A filamentous fungus was isolated from Tuber borchii Vitt. fruiting bodies, and it was identified as an Arthrinium phaeospermum (Corda) M.B. Ellis strain, an “endophyte” that forms various associations with healthy leaves, stems, and roots of plants. Molecular analysis confirmed the association of this filamentous fungus with the ascocarps of all collection sites in Salento, Apulia (South Italy). An in vitro symbiosis system between Cistus creticus L. and T. borchii was set up; A. phaeospermum appears to be able to promote mycorrhiza formation in Cistus seedlings, inducing primary root shortening and an increase of secondary roots, similar to the effect of Mycorrhization Helper Bacteria (MHB). Compartmented and uncompartmented bioassays were carried out to investigate the effects of exudates/volatiles released by the truffle-hosted fungus on root architecture; the results showed root shortening in compartmented bioassay suggesting that volatiles released by the fungus alone are sufficient to alter root morphology in early phase of interaction before the mycorrhiza formation. The first evidence for an influence of a truffle-hosted fungus on ectomycorrhizal symbiosis establishment is reported.

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.

The sour taste of lemons (Citrus limon (L.) Burm.) is determined by the amount of citric acid in vacuoles of juice sac cells. Faris is a “sweet” lemon variety since it accumulates low levels of citric acid. The University of California Riverside Citrus Variety Collection includes a Faris tree that produces sweet (Faris non-acid; FNA) and sour fruit (Faris acid; FA) on different branches; it is apparently a graft chimera with layer L1 derived from Millsweet limetta and layer L2 from a standard lemon. The transcription profiles of Faris sweet lemon were compared with Faris acid lemon and Frost Lisbon (L), which is a standard sour lemon genetically indistinguishable from Faris in prior work with SSR markers. Analysis of microarray data revealed that the transcriptomes of the two sour lemon genotypes were nearly identical. In contrast, the transcriptome of Faris sweet lemon was very different from those of both sour lemons. Among about 1,000 FNAspecific, presumably pH-related genes, the homolog of Arabidopsis H+-ATPase proton pump AHA10 was not expressed in FNA, but highly expressed in FA and L. Since Arabidopsis AHA10 is involved in biosynthesis and acidification of vacuoles, the lack of expression of the AHA10 citrus homolog represents a very conspicuous molecular feature of the FNA sweet phenotype. In addition, high expression of several 2-oxoglutarate degradationrelated genes in FNA suggests activation of the GABA shunt and degradation of valine and tyrosine as components of the mechanism that reduces the level of citric acid in sweet lemon.

Citrus fruits are characterized by the accumulation of high levels of citric acid, which account for 90% of the total organic acids. Citric acid is an important determinant of fruit taste because it affects the sourness of the fruit as well as the perceived sweetness, by masking the taste of sugars. To study the role of gene expression in citric acid accumulation we compared the transcriptomes of juice sac cells in acid and acidless lemon genotypes using an Affymetrix microarray. Three genotypes were used for the transcriptome analysis: ‘Frost Lisbon’ (L), a standard sour lemon, and two ‘Faris’ genotypes obtained from a chimera that produces on the same tree both sweet (‘Faris’ non-acid, FNA) and sour fruit (‘Faris’ acid, FA). After the transcriptome comparison was completed, we discovered that ‘Faris’ is a graft chimera. Here we present SSR marker data showing that layer 1 of ‘Faris’ is derived from ‘Millsweet’ lime or a very similar cultivar, whereas layer 2 is derived from an acid lemon similar to ‘Eureka’ or ‘Lisbon’. Transcriptome comparisons among the three genotypes during two ripening stages showed high similarity between the two acid fruits and considerable divergence of FNA. Among the more interesting differences are genes involved in the GABA shunt pathway and a plasma-membrane type H+-ATPase.

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.

Soil qualities and rootstocks are among the main factors that have been acknowledged to influence grape development as well as fruit and wine composition. Despite the role of soil and rootstock in establishing a successful vineyard in terms of grape quality, almost no molecular evidence linking soil and rootstock properties to the gene expression have been reported. The transcriptome variation in response to different soils and rootstocks was investigated through microarray technology. The cv. Pinot Noir was grown on different soils: sand, turf, and vineyard soil. The plants were grafted on the contrasting 101-14 and 1103 Paulsen rootstocks. The modulation of genes' expression in response to different soils and rootstocks was evaluated considering their potential impact on primary (carbohydrate) and secondary (phenylpropanoid) metabolisms. This study highlights a link among soil composition, rootstock, and gene expression. The results open a perspective for a molecular interpretation of the interaction between soil and grapevine.

