Archaeobotanical evidence supports indigenous cucurbit long-term use in the Mesoamerican Neotropics.

The squash family (Cucurbitaceae) contains some of the most important crops cultivated worldwide and has played an important ecological, economic, and cultural role for millennia. In the American tropics, squashes were among the first cultivated crop species, but little is known about how their domestication unfolded. Here, we employ direct radiocarbon dating and morphological analyses of desiccated cucurbit seeds, rinds, and stems from El Gigante Rockshelter in Honduras to reconstruct human practices of selection and cultivation of Lagenaria siceraria, Cucurbita pepo, and Cucurbita moschata. Direct radiocarbon dating indicates that humans started using Lagenaria and wild Cucurbita starting ~ 10,950 calendar years before present (cal B.P.), primarily as watertight vessels and possibly as cooking and drinking containers. A rind directly dated to 11,150-10,765 cal B.P. represents the oldest known bottle gourd in the Americas. Domesticated C. moschata subsequently appeared ~ 4035 cal B.P., followed by domesticated C. pepo ~ 2190 cal B.P. associated with increasing evidence for their use as food crops. Multivariate statistical analysis of seed size and shape show that the archaeological C. pepo assemblage exhibits significant variability, representing at least three varieties: one similar to present-day zucchini, another like present-day vegetable marrow, and a native cultivar without modern analogs. Our archaeobotanical data supports the hypothesis that Indigenous cucurbit use started in the Early Holocene, and that agricultural complexity during the Late Holocene involved selective breeding that encouraged crop diversification.

So similar yet so different: The distinct contributions of extrafascicular and fascicular phloem to transport and exudation in cucumber plants.

Cucurbits have been used as phloem research models for many decades because their exudates can be accessed with ease. However, cucurbit plants possess two distinct phloem systems known as the fascicular phloem (FP) and extrafascicular phloem (EFP). Therefore, the molecular composition and function of certain exudates can be misinterpreted due to their unclear origin. To characterize the anatomy and function of the different phloem systems more clearly, we generated specific antibodies against marker proteins (PP1 homologs) allowing the clear identification of the EFP at the organ, tissue and cellular levels by immunological staining. We also used detailed microscopy to determine common and unique anatomical features of the FP and EFP sieve elements (SEs) in cucumber (Cucumis sativus). The comparison of exudation rates and the dynamic viscosity, density and sugar content of the exudates from plants grown in the light and dark revealed the consistent composition and behavior of the EFP exudate even when photosynthesis was prevented, thus differing from the properties of the FP exudate. Furthermore, the analysis of phloem transport using a fluorescein disodium salt showed only wound-induced exudation of dye from the EFP, indicating the absence of transport in this tissue. Our results show that it is important to distinguish between the EFP and FP in cucurbits, particularly their differing behaviors in response to wounding.

Genetic Mapping and Identification of the Candidate Gene for White Seed Coat in Cucurbita maxima.

Seed coat color is an important agronomic trait of edible seed pumpkin in Cucurbita maxima. In this study, the development pattern of seed coat was detected in yellow and white seed coat accessions Wuminglv and Agol. Genetic analysis suggested that a single recessive gene white seed coat (wsc) is involved in seed coat color regulation in Cucurbita maxima. An F2 segregating population including 2798 plants was used for fine mapping and a candidate region containing nine genes was identified. Analysis of 54 inbred accessions revealed four main Insertion/Deletion sites in the promoter of CmaCh15G005270 encoding an MYB transcription factor were co-segregated with the phenotype of seed coat color. RNA-seq analysis and qRT-PCR revealed that some genes involved in phenylpropanoid/flavonoid metabolism pathway displayed remarkable distinction in Wuminglv and Agol during the seed coat development. The flanking InDel marker S1548 was developed to predict the seed coat color in the MAS breeding with an accuracy of 100%. The results may provide valuable information for further studies in seed coat color formation and structure development in Cucurbitaceae crops and help the molecular breeding of Cucurbita maxima.

