Reduction in transport in wheat (Triticum aestivum) is caused by sustained phloem feeding by the Russian wheat aphid (Diuraphis noxia)

The Russian wheat aphid (Diuraphis noxia) feeds preferentially from the phloem of longitudinal veins of nonresistant wheat leaves. The xenobiotic, 5,6-CFDA was applied to exposed leaf blade mesophyll cells in control and aphid-infested plants. In control plants, the fluorophore moved approximately 5–6cm from the point of application of 5,6-CFDA within 3h of application. The fluorochrome was transported in the sieve tube companion cell complex, including those in the numerous interconnecting transverse veins. Leaf blades on which the Russian wheat aphid had been feeding demonstrated a marked decrease in 5,6-CF transport. Aphids feeding on the leaves formed local sinks and redirected the fluorophore (and presumably associated assimilate) to the aphids’ gut, with little longitudinal translocation of 5,6-CF below inserted stylets or aphid feeding areas. Aniline blue staining revealed massive deposits of wound and reaction callose caused by the aphids, with callose associated with the sieve plates, pore-plasmodesma between the companion cells and their associated sieve tubes, as well as with plasmodesmal aggregates in parenchymatous elements within the vascular bundles. Leaves that had been colonised by aphids but from which the aphids were removed, showed extensive wound callose deposits, which persisted for up to 48h after removal of aphid colonies, suggesting that the damage caused by aphid feeding is a long-term, non-transient event in non-resistant plants

Aphid (Sitobion yakini) investigation shows thin-walled sieve tubes in barley (Hordeum vulgare L) to be more functional than thick-walled sieve tubes

Barley, like most other grasses that have been studied, contains two kinds of sieve tube. The first formed are called thinwalled sieve tubes because of their thin wall compared to the late-formed, and are associated with companion cells. The late-formed are thick-walled sieve tubes, which differentiate next to the metaxylem vessels and lack companion cells. Aphid (Sitobion yakini (Eastop) feeding was studied using light microscopy to determine if they preferentially feed from thin- or thick-walled sieve tubes in the barley leaf. Penetration of the stylets through the leaf epidermis and mesophyll was largely intercellular, becoming partly intercellular and, partly, intracellular inside the vascular bundle. Sixteen of 19 pairs of stylets (84%), and 293 of 317 (92%) stylet tracks terminated at the thin-walled sieve tubes, suggesting that Sitobion yakini feeds preferentially on the thin-walled sieve tubes which seem to be more attractive to the aphid. These thin-walled sieve tubes are thus probably the most functional in terms of phloem loading and transport

Resistance or tolerance: an examination of aphid (Sitobion yakini) phloem feeding on Betta and Betta-Dn wheat (Triticum aestivum)

Engineering pest resistance into crops is important. However, the mechanisms of resistance are not clearly understood. In this study, we examined the effects of aphid feeding on Russian wheat aphid-resistant and -susceptible cultivars of wheat (Triticum aestivum L.); Betta-Dn and Betta, respectively, by the grass aphid, Sitobion yakini (Eastop). These cultivars were grown with or without aphid colonies. In each case, we examined the plants specifically for the formation of wound callose associated with the phloem, using aniline blue and fluorescence microscopy. We observed that aphid feeding stimulated the formation of wound callose in the susceptible cultivar, but that callose was comparatively reduced in the resistant cultivar of wheat. In a separate series of experiments, the xenobiotic, 5, 6-carboxyfluorescein diacetate was applied to attached sink leaves, distal to feeding aphids. When leaf segments were examined four hours after application, little evidence of phloem transport of the fluorescent cleavage product, 5, 6-carboxyfluorescein (5, 6-CF), was evident below known aphid-probed sieve tubes. Low levels or absence of 5, 6-CF indicates that either the aphids have successfully redirected sap to themselves, or that the phloem is no longer functional. In contrast, 5, 6-CF transport was evident below sites of aphid probing in Betta-Dn, suggesting that the phloem was still capable of long-distance transport. In addition, callose deposition was reduced in Betta-Dn leaf phloem and it is surmised that transport was not as affected by aphid feeding in the resistant cultivar. This indicates that the ‘resistant’ wheat cultivar may in fact be tolerant to aphid feeding by successfully overcoming the nutrient drain that feeding aphids imposed on the phloem transport system

Towards reconciliation of structure with function in plasmodesmata—who is the gatekeeper?

