Costas Delis

Induction and spatial organization of polyamine biosynthesis during nodule development in Lotus japonicus

Putrescine and other polyamines are produced by two alternative pathways in plants. One pathway starts with the enzyme arginine decarboxylase (ADC; EC 4.1.1.19), the other with ornithine decarboxylase (ODC; EC 4.1.1.17). Metabolite profiling of nitrogen-fixing Lotus japonicus nodules, using gas chromatography coupled to mass spectrometry, revealed a two- to sixfold increase in putrescine levels in mature nodules compared with other organs. Genes involved in polyamine biosynthesis in L japonicus nodules were identified by isolating cDNA clones encoding ADC (LjADC1) and ODC (LjODC) from a nodule library. Searches of the public expressed sequence tag databases revealed the presence of a second gene encoding ADC (LjADC2). Real-time reverse-transcription-polymerase chain reaction analysis showed that LjADC1 and LjADC2 were expressed throughout the plant, while LjODC transcripts were detected only in nodules and roots. Induction of LjODC and LjADC gene expression during nodule development preceded symbiotic nitrogen fixation. Transcripts accumulation was maximal at 10 days postinfection, when a 6.5-fold increase in the transcript levels of LjODC was observed in comparison with the uninfected roots, while a twofold increase in the transcript levels of LjADC1 and LjADC2 was detected. At later stages of nodule development, transcripts for ADC drastically declined, while in the case of ODC, transcript accumulation was higher than that in roots until after 21 days postinfection. The expression profile of genes involved in putrescine biosynthesis correlated well with the expression patterns of genes involved in cell division and expansion, including a L. japonicus Cyclin D3 and an alpha-expansin gene. Spatial localization of LjODC and LjADC1 gene transcripts in developing nodules revealed that both transcripts were expressed in nodule inner cortical cells and in the central tissue. High levels of LjADC1 transcripts were also observed in both nodule and connecting root vascular tissue, suggesting that putrescine and other polyamines may be subject to long-distance transport. Our results indicate that polyamines are primarily involved in physiological and cellular processes involved in nodule development, rather than in processes that support directly symbiotic nitrogen fixation and assimilation.

l -Ascorbic acid metabolism in parthenocarpic and seeded cherry tomatoes

The auxin treatment in tomato plants during anthesis has been extensively used for setting fruits in adverse climatic conditions (e.g., low temperatures and inadequate light), which is well known that reduces pollen availability and fertility. Since auxin application may affect fruit composition and quality, we examined l-ascorbic acid metabolism in seeded fruit (set by natural pollination) and parthenocarpic fruit (set by auxin) in cherry tomato cv. Conchita. Specifically, we studied the oxidized and total ascorbic acid contents, the expression of all characterized genes of l-ascorbic acid metabolism, the activity of ascorbate peroxidase and dehydroascorbate reductase and the immunolocalization of ascorbate peroxidase. Differences were detected between seeded and parthenocarpic fruits, in the expression of some of the genes of ascorbic acid metabolism. However, strong presence of l-ascorbic acid peroxidase protein was detected on the developing seeds. Our data indicate that induced parthenocarpy in auxin treated plants has a significant influence in ascorbic acid metabolism comparing to seeded tomato fruits.

Induced parthenocarpic cherry tomato fruits did not shown significant differences in l -ascorbate content but showed different pattern in GalLDH and GME expression

Parthenocarpy in tomato is often induced by auxins to overcome fertilization problems due to low temperatures. To estimate the effect of this agronomical practice on the physiology and dietary value of cherry tomato fruits we determined l-ascorbic acid, the expression and immunolocalization of galactono 1,4 lactone dehydrogenase and the expression of GDP-mannose 3′,5′-epimerase, key genes in l-ascorbic acid biosynthesis. The levels of l-ascorbic acid did not differ between seeded and parthenocarpic fruits while the relative expression of galactono 1,4 lactone dehydrogenase and GDP-mannose 3′,5′-epimerase gene transcripts showed some significant differences between seeded and parthenocarpic fruits. The galactono 1,4 lactone dehydrogenase immunohistolocalization signal was stronger in the ovules and mature embryos of seed-containing fruits. Our data suggest that although there were differences in the expression of the studied genes and in enzyme localization, these did not cause differences in the l-ascorbic acid content of parthenocarpic fruits produced by auxin application.

Seeded and Parthenocarpic Cherry Tomato Fruits Exhibit Similar Sucrose, Glucose, and Fructose Levels, Despite Dissimilarities in UGPase and SPS Gene Expression and Enzyme Activity

Farmers often resort to the production of auxin-induced parthenocarpic tomato fruits to overcome the adverse climatic conditions which are unfavorable for sufficient fertilization and to lengthen the production period. Even though this practice improves the out-of-season yield, it often undermines fruit quality. In the current study, we examined the effect of induced parthenocarpy in cherry tomato cv. Conchita on the concentration of the main sugars, and on the expression, the activity and the localization of UDP-glucose pyrophosphorylase (UGPase) and sucrose phosphate synthase (SPS), enzymes that are actively involved in sucrose metabolism. Minor differences were detected in sugar levels of seeded and parthenocarpic fruits, whereas considerable variances were found in transcript accumulation and enzyme activities of both UGPase and SPS. Moreover, both proteins were present in the developing seeds. Our data indicate that although induced parthenocarpy significantly influences sucrose metabolism, it has a negligible effect on sugar levels.

In vitro studies of ABA and ethephon induced abscission in olive organs

SummaryDetached reproductive shoots of olive (Olea europaea L. cv. Konservolia) were basally fed continuously with solutions of 25 ppm (0.10 mM) or 50 ppm (0.19 mM) abscisic acid (ABA) and 25 ppm (0.17 mM) or 50 ppm (0.32.mM) (2-chloroethyl)phosphonic acid (ET) individually or in combination, four weeks before full bloom. Leaf and flower abscission were counted along shoots at 24 h intervals over a 120 h period. A lag period of 72 h was needed for the first abscission of both organs in the control (water). By 120 h leaf abscission in control remained low (about 15%), although flower abscission was four-fold higher (about 60%). All treatments with growth regulators reduced the lag period for the first leaf and flower abscission to 48.h. Individual ABA or ET applications resulted in a very low leaf abscission at 48 h treatment. The effect was increased by increase in both concentration and application period, attaining maximum values after 120.h (60–95%). The combination of ABA and ET at both levels showed ...

Co-expression of two sugar transporters in primary and lateral roots of etiolated Glycine max seedlings*

Two Glycine max cDNA clones were characterized and designated as GmSUC2 and GmSTP. The encoded proteins were classified, by prediction of membrane topology and sequence homology, as members of the plant sugar porter family of the Major Facilitator Superfamily. The studies on the temporal and spatial accumulation of the corresponding transcripts, using semi-quantitative RT-PCR and in situ hybridization methods, revealed a differing but overlapping expression pattern at various source and sink organs of soybean plants. These results, in accordance to the structural variations apparent from the deduced protein sequences imply that the corresponding proteins may possess diverse roles in source and sink organs of soybean plants, as well as in various tissues, during primary and lateral root development.