Pepper fruit is one of the highest vitamin C sources of plant origin for our diet. In plants, ascorbic acid is mainly synthesized through the L-galactose pathway, being the L-galactono-1,4-lactone dehydrogenase (GalLDH) the last step. Using pepper fruits, the full GalLDH gene was cloned and the protein molecular characterization accomplished. GalLDH protein sequence (586 residues) showed a 37 amino acids signal peptide at the N-terminus, characteristic of mitochondria. The hydrophobic analysis of the mature protein displayed one transmembrane helix comprising 20 amino acids at the N-terminus. By using a polyclonal antibody raised against a GalLDH internal sequence and immunoblotting analysis, a 56 kDa polypeptide cross-reacted with pepper fruit samples. Using leaves, flowers, stems and fruits, the expression of GalLDH by qRT-PCR and the enzyme activity were analyzed, and results indicate that GalLDH is a key player in the physiology of pepper plants, being possibly involved in the processes which undertake the transport of ascorbate among different organs.We also report that an NO (nitric oxide)-enriched atmosphere enhanced ascorbate content in pepper fruits about 40% parallel to increased GalLDH gene expression and enzyme activity. This is the first report on the stimulating effect of NO treatment on the vitamin C concentration in plants. Accordingly, the modulation by NO of GalLDH was addressed. In vitro enzymatic assays of GalLDH were performed in the presence of SIN-1 (peroxynitrite donor) and S-nitrosoglutahione (NO donor). Combined results of in vivo NO treatment and in vitro assays showed that NO provoked the regulation of GalLDH at transcriptional and post-transcriptional levels, but not post-translational modifications through nitration or S-nitrosylation events promoted by reactive nitrogen species (RNS) took place. These results suggest that this modulation point of the ascorbate biosynthesis could be potentially used for biotechnological purposes to increase the vitamin C levels in pepper fruits.
The conductances which determine the resting membrane potential of rat superior cervical ganglia (SCG) neurons were investigated using perforated voltage- and current-clamp whole-cell techniques. The resting potential of SCG cells varied from -47 to -80 mV (-58.3 +/- 0.8 mV, n = 55). Blockade of M and h currents induced a depolarisation (7.4 +/- 0.7 mV, n = 22) and a hyperpolarisation (7.2 +/- 0.7 mV, n = 20) respectively; however, no correlation between the amplitude of these currents and the resting potential was found. The inhibition of the Na/K pump also induced membrane depolarisation (3.2 +/- 0.2 mV, n = 8). Inhibition of voltage-gated currents unmasked a voltage-independent resting conductance reversing at -50 mV. The reversal potential of the voltage-independent conductance, which included the electrogenic contribution of the Na/K pump, was strongly correlated with the resting potential (R = 0.87, p < 0.0001, n = 30). Ionic substitution experiments confirmed the existence of a voltage-independent conductance (leakage) with four components, a main potassium conductance, two minor sodium and chloride conductances and a small contribution of the Na/K pump. It is concluded that the resting potential of SCG cells strongly depends on the reversal potential of the voltage-independent conductance, with voltage-activated M and h currents playing a prominent stabilising role.
Summary Reproductive phenology was studied in a Pinus radiata seed orchard, located in northern Spain. Timing of flowering was determined on the basis of data recorded by visual observations made in 2000, 2001 and 2002. The genetic and environmental factors affecting female and male phenology, as well as reproductive synchronization, were studied. The dates of beginning of the receptive phase and pollen shedding varied greatly from year to year but the variation on the sum of degree-days was low. In general, the flowering periods of the different clones overlapped. The clonal differences in the phenology of receptivity and pollen shedding were in most cases statistically significant. The time needed to reach flowering stages was under strong genetic control. Genetic control was stronger for the female than the male flowering process. However, correlations between years were stronger for male than for female flowering phenology. The male flowering clones that best synchronized with the females appeared to be those that started flowering earlier. The phenological overlap index varied greatly among clones, whether male or female, and also among years.
Adventitious rooting of cuttings is a complex developmental process in forest species, with several exogenous and endogenous factors influencing the outcome of the process. In this study we applied an in vitro working system, comprising two lines of microshoots with the same genotype but at a different ontogenetic stages, in two different tree species (chestnut and oak). We analyzed the expression of a gene encoding an AP2/ERF transcription factor from group VII in the initial hours of the adventitious rooting induction, both in rooting competent and incompetent microshoots. The analysis revealed that expression of this gene is related to wounding, ontogenetic stage and auxin in a complex and species-specific manner. Putative induction of the gene by auxin was also analyzed in the presence of naphthyl-phthalamic acid (NPA), an auxin transport inhibitor. In situ expression analysis in chestnut relates the gene activity to cambial divisions and root primordia in rooting competent microshoots, as well as in the root apex. The putative role of the gene during adventitious roots formation is discussed.
Individual and family heritabilities, as well as phenotypic and genetic correlations were estimated 2 and 3 years after plantation, in progeny tests carried out in plantations of open-pollinated Pinus radiata D.Don progeny located at three different sites in Galicia (NW Spain). The following factors were considered: survival, growth (free growth, height, diameter, volume, height/diameter ratio), form (number of branches, number of whorls, number of branches per whorl, number of branches per tree height meter, number of ramicorn branches, diameter of the thickest branch, internode length, branch angle, stem straightness, stem sinuosity, number of forks) and resistance (frost and insect resistance). Each plantation contained 50 female parents that had been selected as plus trees in genetically unimproved plantations in Galicia, as well as 6 female parents selected from the radiata pine breeding programme in the Basque Country (northern Spain) and two commercial lots of seed from Galicia, as controls. Individual heritabilities varied from low to high levels for each trait studied. Family heritabilities followed similar trends for individual heritabilities, but were of much greater magnitude. The results suggested that all traits except the number of branches per whorl and insect resistance could be expected to respond to selection based on individual performance. The less heritable traits could be improved by progeny testing. Height and diameter were strongly and positively correlated, and both traits were also correlated with the number of branches, the number of whorls and the branch diameter. The results are compared with previously published data and discussed in relation to the implications for breeding activities.
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