Brazilian germplasm of winter squash (Cucurbita moschata D.) displays vast genetic variability, allowing identification of promising genotypes for agro-morphological traits.

Winter squash fruits (Cucurbita moschata D.) are among the best sources of vitamin A precursors and constitute sources of bioactive components such as phenolic compounds and flavonoids. Approximately 70% of C. moschata seed oil is made up of unsaturated fatty acids, with high levels of monounsaturated fatty acids and components such as vitamin E and carotenoids, which represent a promising nutritional aspect in the production of this vegetable. C. moschata germplasm expresses high genetic variability, especially in Brazil. We assessed 91 C. moschata accessions, from different regions of Brazil, and maintained at the Federal University of Viçosa (UFV) Vegetable Germplasm Bank, to identify early-flowering accessions with high levels of carotenoids in the fruit pulp and high yields of seed and seed oil. Results showed that the accessions have high variability in the number and mass of seeds per fruit, number of accumulated degree-days for flowering, total carotenoid content, and fruit productivity, which allowed selection for considerable gains in these characteristics. Analysis of the correlation between these characteristics provided information that will assist in selection to improve this crop. Cluster analysis resulted in the formation of 16 groups, confirming the variability of the accessions. Per se analysis identified accessions BGH-6749, BGH-5639, and BGH-219 as those with the earliest flowering. Accessions BGH-5455A and BGH-5598A had the highest carotenoid content, with averages greater than 170.00 µg g-1 of fresh mass. With a productivity of 0.13 t ha-1, accessions BGH-5485A, BGH-4610A, and BGH-5472A were the most promising for seed oil production. These last two accessions corresponded to those with higher seed productivity, averaging 0.58 and 0.54 t ha-1, respectively. This study confirms the high potential of this germplasm for use in breeding for promotion of earlier flowering and increase in total content of fruit pulp carotenoids and in seed and seed oil productivity.

Complementarity in root architecture for nutrient uptake in ancient maize/bean and maize/bean/squash polycultures.

BACKGROUND AND AIMS: During their domestication, maize, bean and squash evolved in polycultures grown by small-scale farmers in the Americas. Polycultures often overyield on low-fertility soils, which are a primary production constraint in low-input agriculture. We hypothesized that root architectural differences among these crops causes niche complementarity and thereby greater nutrient acquisition than corresponding monocultures. METHODS: A functional-structural plant model, SimRoot, was used to simulate the first 40 d of growth of these crops in monoculture and polyculture and to determine the effects of root competition on nutrient uptake and biomass production of each plant on low-nitrogen, -phosphorus and -potassium soils. KEY RESULTS: Squash, the earliest domesticated crop, was most sensitive to low soil fertility, while bean, the most recently domesticated crop, was least sensitive to low soil fertility. Nitrate uptake and biomass production were up to 7 % greater in the polycultures than in the monocultures, but only when root architecture was taken into account. Enhanced nitrogen capture in polycultures was independent of nitrogen fixation by bean. Root competition had negligible effects on phosphorus or potassium uptake or biomass production. CONCLUSIONS: We conclude that spatial niche differentiation caused by differences in root architecture allows polycultures to overyield when plants are competing for mobile soil resources. However, direct competition for immobile resources might be negligible in agricultural systems. Interspecies root spacing may also be too large to allow maize to benefit from root exudates of bean or squash. Above-ground competition for light, however, may have strong feedbacks on root foraging for immobile nutrients, which may increase cereal growth more than it will decrease the growth of the other crops. We note that the order of domestication of crops correlates with increasing nutrient efficiency, rather than production potential.

High-throughput SNP genotyping in Cucurbita pepo for map construction and quantitative trait loci mapping.

