Publications by authors named "Patrizia Rampino"

16 Publications

  • Page 1 of 1

New gene functions are involved in the thermotolerance of the wild wheat relative Aegilops umbellulata.

Plant Physiol Biochem 2020 Nov 3;156:115-124. Epub 2020 Sep 3.

Department of Chemistry, Life Sciences and Environmental Sustainability, University of Parma, Parco Area delle Scienze, 11/A, 43124, Parma, Italy.

Wheat is one of the most important food crops in the world for human consumption, like all plants it is exposed to environmental stresses including high temperatures. The deleterious effect of high temperatures negatively affects plant growth and development, leading to reduced viability and yield. These effects can be reduced by improvement of thermotolerance through innovative breeding strategies, based on the expansion of the genetic pool available, by exploring important genetic functions from wheat wild progenitors. Improving the genetic thermotolerance characteristics of wheat requires greater understanding of genetic bases of thermotolerance, through identification of high temperature stress related genes. A good source of new useful alleles is given by Aegilops species characterized by thermotolerant habits. In this study we have classified as thermotolerant or thermosensitive, on the basis of physiologic tests, some accessions of wheat wild relative species belonging to Aegilops and Triticum genera. A thermotolerant accession of Aegilops umbellulata (AUM5) was selected, subjected to different thermal treatments and analyzed at transcriptional level. By differential display reverse transcriptase polymerase chain reaction (DDRT-PCR), we investigated modulation of gene expression elicited by heat treatments. This approach allowed the identification of various transcript-derived fragments (TDFs) produced by AUM5 in response to different thermal treatments. The functions of the inducible unique genes in the molecular determination of thermotolerance process are discussed.
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http://dx.doi.org/10.1016/j.plaphy.2020.09.005DOI Listing
November 2020

Molecular Effects of and Drought Combined Stress in Olive Trees.

Plants (Basel) 2019 Oct 23;8(11). Epub 2019 Oct 23.

Department of Biological and Environmental Sciences and Technologies, University of Salento, 73100 Lecce, Italy.

Due to global climate change, complex combinations of stresses are expected to occur, among which the interaction between pathogens and drought stress may have a significant effect on growth and yield. In this study, the ()-resistant Leccino and the susceptible one Cellina di Nardò were subjected to (a) individual drought stress, (b) infection and (c) combination of both stress conditions. Here we report the physiological response to stresses in water content in leaves and the modulation in the expression level of seven genes responsive to plant water status and pathogen infection. In -resistant plants, higher expression levels are reported for genes belonging to ROS-scavenging systems and for genes involved in pathogen stress (pathogenesis-related, , and leucine-rich repeat genes, ). However, and were not further induced by water deficit. Interestingly, the genes related to drought response (aquaporin, , dehydration responsive element binding, , and dehydrin, ), which induction was higher in Cellina di Nardò compared to Leccino during drought stress, was poorly induced in -susceptible plants when occur. Conversely, was induced by presence in Leccino. These results were consistent with observations on water content. Indeed, response was similar in Leccino regardless kind of stress or combination, whereas a strong reduction was observed in -susceptible plants infected by or in presence of combined stresses. Thus, the reported findings indicate that resistance of Leccino to could be linked to its lower resistance to water stress, probably leading to the activation of alternative defense pathways that support the plant in response.
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http://dx.doi.org/10.3390/plants8110437DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6918294PMC
October 2019

Exploitation of Prunus mahaleb fruit by fermentation with selected strains of Lactobacillus plantarum and Saccharomyces cerevisiae.

Food Microbiol 2019 Dec 8;84:103262. Epub 2019 Jul 8.

