in the walls of the storage parenchyma cells. This suggests that the sucrose transport into, and accumulation by, storage parenchyma of mature stem tissue is under apoplasmic control.
- Full Text:
- Date Issued: 2004
Regulation within the supracellular highway - plasmodesma are the key
- Botha, Christiaan E J, Cross, Robin H M
- Authors: Botha, Christiaan E J , Cross, Robin H M
- Date: 2001
- Language: English
- Type: Article
- Identifier: vital:6500 , http://hdl.handle.net/10962/d1005918
- Description: Plasmodesmal connections are unique, highly dynamic intercellular structures that are lined by the plasmamembrane. They are believed to be a vital intercellular communication channel between living cells, linking numbers of living cells into interconnected, highly specialised cellular domains, thus enabling the plant to act as an integrated organism. Their evolution in the higher plant was inevitable. It is accepted that cell heterogeneity rather than cell divergence pressurised developing plant systems along a route that led to the development of intercellular passages and connections. With time these connections have evolved to allow some degree of regulation and traffic control. This paper explores some of the structure/function relationships in plasmodesmata. Attention is focused on the potential role of the neck region of these remarkable structures and discusses models which may explain the processes involved in regulating the movement of substances from cell to cell.
- Full Text: false
- Date Issued: 2001
- Authors: Botha, Christiaan E J , Cross, Robin H M
- Date: 2001
- Language: English
- Type: Article
- Identifier: vital:6500 , http://hdl.handle.net/10962/d1005918
- Description: Plasmodesmal connections are unique, highly dynamic intercellular structures that are lined by the plasmamembrane. They are believed to be a vital intercellular communication channel between living cells, linking numbers of living cells into interconnected, highly specialised cellular domains, thus enabling the plant to act as an integrated organism. Their evolution in the higher plant was inevitable. It is accepted that cell heterogeneity rather than cell divergence pressurised developing plant systems along a route that led to the development of intercellular passages and connections. With time these connections have evolved to allow some degree of regulation and traffic control. This paper explores some of the structure/function relationships in plasmodesmata. Attention is focused on the potential role of the neck region of these remarkable structures and discusses models which may explain the processes involved in regulating the movement of substances from cell to cell.
- Full Text: false
- Date Issued: 2001
Phloem loading in the sucrose-export-defective (SXD-1) mutant maize is limited by callose deposition at plasmodesmata in bundle sheath-vascular parenchyma interface
- Botha, Christiaan E J, Cross, Robin H M, Van Bel, A J E, Peter, Craig I
- Authors: Botha, Christiaan E J , Cross, Robin H M , Van Bel, A J E , Peter, Craig I
- Date: 2000
- Language: English
- Type: Article
- Identifier: vital:6503 , http://hdl.handle.net/10962/d1005926
- Description: Using Lucifer Yellow we have demonstrated that the phloem-loading pathway from the mesophyll to the bundle sheath-vascular parenchyma interface in Zea mays source leaves follows a symplasmic route in small and intermediate vascular bundles in control as well as in the green sections of mutant sucrose-export-defective (SXD-1) plants. In the anthocyanin-rich mutant leaf sections, Lucifer Yellow transport was prohibited along the same path, at the bundle sheath-vascular parenchyma interface in particular. Plasmodesmata at the latter interface in SXD-1 anthocyanin-rich leaf sections appear to be structurally altered through callose deposition at the plasmodesmal orifices. We suggest that a transport bottleneck at the bundle sheath-vascular parenchyma interface is thus orchestrated and regulated through callose formation, preventing symplasmic transport across this important loading interface.
- Full Text:
- Date Issued: 2000
- Authors: Botha, Christiaan E J , Cross, Robin H M , Van Bel, A J E , Peter, Craig I
- Date: 2000
- Language: English
- Type: Article
- Identifier: vital:6503 , http://hdl.handle.net/10962/d1005926
- Description: Using Lucifer Yellow we have demonstrated that the phloem-loading pathway from the mesophyll to the bundle sheath-vascular parenchyma interface in Zea mays source leaves follows a symplasmic route in small and intermediate vascular bundles in control as well as in the green sections of mutant sucrose-export-defective (SXD-1) plants. In the anthocyanin-rich mutant leaf sections, Lucifer Yellow transport was prohibited along the same path, at the bundle sheath-vascular parenchyma interface in particular. Plasmodesmata at the latter interface in SXD-1 anthocyanin-rich leaf sections appear to be structurally altered through callose deposition at the plasmodesmal orifices. We suggest that a transport bottleneck at the bundle sheath-vascular parenchyma interface is thus orchestrated and regulated through callose formation, preventing symplasmic transport across this important loading interface.
