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Foreseen climate change is expected to impact on grape composition, both sugar and pigment content. We tested the hypothesis that interactions between main factors associated with climate change (elevated CO 2 , elevated temperature, and water deficit) decouple sugars and anthocyanins, and explored the possible involvement of vegetative area, photosynthesis, and grape C uploading on the decoupling. Tempranillo grapevine fruit-bearing cuttings were exposed to CO 2 (700 vs. 400 ppm), temperature (ambient vs. + 4 °C), and irrigation levels (partial vs. full) in temperature-gradient greenhouses. In a search for mechanistic insights into the underlying processes, experiments 1 and 2 were designed to maximize photosynthesis and enlarge leaf area range among treatments, whereas plant growth was manipulated in order to deliberately down-regulate photosynthesis and control vegetative area in experiments 3 and 4. Towards this aim, treatments were applied either from fruit set to maturity with free vegetation and fully irrigated or at 5-8% of pot capacity (experiments 1 and 2), or from veraison to maturity with controlled vegetation and fully irrigated or at 40% of pot capacity (experiments 3 and 4). Modification of air 13 C isotopic composition under elevated CO 2 enabled the further characterization of whole C fixation period and C partitioning to grapes. Increases of the grape sugars-to-anthocyanins ratio were highly and positively correlated with photosynthesis and grape 13 C labeling, but not with vegetative area. Evidence is presented for photosynthesis, from fruit set to veraison, and grape C uploading, from veraison to maturity, as key processes involved in the establishment and development, respectively, of the grape sugars to anthocyanins decoupling.

The increase in grape berry ripening rates associated to climate change is a growing concern for wine makers as it rises the alcohol content of the wine. The present work studied the combined effects of elevated CO2, temperature and UV-B radiation on leaf physiology and berry ripening rates. Three doses of UV-B: 0, 5.98, 9.66 kJm(-2)d(-1), and two CO2-temperature regimes: ambient CO2-24/14 °C (day/night) (current situation) and 700 ppm CO2-28/18 °C (climate change) were imposed to grapevine fruit-bearing cuttings from fruit set to maturity under greenhouse-controlled conditions. Photosynthetic performance was always higher under climate change conditions. High levels of UV-B radiation down regulated carbon fixation rates. A transient recovery took place at veraison, through the accumulation of flavonols and the increase of antioxidant enzyme activities. Interacting effects between UV-B and CO2-temperature regimes were observed for the lipid peroxidation, which suggests that UV-B may contribute to palliate the signs of oxidative damage induced under elevated CO2-temperature. Photosynthetic and ripening rates were correlated. Thereby, the hastening effect of climate change conditions on ripening, associated to higher rates of carbon fixation, was attenuated by UV-B radiation., (Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.)

This work aims to characterize the physiological response of grapevine (Vitis vinifera L.) cv. Tempranillo to UV-B radiation under water deficit conditions. Grapevine fruit-bearing cuttings were exposed to three levels of supplemental biologically effective UV-B radiation (0, 5.98 and 9.66kJm(-2)day(-1)) and two water regimes (well watered and water deficit), in a factorial design, from fruit-set to maturity under glasshouse-controlled conditions. UV-B induced a transient decrease in net photosynthesis (Anet), actual and maximum potential efficiency of photosystem II, particularly on well watered plants. Methanol extractable UV-B absorbing compounds (MEUVAC) concentration and superoxide dismutase activity increased with UV-B. Water deficit effected decrease in Anet and stomatal conductance, and did not change non-photochemical quenching and the de-epoxidation state of xanthophylls, dark respiration and photorespiration being alternative ways to dissipate the excess of energy. Little interactive effects between UV-B and drought were detected on photosynthesis performance, where the impact of UV-B was overshadowed by the effects of water deficit. Grape berry ripening was strongly delayed when UV-B and water deficit were applied in combination. In summary, deficit irrigation did not modify the adaptive response of grapevine to UV-B, through the accumulation of MEUVAC. However, combined treatments caused additive effects on berry ripening., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

