Relationships between feeding of Philaenus spumarius (L.) and the amino acid concentration in the xylem sap

Summary. 1. P.spwnarius nymphs selected young, upper leaves of Xanthium strumarium plants, which have a relatively high amino acid concentration in the xylem sap.
2. Nymphs selected or rejected a host leaf as a feeding site after a test ingestion of plant sap.
3. Nymphs fed on detached leaves kept with their petioles in a solution of amino acids, in preference to leaves with petioles in a solution containing no amino acids.
4. Nymphs caged on leaves with relatively high amino acid concentrations in the xylem sap suffered a lower mortality after 7 days than nymphs caged on leaves with low amino acid concentrations in the xylem sap.
5. Mean excretory rate increased with the increase in the amino acid concentration in the xylem sap for third and fourth instar nymphs, and adults, but not in fifth instars.     

Seasonal variation of amino acids in the xylem sap of “Populus x canadensis” and its relation to protein body mobilization

Summary The seasonal changes in the pattern of 21 amino acids occurring in the xylem vessels of Populus twigs have been studied in connection to the mobilization of protein bodies in ray parenchyma cells at the electron microscopic level. Hydrolysis of protein bodies in spring and movement of amino acids into vessels are found to be closely linked. Comprising more than 75% of total amino acid content, glutamine (Gln) is by far the dominant N-constituent of the sap. Gln reaches up to 11 mol ml^-1 in the spring sap while other amino acids only show 1/20 to 1/100 of this amount. From the measured Gln accumulation rates in the vessels in nature and in the vessels of isolated shoots, a minimum flux rate for Gln of 5.6 pmol cm^-2 min^-1 is calculated for the ray contact cell/vessel interface. Furthermore, because Gln constitutes 75% of the amino acid content of the sap but only 1.3% of the amino acids in the 32 kDa storage protein of the ray cells in the wood (Clausen and Apel 1991), it becomes evident that most amino acids originating from protein body mobilization do not enter the vessels but are used for Gln synthesis preceding Gln release into the vessels.     

Induction of xylem sap methylglycine by a drought and rewatering treatment and its inhibitory effects on the growth and development of plant organs

In higher plants, the xylem vessels functionally connect the roots with the above-ground organs. The xylem sap transports various organic compounds, such as proteins and amino acids. We examined drought and rewatering-inducible changes in the amino acid composition of root xylem sap collected from Cucurbita maxima roots. The major free amino acids in C . maxima root xylem sap were methylglycine (MeGly; sarcosine) and glutamine (Gln), but MeGly was not detected in the xylem sap of cucumber. MeGly is an intermediate compound in the metabolism of trimethylglycine (TMG; betaine), but its physiological effects in plants are unknown. Drought and rewatering treatment resulted in an increase in the concentration of MeGly in root xylem sap to 2.5 m M . After flowering, the MeGly concentration in the xylem sap dropped significantly, whereas the concentration of Gln decreased only after fruit ripening. One milli molar MeGly inhibited the formation of adventitious roots and their elongation in C . maxima , but glycine, dimethylglycine, or TMG had no effect. Similar effects and the inhibition of stem elongation were observed in shoot cuttings of cucumber and Phaseolus angularis . These observations seem to imply a possible involvement of xylem sap MeGly in the physiological responses of C . maxima plants to drought stress.     

Copper uptake and translocation in chicory (Cichorium intybus L. cv Grasslands Puna) and tomato (Lycopersicon esculentum Mill. cv Rondy) plants grown in NFT system. II. The role of nicotianamine and histidine in xylem sap copper transport

The effect of rooting media Cu concentration (0.05–20 mg Cu L^-1) on amino acid concentrations and copper speciation in the xylem sap of chicory and tomato plants was measured using 6 week old plants grown in a nutrient film technique system (NFT). Irrespective of the Cu concentration in the nutrient solutions, more than 99.68% and 99.74% of total Cu in tomato and chicory xylem sap was in a bound form. When exposed to high Cu concentrations in the rooting media, amino acid concentrations in the sap increased. Relative to other amino acids, the concentrations of glutamine (Gln), histidine (His), asparagine (Asn), valine (Val), nicotianamine (NA) and proline (Pro) in tomato xylem saps, and His, γ-aminobutyric acid (Gaba), glutamic acid (Glu), leucine (Leu), NA and phenylalanine (Phe) in chicory xylem saps showed the greatest increases. The data indicate that induced synthesis of some free amino acids as a specific and proportional response to Cu treatment. For a single complexation amino acid, the solution Cu²⁺concentration vs pH titration curve for NA at 0.06–0.07 mM was most similar, closely followed by His at 0.5–0.6 mM, to the solution Cu²⁺concentration behaviour in both tomato and chicory xylem sap. It is concluded that increased Cu concentrations in the rooting media induced selective synthesis of certain amino acid which include NA, His, Asn and Gln which have high stability constants with Cu. NA and His have the highest binding constants for Cu and the concentrations of NA and His in chicory and tomato xylem saps can account for all the bound Cu carried in the sap. This revised version was published online in June 2006 with corrections to the Cover Date.     