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

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

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

A re-evaluation of plastochron index determination in peas - a case for using leaflet length

The plastochron index (PI) is a measure of plant growth reports our findings on PI using the average length of and can be used to determine growth rate, based upon the first pair of leaflets on each node. Early leaflet appearance of successive leaves on the axis of the growth in peas occurs exponentially and the early plant. PI should under ideal growth conditions be a stages of growth of successive pairs of leaflets occur at regular event and should be predictable with a relatively the same relative growth rate. Given that growth of small error of a few hours. PI has been variously leaflets during early development can be measured calculated in peas, and each method reported has had successfully, we propose the use of leaflet growth as a with it a number of problems that do not allow for measure of the plastochron index in peas. Our results reasonable prediction of PI. Internode length varies suggest that plant age is best expressed using the greatly and is dependent upon the variety, which may be plastochron index, which is a measure of the time short- or long-stemmed; thus this parameter is not ideal interval between the initiations of successive events — for determining growth rate or plant age. This paper in the case of peas, of successive pairs of leaflets

The microstructure of plasmodesmata in internodal stem tissue of the Saccharum hybrid var. NCo376 : evidence for an apoplasmic loading pathway

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

Comparative ultrastructure of plasmodesmata of Chara and selected bryophytes: towards an elucidation of the evolutionary origin of plant plasmodesmata

We have used transmission electron microscopy to examine plasmodesmata of the charophycean green alga Chara zeylanica, and of the putatively early divergent bryophytes Monoclea gottschei (liverwort), Notothylas orbicularis (hornwort), and Sphagnum fimbriatum (moss), in an attempt to learn when seed plant plasmodesmata may have originated. The three bryophytes examined have desmotubules. In addition, Monoclea was found to have branched plasmodesmata, and plasmodesmata of Sphagnum displayed densely staining regions around the neck region, as well as ring-like wall specializations. In Chara, longitudinal sections revealed endoplasmic reticulum (ER) that sometimes appeared to be associated with plasmodesmata, but this was rare, despite abundant ER at the cell periphery. Across all three fixation methods, cross-sectional views showed an internal central structure, which in some cases appeared to be connected to the plasma membrane via spoke-like structures. Plasmodesmata were present even in the incompletely formed reticulum of forming cell plates, from which we conclude that primary plasmodesmata are formed at cytokinesis in Chara zeylanica. Based on these results it appears that plasmodesmata of Chara may be less specialized than those of seed plants, and that complex plasmodesmata probably evolved in the ancestor of land plants before extant lineages of bryophytes diverged

Phloem loading in the sucrose-export-defective (SXD-1) mutant maize is limited by callose deposition at plasmodesmata in bundle sheath-vascular parenchyma interface

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

Simulation of the Einstein-de Haas effect combining molecular and spin dynamics

The spin and lattice dynamics of a ferromagnetic nanoparticle are studied via molecular dynamics and with semi-classical spin dynamics simulations where spin and lattice degrees of freedom are coupled via a dynamic uniaxial anisotropy term. We show that this model conserves total angular momentum, whereas spin and lattice angular momentum are not conserved. We carry out simulations of the the Einstein-de Haas effect for a Fe nanocluster with more than 500 atoms that is free to rotate, using a modified version of the open-source spinlattice dynamics code (SPILADY). We show that the rate of angular momentum transfer between spin and lattice is proportional to the strength of the magnetic anisotropy interaction. The addition of the anisotropy allows full spin-lattice relaxation to be achieved on previously reported timescales of \sim 100 ps and for tight-binding magnetic anisotropy energies comparable to those of small Fe nanoclusters.Comment: 23 pages, 3 figure