Although plant performance under elevated CO2 has been extensively studied in the past little is known about photosynthetic performance changing simultaneously CO2, water availability and temperature conditions. Moreover, despite of its relevancy in crop responsiveness to elevated CO2 conditions, plant level C balance is a topic that, comparatively, has received little attention. In order to test responsiveness of grapevine photosynthetic apparatus to predicted climate change conditions, grapevine (Vitis vinifera L. cv. Tempranillo) fruit-bearing cuttings were exposed to different CO2 (elevated, 700ppm vs. ambient, ca. 400ppm), temperature (ambient vs. elevated, ambient +4°C) and irrigation levels (partial vs. full irrigation). Carbon balance was followed monitoring net photosynthesis (AN, C gain), respiration (RD) and photorespiration (RL) (C losses). Modification of environment (13)C isotopic composition (δ(13)C) under elevated CO2 (from -10.30 to -24.93‰) enabled the further characterization of C partitioning into roots, cuttings, shoots, petioles, leaves, rachides and berries. Irrespective of irrigation level and temperature, exposure to elevated CO2 induced photosynthetic acclimation of plants. C/N imbalance reflected the inability of plants grown at 700ppm CO2 to develop strong C sinks. Partitioning of labeled C to storage organs (main stem and roots) did not avoid accumulation of labeled photoassimilates in leaves, affecting negatively Rubisco carboxylation activity. The study also revealed that, after 20 days of treatment, no oxidative damage to chlorophylls or carotenoids was observed, suggesting a protective role of CO2 either at current or elevated temperatures against the adverse effect of water stress., (Copyright © 2014 Elsevier GmbH. All rights reserved.)

UV-B radiation and water deficit may trigger flavonol and anthocyanin biosynthesis in plant tissues. In addition, previous research has showed strong qualitative effects on grape berry skin flavonol and anthocyanin profiles in response to UV-B and water deficit. The aim of this study is to identify the mechanisms leading to quantitative and qualitative changes in flavonol and anthocyanin profiles, in response to separate and combined UV-B and water deficit. Grapevines (Vitis vinifera L. cv. Tempranillo) were exposed to three levels of UV-B radiation (0, 5.98 and 9.66 kJ m(-2) day(-1)) and subjected to two water regimes. A strong effect of UV-B on flavonol and anthocyanin biosynthesis was found, resulting in an increased anthocyanin concentration and a change in their profile. Concomitantly, two key biosynthetic genes (FLS1 and UFGT) were up-regulated by UV-B, leading to increased flavonol and anthocyanin skin concentration. Changes in flavonol and anthocyanin composition were explained to a large extend by transcript levels of F3'H, F3'5'H and OMT2. A significant interaction between UV-B and water deficit was found in the relative abundance of 3'4' and 3'4'5' substituted flavonols, but not in their anthocyanin homologues. The ratio between 3'4'5' and 3'4' substituted flavonols was linearly related to the ratios of F3'5'H and FLS1 transcription, two steps up-regulated independently by water deficit and UV-B radiation, respectively. Our results indicate that changes in flavonol profiles in response to environmental conditions are not only a consequence of changes in the expression of flavonoid hydroxylases; but also the result of the competition of FLS, F3'5'H and F3'H enzymes for the same flavonol substrates., (© The Author 2014. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Continued emissions of CO2, derived from human activities, increase atmospheric CO2 concentration. The CO2 rise stimulates plant growth and affects yield quality. Effects of elevated CO2 on legume quality depend on interactions with N2-fixing bacteria and mycorrhizal fungi. Growth at elevated CO2 increases photosynthesis under short-term exposures in C3 species. Under long-term exposures, however, plants generally acclimate to elevated CO2 decreasing their photosynthetic capacity. An updated survey of the literature indicates that a key factor, perhaps the most important, that characteristically influences this phenomenon, its occurrence and extent, is the plant source-sink balance. In legumes, the ability of exchanging C for N at nodule level with the N2-fixing symbionts creates an extra C sink that avoids the occurrence of photosynthetic acclimation. Arbuscular mycorrhizal fungi colonizing roots may also result in increased C sink, preventing photosynthetic acclimation. Defoliation (Anthyllis vulneraria, simulated grazing) or shoot cutting (alfalfa, usual management as forage) largely increases root/shoot ratio. During re-growth at elevated CO2, new shoots growth and nodule respiration function as strong C sinks that counteracts photosynthetic acclimation. In the presence of some limiting factor, the legumes response to elevated CO2 is weakened showing photosynthetic acclimation. This survey has identified limiting factors that include an insufficient N supply from bacterial strains, nutrient-poor soils, low P supply, excess temperature affecting photosynthesis and/or nodule activity, a genetically determined low nodulation capacity, an inability of species or varieties to increase growth (and therefore C sink) at elevated CO2 and a plant phenological state or season when plant growth is stopped., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

