Evaluating the potential use of Cu-contaminated soils for giant reed (Arundo donax, L.) cultivation as a biomass crop.

Over the past decades, the important topic of environmental sustainability, impact, and security of the fossil fuel supply has stimulated interest in using lignocellulosic feedstocks as biofuel to partially cover energy demands. Among energy no-food crops, giant reed (Arundo donax, L.), a perennial rhizomatous grass has been identified as a leading candidate crop for lignocellulosic feedstock, due to its positive energy balance, and low ecological/agro-management demands. The aim of the present study was to characterize the physiological response of Arundo donax (L.) to artificial soil contamination with three different Cu levels (200, 400, and 800 ppm), and to assess the relationship between plant Cu tolerance and S assimilation rate. The present study not only confirms the ability of Arundo donax L. to cope with Cu stress and therefore to grow in marginal, degraded lands abandoned by mainstream agricultural, but also shows that plant performance might be likely ascribed to a modulation of sulfate metabolism resulting in increased thiols content.

Novel phenotypes related to the breeding of purple-fruited tomatoes and effect of peel extracts on human cancer cell proliferation.

The production of anthocyanins in the tomato (Solanum lycopersicum L.) fruit is normally absent or poor, but a number of mutants or introgression lines are known to increase anthocyanin levels in vegetative and reproductive tissues. Through conventional breeding, a genetic combination was obtained with the remarkable phenotype of a deep purple fruit pigmentation, due to an accumulation of anthocyanins on the peel. Such a genotype was named Sun Black (SB) as a consequence of its sensitivity to light induction. When characterized for morpho-agronomic traits, SB plants showed increased fertility. Purple fruits displayed an arrangement of the epicarp cells different from normal tomatoes, a feature that could account for different mechanical properties and shelf-life potential. The SB genotype and, to a lesser extent, its single mutant parents showed the capacity to accumulate anthocyanins in the seedling root when grown under light. This phenotype, which was greatly improved by the addition of sucrose to the germination medium, proved to be useful as selection index and gave new insights for in vitro production of anthocyanin extracts. To assess the nutraceutical potential of purple tomatoes, we tested the activity of SB skin extracts on the proliferation of two human cancer cells lines. Cell proliferation was significantly inhibited by SB extract in a dose-dependent manner. When the bioactivity of SB extracts was compared with that of other anthocyanin-containing fruits or vegetables, a significant "Extract*Line" interaction was evidenced, suggesting a crucial role for the extract composition in terms of anthocyanidins and other eventual cell growth-inhibiting compounds.

Constitutive co-suppression of the GA 20-oxidase1 gene in tomato leads to severe defects in vegetative and reproductive development.

To dissect the role of gibberellins in tomato development, we have constitutively down-regulated the gene GA 20-oxidase1 (GA20ox1). Plants co-suppressed for GA20ox1 (referred to as CO-6 plants) showed vegetative defects typical of GA deficiency such as darker and mis-shaped leaves and dwarfism. CO-6 plants flowered as the controls, although their flowers had subtle defects in the pedicel and in organ insertion. Analysis of male development revealed defects before, during and after meiosis, and a final pollen viability of 22%. The development of female organs and gametes appeared normal. Pollination experiments indicated that the pollen produced by CO-6 plants was able to fertilize control ovaries, but the analysis of the progeny showed that the construct was not transmitted. Ovaries of CO-6 plants showed high fruit set and normal fruit development when pollinated with control pollen. However these fruits were completely seedless due to a stenospermocarpic behaviour that was evidenced by callose layering in the endothelium between 7 and 15 days after pollination. We conclude that GA20ox1 in tomato exerts specific developmental roles that are not redundantly shared with other members of this gene family. For reproductive male development, silencing of this gene is detrimental for pollen production and either gametophytically lethal or severely hampering seed germination. In the pistil, the co-suppression construct does not affect the progamic phase, nor fruit set and growth, but it interferes with seed development after fertilization leading to seed abortion.

A H2O2-forming peroxidase rather than a NAD(P)H-dependent O2•- synthase may be the major player in cell death responses controlled by the Pto-Fen complex following fenthion treatment.

Four tomato (Lycopersicon esculentum Mill.) near-isogenic lines were treated by foliar spraying with the insecticide fenthion. Two, Riogrande and Rimone, differed from each other only for the presence in the latter of the Fen gene, conferring propensity to develop foliar symptoms upon exposure to fenthion. The other two, namely RC332 and RC131, were the transgenic versions of Riogrande and Rimone, respectively, harbouring the Gox gene encoding for glucose oxidase of Aspergillus niger. The production of H2O2 as well as the activities of H+-ATPase, NAD(P)H-dependent superoxide synthase, catalase, peroxidase, and Cu,Zn-superoxide dismutase were evaluated in the foliar tissues up to 24 h after exposure to fenthion. The Fen gene conferred sensitivity to fenthion, regardless of the expression of a Gox transgene. A prolonged accumulation of H2O2 was observed in the leaves of Rimone and of RC131, which was instead transient in Riogrande and in RC332. In all the tomato lines, exposure to fenthion induced rapid but transient changes in the activities of most enzymes. The only exception was peroxidase activity in the leaves of Rimone and of RC131, which steadily increased until the end of the sampling period. It is suggested that the sensitivity of Rimone to fenthion might be due to the sustained activity of a H2O2-forming peroxidase.

The Cf-2 / Rcr3esc gene interaction in tomato (Lycopersicon esculentum) induces autonecrosis and triggers biochemical markers of oxidative burst at cellular level.

A tomato plant (Lycopersicon esculentum Mill.) with necrotic leaf spots mimicking disease lesions was singled out in progeny under selection in Moscow (breeding material of Ignatova Svetlana). The progeny from spontaneous selfing of such a plant (V20368), in the presence of increasing temperature and high light intensity, exhibited spontaneous necrotic lesions on the leaves, with acropetal progression (autonecrosis). A similar phenotype, described in 1948 by Langford, appeared to be associated with the Cf-2 resistance gene, introgressed from L. pimpinellifolium. Recently, Kruger et al. (2002) demonstrated that the Cf-2 effect depends on a second gene (Rcr3pim) encoding a cysteine protease, and that autonecrosis is activated by the contemporary presence of Cf-2 and the L. esculentum allele Rcr3esc. In this work we characterised the V20368 mutant and verified that autonecrosis is caused by the presence of an interaction between Cf-2 and Rcr3esc. When the environmental conditions are favourable, this interaction triggers an oxidative burst, as evidenced by a strong increase in H2O2 production and activities of catalase (EC 1.11.1.6), peroxidase (EC 1.11.1.7) and ATPase (EC 3.6.1.3). In addition, by grafting the necrotic mutant on the cv. Riogrande and vice versa, we proved that the necrotic phenotype is not associated with the movement of a signal molecule, since the autonecrosis was not transmitted across the grafting point. Finally, the interaction between Cf-2 and Rcr3esc appeared to lower the threshold of stress perception, as evidenced by an increased sensitivity to the insecticide Fenthion.