Apoplastic expression of yeast-derived invertase in potato : effects on photosynthesis, leaf solute composition, water relations, and tuber composition

In potato plants (Solanum tuberosum), a chimeric yeast-derived invertase gene fused to a 35S cauliflower mosaic virus promoter has been expressed. The protein was targeted to the cell wall by using the signal peptide of proteinase inhibitor II fused to the amino terminus of the yeast invertase. The transformed plants had crinkled leaves, showed a reduced growth rate, and produced fewer tubers. Although in the apoplast of the leaves of the transformed plants the content of glucose and fructose rose by a factor of 20, and that of sucrose declined 20-fold, 98% of the carbohydrate in the phloem sap consisted of sucrose, demonstrating the strong specificity of phloem loading. In the leaf cells of the transformed plants, glucose, fructose, and amino acids, especially proline, were accumulated. Consequently, the osmolality of the cell sap rose from 250 to 350 mosmol/kg. Our results show that the observed 75% decrease of photosynthesis is not caused by a feedback regulation of sucrose synthesis and is accompanied by an increase in the osmotic pressure in the leaf cells. In the transformed plants, not only the amino acid to sucrose ratio in the phloem sap, but also the amino acid and protein contents in the tubers were found to be elevated. In the tubers of the transformed plants, the protein to starch ratio increased.     

Composition of Bleeding Sap in Vigna radiata

Bleeding sap and nodules from Vigna radiata were analysed for their free amino nitrogen content and amino acid composition at different stages of growth and development. The bleeding sap contained mostly basic amino acids, whereas the nodules contained both acidic and basic amino acids. The amino nitrogen content of the bleeding sap increased during growth and then declined appreciably during fruit development. In contrast, nodule amino nitrogen declined from seedling stage onwards till flowering, increased during fruit development and then declined again. Nitrate reductase activity in the leaves examined at different stages of development increased from seedling stage onwards and was maximum during early fruit-development stage. It declined during pod-filling stage. The study suggests that the amount of nitrogen fixed from the atmosphere is insufficient, so that the plant has to draw upon soil nitrogen as well. This may be necessary due to the high demand of nitrogen during pod filling.     

Plant nitrogen status rapidly alters amino acid metabolism and excretion in Bemisia tabaci

The effect of plant nitrogen (N) status on the content and distribution of free amino acids in the bodies and honeydew of silverleaf whiteflies Bemisia tabaci (Gennadius) Biotype B (= B. argentifolii Bellows and Perring) was determined. Whiteflies fed for 4 days on cotton leaves that received high or low N fertility. For low-N plants, photosynthesis and leaf total N levels were decreased, and a much-reduced amount of free amino acids was recovered in phloem sap. Low N fertility did not affect whitefly total N content, but did markedly decrease the free amino acid content. Glutamine, alanine and proline accounted for over half of the insect free amino acid pool for both N treatments. On a relative basis, adjustments in glutamine levels in response to plant N status were much larger compared to the other amino acids. Large amounts of amino N, especially asparagine, were excreted from whiteflies fed on high-N plants whereas amino N excretion essentially ceased for whiteflies fed on low N plants. The distribution of amino acids in the insects and honeydew was not closely related to the phloem sap amino acids. However, total amino acid excretion was quite indicative of the plant N status and the quality of the insect diet. The results indicated that whitefly free amino acid pools and excretion of amino N were rapidly altered by plant N status.     