- Full Text:
- Date Issued: 2000
Towards reconciliation of structure with function in plasmodesmata—who is the gatekeeper?
- Botha, Christiaan E J, Cross, Robin H M
- Authors: Botha, Christiaan E J , Cross, Robin H M
- Date: 2000
- Language: English
- Type: Article
- Identifier: vital:6497 , http://hdl.handle.net/10962/d1004493
- Description: Whilst the structure of higher plant plasmodesmata was first described by Robards (1963. Desmotubule—a plasmodesmatal substructure. Nature 218, 784), and despite many subsequent intensive investigations, there is still much that remains unclear relating to their ultrastructure and functioning in higher plants. We have examined chemically fixed plant material, and suggest that the conformational changes seen in plasmodesmatal substructure, particularly the deposition of electron-dense extra-plasmodesmal material, is linked to either manipulation of the hormonal balance (as in Avocado fruit), or of osmotic potential in leaf blade material. These changes result in the deposition of β 1,3-glucan (callose) at the neck region of these plasmodesmata. This electron-dense material is deposited at the neck region of plasmodesmata, and forms a collar-like structure. The formation of a collar is shown to be coupled with loss of lucence within the cytoplasmic sleeve. The formation of a collar at the plasmodesmatal orifice thus results in encapsulation and closure of the plasmodesmatal orifice. Closure of the orifice coincides with a loss of electron-lucence and a lack of resolution of the desmotubule. These ultrastructural changes are potentially significant and could contribute to, result in, or assist in the down-regulation of cell to cell trafficking via plasmodesmata.
- Full Text:
- Date Issued: 2000
- Authors: Botha, Christiaan E J , Cross, Robin H M
- Date: 2000
- Language: English
- Type: Article
- Identifier: vital:6497 , http://hdl.handle.net/10962/d1004493
- Description: Whilst the structure of higher plant plasmodesmata was first described by Robards (1963. Desmotubule—a plasmodesmatal substructure. Nature 218, 784), and despite many subsequent intensive investigations, there is still much that remains unclear relating to their ultrastructure and functioning in higher plants. We have examined chemically fixed plant material, and suggest that the conformational changes seen in plasmodesmatal substructure, particularly the deposition of electron-dense extra-plasmodesmal material, is linked to either manipulation of the hormonal balance (as in Avocado fruit), or of osmotic potential in leaf blade material. These changes result in the deposition of β 1,3-glucan (callose) at the neck region of these plasmodesmata. This electron-dense material is deposited at the neck region of plasmodesmata, and forms a collar-like structure. The formation of a collar is shown to be coupled with loss of lucence within the cytoplasmic sleeve. The formation of a collar at the plasmodesmatal orifice thus results in encapsulation and closure of the plasmodesmatal orifice. Closure of the orifice coincides with a loss of electron-lucence and a lack of resolution of the desmotubule. These ultrastructural changes are potentially significant and could contribute to, result in, or assist in the down-regulation of cell to cell trafficking via plasmodesmata.
- Full Text:
- Date Issued: 2000
Symplastic solute transport and avocado fruit development : a decline in cytokinin/ABA ratio is related to appearance of the Hass small fruit variant
- Moore-Gordon, Clive S, Cowan, Keith A, Bertling, Isa, Botha, Christiaan E J, Cross, Robin H M
- Authors: Moore-Gordon, Clive S , Cowan, Keith A , Bertling, Isa , Botha, Christiaan E J , Cross, Robin H M
- Date: 1998
- Language: English
- Type: Article
- Identifier: vital:6525 , http://hdl.handle.net/10962/d1005959
- Description: Studies on the effect of fruit size on endogenous ABA and isopentenyladenine (iP) in developing avocado (Persea americana Mill. cv. Hass) fruit revealed that ABA content was negatively correlated with fruit size whilst the iP/ABA ratio showed a linear relationship with increasing size of fruit harvested 226 d after full bloom. The effect of this change in hormone balance on the relationship between symplastic solute transport and appearance of the small fruit variant was examined following manipulation of the endogenous cytokinin (CK)/ABA ratio. Application of ABA caused seed coat senescence and retarded fruit growth but these effects were absent in fruit treated with equal amounts of ABA plus iP. Thus, the underlying physiological mechanisms associated with ABA-induced retardation of Hass avocado fruit growth appeared to be inextricably linked to a decline in CK content and included: diminution of mesocarp and seed coat plasmodesmatal branching, gating of mesocarp and seed coat plasmodesmata by deposition of electron dense material in the neck region, abolishment of the electrochemical gradient between mesocarp and seed coat parenchyma, and arrest of cell-to-cell chemical communication.