Human activities are increasing atmospheric CO2 concentration and temperature. Related to this global warming, periods of low water availability are also expected to increase. Thus, CO2 concentration, temperature and water availability are three of the main factors related to climate change that potentially may influence crops and ecosystems. In this report, we describe the use of growth chamber - greenhouses (GCG) and temperature gradient greenhouses (TGG) to simulate climate change scenarios and to investigate possible plant responses. In the GCG, CO2 concentration, temperature and water availability are set to act simultaneously, enabling comparison of a current situation with a future one. Other characteristics of the GCG are a relative large space of work, fine control of the relative humidity, plant fertirrigation and the possibility of light supplementation, within the photosynthetic active radiation (PAR) region and/or with ultraviolet-B (UV-B) light. In the TGG, the three above-mentioned factors can act independently or in interaction, enabling more mechanistic studies aimed to elucidate the limiting factor(s) responsible for a given plant response. Examples of experiments, including some aimed to study photosynthetic acclimation, a phenomenon that leads to decreased photosynthetic capacity under long-term exposures to elevated CO2, using GCG and TGG are reported., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

Grapevine cv. Tempranillo fruit-bearing cuttings were exposed to supplemental ultraviolet-B (UV-B) radiation under controlled conditions, in order to study its effect on grape traits, ripening, amino acids and flavonoid profile. The plants were exposed to two doses of UV-B biologically effective (5.98 and 9.66kJm(-2)d(-1)), applied either from fruit set to ripeness or from the onset of veraison to ripeness. A 0kJm(-2)d(-1) treatment was included as a control. UV-B did not significantly modify grape berry size, but increased the relative mass of berry skin. Time to reach ripeness was not affected by UV-B, which may explain the lack of changes in technological maturity. The concentration of must extractable anthocyanins, colour density and skin flavonols were enhanced by UV-B, especially in plants exposed from fruit set. The quantitative and qualitative profile of grape skin flavonols were modified by UV-B radiation. Monosubstituted flavonols relative abundance increased proportionally to the accumulated UV-B doses. Furthermore, trisubstituted forms, which where predominant in non-exposed berries, were less abundant as UV-B exposure increased. Although total free amino acid content remained unaffected by the treatments, the increased levels of gamma-aminobutyric acid (GABA), as well as the decrease in threonine, isoleucine, methionine, serine and glycine, revealed a potential influence of UV-B on the GABA-mediated signalling and amino acid metabolism. UV-B had an overall positive impact on grape berry composition., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

The present study aimed at evaluating the short- and long-term effects of UV-B radiation on leaves of grapevine Vitis vinifera (cv. Tempranillo). Grapevine fruit-bearing cuttings were exposed to two doses of supplemental biologically effective UV-B radiation (UV-BBE) under glasshouse-controlled conditions: 5.98 and 9.66kJm(-2)d(-1). The treatments were applied either for 20d (from mid-veraison to ripeness) or 75d (from fruit set to ripeness). A 0kJm(-2)d(-1) UV-B treatment was included as control. The main effects of UV-B were observed after the short-term exposure (20d) to 9.66kJm(-2)d(-1). Significant decreases in net photosynthesis, stomatal conductance, sub-stomatal CO2 concentration, the actual photosystem II (PSII) efficiency, total soluble proteins and de-epoxidation state of the VAZ cycle were observed, whereas the activities of several antioxidant enzymes increased significantly. UV-B did not markedly affect dark respiration, photorespiration, the maximum potential PSII efficiency (Fv/Fm), non-photochemical quenching (NPQ), as well as the intrinsic PSII efficiency. However, after 75d of exposure to 5.98and 9.66kJm(-2)d(-1) UV-B most photosynthetic and biochemical variables were unaffected and there were no sign of oxidative damage in leaves. The results suggest a high long-term acclimation capacity of grapevine to high UV-B levels, associated with a high accumulation of UV-B absorbing compounds in leaves, whereas plants seemed to be tolerant to moderate doses of UV-B., (Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.)

