The composition of pomegranate peel, the main by-product during pomegranate processing, and some of the characteristics of the water-soluble pectins were investigated. Four tunisian pomegranate peels were subjected to hot aqueous extractions (86°C, 80min, 20mM nitric acid). Pomegranate peels yielded between 6.8% and 10.1% pectins. The extracted pectins were low methylated and were characterized by the predominance of homogalacturonan regions. Principal component analysis applied on FT-IR spectral data in the region between 4000 and 650cm(-1) differentiated the samples according to their degree of methylation. At pH 3, in the presence of 0.7% pectin, all solutions showed a rapid gel formation with G'>G″. With decreasing temperature from 90°C to 10°C, G' increased to reach a plateau at 10°C. The variation in the pectin gel formation between varieties was attributed to difference in pectin characteristics particularly the hydrodynamic volume and the neutral sugar content.
WRKY transcription factors belong to a large family of plant transcriptional regulators whose members have been reported to be involved in a wide range of biological roles including plant development, adaptation to environmental constraints and response to several diseases. However, little or poor information is available about WRKY's in Citrus. The recent release of completely assembled genomes sequences of Citrus sinensis and Citrus clementina and the availability of ESTs sequences from other citrus species allowed us to perform a genome survey for Citrus WRKY proteins. In the present study, we identified 100 WRKY members from C. sinensis (51), C. clementina (48) and Citrus unshiu (1), and analyzed their chromosomal distribution, gene structure, gene duplication, syntenic relation and phylogenetic analysis. A phylogenetic tree of 100 Citrus WRKY sequences with their orthologs from Arabidopsis has distinguished seven groups. The CsWRKY genes were distributed across all ten sweet orange chromosomes. A comprehensive approach and an integrative analysis of Citrus WRKY gene expression revealed variable profiles of expression within tissues and stress conditions indicating functional diversification. Thus, candidate Citrus WRKY genes have been proposed as potentially involved in fruit acidification, essential oil biosynthesis and abiotic/biotic stress tolerance. Our results provided essential prerequisites for further WRKY genes cloning and functional analysis with an aim of citrus crop improvement.
A central composite design was employed to determine the influence of extraction conditions on production yield and chemical composition of pectin from pomegranate peels. Response surface methodology (RSM) was used to quantify the integral effect of the three processing parameters (extraction duration, temperature and pH) on yield. A second-order polynomial model was developed for predicting the yield of pomegranate peels pectin based on the composite design. Yields ranged from 6.4 to 11.0±0.2%. Optimal temperature, duration and pH value of the extraction were 86°C, 80min and 1.7, respectively. The uronic acid and the total neutral sugar content of the extracted pectins ranged from 377 to 755mg/g and from 161 to 326mg/g, respectively. Moreover, the degree of methylation varied with the extraction conditions and the extracted pectins were low methylated. On high pressure size exclusion chromatography (HPSEC), the elution pattern of the acid-extracted pectins showed that severe conditions were associated with lower hydrodynamic volume.
Our study aims to assess the implication of WRKY transcription factor in the molecular mechanisms of grapevine adaptation to salt and water stresses. In this respect, a full-length cDNA, isolated from a grape berry cDNA library, was constitutively over-expressed in seedlings. Our results showed that transgenic tobacco plants exhibited higher seed germination rates and better growth, under both salt and osmotic stress treatments, when compared to wild type plants. Furthermore, our analyses demonstrated that, under stress conditions, transgenic plants accumulated more osmolytes, such as soluble sugars and free proline, while no changes were observed regarding electrolyte leakage, HO, and malondialdehyde contents. The improvement of osmotic adjustment may be an important mechanism underlying the role of 2 in promoting tolerance and adaptation to abiotic stresses. Principal component analysis of our results highlighted a clear partition of plant response to stress. On the other hand, we observed a significant adaptation behaviour response for transgenic lines under stress. Taken together, all our findings suggest that over-expression of gene has a compelling role in abiotic stress tolerance and, therefore, would provide a useful strategy to promote abiotic stress tolerance in grape molecular-assisted breeding and/or new biotechnology tools.
Plant NHX antiporters are responsible for monovalent cation/H+ exchange across cellular membranes and play therefore a critical role for cellular pH regulation, Na+ and K+ homeostasis, and salt tolerance. Six members of grapevine NHX family (VvNHX1-6) have been structurally characterized. Phylogenetic analysis revealed their organization in two groups: VvNHX1-5 belonging to group I (vacuolar) and VvNHX6 belonging to group II (endosomal). Conserved domain analysis of these VvNHXs indicates the presence of different kinds of domains. Out of these, two domains function as monovalent cation-proton antiporters and one as the aspartate-alanine exchange; the remaining are not yet with defined function. Overall, VvNHXs proteins are typically made of 11-13 putative transmembrane regions at their N-terminus which contain the consensus amiloride-binding domain in the 3rd TM domain and a cation-binding site in between the 5th and 6th TM domain, followed by a hydrophilic C-terminus that is the target of several and diverse regulatory posttranslational modifications. Using a combination of primary structure analysis, secondary structure alignments, and the tertiary structural models, the VvNHXs revealed mainly 18 α helices although without β sheets. Homology modeling of the 3D structure showed that VvNHX antiporters are similar to the bacterial sodium proton antiporters MjNhaP1 (Methanocaldococcus jannaschii) and PaNhaP (Pyrococcus abyssi).
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