- Full Text:
- Date Issued: 1998
- Authors: Moore-Gordon, Clive S , Cowan, Keith A , Bertling, Isa , Botha, Christiaan E J , Cross, Robin H M
- Date: 1998
- Language: English
- Type: Article
- Identifier: vital:6525 , http://hdl.handle.net/10962/d1005959
- Description: Studies on the effect of fruit size on endogenous ABA and isopentenyladenine (iP) in developing avocado (Persea americana Mill. cv. Hass) fruit revealed that ABA content was negatively correlated with fruit size whilst the iP/ABA ratio showed a linear relationship with increasing size of fruit harvested 226 d after full bloom. The effect of this change in hormone balance on the relationship between symplastic solute transport and appearance of the small fruit variant was examined following manipulation of the endogenous cytokinin (CK)/ABA ratio. Application of ABA caused seed coat senescence and retarded fruit growth but these effects were absent in fruit treated with equal amounts of ABA plus iP. Thus, the underlying physiological mechanisms associated with ABA-induced retardation of Hass avocado fruit growth appeared to be inextricably linked to a decline in CK content and included: diminution of mesocarp and seed coat plasmodesmatal branching, gating of mesocarp and seed coat plasmodesmata by deposition of electron dense material in the neck region, abolishment of the electrochemical gradient between mesocarp and seed coat parenchyma, and arrest of cell-to-cell chemical communication.
- Full Text:
- Date Issued: 1998
Plasmodesmatal frequency in relation to short-distance transport and phloem loading in leaves of barley (Hordeum vulgare). Phloem is not loaded directly from the symplast
- Botha, Christiaan E J, Cross, Robin H M
- Authors: Botha, Christiaan E J , Cross, Robin H M
- Date: 1997
- Language: English
- Type: Article
- Identifier: vital:6504 , http://hdl.handle.net/10962/d1005928
- Description: We investigated the phloem loading pathway in barley, by determining plasmodesmatal frequencies at the electron microscope level for both intermediate and small blade bundles of mature barley leaves. Lucifer yellow was injected intercellularly into bundle sheath, vascular parenchyma, and thin-walled sieve tubes. Passage of this symplastically transported dye was monitored with an epifluorescence microscope under blue light. Low plasmodesmatal frequencies endarch to the bundle sheath cells are relatively low for most interfaces terminating at the thin- and thick-walled sieve tubes within this C3 species. Lack of connections between vascular parenchyma and sieve tubes, and low frequencies (0.5% plasmodesmata per μm cell wall interface) of connections between vascular parenchyma and companion cells, as well as the very low frequency of pore-plasmodesmatal connections between companion cells and sieve tubes in small bundles (0.2% plasmodesmata per μm cell wall interface), suggest that the companion cell-sieve tube complex is symplastically isolated from other vascular parenchyma cells in small bundles. The degree of cellular connectivity and the potential isolation of the companion cell-sieve tube complex was determined electrophysiologically, using an electrometer coupled to microcapillary electrodes. The less negative cell potential {average -52 mV) from mesophyll to the vascular parenchyma cells contrasted sharply with the more negative potential (-122.5 mV) recorded for the companion cell-thin-walled sieve tube complex. Although intercellular injection of lucifer yellow clearly demonstrated rapid (0.75 μm s-1) longitudinal and radial transport in the bundle sheath-vascular parenchyma complex, as well as from the bundle sheath through transverse veins to adjacent longitudinal veins, we were neither able to detect nor present unequivocal evidence in support of the symplastic connectivity of the sieve tubes to the vascular parenchyma. Injection of the companion cell-sieve tube complex, did not demonstrate backward connectivity to the bundle sheath. We conclude that the low plasmodesmatal frequencies, coupled with a two-domain electropotential zonation configuration, and the negative transport experiments using lucifer yellow, precludes symplastic phloem loading in barley leaves.
