C4 photosynthesis: a unique elend of modified biochemistry, anatomy and ultrastructure

. Plant penetration by Aphis fabae (Scopoli) was recorded by the electrical penetration graph (EPG) technique and followed by stylectomy during wave-forms that were suspected of indicating sieve element punctures. The severed stylets in the plant tissue were subsequently processed for transmission electron microscopy (TEM) and sectioned either transverse or longitudinal to the stylets. Two completely serially sectioned probes from the epidermis to the phloem were reconstructed.



In one probe the stylet pathway went to a sieve element and showed many empty branches of salivary sheath material. Breaks in cell walls filled with sheath material demonstrated that the majority of cells bordering the track had been punctured, which supports earlier evidence from EPGs. All types of cells showed punctures and the highest number was found inside the vascular bundle. Very few cells died, which would appear to be important for virus transmission, and in others cellular reactions remained limited to some callose formation. The route of the stylets was intercellular and passed through the secondary wall material. The role of pectinase in intercellular penetration, and previous evidence for intracellular tracks are discussed. Most sieve elements had been punctured but only one was eventually accepted. Thus, reaching a sieve element in a host plant does not automatically imply its acceptance though the reason remains unclear. Gelation of phloem proteins was shown in the stylet canal.



In a second probe, plant cytological and morphological correlations with the EPG were emphasized. Probes by other aphid-plant combinations showed great similarity.

https://onlinelibrary.wiley.com/doi/pdf/10.1111/j.1365-3032.1993.tb00604.x

Fine structure of aphid stylet routes in plant tissues in correlation with EPG signals.

STRUCTURAL MATURATION OF LEAVES ... Basipetal Maturation .. ... . ... . . .. . .... . ... . . .. . Timing of the Sink-Source Transition . Development of Minor Veins . Bidirectional Transport . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

The Sink-Source Transition in Leaves

The phloem transports nutrients, defensive compounds, and informational signals throughout vascular plants. Sampling the complex components of mobile phloem sap is difficult because of the damage incurred when the pressurized sieve tubes are breached. In this review we discuss sampling methods, the artifacts that can be introduced by different sampling procedures, the intricate pathways by which materials enter and exit the phloem, and the major types of compounds transported. Loading and unloading patterns are largely determined by the conductivity and number of plasmodesmata and the position-dependent function of solute-specific, plasma membrane transport proteins. Recent evidence indicates that mobile proteins and RNA are part of the plant’s longdistance communication signaling system. Evidence also exists for the directed transport and sorting of macromolecules as they pass through plasmodesmata. A future challenge is to dissect the molecular and cellular aspects of long-distance macromolecular trafficking in the signal transduction pathways of the whole plant.

Phloem Transport: Cellular Pathways and Molecular Trafficking

It has only recently become apparent that the apoplast plays a major role in a diverse range of processes, including intercellular signalling, plant–microbe interactions and both water and nutrient transport. Broadly defined, the apoplast constitutes all compartments beyond the plasmalemma – the interfibrillar and intermicellar space of the cell walls, and the xylem, including its gas- and water-filled intercellular space – extending to the rhizoplane and cuticle of the outer plant surface. The physico-chemical properties of cell walls influence plant mineral nutrition, as nutrients do not simply pass through the apoplast to the plasmalemma but can also be adsorbed or fixed to cell-wall components. Here, current progress in understanding the significance of the apoplast in plant mineral nutrition is reviewed. The contribution of the root apoplast to short-distance transport and nutrient uptakes is examined particularly in relation to Na+ toxicity and Al3+ tolerance. The review extends to long-distance transport and the role of the apoplast as a habitat for microorganisms. In the leaf, the apoplast might have benefits over the vacuole as a site for short-term nutrient storage and solute exchange with the atmosphere.