CNR, Institute of Sciences of Food Production (ISPA), via Prov.le Lecce-Monteroni, 73100, Lecce, Italy. Electronic address:

The organoleptic attributes of Prunus mahaleb, a fruit representing a new source of bioactive compounds, are so pronounced that it can be consider non-edible. This study was designed to evaluate the acceptance of P. mahaleb fruits after fermentation with different Saccharomyces cerevisiae and Lactobacillus plantarum protechnological strains. Four different bacterial and one yeast strains, as single or mixed starter formulation, were used to inoculate an aqueous suspension of P. mahaleb fruits. The fermented fruits and fermentation broths were subjected to physico-chemical characterization and the organoleptic properties of both samples were also assessed by a hedonic panel. The obtained results indicated that all the employed strains were able to grow and to ferment the matrix. However, the mixed starter FG69 + Li180-7 (L. plantarum/S. cerevisiae) had the best impact on sensory characteristics of P. mahaleb fruit and fermented medium. The adopted protocol allowed us to attain edible fruits and a new fermented non-dairy drink with valuable probiotic health-promoting properties. In our knowledge, this is the first study concerning the exploitation of P. mahaleb fruits. This investigation confirmed the potential of yeasts and lactic acid bacteria co-inoculation in the design of starter tailored for this kind of food applications.
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http://dx.doi.org/10.1016/j.fm.2019.103262DOI Listing
December 2019

Activation of a gene network in durum wheat roots exposed to cadmium.

BMC Plant Biol 2018 Oct 16;18(1):238. Epub 2018 Oct 16.

Department of Biological and Environmental Sciences and Technologies, University of Salento, via Prov.le Monteroni 165, 73100, Lecce, Italy.

Background: Among cereals, durum wheat (Triticum turgidum L. subsp. durum) accumulates cadmium (Cd) at higher concentration if grown in Cd-polluted soils. Since cadmium accumulation is a risk for human health, the international trade organizations have limited the acceptable concentration of Cd in edible crops. Therefore, durum wheat cultivars accumulating low cadmium in grains should be preferred by farmers and consumers. To identify the response of durum wheat to the presence of Cd, the transcriptomes of roots and shoots of Creso and Svevo cultivars were sequenced after a 50-day exposure to 0.5 μM Cd in hydroponic solution.

Results: No phytotoxic effects or biomass reduction was observed in Creso and Svevo plants at this Cd concentration. Despite this null effect, cadmium was accumulated in root tissues, in shoots and in grains suggesting a good cadmium translocation rate among tissues. The mRNA sequencing revealed a general transcriptome rearrangement after Cd treatment and more than 7000 genes were found differentially expressed in root and shoot tissues. Among these, the up-regulated genes in roots showed a clear correlation with cadmium uptake and detoxification. In particular, about three hundred genes were commonly up-regulated in Creso and Svevo roots suggesting a well defined molecular strategy characterized by the transcriptomic activation of several transcription factors mainly belonging to bHLH and WRKY families. bHLHs are probably the activators of the strong up-regulation of three NAS genes, responsible for the synthesis of the phytosiderophore nicotianamine (NA). Moreover, we found the overall up-regulation of the methionine salvage pathway that is tightly connected with NA synthesis and supply the S-adenosyl methionine necessary for NA biosynthesis. Finally, several vacuolar NA chelating heavy metal transporters were vigorously activated.

Conclusions: In conclusion, the exposure of durum wheat to cadmium activates in roots a complex gene network involved in cadmium translocation and detoxification from heavy metals. These findings are confident with a role of nicotianamine and methionine salvage pathway in the accumulation of cadmium in durum wheat.
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http://dx.doi.org/10.1186/s12870-018-1473-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6192290PMC
October 2018

Early Ongoing Speciation of Within the Grape Ecosystem Revealed by the Internal Variability Among the rDNA Operon Repeats.

Front Microbiol 2018 3;9:1687. Epub 2018 Aug 3.

Institute of Sciences of Food Production (ISPA), National Research Council (CNR), Lecce, Italy.

A yeast strain was isolated during a study on vineyard-associated yeast strains from Apulia in Southern Italy. ITS and LSU D1/D2 rDNA sequences showed this strain not to belong to any known species and was described as the type strain of , a close relative of . Several secondary peaks appeared in the sequences, suggesting internal heterogeneity among the copies of the rDNA. This hypothesis was tested by sequencing single clones of the marker region. The analyses showed different levels of variability throughout the operon with differences between the rRNA encoding genes and the internally transcribed regions. and share high frequency variants, i.e., variants frequently found in many clones, whereas there is a large variability of the low frequency polymorphisms, suggesting that the mechanism of homogenization is more active with the former than with the latter type of variation. These findings indicate that low frequency variants are detected in Sanger sequencing as secondary peaks whereas in Next Generation Sequencing (NGS) of metagenomics DNA would lead to an overestimate of the alpha diversity. For the first time in our knowledge, this investigation shed light on the variation of the copy number of the rDNA cistron during the yeast speciation process. These polymorphisms can be used to investigate on the processes occurring in these taxonomic markers during the separation of fungal species, it being a genetic process highly frequent in the complex microbial ecosystem existing in grape, must and wine.
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http://dx.doi.org/10.3389/fmicb.2018.01687DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6085423PMC
August 2018

Salvia clandestina L.: unexploited source of danshensu.