The plant-specific VIMAG domain of Glutamine Dumper1 is necessary for the function of the protein in Arabidopsis

The over-expression of the arabidopsis GLUTAMINE DUMPER1 gene (GDU1) leads to increased amino acid content and transport. In a screening for mutations suppressing this phenotype, a mutant was isolated. The mutation leads to a glycine to arginine substitution in one of the two conserved domains of the protein, the VIMAG domain. More detailed structure function relationship analyses showed that the presence of this domain and the membrane localisation are both necessary for the function of the GDU1 protein. These results shed light on the function of the GDU1 protein whose family is specific to plants.     

Seasonal changes in the free amino acids of oat and barley phloem sap in relation to plant growth stage and growth of Rhopalosiphum padi

The concentrations and composition of free amino acids in phloem sap from two cultivars of oats and barley, both susceptible to the aphid Rhopalosiphum padi, were determined by means of high performance liquid chromatography. Sap was collected from excised aphid stylets at three developmental stages (seedlings, tillering plants and plants undergoing stem elongation) from plants given or not given fertiliser and grown outdoors. In connection, the growth of individual R. padi nymphs was estimated at the same phenological stages on plants grown in the greenhouse. The content of free amino acids was consistently higher in seedlings than in plants at the early tillering stage. Only in seedlings did the addition of fertiliser increase amino acid levels. Barley phloem sap contained more free amino acids than that of oats when fertiliser was added and at later developmental stages. Phloem sap of oats and barley showed similar patterns in their composition of free amino acids at the seedling stage, but as the plants grew older the patterns became increasingly different. Plants given fertiliser had higher amounts of dicarboxylic amino acids (glutamic and aspartic acid) than unfertilised plants. The concentrations of γ-amino butyric acid, glycine, histidine, and methionine were very low in all treatments. The relative growth rates of R. padi nymphs were low when amino acid content was low and vice versa. The results are discussed in relation to host plant suitability and plant resistance mechanisms.     

Changes in the amide and amino acid composition of xylem exudate from perennial ryegrass (Lolium perenne L.) during regrowth after defoliation

The paper presents the results of amino acid analyses in xylem sap during leaf regrowth of ryegrass plants defoliated firstly at the 8th and secondly at the 12th week of culture. The free amino acid composition of leaves, stubble and roots was also determined and some of the results are reported. Prior to defoliation, xylem sap contained a high proportion of amides, particularly glutamine. During regrowth after defoliation, the proportion of asparagine in the xylem sap increased until the third day when the highest ratios of asparagine/glutamine appeared. The results are compared with relative amounts of free amino acids in the different plant parts and discussed in relation to source-sink nitrogen transfer.     

Variation in Eastern cottonwood (Populus deltoides Bartr.) phloem sap content caused by leaf development may affect feeding site selection behavior of the aphid, Chaitophorous populicola Thomas (Homoptera: Aphididae)

Apterous populations of Chaitophorous populicola Thomas (Homoptera: Aphididae) appear to track Eastern cottonwood (Populus deltoides Bartr.) leaf development. Few aphids occur on mature leaves. Marked individual aphids on leaves of different developmental stages were observed through a period of new leaf initiation. Nymph and adult C. populicola frequently track leaf development by moving up to younger leaves. A comparison of phloem sap constituents and leaf toughness among leaf developmental stages revealed some differences that could be used by C. populicola to determine leaf age. Phloem sap exudates, collected from P. deltoides leaves of different developmental stages, were analyzed by high-performance liquid chromatography for free amino acids and the phenolic glycoside salicin. Sucrose concentration in exudates, indicative of phloem sap exudation rate, was uniform among leaf stages. Of 20 amino acids examined, only aspartic acid and gamma-amino-n-butyric acid (GABA) concentrations differed significantly between leaf stages. Forward stepwise discriminant function analysis showed that seven of the amino acids analyzed are useful for classifying leaf maturity groupings. Aphid-infested cottonwoods had lower cystine concentrations in phloem sap than aphid-free plants. Salicin concentration was significantly higher in new leaves. Leaf toughness was assessed by lignin density and distance measurements in petiole cross-sections. Rapidly expanding leaves had significantly less lignification and new leaves had shorter distances to the vascular bundles than senescent leaves. These physiological and phytochemical differences among P. deltoides leaf developmental stages may contribute to the leaf stage selection patterns exhibited by the aphid, C. populicola.     