BACKGROUND: Cucurbita pepo is a member of the Cucurbitaceae family, the second- most important horticultural family in terms of economic importance after Solanaceae. The "summer squash" types, including Zucchini and Scallop, rank among the highest-valued vegetables worldwide. There are few genomic tools available for this species.The first Cucurbita transcriptome, along with a large collection of Single Nucleotide Polymorphisms (SNP), was recently generated using massive sequencing. A set of 384 SNP was selected to generate an Illumina GoldenGate assay in order to construct the first SNP-based genetic map of Cucurbita and map quantitative trait loci (QTL). RESULTS: We herein present the construction of the first SNP-based genetic map of Cucurbita pepo using a population derived from the cross of two varieties with contrasting phenotypes, representing the main cultivar groups of the species' two subspecies: Zucchini (subsp. pepo) × Scallop (subsp. ovifera). The mapping population was genotyped with 384 SNP, a set of selected EST-SNP identified in silico after massive sequencing of the transcriptomes of both parents, using the Illumina GoldenGate platform. The global success rate of the assay was higher than 85%. In total, 304 SNP were mapped, along with 11 SSR from a previous map, giving a map density of 5.56 cM/marker. This map was used to infer syntenic relationships between C. pepo and cucumber and to successfully map QTL that control plant, flowering and fruit traits that are of benefit to squash breeding. The QTL effects were validated in backcross populations. CONCLUSION: Our results show that massive sequencing in different genotypes is an excellent tool for SNP discovery, and that the Illumina GoldenGate platform can be successfully applied to constructing genetic maps and performing QTL analysis in Cucurbita. This is the first SNP-based genetic map in the Cucurbita genus and is an invaluable new tool for biological research, especially considering that most of these markers are located in the coding regions of genes involved in different physiological processes. The platform will also be useful for future mapping and diversity studies, and will be essential in order to accelerate the process of breeding new and better-adapted squash varieties.

A statistical model for mapping morphological shape.

BACKGROUND: Living things come in all shapes and sizes, from bacteria, plants, and animals to humans. Knowledge about the genetic mechanisms for biological shape has far-reaching implications for a range spectrum of scientific disciplines including anthropology, agriculture, developmental biology, evolution and biomedicine. RESULTS: We derived a statistical model for mapping specific genes or quantitative trait loci (QTLs) that control morphological shape. The model was formulated within the mixture framework, in which different types of shape are thought to result from genotypic discrepancies at a QTL. The EM algorithm was implemented to estimate QTL genotype-specific shapes based on a shape correspondence analysis. Computer simulation was used to investigate the statistical property of the model. CONCLUSION: By identifying specific QTLs for morphological shape, the model developed will help to ask, disseminate and address many major integrative biological and genetic questions and challenges in the genetic control of biological shape and function.

Divergent metabolome and proteome suggest functional independence of dual phloem transport systems in cucurbits.

Cucurbitaceous plants (cucurbits) have long been preferred models for studying phloem physiology. However, these species are unusual in that they possess two different phloem systems, one within the main vascular bundles [fascicular phloem (FP)] and another peripheral to the vascular bundles and scattered through stem and petiole cortex tissues [extrafascicular phloem (EFP)]. We have revisited the assumption that the sap released after shoot incision originates from the FP, and also investigated the long-standing question of why the sugar content of this sap is ~30-fold less than predicted for requirements of photosynthate delivery. Video microscopy and phloem labeling experiments unexpectedly reveal that FP very quickly becomes blocked upon cutting, whereas the extrafascicular phloem bleeds for extended periods. Thus, all cucurbit phloem sap studies to date have reported metabolite, protein, and RNA composition and transport in the relatively minor extrafascicular sieve tubes. Using tissue dissection and direct sampling of sieve tube contents, we show that FP in fact does contain up to 1 M sugars, in contrast to low-millimolar levels in the EFP. Moreover, major phloem proteins in sieve tubes of FP differ from those that predominate in the extrafascicular sap, and include several previously uncharacterized proteins with little or no homology to databases. The overall compositional differences of the two phloem systems strongly indicate functional isolation. On this basis, we propose that the fascicular phloem is largely responsible for sugar transport, whereas the extrafascicular phloem may function in signaling, defense, and transport of other metabolites.