Nat Prod Res 2019 Feb 19;33(3):439-442. Epub 2018 Mar 19.

a Department of Biological and Environmental Sciences and Technologies, University of Salento , Lecce , Italy.

We report the characterisation of Salvia clandestina L. shoots and roots aqueous extract using HPLC-ESI/MS-TOF. Among the 29 compounds detected, we observed the presence of danshensu (3-(3,4-dihydroxy-phenyl) 2-hydroxy-propinic acid), a powerful antioxidant and a cardio-protective agent. The danshensu content found in the shoots of S. clandestina was considerable (4.96 mg g DW) if compared with previous studies on S. miltiorrhiza Bunge, the main source of this compound. We also determined the total phenolic concentration and we evaluated their antioxidant activity by ABTS, FRAP and Superoxide anion scavenging methods. All the three assays confirmed a greater antioxidant activity for the shoots in comparison to roots. S. clandestina shoots may represent a valuable and natural unexploited source of danshensu and other phenolic compounds, so that it may be useful for future applications in functional foods and pharmaceutical industries.
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http://dx.doi.org/10.1080/14786419.2018.1452015DOI Listing
February 2019

Td4IN2: A drought-responsive durum wheat (Triticum durum Desf.) gene coding for a resistance like protein with serine/threonine protein kinase, nucleotide binding site and leucine rich domains.

Plant Physiol Biochem 2017 Nov 16;120:223-231. Epub 2017 Oct 16.

Dipartimento di Scienze e Tecnologie Biologiche ed Ambientali, Università del Salento, Lecce, Italy.

Wheat, the main food source for a third of world population, appears strongly under threat because of predicted increasing temperatures coupled to drought. Plant complex molecular response to drought stress relies on the gene network controlling cell reactions to abiotic stress. In the natural environment, plants are subjected to the combination of abiotic and biotic stresses. Also the response of plants to biotic stress, to cope with pathogens, involves the activation of a molecular network. Investigations on combination of abiotic and biotic stresses indicate the existence of cross-talk between the two networks and a kind of overlapping can be hypothesized. In this work we describe the isolation and characterization of a drought-related durum wheat (Triticum durum Desf.) gene, identified in a previous study, coding for a protein combining features of NBS-LRR type resistance protein with a S/TPK domain, involved in drought stress response. This is one of the few examples reported where all three domains are present in a single protein and, to our knowledge, it is the first report on a gene specifically induced by drought stress and drought-related conditions, with this particular structure.
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http://dx.doi.org/10.1016/j.plaphy.2017.10.010DOI Listing
November 2017

Cadmium Concentration in Grains of Durum Wheat (Triticum turgidum L. subsp. durum).

J Agric Food Chem 2017 Aug 19;65(30):6240-6246. Epub 2017 Jul 19.

Department of Biological and Environmental Sciences and Technologies, University of Salento , via Prov.le Monteroni 165, 73100 Lecce, Italy.

Heavy metal excess in soil represents a critical problem for crop productivity. Among these pollutants, cadmium (Cd) is one of the most dangerous in terms of food-chain contamination. Two durum wheat near-isogenic lines (NILs) and 12 commercial varieties (cultivars Arcangelo, Aureo, Aziziah, Cappelli, Cirillo, Creso, Iride, Maestrale, Parsifal, Russello, Strongfield, and Svevo) of durum wheat were exposed to a nontoxic level of Cd to evaluate its concentration in grains, roots, and shoots, as well as effects on biomass production. Cultivar Iride showed the most interesting behavior because it stored large amounts of Cd in the roots, preventing its translocation to grains. On the contrary, Cirillo and Svevo genotypes were characterized by a high Cd concentration in the grains. Furthermore, a molecular characterization employing the ScOPC20 marker associated with the Cd uptake locus has shown the absence of the expected fragment in the Iride variety and in other varieties characterized by low Cd concentration, as well as the presence of it in high Cd-accumulating cultivars.
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http://dx.doi.org/10.1021/acs.jafc.7b01946DOI Listing
August 2017

Drought and Heat Differentially Affect XTH Expression and XET Activity and Action in 3-Day-Old Seedlings of Durum Wheat Cultivars with Different Stress Susceptibility.