Changes in the composition of xylem sap during development of the spadix of skunk cabbage (Symplocarpus foetidus)

The spadix of skunk cabbage, Symplocarpus foetidus, is thermogenic and maintains an internal temperature of around 20 degrees C even when the ambient air temperature drops below freezing. This homeothermic heat production is observed only during the stigma stage, and thereafter ceases at the male stage when pollen is shed. To clarify the regulatory mechanism by which the stigma stage-specific heat production occurs in the spadix, sugars, organic acids, and amino acids in xylem sap were analyzed and compared with those of post-thermogenic plants. Interestingly, no significant difference was observed in the total volume of xylem sap per fresh weight of the spadix between thermogenic (31.2+/-24.7 microl h(-1) g(-1)) and post-thermogenic (50.5+/-30.4 microl h(-1) g(-1)) plants. However, concentrations of sugars (sucrose, glucose, and fructose), organic acids (malate and succinate), and amino acids (Asp, Asn, Glu, Gln, Gly, and Ala) in xylem sap decreased remarkably in post-thermogenic plants. Our results indicate that the composition of the xylem sap differs during the development of the spadix of S. foetidus.     

N content of phloem and xylem exudates during the transition to flowering in Sinapis alba and Arabidopsis thaliana

The involvement of nitrogenous substances in the transition to flowering was investigated in Sinapis alba and Arabidopsis thaliana (Columbia). Both species grown in short days (SD) are induced to flower by one long day (LD). In S. alba, the phloem sap (leaf and apical exudates) and the xylem sap (root exudate) were analysed in LD versus SD. In A. thaliana, only the leaf exudate could be analysed but an alternative system for inducing flowering without day‐length extension was used: the displaced SD (DSD). Significant results are: (i) in both species, the leaf exudate was enriched in Gln during the inductive LD, at a time compatible with export of the floral stimulus; (ii) in S. alba, the root export of amino acids decreased in LD, whereas the nitrate remained unchanged – thus the extra‐Gln found in the leaf exudate should originate from the leaves; (iii) extra‐Gln was also found very early in the apical exudate of S. alba in LD, together with more Glu; (iv) in A. thaliana induced by one DSD, the leaf export of Asn increased sharply, instead of Gln in LD. This agrees with Asn prevalence in C‐limited plants. The putative role of amino acids in the transition to flowering is discussed.     

Dependence of amino acid composition upon nitrogen availability in birch (Betula pendula)

Small birch plants ( Betula pendula Roth .) were grown at different rates of exponentially increasing nitrogen supply. This resulted in plants with different relative growth rates and different internal nitrogen concentrations. Within a nitrogen treatment, both of these variables remained constant with time.
Free amino acids were measured in leaves and roots of the seedlings at two different harvests. At greater nitrogen supply, higher concentrations of total amino acid nitrogen were found in roots and leaves. The ratio of amino acid nitrogen to total nitrogen was low albeit greater at higher nitrogen supply. Higher concentrations of amino acid nitrogen were mainly due to high concentrations of citrulline, glutamine, γ-aminobuitric acid and arginine.
Greater leaf concentrations of amino acid nitrogen at higher nitrogen supply may be related lo increased concentrations in the xylem sap and/or may be indicative of small excesses of nitrogen with respect to current nitrogen usage in protein synthesis.     

Metabolomic analysis reveals the potential metabolites and pathogenesis involved in mulberry yellow dwarf disease

To analyse the molecular mechanisms of phytoplasma pathogenicity, the comprehensive metabolomic changes of mulberry leaf and phloem sap in response to phytoplasma infection were examined using gas chromatography‐mass spectrometry. The metabolic profiles obtained revealed that the metabolite compositions of leaf and phloem sap were different, and phytoplasma infection has a greater impact on the metabolome of phloem sap than of leaf. Phytoplasma infection brought about the content changes in various metabolites, such as carbohydrates, amino acids, organic acids, etc. Meanwhile, the results of biochemical analysis showed that the degradation of starch was repressed, and the starch content was increased in the infected leaves. In addition, we found that phytoplasma infection changed the levels of abscisic acid and cytokinin and break phytohormone balance. Interestingly, our data showed that the contents of H₂O₂ and superoxide were increased in the infected leaves, but not in the phloem saps. Based on the results, the expression levels of the genes involved in the metabolism of some changed metabolites were examined, and the potential molecular mechanisms of these changes were discussed. It can be concluded that both the leaf and phloem saps have a complicated metabolic response to phytoplasma infection, but their response mechanisms were different.     