Elevated CO₂ leads to a decrease in potential net photosynthesis in long-term experiments and thus to a reduction in potential growth. This process is known as photosynthetic downregulation. There is no agreement on the definition of which parameters are the most sensitive for detecting CO₂ acclimation. In order to investigate the most sensitive photosynthetic and molecular markers of CO₂ acclimation, the effects of elevated CO₂, and associated elevated temperature were analyzed in alfalfa plants inoculated with different Sinorhizobium meliloti strains. Plants (Medicago sativa L. cv. Aragón) were grown in summer or autumn in temperature gradient greenhouses (TGG). At the end of the experiment, all plants showed acclimation in both seasons, especially under elevated summer temperatures. This was probably due to the lower nitrogen (N) availability caused by decreased N₂-fixation under higher temperatures. Photosynthesis measured at growth CO₂ concentration, rubisco in vitro activity and maximum rate of carboxylation were the most sensitive parameters for detecting downregulation. Severe acclimation was also related with decreases in leaf nitrogen content associated with declines in rubisco content (large and small subunits) and activity that resulted in a drop in photosynthesis. Despite the sensitivity of rubisco content as a marker of acclimation, it was not coordinated with gene expression, possibly due to a lag between gene transcription and protein translation., (© 2013 Institute of Botany, Chinese Academy of Sciences.)

Berry organoleptic properties are highly influenced by ripening environmental conditions. In this study, we used grapevine fruiting cuttings to follow berry ripening under different controlled conditions of temperature and irradiation intensity. Berries ripened at higher temperatures showed reduced anthocyanin accumulation and hastened ripening, leading to a characteristic drop in malic acid and total acidity. The GrapeGen GeneChip® combined with a newly developed GrapeGen 12Xv1 MapMan version were utilized for the functional analysis of berry transcriptomic differences after 2 week treatments from veraison onset. These analyses revealed the establishment of a thermotolerance response in berries under high temperatures marked by the induction of heat shock protein (HSP) chaperones and the repression of transmembrane transporter-encoding transcripts. The thermotolerance response was coincident with up-regulation of ERF subfamily transcription factors and increased ABA levels, suggesting their participation in the maintenance of the acclimation response. Lower expression of amino acid transporter-encoding transcripts at high temperature correlated with balanced amino acid content, suggesting a transcriptional compensation of temperature effects on protein and membrane stability to allow for completion of berry ripening. In contrast, the lower accumulation of anthocyanins and higher malate metabolization measured under high temperature might partly result from imbalance in the expression and function of their specific transmembrane transporters and expression changes in genes involved in their metabolic pathways. These results open up new views to improve our understanding of berry ripening under high temperatures.

Elevated CO(2) may decrease alfalfa forage quality and in vitro digestibility through a drop in crude protein and an enhancement of fibre content. The aim of the present study was to analyse the effect of elevated CO(2), elevated temperature and Sinorhizobium meliloti strains (102F78, 102F34 and 1032 GMI) on alfalfa yield, forage quality and in vitro dry matter digestibility. This objective is in line with the selection of S. meliloti strains in order to maintain high forage yield and quality under future climate conditions. Plants inoculated with the 102F34 strain showed more DM production than those inoculated with 1032GMI; however, these strains did not show significant differences with 102F78 plants. Neutral or acid detergent fibres were not enhanced in plants inoculated with the 102F34 strain under elevated CO(2) or temperature and hence, in vitro dry matter digestibility was unaffected. Crude protein content, an indicator of forage quality, was negatively related to shoot yield. Plants inoculated with 102F78 showed a similar shoot yield to those inoculated with 102F34, but had higher crude protein content at elevated CO(2) and temperature. Under these climate change conditions, 102F78 inoculated plants produced higher quality forage. However, the higher digestibility of plants inoculated with the 102F34 strain under any CO(2) or temperature conditions makes them more suitable for growing under climate change conditions. In general, elevated CO(2) in combination with high temperature (Climate Change scenario) reduced IVDMD and CP content and enhanced fibre content, which means that animal production will be negatively affected., (Copyright © 2012 Elsevier GmbH. All rights reserved.)

Photosynthetic carbon fixation (A(N) ) and photosynthetic electron transport rate (ETR) are affected by different environmental stress factors, such as those associated with climate change. Under stress conditions, it can be generated an electron excess that cannot be consumed, which can react with O₂, producing reactive oxygen species. This work was aimed to evaluate the influence of climate change (elevated CO₂, elevated temperature and moderate drought) on the antioxidant status of grapevine (Vitis vinifera) cv. Tempranillo leaves, from veraison to ripeness. The lowest ratios between electrons generated (ETR) and consumed (A(N) + respiration + photorespiration) were observed in plants treated with elevated CO₂ and elevated temperature. In partially irrigated plants under current ambient conditions, electrons not consumed seemed to be diverted to alternative ways. Oxidative damage to chlorophylls and carotenoids was not observed. However, these plants had increases in thiobarbituric acid reacting substances, an indication of lipid peroxidation. These increases matched well with an early rise of H₂O₂ and antioxidant enzyme activities, superoxide dismutase (EC 1.15.1.1), ascorbate peroxidase (EC 1.11.1.11) and catalase (EC 1.11.1.6). Enzymatic activities were maintained high until ripeness. In conclusion, plants grown under current ambient conditions and moderate drought were less efficient to cope with oxidative damage than well-irrigated plants, and more interestingly, plants grown under moderate drought but treated with elevated CO₂ and elevated temperature were not affected by oxidative damage, mainly because of higher rates of electrons consumed in photosynthetic carbon fixation., (Copyright © Physiologia Plantarum 2011.)