Front Plant Sci 2016 10;7:1686. Epub 2016 Nov 10.

Dipartimento di Scienze e Tecnologie Biologiche ed Ambientali, Università del Salento Lecce, Italy.

Heat and drought stress have emerged as major constraints for durum wheat production. In the Mediterranean area, their negative effect on crop productivity is expected to be exacerbated by the occurring climate change. Xyloglucan endotransglucosylase/hydrolases (XTHs) are chief enzymes in cell wall remodeling, whose relevance in cell expansion and morphogenesis suggests a central role in stress responses. In this work the potential role of XTHs in abiotic stress tolerance was investigated in durum wheat. The separate effects of dehydration and heat exposure on XTH expression and its endotransglucosylase (XET) activity and action have been monitored, up to 24 h, in the apical and sub-apical root regions and shoots excised from 3-day-old seedlings of durum wheat cultivars differing in stress susceptibility/tolerance. Dehydration and heat stress differentially influence the XTH expression profiles and the activity and action of XET in the wheat seedlings, depending on the degree of susceptibility/tolerance of the cultivars, the organ, the topological region of the root and, within the root, on the gradient of cell differentiation. The root apical region was the zone mainly affected by both treatments in all assayed cultivars, while no change in XET activity was observed at shoot level, irrespective of susceptibility/tolerance, confirming the pivotal role of the root in stress perception, signaling, and response. Conflicting effects were observed depending on stress type: dehydration evoked an overall increase, at least in the apical region of the root, of XET activity and action, while a significant inhibition was caused by heat treatment in most cultivars. The data suggest that differential changes in XET action in defined portions of the root of young durum wheat seedlings may have a role as a response to drought and heat stress, thus contributing to seedling survival and crop establishment. A thorough understanding of the mechanisms underlying these variations could represent the theoretical basis for implementing breeding strategies to develop new highly productive hybrids adapted to future climate scenarios.
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http://dx.doi.org/10.3389/fpls.2016.01686DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5102909PMC
November 2016

Different stress responsive strategies to drought and heat in two durum wheat cultivars with contrasting water use efficiency.

BMC Genomics 2013 Nov 22;14:821. Epub 2013 Nov 22.

Department of Biological and Environmental Sciences and Technologies, University of Salento, Prov,le Lecce Monteroni, I-73100 Lecce, Italy.

Background: Durum wheat often faces water scarcity and high temperatures, two events that usually occur simultaneously in the fields. Here we report on the stress responsive strategy of two durum wheat cultivars, characterized by different water use efficiency, subjected to drought, heat and a combination of both stresses.

Results: The cv Ofanto (lower water use efficiency) activated a large set of well-known drought-related genes after drought treatment, while Cappelli (higher water use efficiency) showed the constitutive expression of several genes induced by drought in Ofanto and a modulation of a limited number of genes in response to stress. At molecular level the two cvs differed for the activation of molecular messengers, genes involved in the regulation of chromatin condensation, nuclear speckles and stomatal closure. Noteworthy, the heat response in Cappelli involved also the up-regulation of genes belonging to fatty acid β-oxidation pathway, glyoxylate cycle and senescence, suggesting an early activation of senescence in this cv. A gene of unknown function having the greatest expression difference between the two cultivars was selected and used for expression QTL analysis, the corresponding QTL was mapped on chromosome 6B.