- Full Text:
- Date Issued: 1997
- Authors: Botha, Christiaan E J , Cross, Robin H M
- Date: 1997
- Language: English
- Type: Article
- Identifier: vital:6504 , http://hdl.handle.net/10962/d1005928
- Description: We investigated the phloem loading pathway in barley, by determining plasmodesmatal frequencies at the electron microscope level for both intermediate and small blade bundles of mature barley leaves. Lucifer yellow was injected intercellularly into bundle sheath, vascular parenchyma, and thin-walled sieve tubes. Passage of this symplastically transported dye was monitored with an epifluorescence microscope under blue light. Low plasmodesmatal frequencies endarch to the bundle sheath cells are relatively low for most interfaces terminating at the thin- and thick-walled sieve tubes within this C3 species. Lack of connections between vascular parenchyma and sieve tubes, and low frequencies (0.5% plasmodesmata per μm cell wall interface) of connections between vascular parenchyma and companion cells, as well as the very low frequency of pore-plasmodesmatal connections between companion cells and sieve tubes in small bundles (0.2% plasmodesmata per μm cell wall interface), suggest that the companion cell-sieve tube complex is symplastically isolated from other vascular parenchyma cells in small bundles. The degree of cellular connectivity and the potential isolation of the companion cell-sieve tube complex was determined electrophysiologically, using an electrometer coupled to microcapillary electrodes. The less negative cell potential {average -52 mV) from mesophyll to the vascular parenchyma cells contrasted sharply with the more negative potential (-122.5 mV) recorded for the companion cell-thin-walled sieve tube complex. Although intercellular injection of lucifer yellow clearly demonstrated rapid (0.75 μm s-1) longitudinal and radial transport in the bundle sheath-vascular parenchyma complex, as well as from the bundle sheath through transverse veins to adjacent longitudinal veins, we were neither able to detect nor present unequivocal evidence in support of the symplastic connectivity of the sieve tubes to the vascular parenchyma. Injection of the companion cell-sieve tube complex, did not demonstrate backward connectivity to the bundle sheath. We conclude that the low plasmodesmatal frequencies, coupled with a two-domain electropotential zonation configuration, and the negative transport experiments using lucifer yellow, precludes symplastic phloem loading in barley leaves.
- Full Text:
- Date Issued: 1997
The ultrastructure and computer-enhanced digital image analysis of plasmodesmata at the Kranz mesophyll-bundle sheath interface of Themeda triandra var. imberbis (Retz) A. Camus in conventionally-fixed leaf blades
- Botha, Christiaan E J, Hartley, B J, Cross, Robin H M
- Authors: Botha, Christiaan E J , Hartley, B J , Cross, Robin H M
- Date: 1993
- Language: English
- Type: Article
- Identifier: vital:6502 , http://hdl.handle.net/10962/d1005925
- Description: The ultrastructure of the plasmodesmata at the Kranz mesophyll-bundle sheath (KMS-BS) interface in Themeda triandra, and the substructures within the plasmodesmata were investigated, using conventionally fixed leaf-blade material, enhanced by the addition of 0.1% tannic acid to the primary fixative. Examination of high-resolution electronmicrographs, and computer-enhanced digital images suggests that these plasmodesmata are complex structures, comprised of helically-arranged particulate material. The electron-dense particles are between 2.5 and 3.0nm in diameter. These particles are specifically associated with the inner face of the inner plasmalemma membrane leaflet, and the outer region of the desmotubule wall. The electron-dense particles are presumably proteinaceous and embedded in a lipid matrix. In the constricted median portion of the KMS-BS plasmodesmata, the space between the desmotubule and the inner plasmalemma membrane leaflet and areas surrounding the proteinaceous particles thereof (the cytoplasmic sleeve) is about 3nm in cross-section, and constitutes what we believe to be the space through which intercellular transport takes place.
- Full Text: false
- Date Issued: 1993
- Authors: Botha, Christiaan E J , Hartley, B J , Cross, Robin H M
- Date: 1993
- Language: English
- Type: Article
- Identifier: vital:6502 , http://hdl.handle.net/10962/d1005925
- Description: The ultrastructure of the plasmodesmata at the Kranz mesophyll-bundle sheath (KMS-BS) interface in Themeda triandra, and the substructures within the plasmodesmata were investigated, using conventionally fixed leaf-blade material, enhanced by the addition of 0.1% tannic acid to the primary fixative. Examination of high-resolution electronmicrographs, and computer-enhanced digital images suggests that these plasmodesmata are complex structures, comprised of helically-arranged particulate material. The electron-dense particles are between 2.5 and 3.0nm in diameter. These particles are specifically associated with the inner face of the inner plasmalemma membrane leaflet, and the outer region of the desmotubule wall. The electron-dense particles are presumably proteinaceous and embedded in a lipid matrix. In the constricted median portion of the KMS-BS plasmodesmata, the space between the desmotubule and the inner plasmalemma membrane leaflet and areas surrounding the proteinaceous particles thereof (the cytoplasmic sleeve) is about 3nm in cross-section, and constitutes what we believe to be the space through which intercellular transport takes place.
- Full Text: false
- Date Issued: 1993
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