[Effects of exogenous Ca2+ on morphological and photosynthetic characteristics and chlorophyll fluorescent parameters of squash seedlings under high temperature and strong light stress].

Taking squash (Cucurbita pepo L.) variety Alan as test object, this paper studied the effects of exogenous Ca2+ on the morphological and photosynthetic characteristics and chlorophyll fluorescent parameters of squash seedlings under the cross-stress of high temperature and strong light. Under the stress, applying 5-20 mmol x L(-1) of Ca2+ increased the plant height, leaf area, chlorophyll and carotenoid contents, photosynthetic rate (Pn), stoma conductance (Gs), transpiration rate (Tr), maximal PS II efficiency (Fv/Fm), actual PS II efficiency (phi(PS II)), and photochemical queching coefficient (q(P)), and decreased the intercellular CO2 concentration (Ci) and non-photochemical fluorescence quenching coefficient (NPQ), suggesting that this application of exogenous Ca2+ could effectively mitigate the damage of high temperature and strong light stress on the squash seedlings leaf, and make it keep more rapid photosynthetic electron transfer rate and higher PS II electron transfer activity. Among the treatments of applying Ca2+, 10 mmol Ca2+ x L(-1) had the best effect. When the Ca2+ application rate exceeded 40 mmol x L(1), no mitigation effect was observed on the high temperature and strong light stress.

Cadmium induced changes in subcellular glutathione contents within glandular trichomes of Cucurbita pepo L.

Plants cope with cadmium (Cd) stress by complexation with phytochelatins (Pc), metallothioneins and glutathione and sequestration within vacuoles. Especially glutathione was found to play a major role in Cd detoxification as Cd shows a high binding affinity towards thiols and as glutathione is a precursor for Pc synthesis. In the present study, we have used an immunohistochemical approach combined with computer-supported transmission electron microscopy in order to measure changes in the subcellular distribution of glutathione during Cd-stress in mesophyll cells and cells of different glandular trichomes (long and short stalked) of Cucurbita pepo L. subsp. pepo var. styriaca GREB: . Even though no ultrastructural alterations were observed in leaf and glandular trichome cells after the treatment of plants with 50 microM cadmium chloride (CdCl(2)) for 48 h, all cells showed a large decrease in glutathione contents. The strongest decrease was found in nuclei and the cytosol (up to 76%) in glandular trichomes which are considered as a major side of Cd accumulation in leaves. The ratio of glutathione between the cytosol and nuclei and the other cell compartments was strongly decreased only in glandular trichomes (more than 50%) indicating that glutathione in these two cell compartments is especially important for the detoxification of Cd in glandular trichomes. Additionally, these data indicate that large amounts of Cd are withdrawn from nuclei during Cd exposure. The present study gives a detailed insight into the compartment-specific importance of glutathione during Cd exposure in mesophyll cells and glandular trichomes of C. pepo L. plants.

Assay-dependent phytotoxicity of nanoparticles to plants.