https://nph.onlinelibrary.wiley.com/doi/epdf/10.1046/j.1469-8137.2001.00034.x

The apoplast and its significance for plant mineral nutrition

In leaf blades of Zea mays L. plasmodesmata between mesophyll cells are aggregated in numerous thickened portions of the walls. The plasmodesmata are unbranched and all are characterized by the presence of electron-dense structures, called sphincters by us, near both ends of the plasmodesmatal canal. The sphincters surround the desmotubule and occlude the cytoplasmic annulus where they occur. Plasmodesmata between mesophyll and bundle-sheath cells are aggregated in primary pit-fields and are constricted by a wide suberin lamella on the sheath-cell side of the wall. Each plasmodesma contains a sphincter on the mesophyll-cell side of the wall. The outer tangential and radial walls of the sheath cells exhibit a continuous suberin lamella. However, on the inner tangential wall only the sites of plasmodesmatal aggregates are consistently suberized. Apparently the movement of photosynthetic intermediates between mesophyll and sheath cells is restricted largely or entirely to the plasmodesmata (symplastic pathway) and transpirational water movement to the cell walls (apoplastic pathway).

Distribution and structure of the plasmodesmata in mesophyll and bundle-sheath cells of Zea mays L.

Portions of the hypocotyls of 16-day-old Cucurbita maxima plants, from which the cotyledons and first foliage leaves had been removed 2 days earlier, were fixed in glutaraldehyde and postfixed in osmium tetroxide for electron microscopy. In well over 90% of the mature sieve elements examined the P-protein was entirely parietal in distribution in both the lumina and sieve-plate pores. In addition to the parietal P-protein, the unoccluded sieve-plate pores were lined by narrow callose cylinders and the plasmalemma. Segments of endoplasmic reticulum also occurred along the margins of the pores.

P-protein distribution in mature sieve elements of Cucurbita maxima.

Penetration of leaves of barley,Hordeum vulgare L., by the corn leaf aphid,Rhopalosiphum maidis (Fitch), was studied with light, phase, and electron microscopes. Penetration of epidermis and mesophyll was largely intercellular, that of vascular bundles or veins largely intracellular. Like other aphids,R. maidis secretes a salivary sheath which surrounds the stylets. When mesophyll cells and parenchymatous elements of the veins were penetrated by stylets, their protoplasts were pushed to one side by intruding sheath material; hence, the protoplasts were not punctured by the stylets, although sometimes the plasmalemma of penetrated cells was ruptured by sheath material. The salivary sheaths ended more or less abruptly outside the walls of sieve elements being fed upon, the maxillary stylets projecting beyond the sheaths and into the sieve elements. Before penetrating a functional sieve element the aphid apparently flushes its stylets in order to clear them for ingestion of food. Salivary and food canals merge near the tips of the maxillary stylets to form a single canal, which ends short of the tips.

Observations on penetration of Barley leaves by the aphid Rhopalosiphum maidis (Fitch)

A steady-state model of solution flow in a tubular semipermeable membrane is developed for an arbitrary distribution of solute sources and sinks along the translocation path. It is demonstrated that the volume-flow mechanism of phloem transport depends only on the two assumptions: 1. that the plasmalemma of the sieve tube is a differentially permeable membrane, and 2. that sugars are actively secreted into and absorbed from the lumen of the sieve tube. It is shown that in the absence of a pressure gradient, there is a negligible concentration gradient over most of the translocation path. However, in the presence of a pressure gradient a small concentration gradient develops as a result of the continually changing chemical potential of water along the direction of solution flow. For Poiseuille flow the concentration gradient is approximately proportional to the mean stream velocity.

On the volume-flow mechanism of phloem transport.

The sieve-plate pores of sieve elements in leaf veins of Hordeum vulgare, fixed in glutaraldehyde with postfixation in osmium tetroxide, were lined by the plasmalemma and variable amounts of callose. All pores were filled with endoplasmic reticulum, which was continuous from cell to cell. Mature sieve elements lacked P-protein.

Walter Eschrich

Papers

Distribution and structure of the plasmodesmata in mesophyll and bundle-sheath cells of Zea mays L.

P-protein distribution in mature sieve elements of Cucurbita maxima.

Observations on penetration of Barley leaves by the aphid Rhopalosiphum maidis (Fitch)

On the volume-flow mechanism of phloem transport.

Sieve-plate pores in leaf veins of Hordeum vulgare.