Future climatic conditions, including rising atmospheric CO(2) and temperature may increase photosynthesis and, consequently, plant production. A larger knowledge of legume performance under the predicted growth conditions will be crucial for safeguarding crop management and extending the area under cultivation with these plants in the near future. N(2) fixation is a key process conditioning plant responsiveness to varying growth conditions. Moreover, it is likely to increase under future environments, due to the higher photosynthate availability, as a consequence of the higher growth rate under elevated CO(2). However, as described in the literature, photosynthesis performance is frequently down-regulated (acclimated) under long-term exposure to CO(2), especially when affected by stressful temperature and water availability conditions. As growth responses to elevated CO(2) are dependent on sink-source status, it is generally accepted that down-regulation occurs in situations with insufficient plant C sink capacity. Alfalfa management involves the cutting of shoots, which alters the source-sink relationship and thus the photosynthetic behaviour. As the growth rate decreases at the end of the pre-cut vegetative growth period, nodulated alfalfa plants show photosynthetic down-regulation, but during regrowth following defoliation, acclimation to elevated CO(2) disappears. The shoot harvest also leads to a drop in mineral N uptake and C translocation to the roots, resulting in a reduction in N(2) fixation due to the dependence on photosynthate supply to support nodule function. Therefore, the production of new shoots during the first days following cutting requires the utilization of reduced C and N compounds that have been stored previously in reserve organs. The stored reserves are mediated by phytohormones such as methyl jasmonate and abscisic acid and in situations where water stress reduces shoot production this potentially enables the enhancement of taproot protein levels in nodulated alfalfa, which may lead to these plants being in better condition in the following cut/regrowth cycle. Furthering our knowledge of legume performance under predicted climate change conditions will be crucial for the development of varieties with better adaptation that will achieve greater and more efficient production values. Furthermore, for this purpose it will be necessary to improve existing methodologies and create new ones for phenotype characterization. Such knowledge will provide key information for future plant breeding programs., (Copyright © 2011 Elsevier GmbH. All rights reserved.)

Greenhouse experiments were conducted to compare stress effects caused by Verticillium dahliae and drought on gas exchange, chlorophyll (Chl) fluorescence and photosynthetic pigments of pepper plants. Three treatments were compared: Verticillium inoculated plants (+V), non-inoculated well-watered plants (-V) and non-inoculated plants subjected to progressive drought (D). Gas exchange, fluorescence and photosynthetic pigments were measured and represented along a gradient of relative water content (RWC) and stomatal conductance (g(s)). Net photosynthesis (A) and electron transport rate (ETR) decreased, as RWC and g(s) declined, similarly in both +V and D plants. However, dark respiration (R(D)) and photorespiration (R(L)) tended to increase in inoculated plants compared to those subjected to soil drought, as g(s) decreased. Photoinhibitory damage was not observed in infected or in droughted plants. Soil drought decreased intrinsic PSII efficiency (Phi(exc.)), which seemed to result in part from enhanced xanthophyll cycle- and/or lutein-related thermal energy dissipation. Nevertheless, the fact that 1-Phi(exc.) increased in D only at high values of the de-epoxidation state of the xanthophyll cycle (DPS) suggests that DeltapH could be the major factor controlling thermal energy dissipation in this treatment. By contrast, antheraxanthin, zeaxanthin and lutein, as well as Phi(exc.), were not markedly affected in +V. Water stress appeared to be the main limitation to photosynthesis in Verticillium infected plants, probably through stomatal closure, together with impaired mesophyll conductance (g(m)). However, our results indicate differential effects of V. dahliae on dark respiration, photorespiration, g(m) and on the capability of thermal energy dissipation under low g(s) values., (Copyright 2010 Elsevier GmbH. All rights reserved.)