The effects of five nanomaterials (multiwalled carbon nanotubes [MWCNTs], Ag, Cu, ZnO, Si) and their corresponding bulk counterparts on seed germination, root elongation, and biomass of Cucurbita pepo (zucchini) were investigated. The plants were grown in hydroponic solutions amended with nanoparticles or bulk material suspensions at 1000 mg/L. Seed germination was unaffected by any of the treatments, but Cu nanoparticles reduced emerging root length by 77% and 64% relative to unamended controls and seeds exposed to bulk Cu powder, respectively. During a 15-day hydroponic trial, the biomass of plants exposed to MWCNTs and Ag nanoparticles was reduced by 60% and 75%, respectively, as compared to control plants and corresponding bulk carbon and Ag powder solutions. Although bulk Cu powder reduced biomass by 69%, Cu nanoparticle exposure resulted in 90% reduction relative to control plants. Both Ag and Cu ion controls (1-1000 mg/L) and supernatant from centrifuged nanoparticle solutions (1000 mg/L) indicate that half the observed phytotoxicity is from the elemental nanoparticles themselves. The biomass and transpiration volume of zucchini exposed to Ag nanoparticles or bulk powder at 0-1000 mg/mL for 17 days was measured. Exposure to Ag nanoparticles at 500 and 100 mg/L resulted in 57% and 41% decreases in plant biomass and transpiration, respectively, as compared to controls or to plants exposed to bulk Ag. On average, zucchini shoots exposed to Ag nanoparticles contained 4.7 greater Ag concentration than did the plants from the corresponding bulk solutions. These findings demonstrate that standard phytotoxicity tests such as germination and root elongation may not be sensitive enough or appropriate when evaluating nanoparticle toxicity to terrestrial plant species.

The effects of planting methods and head pruning on seed yield and yield components of medicinal pumpkin (Cucurbita pepo subsp. Pepo convar. Pepo var. styriaca) at low temperature areas.

This experiment carried out to evaluate the effects of planting methods (seed sowing and transplanting) and head pruning (no pruning, pruning after 12th node and pruning after 16th node) on yield and yield components such as number of branches (sub-branches) per plant, fruits per plant, growth, fruit size, weight of fresh fruit, weight of seeds per fruit, number of seeds per fruit and seed yield of medicinal pumpkin. The experiment was carried out based of factorial experiment with Randomized Completely Blocks Design (RCBD) by three replications in Ardabil Agricultural and Natural Resources Researches Station at 2007. Seedlings were grown in heated greenhouse. When the climatic condition became suitable and seedlings were at the four leaves stage, both seeds and seedlings were planted at the same time in the farm. Maintenance operations were done during the growth season. Head pruning treatments were done the forecast time. The results showed that the planting methods had significant effect on the number of ripen fruits per plant, fruits diameter, weight of seeds per fruit, weight of 1000 seeds and seed yield and had no significant effect on the other traits. Also the results indicated that head pruning treatments had significant effects on the number of branches per plant, growth and seed yield and no significant on the other traits. In this experiment the most seed yield (997.8 kg ha(-1)) obtained from transplanting method with head pruning after 12th node and the least seed yield obtained from control.

Analysis of the pumpkin phloem proteome provides insights into angiosperm sieve tube function.

Increasing evidence suggests that proteins present in the angiosperm sieve tube system play an important role in the long distance signaling system of plants. To identify the nature of these putatively non-cell-autonomous proteins, we adopted a large scale proteomics approach to analyze pumpkin phloem exudates. Phloem proteins were fractionated by fast protein liquid chromatography using both anion and cation exchange columns and then either in-solution or in-gel digested following further separation by SDS-PAGE. A total of 345 LC-MS/MS data sets were analyzed using a combination of Mascot and X!Tandem against the NCBI non-redundant green plant database and an extensive Cucurbit maxima expressed sequence tag database. In this analysis, 1,209 different consensi were obtained of which 1,121 could be annotated from GenBank and BLAST search analyses against three plant species, Arabidopsis thaliana, rice (Oryza sativa), and poplar (Populus trichocarpa). Gene ontology (GO) enrichment analyses identified sets of phloem proteins that function in RNA binding, mRNA translation, ubiquitin-mediated proteolysis, and macromolecular and vesicle trafficking. Our findings indicate that protein synthesis and turnover, processes that were thought to be absent in enucleate sieve elements, likely occur within the angiosperm phloem translocation stream. In addition, our GO analysis identified a set of phloem proteins that are associated with the GO term "embryonic development ending in seed dormancy"; this finding raises the intriguing question as to whether the phloem may exert some level of control over seed development. The universal significance of the phloem proteome was highlighted by conservation of the phloem proteome in species as diverse as monocots (rice), eudicots (Arabidopsis and pumpkin), and trees (poplar). These results are discussed from the perspective of the role played by the phloem proteome as an integral component of the whole plant communication system.