Conclusion: Ofanto and Cappelli are characterized by two opposite stress-responsive strategies. In Ofanto the combination of drought and heat stress led to an increased number of modulated genes, exceeding the simple cumulative effects of the two single stresses, whereas in Cappelli the same treatment triggered a number of differentially expressed genes lower than those altered in response to heat stress alone. This work provides clear evidences that the genetic system based on Cappelli and Ofanto represents an ideal tool for the genetic dissection of the molecular response to drought and other abiotic stresses.
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http://dx.doi.org/10.1186/1471-2164-14-821DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4046701PMC
November 2013

Role of the C-terminus of Pleurotus eryngii Ery4 laccase in determining enzyme structure, catalytic properties and stability.

Protein Eng Des Sel 2013 Jan 20;26(1):1-13. Epub 2012 Sep 20.

CNR-Istituto di Scienze delle Produzioni Alimentari (ISPA), Lecce, Italy.

The ERY4 laccase gene of Pleurotus eryngii is not biologically active when expressed in yeast. To explain this finding, we analysed the role of the C-terminus of Ery4 protein by producing a number of its different mutant variants. Two different categories of ERY4 mutant genes were produced and expressed in yeast: (i) mutants carrying C-terminal deletions and (ii) mutants carrying different site-specific mutations at their C-terminus. Investigation of the catalytic properties of the recombinant enzymes indicated that each novel variant acquired different affinities and catalytic activity for various substrates. Our results highlight that C-terminal processing is fundamental for Ery4 laccase enzymatic activities allowing substrate accessibility to the enzyme catalytic core. Apparently, the last 18 amino acids in the C-terminal end of the Ery4 laccase play a critical role in enzyme activity, stability and kinetic and, in particular biochemical and structural data indicate that the K532 residue is fundamental for enzyme activation. These studies shed light on the structure/function relationships of fungal laccases and will enhance the development of biotechnological strategies for the industrial exploitation of these enzymes.
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http://dx.doi.org/10.1093/protein/gzs056DOI Listing
January 2013

Novel durum wheat genes up-regulated in response to a combination of heat and drought stress.

Plant Physiol Biochem 2012 Jul 21;56:72-8. Epub 2012 Apr 21.

Dipartimento di Scienze e Tecnologie Biologiche ed Ambientali, Università del Salento, Via Prov. le Monteroni, 73100 Lecce, Italy.

We report the effect of heat, drought and combined stress on the expression of a group of genes that are up-regulated under these conditions in durum wheat (Triticum turgidum subsp. durum) plants. Modulation of gene expression was studied by cDNA-AFLP performed on RNAs extracted from flag leaves. By this approach, we identified several novel durum wheat genes whose expression is modulated under different stress conditions. We focused on a group of hitherto undescribed up-regulated genes in durum wheat, among these, 7 are up-regulated by heat, 8 by drought stress, 15 by combined heat and drought stress, 4 are up-regulated by both heat and combined stress, and 3 by both drought and combined stress. The functional characterization of these genes will provide new data that could help the developing of strategies aimed at improving durum wheat tolerance to field stress.
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http://dx.doi.org/10.1016/j.plaphy.2012.04.006DOI Listing
July 2012

Identification of candidate genes associated with senescence in durum wheat (Triticum turgidum subsp. durum) using cDNA-AFLP.

Mol Biol Rep 2011 Nov 1;38(8):5219-29. Epub 2011 Jan 1.

Di.S.Te.B.A. Università del Salento, via prov.le Monteroni, 73100 Lecce, Italy.

Senescence is an integrated response of plants to various internal (developmental) and external (environmental) signals. It is a highly regulated process leading eventually to the death of cells, single organs such as leaves, or even whole plants. In cereals, which are monocarpic plants, senescence represents the final stage of development. In order to study senescence in durum wheat (Triticum turgidum subsp. durum), a cDNA-AFLP analysis was performed. The transcription profiles of plants at different developmental stages (flowering and senescent) were compared. About 2000 cDNA fragments, ranging in size from 160 to 1900 bp, were reproducibly detected. This allowed the identification of 57 differentially expressed cDNAs corresponding to genes belonging to different functional categories related to cellular metabolism, transcription, maintenance of DNA structure, transport and signal transduction. This paper reports the identification of novel durum wheat candidate genes involved in the senescence process, and provides new information about the senescence programme of this important crop species.
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http://dx.doi.org/10.1007/s11033-010-0673-2DOI Listing
November 2011

Molecular cloning and heterologous expression of a laccase gene from Pleurotus eryngii in free and immobilized Saccharomyces cerevisiae cells.

