Quantification of the xylem-to-phloem transfer of amino acids by use of inulin [14C]carboxylic acid as xylem transport marker

Inulin [¹⁴C] carboxylic acid and ¹⁴C.labelled amino acid (a-aminoisobutyric acid (aib) and valine) solutions were introduced into the transpiration stream through the cut stem bases of young (4-12 leaves) tomato plants. Inulin carboxylic acid (inu) was translocated exclusively by the xylem, whereas the amino acid distribution resulted from both xylem and phloem import. Comparison of the distribution of inu and aib permitted a quantitative assessment of the xylem-to-phloem transfer in the stem. Of aib, 20.6% traversed from xylem to phloem in a plant with 12 leaves. The phloem import was not evenly distributed over the leaves and varied from 0% (first five leaves) to 95% (top leaf) of the aib import per leaf. Doubling the flow rates in the xylem reduced the aib supply to 25% in the top leaf and 55% in the next leaf, which reflects a reduced xylem-to-phloem transfer

Potassium co-transport and antiport during the uptake of sucrose and glutamic acid from the xylem vessels

Perfusion experiments with excised internodes of tomato (Lycopersicon esculentum cv Moneymaker) showed that the uptake of glutamic acid and sucrose from the xylem vessels is accompanied with coupled proton co-transport and potassium antiport at low pH (<5.5). At high pH (5.5) both proton and potassium co-transport accompany the uptake. The results fit into the proton pump concept

Light-stimulated biphasic amino acid uptake by xylem parenchyma cells

The secondary xylem of tomato (Lycopersion esculentum cv Money-maker) internodes was isolated and was shaken in solutions of 14C-labelled amino acids. The uptake showed biphasic Michaelis-Menten kinetics. Light stimulated both uptake systems. During the uptake of amino acids the pH of the medium rose, more in the light than in the dark. Addition of 30 mmol 1−1 K+ to the medium retarded the pH rise. High K+ concentrations (>5 mmol l−1) decreased the uptake, whereas low K+ concentrations (< 2 mmol l−1) stimulated it. The observations indicated amino acid/proton co-transport into xylem parenchyma cells

Molecular sabotage of plant defense by aphid saliva

Aphids, which constitute one of the most important groups of agricultural pests, ingest nutrients from sieve tubes, the photoassimilate transport conduits in plants. Aphids are able to successfully puncture sieve tubes with their piercing mouthparts (stylets) and ingest phloem sap without eliciting the sieve tubes' normal occlusion response to injury. Occlusion mechanisms are calcium-triggered and may be prevented by chemical constituents in aphid saliva injected into sieve tubes before and during feeding. We recorded aphid feeding behavior with the electrical penetration graph (EPG) technique and then experimentally induced sieve tube plugging. Initiation of sieve tube occlusion caused a change in aphid behavior from phloem sap ingestion to secretion of watery saliva. Direct proof of "unplugging" properties of aphid saliva was provided by the effect of aphid saliva on forisomes. Forisomes are proteinaceous inclusions in sieve tubes of legumes that show calcium-regulated changes in conformation between a contracted state (below calcium threshold) that does not occlude the sieve tubes and a dispersed state (above calcium threshold) that occludes the sieve tubes. We demonstrated in vitro that aphid saliva induces dispersed forisomes to revert back to the nonplugging contracted state. Labeling Western-blotted saliva proteins with 45Ca2+ or ruthenium red inferred the presence of calcium-binding domains. These results demonstrate that aphid saliva has the ability to prevent sieve tube plugging by molecular interactions between salivary proteins and calcium. This provides aphids with access to a continuous flow of phloem sap and is a critical adaptation instrumental in the evolutionary success of aphids

Aphid watery saliva counteracts sieve-tube occlusion: a universal phenomenon?

Ca2+-binding proteins in the watery saliva of Megoura viciae counteract Ca2+-dependent occlusion of sieve plates in Vicia faba and so prevent the shut-down of food supply in response to stylet penetration. The question arises whether this interaction between aphid saliva and sieve-element proteins is a universal phenomenon as inferred by the coincidence between sieve-tube occlusion and salivation. For this purpose, leaf tips were burnt in a number of plant species from four different families to induce remote sieve-plate occlusion. Resultant sieve-plate occlusion in these plant species was counteracted by an abrupt switch of aphid behaviour. Each of the seven aphid species tested interrupted its feeding behaviour and started secreting watery saliva. The protein composition of watery saliva appeared strikingly different between aphid species with less than 50% overlap. Secretion of watery saliva seems to be a universal means to suppress sieve-plate occlusion, although the protein composition of watery saliva seems to diverge between species

Aphid watery saliva counteracts sieve-tube occlusion: a universal phenomenon?

Ca2+-binding proteins in the watery saliva of Megoura viciae counteract Ca2+-dependent occlusion of sieve plates in Vicia faba and so prevent the shut-down of food supply in response to stylet penetration. The question arises whether this interaction between aphid saliva and sieve-element proteins is a universal phenomenon as inferred by the coincidence between sieve-tube occlusion and salivation. For this purpose, leaf tips were burnt in a number of plant species from four different families to induce remote sieve-plate occlusion. Resultant sieve-plate occlusion in these plant species was counteracted by an abrupt switch of aphid behaviour. Each of the seven aphid species tested interrupted its feeding behaviour and started secreting watery saliva. The protein composition of watery saliva appeared strikingly different between aphid species with less than 50% overlap. Secretion of watery saliva seems to be a universal means to suppress sieve-plate occlusion, although the protein composition of watery saliva seems to diverge between species

Rapid cooling triggers forisome dispersion just before phloem transport stops

Phloem transport stops transiently within dicot stems that are cooled rapidly, but the cause remains unknown. Now it is known that (1) rapid cooling depolarizes cell membranes giving a transient increase in cytoplasmic Ca(2+), and (2) a rise of free calcium triggers dispersion of forisomes, which then occlude sieve elements (SEs) of fabacean plants. Therefore, we compared the effects of rapid chilling on SE electrophysiology, phloem transport and forisomes in Vicia faba. Forisomes dispersed after rapid cooling with a delay that was longer for slower cooling rates. Phloem transport stopped about 20 s after forisome dispersion, and then transport resumed and forisomes re-condensed within similar time frames. Transport interruption and forisome dispersion showed parallel behaviour--a cooling rate-dependent response, transience and desensitization. Chilling induced both a fast and a slow depolarization of SE membranes, the electrical signature suggesting strongly that the cause of forisome dispersion was the transient promotion of SE free calcium. This apparent block of SEs by dispersed forisomes may be assisted by other Ca(2+)-dependent sealing proteins that are present in all dicots