The effect of stem pruning and nitrogen levels of on some physico-chemical characteristics of pumpkin seed (Cucurbita pepo L.).

To investigate the effects of stem pruning (No heading, head pruning of stem after formation of 10 and 14 nodes) and nitrogen levels (0, 50, 100 and 200 kg ha(-1)) on physical and chemical characteristic of pumpkin seed a Factorial experiment based on randomized complete block design with three replication was carried out in Gorgan at 2003 and repeated in 2004 years. Results showed that the stem pruning has significant effect on traits such as seed oil, linoleic acid and oleic acid content. Nitrogen levels also have significant effect on seed dimension, seed oil, linoleic acid and oleic acid content. The largest amount of oil and linoleic acid content was obtained by stem pruning after forming 14 node and 100 kg ha(-1) nitrogen in separately, but the interaction of treatments were not significant difference for all of traits.

First known image of Cucurbita in Europe, 1503-1508.

BACKGROUND: The genus Cucurbita (pumpkin, squash, gourd) is native to the Americas and diffused to other continents subsequent to the European contact in 1492. For many years, the earliest images of this genus in Europe that were known to cucurbit specialists were the two illustrations of C. pepo pumpkins that were published in Fuchs' De Historia Stirpium, 1542. Images of fruits of two Cucurbita species, drawn between 1515 and 1518, were recently discovered in the Villa Farnesina in Rome. FINDINGS: An even earlier image of Cucurbita exists in the prayer book, Grandes Heures d'Anne de Bretagne, illustrated by Jean Bourdichon in Touraine, France, between 1503 and 1508. This image, which shows a living branch bearing flowers and fruits, had not been examined and analysed by cucurbit specialists until now. The image is identified as depicting Cucurbita pepo subsp. texana. Unlike some of the fruits of Cucurbita depicted in the Villa Farnesina a decade later, this image does not depict an esculent and does not constitute evidence of early European contact with New World agriculture. Based on the descriptive, ecological and geographical accounts of C. pepo subsp. texana in the wild, the idea is considered that the image was based on an offspring of a plant found growing along the Gulf Coast of what is now the United States.

Cucurbit phloem serpins are graft-transmissible and appear to be resistant to turnover in the sieve element-companion cell complex.

Serpins are unique inhibitors of serine proteases that are located in various plant tissues and organs. An orthologue of the pumpkin (Cucurbita maxima) phloem serpin CmPS-1 was amplified from cucumber (Cucumis sativus) RNA by RT-PCR, cloned, and designated as CsPS-1 (GenBank accession no. AJ866989). Alternative amino acid sequences in the reactive centre loop suggest distinct inhibitory specificity between CmPS-1 and CsPS-1. A difference in the electrophoretic mobility of these serpins was used in heterografts to establish that serpins are phloem-mobile. Immuno light microscopy revealed that the phloem serpins are localized exclusively to sieve elements (SE), while the phloem filament protein CmPP1, used as a reference, is localized to both SEs and companion cells (CCs). Similar to CmPS-1, CsPS-1 accumulates over time in phloem exudates, indicating that serpins differ from other phloem-mobile proteins whose concentrations appear to be stable in phloem exudates. These differences could reflect alternative mechanisms regulating protein turnover and/or inaccessibility of protein degradation. The functionality of the pore/plasmodesma units connecting SEs and CCs was tested with graft-transmitted CmPP1 as a transport marker. The occurrence of CmPP1 in the CCs of the Cucumis graft partner shows that translocated 88 kDa phloem filament protein monomers can symplasmically exit the SE and accumulate in the CC. By contrast, serial sections probed with the serpin antibody demonstrate that the 43 kDa serpin does not enter CCs. Collectively, these data indicate that CCs play a decisive role in homeostasis of exudate proteins; proteins not accessing the CCs accumulate in SEs and display a time-dependent increase in concentration.

