Here, we report the aluminum (Al)-induced organ-specific expression of a WAK1 (cell wall-associated receptor kinase 1) gene and cell type-specific localization of WAK proteins in Arabidopsis. WAK1 -specific reverse transcriptase-polymerase chain reaction analysis revealed an Al-induced WAK1 gene expression in roots. Short- and long-term analysis of gene expression in root fractions showed a typical “on” and “off” pattern with a first peak at 3 h of Al exposure followed by a sharp decline at 6 h and a complete disappearance after 9 h of Al exposure, suggesting the WAK1 is a further representative of Al-induced early genes. In shoots, upon root Al exposure, an increased but stable WAK1 expression was observed. Using confocal microscopy, we visualized Al-induced closure of leaf stomata, consistent with previous suggestions that the Al stress primarily experienced in roots associated with the transfer of root-shoot signals. Elevated levels of WAK protein in root cells were observed through western blots after 6 h of Al exposure, indicating a lag time between the Al-induced WAK transcription and translation. WAK proteins are localized abundantly to peripheries of cortex cells within the elongation zone of the root apex. In these root cells, disintegration of cortical microtubules was observed after Al treatment but not after the Al analog lanthanum treatments. Tip-growing control root hairs, stem stomata, and leaf stomatal pores are characterized with high amounts of WAKs, suggesting WAKs are accumulating at plasma membrane domains, which suffer from mechanical stress and lack dense arrays of supporting cortical microtubules. Further, transgenic plants overexpressing WAK1 showed an enhanced Al tolerance in terms of root growth when compared with the wild-type plants, making the WAK1 one of the important candidates for plant defense against Al toxicity.

/pdf/aluminum-induced-gene-expression-and-protein-localization-of-4nfdu5lv8s.pdf

Aluminum-Induced Gene Expression and Protein Localization of a Cell Wall-Associated Receptor Kinase in Arabidopsis

The transport of ions and metabolites through plasmodesmata has been thought to be controlled at the neck region where the cytoplasmic annulus is constricted and where callose has also been localised. In order to determine the possible structural and functional effects of callose, its deposition was inhibited through incubation of the plant tissue with 2-deoxy-D-glucose (DDG) for 1 h prior to fixation in 2.5% glutaraldehyde. The inhibition of callose formation was monitored through aniline blue-induced fluorescence of callose. The neck region of the plasmodesmata fromAllium cepa L. roots treated with DDG exhibited a funnel-shaped configuration. This is in contrast to the plasmodesmata from tissue not incubated with DDG, which exhibited constricted necks similar to those previously reported. Both initial dissection and glutaraldehyde fixation induced neck constriction in plasmodesmata, however, dissection of tissue increased the frequency of constrictions. The inhibition of callose formation by chemical means showed that the neck constrictions and raised collars in this area are artefacts due to physical wounding and glutaraldehyde fixation. The external electron-dense material observed when tannic acid is included in the primary fixative appears to be unrelated to the deposition of callose at the neck region.

Callose deposition at plasmodesmata

Ultrastructural and Temporal Observations of the Potyvirus Cylindrical Inclusions (CIs) Show That the CI Protein Acts Transiently in Aiding Virus Movement

Interactions of plant cells with pathogens or other biotic or abiotic environmental factors can give rise to systemic defense responses that rely upon the cell-to-cell and systemic transport of specific signals. A novel type of systemic signaling was revealed by recent evidence indicating the existence of RNA species that travel cell to cell and through the vasculature. The most compelling evidence for intercellular and systemic transport of RNA in plants is provided by viroids and viruses that apparently use the endogenous transport machinery to spread infection. The cell to cell movement of plant viruses occurs through small pores in the cell wall known as plasmodesmata and depends on virus-encoded 'movement proteins'. This review summarizes current knowledge of Tobacco mosaic virus infection with emphasis on the mechanism by which this virus targets its RNA genome from sites of replication to plasmodesmata to achieve intercellular spread.

The spread of tobacco mosaic virus infection: insights into the cellular mechanism of RNA transport.

The key tissue for the integration of growth processes within a plant is the phloem. Its highly specialised sieve elements (SEs) form an intimate symplasmic domain for the long-distance transport of assimilates from green source organs to the assimilate consuming or storing sink organs. When functional, the SEs have a greatly reduced cytoplasm and are interconnected by wide sieve pores originating in plasmodesmata, thus forming a low-resistance pathway for translocates. In contrast to the xylem, the driving force for the long distance transport is produced endogenously within the phloem tissue and the conducting cells are vital when functioning, so that changes in assimilate demand can rapidly and flexibly be met.

Phloem. Structure Related to Function

Initial study indicated that much callose is produced in pea root tissues during the preparation of fresh hand-cut sections or during conventional fixation in formalin-acetic acid-alcohol, glutaraldehyde or acrolein. In contrast, there is little callose in freeze-substituted tissues and this is mostly in sieve tubes and considered endogenous. Freeze-substitution was subsequently used to monitor wound-induced callose development in the various tissues of pea roots. This development was fastest in the phloem, first detected 1 minute post-wounding and complete by 3 hours. In some parenchyma cells full development was delayed by 20 hours. By 100 hours, wound callose was no longer detectable in parenchyma cells but remained undiminished in the phloem. The implications of these results for all studies involving callose localization are discussed.

The time course of the induction of callose in wounded pea roots

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The time course of the induction of callose in wounded pea roots