Appl Microbiol Biotechnol 2008 Jul 29;79(5):731-41. Epub 2008 Apr 29.

Istituto di Scienze delle Produzioni Alimentari del CNR, Unità di Lecce, via Prov le Lecce-Monteroni, Lecce, Italy.

A full length cDNA encoding an extracellular laccase was isolated by reverse transcription polymerase chain reaction from the mycelia of the mushroom Pleurotus eryngii. The isolated sequence, denoted Ery3, encodes for a mature laccase isoenzyme of 531 amino acid residues with a predicted molecular weight of 56.6 kDa. All sequence motifs, being the signature sequences used to identify the laccases, were found in the Ery3 protein sequence. The Ery3 cDNA was expressed in Saccharomyces cerevisiae and the effects of copper concentration and cultivation temperature were investigated. S. cerevisiae cells were immobilized in calcium alginate gel and the optimal immobilization parameters for the enhanced production of laccase were determined. The immobilization was most effective with 3% sodium alginate, 0.1 M calcium chloride and an initial biomass of 4.5 x 10(8) cells. The enzyme yield obtained with immobilized cells (139 mU ml(-1)) showed a 1.6-fold increase compared to the highest yield obtained with free cells. The alginate beads showed good stability and retained 84% capacity of enzyme production after seven repeated cycles of batch fermentation. The immobilization system proved to increase the proteolytic stability of the recombinant Ery3 protein. To our knowledge, this is the first report on S. cerevisiae whole-cell immobilization for recombinant laccase production.
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http://dx.doi.org/10.1007/s00253-008-1479-1DOI Listing
July 2008

Four members of the HSP101 gene family are differently regulated in Triticum durum Desf.

FEBS Lett 2007 Oct 14;581(25):4841-9. Epub 2007 Sep 14.

Dipartimento di Scienze Ambientali, Sez. Genetica e Biotecnologie Ambientali, Università di Parma, Italy.

Heat shock proteins play an essential role in preventing deleterious effects of high temperatures. In many plants, HSP101 has a central role in heat stress survival. We report the isolation and characterization of four cDNAs corresponding to different members of the durum wheat HSP101 gene family. Expression analysis revealed differences in their induction. Accordingly, durum wheat HSP101 genes are differently regulated, therefore having distinct roles in stress response and thermotolerance acquisition. These findings are important for further dissection of the molecular mechanisms underlying the stress response and for understanding the functions of the HSP101 family members. This information could be important for the exploitation of specific alleles in marker assisted selection for abiotic stress resistance.
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http://dx.doi.org/10.1016/j.febslet.2007.09.010DOI Listing
October 2007

Drought stress response in wheat: physiological and molecular analysis of resistant and sensitive genotypes.

Plant Cell Environ 2006 Dec;29(12):2143-52

Dipartimento di Scienze e Tecnologie Biologiche ed Ambientali, Università di Lecce, via prov. le Monteroni, Lecce, Italy.

Water deficit is a severe environmental stress and the major constraint on plant productivity with an evident effect on plant growth. The aim of this work was to study Triticum and Aegilops seedlings differing in their response to drought stress at the physiological and molecular levels. The identification of resistant and sensitive genotypes was firstly based on the relative water content (RWC) measurement. Further characterization of genotypes contrasting in their response to water stress was performed at the physiological level by determination of RWC, water loss rate (WLR) and free proline content after different hours of dehydration. Modification in the expression level of five dehydrin (DHN) genes was also analysed by reverse transcription-polymerase chain reaction (RT-PCR). Five cDNAs coding for different DHNs were identified and characterized. These genes are not expressed in the well-watered plants, but only in the stressed plants. Four of these cDNAs are related to novel DHN sequences. The results obtained clearly indicate a relation between the expression of these genes and tissue water content. In particular, in the resistant genotypes the expression of DHN genes is initiated even though tissue hydration levels are still high, indicating also in wheat the involvement of these proteins in water retention.
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http://dx.doi.org/10.1111/j.1365-3040.2006.01588.xDOI Listing
December 2006