Genetic diversity of a germplasm collection of Cucurbita pepo using SRAP and AFLP markers.

Cucurbita pepo is a highly polymorphic species. The cultivars can be grouped into eight morphotypes in two subspecies, ssp. pepo and ssp. ovifera. A collection of 69 accessions representative of the morphotypes and some unclassified types was used for analysing the morphological and molecular diversity of this species. This collection includes commercial cultivars and Spanish landraces, which represent the great diversification of types that have arisen in Europe after this species arrived from America. For the molecular variability studies, two PCR-based systems were employed, AFLP and SRAP, which preferentially amplify ORFs. Principal coordinates analysis and cluster analysis using the UPGMA method clearly separate the accessions into the two subspecies through the use of both markers. However, the gene diversity and the genetic identity values among morphotypes and subspecies varied between the two marker systems. The information given by SRAP markers was more concordant to the morphological variability and to the evolutionary history of the morphotypes than that of AFLP markers. In ssp. ovifera, the accessions of the different morphotypes were basically grouped according to the fruit colour. This may indicate different times of development and also the extent of breeding in the accessions used. This study has allowed identification of new types that can be employed for the development of new cultivars. The landraces of the spp. ovifera, used as ornamental in Europe, have proved to be of great interest for preserving the diversity of C. pepo.

Metabolic networks of Cucurbita maxima phloem.

Metabolomic analysis aims at a comprehensive characterization of biological samples. Yet, biologically meaningful interpretations are often limited by the poor spatial and temporal resolution of the acquired data sets. One way to remedy this is to limit the complexity of the cell types being studied. Cucurbita maxima Duch. vascular exudates provide an excellent material for metabolomics in this regard. Using automated mass spectral deconvolution, over 400 components have been detected in these exudates, but only 90 of them were tentatively identified. Many amino compounds were found in vascular exudates from leaf petioles at concentrations several orders of magnitude higher than in tissue disks from the same leaves, whereas hexoses and sucrose were found in far lower amounts. In order to find the expected impact of assimilation rates on sugar levels, total phloem composition of eight leaves from four plants was followed over 4.5 days. Surprisingly, no diurnal rhythm was found for any of the phloem metabolites that was statistically valid for all eight leaves. Instead, each leaf had its own distinct vascular exudate profile similar to leaves from the same plant, but clearly different from leaves harvested from plants at the same developmental stage. Thirty to forty per cent of all metabolite levels of individual leaves were different from the average of all metabolite profiles. Using metabolic co-regulation analysis, similarities and differences between the exudate profiles were more accurately characterized through network computation, specifically with respect to nitrogen metabolism.

Evidence for the presence and activity of a complete antioxidant defence system in mature sieve tubes.

The phloem is the major route for the transport of solutes and nutrients from source to sink organs in plants. The functional transport phloem consists of parenchymal tissue, enucleate sieve elements, and the intimately connected companion cells. The general absence of a nucleus and functional ribosomes in sieve tubes poses problems especially for damage avoidance and repair of sieve element components. To examine how sieve tubes can remain functional during oxidative stress, we analysed phloem sap of cucumber and pumpkin plants with respect to the presence of antioxidant defence enzymes, their enzymatic activity, and activity changes after exposure to drought stress. Using 1D SDS-PAGE and nano ESI MS/MS, the presence of proteins such as cytosolic Cu/Zn superoxide dismutase, monodehydroascorbate reductase, and peroxidase could be shown. Moreover, activities for several antioxidant enzymes (superoxide dismutase, dehydroascorbate reductase, peroxidase) in phloem exudate could be demonstrated. The activity of these enzymes in phloem sap from cucumber and pumpkin plants increased in response to drought stress. The presented results together with earlier findings provide evidence supporting the presence of a complete machinery of antioxidant defence enzymes and detoxifying metabolites important for avoiding damage to essential components of the sieve elements due to oxidative stress.