We report a series of microarray-based leaf and crown transcriptome comparisons involving three barley cultivars (cvs. Luxor, Igri and Atlas 68) which express differing degrees of frost tolerance. The transcripts were obtained following the exposure of seedlings to low (above and below zero) temperatures, aiming to identify those genes and signalling/metabolic pathways which are associated with frost tolerance. Both the leaves and the crowns responded to low temperature by the up-regulation of a suite of abscisic acid (ABA)-responsive genes, most of which have already been recognized as components of the plant low temperature response. The inter-cultivar comparison indicated that genes involved in maintaining the leaf's capacity to synthesize protein and to retain chloroplast activity were important for the expression of frost tolerance. In the crown, the repression of genes associated with nucleosome assembly and transposon regulation were the most relevant transcriptional changes associated with frost tolerance, highlighting the role of gene repression in the cold acclimation response.

Heavy metals (HMs), such as copper, zinc, lead, mercury and cadmium, are the most abundant and dangerous inorganic environmental pollutants. Growing pieces of evidence suggest that mycorrhizal fungi can alleviate metal toxicity in plants. In this study, we focused attention on the ectomycorrhizal (ECM) fungus Tuber borchii Vitt., which is widespread in Italy and is of great ecological interest because of the mutualistic associations and the advantages it provides to host plants. Seedlings of the Mediterranean shrub Cistus creticus L., mycorrhized and non mycorrhized with the ECM fungus 7: borchii, were treated with HMs (zinc, lead and chromium). HMs induced leaves' chlorosis in non mycorrhized seedlings; while no significant differencewas observed impigmentation of mycorrhized seedlings' leaves. This observation was confirmed by Euclidean Distance of color measurements in L*a*b* units from RGB digital images of leaves. The decrease in leaves pigmentation observed in HM treated non mycorrhized seedlings strongly correlated with a reduced expression of key genes associated with chlorophyll biosynthesis; instead, no significant variation of gene expression was detected in mycorrhized seedlings treated with HMs.

Glyphosate is a non-selective herbicide that inhibits the shikimate pathway’s enzyme EPSPS (5-enolpyruvylshikimate-3-phosphate synthase) preventing the production of aromatic amino acids. This herbicide is largely used and appreciated because it controls a wide range of annual and perennial weeds but it has a minimal environmental impact when compared with other herbicides. Initially it was thought that resistance to glyphosate was not easy to evolve but the continuous applications, as it happened for other herbicides, have induced the development of several glyphosate-resistant weeds. Glyphosate resistance can be developed as target-site and non-target-site mechanisms. In the target-site mechanism of resistance, either a mutation on the EPSPS enzyme (enzyme modification) or the overexpression of the EPSPS enzyme have been found to confer resistance. In the non-target-site mechanism of glyphosate resistance, the herbicide translocation and neutralization is observed. Pumping glyphosate into vacuoles via membrane transporters has been suggested as a possible process involved in the restricted glyphosate translocation. As a consequence, a different vacuolar organization or plasticity could be an interesting character or marker to correlate to glyphosate resistance. Vacuolar markers AleuGFP (Sar1 dependent sorting) or GFPChi (Sar1 independent sorting) respectively can be used to monitor independent vacuolar sorting mechanisms during glyphosate induced stress. We observed that the adaptive reaction of tobacco protoplasts vacuolar system to the treatment with glyphosate, can be mimicked by the overexpression of a Triticum durum TdGST gene. Previous analysis evidenced that the herbicide glyphosate increased TdGST expression, confirming the role of GST in the protection against xenobiotics. Non-target-site glyphosate resistance mechanisms may correlate with an independent regulation of cell compartmentalization and herbicide induced genes may have a direct effect on it.

Recently, Xylella fastidiosa was reported in Italy, associated with the "Olive Quick Decline Syndrome". The cv. Leccino exhibits an evident tolerance with a slow disease progression compared with the other cultivars. Between the mechanisms proposed to explain the putative tolerance of some hosts to X. fastidiosa diseases, lignin deposition plays an important role. Analysis of phenolic compounds in healthy and infected Leccino and Cellina di Nardò leaves showed, in the two cultivars, a reduction of hydroxytyrosol glucoside (usually associated with drought and cold stress) and, only in Leccino, an increase of quinic acid, precursor of lignin. To determine if lignin biosynthesis is involved in defence response, we investigated the expression of genes coding for entry-point enzymes in different branches of the phenylpropanoid pathway. In stems of Cellina di Nardò infected plants, Cinnamate-4-Hydroxylase (C4H) and 4-Coumarate:CoA Ligase (4CL) resulted strongly down-regulated, indicating a plant disease response since the inhibition of C4H is reported to promote the accumulation of benzoic acid and salicylic acid as defence signals. Instead, in the cv. Leccino, Cinnamoyl-CoA Reductase (CCR, reported to be strongly induced during the formation of lignin defence response associated) was up-regulated in the stem of infected plants; moreover, Polyphenol oxidase (PPO), coding for an enzyme involved in the hydroxytyrosol biosynthesis, was down-regulated. The quantification of lignin in healthy and infected branches of both cultivars, showed a significant increase of total lignin in infected Leccino compared with the sensitive cultivar; moreover, histochemical observations of stem sections exhibited a different lignin distribution in the sclerenchyma and in the xylem tissue of infected Leccino plants compared to sections of healthy ones. Results suggest a critical role for lignin in X. fastidiosa tolerance of cv. Leccino.