Magnesium deficiency results in accumulation of carbohydrates and amino acids in source and sink leaves of spinach

Accumulation of assimilates in source leaves of magnesium‐deficient plants is a well‐known feature. We had wished to determine whether metabolite concentrations in sink leaves and roots are affected by magnesium nutrition. Eight‐week‐old spinach plants were supplied either with a complete nutrient solution (control plants) or with one lacking Mg (deficient plants) for 12 days. Shoot and root fresh weights and dry weights were lower in deficient than in control plants. Mg concentrations in deficient plants were 11% of controls in source leaves, 12% in sink leaves and 26% in roots, respectively. As compared with controls, increases were found in starch and amino acids in source leaves and in sucrose, hexoses, starch and amino acids in sink leaves, whereas they were only slightly enhanced in roots. In phloem sap of magnesium‐deficient and control plants no differences in sucrose and amino acid concentrations were found. To prove that sink leaves were the importing organs they were shaded, which did not alter the response to magnesium deficiency as compared with that without shading. Since in the shaded sink leaves the photosynthetic production of metabolites could be excluded, those carbohydrates and amino acids that accumulated in the sink leaves of the deficient plants must have been imported from the source leaves. It is concluded that in magnesium‐deficient spinach plants the growth of sink leaves and roots was not limited by carbohydrate or amino acid supply. It is proposed that the accumulation of assimilates in the source leaves of Mg‐deficient plants results from a lack of utilization of assimilates in the sink leaves.     

Short-term changes in xylem N compounds in Lolium perenne following defoliation

Previous studies have indicated that an increased asparagine to glutamine ratio (Asn : Gln) occurs in the xylem fluid of Lolium perenne 24 h after defoliation. However, the absolute changes in Asn and Gln leading to the increased Asn : Gln ratio are unknown. The present study tested the hypotheses that: (1) defoliation-induced changes in xylem amino acid composition occur in L perenne within the first 24 h following defoliation, irrespective of phasing with respect to the diurnal light/dark cycle; and (2) the increase in Asn : Gln ratio in the xylem fluid of L perenne following defoliation is due to an increase in Asn content. Plants of L perenne L. 'Aurora' were grown in flowing solution culture for 40 d. Plants were then either left intact, defoliated at the end of the light period or defoliated at the end of the dark period. 15N-labelled NO3- was supplied following defoliation to discriminate between the recovery of N absorbed prior to, and following, defoliation. Xylem samples were collected over the subsequent 24 h period with amino acids speciated by GC-MS. There was support for the first hypothesis: increased Asn : Gln ratios occurred within the first 24 h, irrespective of the phasing of defoliation with respect to light/dark cycles. The second hypothesis was not supported: the concentration of all amino acids in the xylem exudate declined after defoliation, and the increased Asn : Gln ratio was accounted for by a disproportionately large reduction in Gln levels. Low concentrations of amino acids in the xylem of defoliated plants precluded accurate discrimination of their nitrogen content into pre- and post-defoliation sources.     

Assimilation of gaseous ammonia and the transport of its products in barley and spinach leaves

The interactions between the assimilation and transport of nitrogenand carbon were investigated in barley and spinach leaves. Bothplants were fumigated with NH₃ (1 mg m^–3 and the contentof amino acids, sucrose and carbon intermediates of amino acidmetabolism were analysed in the leaves, apoplast and phloemsap. The following changes took place in the C- and N-metabolismof barley leaves during 5 h of fumigation with NH₃ (a) The contentsof amino acids, especially glutamine, largely increased andthe contents of sucrose, 2-oxoglutarate, phosphoenolpyruvate,and glycerate-3-phosphate declined. (b) A decrease in the phophoenolpyruvatecontent was accompanied by an increased activity of phosphoenolpyruvatecarboxylase. (c) The altered cytosolic concentrations of aminoacids and sucrose during NH₃ fumigation correlated with similarchanges in the apoplast and phloem sap. The altered percentageof each amino acid relative to the total amino acid concentrationin the cytosol, caused by NH₃ fumigation, is reflected in theapoplast and the phloem sap. The results indicate that the concentrations of amino acids in the cytosol determine their concentrationsin the phloem. Key words: Amino acids, ammonia fumigation, barley leaves, C: N partitioning, phosphoenolpyruvate carboxylase, phloem sap, spinach leaves