Organic wastes such as sewage sludge have been successfully used to increase crop productivity of horticultural soils. Nevertheless, considerations of the impact of sludges on vegetable and fruit quality have received little attention. Therefore, the objective of the present work was to investigate the impact of two sanitized sewage sludges, autothermal thermophilic aerobic digestion (ATAD) and compost sludge, on the growth, yield, and fruit quality of pepper plants ( Capsicum annuum L. cv. Piquillo) grown in the greenhouse. Two doses of ATAD (15 and 30% v/v) and three of composted sludge (15, 30, and 45%) were applied to a peat-based potting mix. Unamended substrate was included as control. ATAD and composted sludge increased leaf, shoot, and root dry matter, as well as fruit yield, mainly due to a higher number of fruits per plant. There was no effect of sludge on fruit size (dry matter per fruit and diameter). The concentrations of Zn and Cu in fruit increased with the addition of sewage sludges. Nevertheless, the levels of these elements remained below toxic thresholds. Pepper fruits from sludge-amended plants maintained low concentrations of capsaicin and dihydrocapsaicin, thus indicating low pungency level, in accordance with the regulations prescribed by the Control Board of "Lodosa Piquillo peppers" Origin Denomination. The application of sludges did not modify the concentration of vitamin C (ASC) in fruit, whereas the highest doses of composted sludge tended to increase the content of reduced (GSH) and oxidized (GSSG) glutathione, without change in the GSH/GSSG ratio. There were no effects of sludge on the transcript levels of enzymes involved in the synthesis of vitamin C, l-galactono-1,4-lactone dehydrogenase (GLDH) or in the ascorbate-glutathione cycle, ascorbate peroxidase (APX), monodehydroascorbate reductase (MDAR), and glutathione reductase (GR). Results suggest that the synthesis and degradation of ASC and GSH were compensated for in most of the treatments assayed. The application of sanitized sludges to pepper plants can improve pepper yield without loss of food nutritional quality, in terms of fruit size and vitamin C, glutathione, and capsaicinoid contents.

Previous studies have shown that the arbuscular mycorrhizal fungus (AMF) Glomus deserticola (Trappe, Bloss and Menge) can diminish the negative effect of Verticillium dahliae Kleb. on pepper yield. On the other hand, it is known that AMF can be more beneficial for plant growth and physiology under dry conditions than when soil moisture is plentiful. Therefore, our objective was to assess if a moderate water deficit imposed on pepper plants before their inoculation with V. dahliae could improve the effectiveness of G. deserticola as biocontrol agent. In the present experiment, the delay in disease development in Verticillium-inoculated plants associated with AMF did not occur under well watered conditions. In addition, the establishment of mycorrhizal symbiosis and the development of structures by AMF were delayed when both symbiotic and pathogenic fungi infected the same root. Therefore, it is suggested that the equilibrium between pepper plant, G. deserticola and V. dahliae is so complex that small changes in competition between symbiotic and pathogenic fungi for host resources can modify the efficiency of AMF as a biocontrol agent. On the other hand, water deficit enhanced the deleterious effect of V. dahliae on fruit set and yield only when pepper plants were not associated with G. deserticola, which reinforces the idea that AMF may be more important for host plants subjected to stressful conditions. However, comparing well watered non-mycorrhizal and predroughted mycorrhizal plants, we found that moderate water deficit imposed before inoculation with V. dahliae did not improve the effectiveness of G. deserticola as a biocontrol agent.

Identification of molecular markers of monoterpenoid indole alkaloid (MIA) accumulation in cell-suspension cultures of Madagascar periwinkle (Catharanthus roseus (L.) G. Don) was performed by two-dimensional polyacrylamide gel electrophoresis. Comparison of the protein patterns from alkaloid-producing and non-producing cells showed the specific occurrence of a 28 kDa polypeptide restricted to cells accumulating MIAs. The polypeptide was purified by preparative two-dimensional gel electrophoresis, digested with trypsin, and microsequenced by the Edman degradation method. Cloning of the corresponding cDNA revealed that the protein which has been named CrPS (Catharanthus roseus Protein S) is a member of the alpha/beta hydrolase superfamily. Time-course expression studies by northern blot analysis confirmed that CrPS gene expression was associated with MIA accumulation in cell suspension cultures. In the whole plant, multicellular compartmentation is required for alkaloid biosynthesis. In situ mRNA hybridization on developing leaves revealed that CrPS mRNA and transcripts encoding the first enzymes of the MIA pathway were co-localized in internal phloem parenchyma cells. The possible implication of the alkaloid-accumulation associated protein CrPS in the signal transduction pathway leading to MIA production is discussed.