Rubén Bottini

Gibberellin production by bacteria and its involvement in plant growth promotion and yield increase

This review focuses on studies with bacteria for which biosynthesis/production of the plant hormones gibberellins have been demonstrated. Actual data on gibberellin metabolism by bacteria are analyzed in comparison with the biosynthetic pathways known for vascular plants and fungi. The potential involvement of gibberellins produced by symbiotic and soil-endophytic microorganisms in plant growth promotion and yield increase is also discussed.

Production of indole-3-acetic acid and gibberellins A1 and A3 by Acetobacter diazotrophicus and Herbaspirillum seropedicae in chemically-defined culture media

The characterization by capillary gas chromatography-mass spectrometry of the plant hormones indole-3-acetic acid and the gibberellins GA1 and GA3 from chemically-defined cultures of Acetobacter diazotrophicus and Herbaspirillum seropedicae is reported. Both bacteria are endophytic in gramineae species where they promote growth and yield. Quantification was also done by selected ion monitoring with [17,17-2H2]-Gibberellin A1, [17,17-2H2]-Gibberellin A3 and [13C6]-indole-3-acetic acid as internal standards. The results presented show the importance of studying phytohormonal production when the interrelationships between plants and microorganisms are analyzed and may help explain the beneficial effects of endophytic bacteria to the host plant, as has been demonstrated previously for Azospirillum spp.

Grape pomace as a sustainable source of bioactive compounds: extraction, characterization, and biotechnological applications of phenolics.

Grape pomaces (GPs) are characterized by high contents of phenolics due to an incomplete extraction during the winemaking process. These phenolics are secondary plant metabolites with potential beneficial effects on human health because of their antioxidant activity and antimicrobial, antiviral, and anti-inflammatory properties. Therefore, GP constitutes an inexpensive source for the extraction of phytochemicals that can be used in the pharmaceutical, cosmetic, and food industries. As a result of the increased attention to sustainability of agricultural practices, efforts have been made to use GP in different fields of industry. Thus, it is necessary to have efficient extraction techniques to achieve good recoveries of compounds. In this respect, sensitive and selective analytical methods have been tried for the characterization of phenolic extracts. This review summarizes the most recent developments in the extraction of polyphenols from GPs. Furthermore, the techniques used for characterization of extracts are explained, with emphasis on sample preparation, separation, and analysis of phenolics. Finally, the possible applications of GP extracts in diverse biotechnological fields are also discussed.

Abscisic acid is involved in the response of grape (Vitis vinifera L.) cv. Malbec leaf tissues to ultraviolet-B radiation by enhancing ultraviolet-absorbing compounds, antioxidant enzymes and membrane sterols.

We investigated the interactions of abscisic acid (ABA) in the responses of grape leaf tissues to contrasting ultraviolet (UV)-B treatments. One-year-old field-grown plants of Vitis vinifera L. were exposed to photosynthetically active radiation (PAR) where solar UV-B was eliminated by using polyester filters, or where PAR was supplemented with UV-B irradiation. Treatments combinations included weekly foliar sprays of ABA or a water control. The levels of UV-B absorbing flavonols, quercetin and kaempferol were significantly decreased by filtering out UV-B, while applied ABA increased their content. Concentration of two hydroxycinnamic acids, caffeic and ferulic acids, were also increased by ABA, but not affected by plus UV-B (+UV-B) treatments. Levels of carotenoids and activities of the antioxidant enzymes, catalase, ascorbate peroxidase and peroxidase were elevated by +ABA treatments, but only if +UV-B was given. Cell membrane beta-sitosterol was enhanced by ABA independently of +UV-B. Changes in photoprotective compounds, antioxidant enzymatic activities and sterols were correlated with lessened membrane harm by UV-B, as assessed by ion leakage. Oxidative damage expressed as malondialdehyde content was increased under +UV-B treatments. Our results suggest that the defence system of grape leaf tissues against UV-B is activated by UV-B irradiation with ABA acting downstream in the signalling pathway.

Transcriptome changes in grapevine (Vitis vinifera L.) cv. Malbec leaves induced by ultraviolet-B radiation

BackgroundUltraviolet-B radiation (UV-B, 280-315 nm) is a natural component of sunlight, which has numerous regulatory effects on plant physiology. The nature of the response to UV-B is dependent on fluence rate, dose, duration and wavelength of the UV-B treatment. Some reports have analyzed the changes in gene expression caused by UV-B light on several plant species using microarray technology. However, there is no information on the transcriptome response triggered by UV-B in grapevine. In this paper we investigate the gene expression responses of leaves from in vitro cultured Vitis vinifera cv. Malbec plants subjected to the same dose of biologically effective UV-B radiation (4.75 kJ m-2 d-1) administered at two different fluence rates (16 h at ≅ 8.25 μW cm-2, 4 h at ≅ 33 μW cm-2) using a new custom made GrapeGen Affymetrix GeneChip®.ResultsThe number of genes modulated by high fluence rate UV-B doubled the number of genes modulated by low fluence UV-B. Their functional analyses revealed several functional categories commonly regulated by both UV-B treatments as well as categories more specifically modulated depending on UV-B fluence rate. General protective responses, namely the induction of pathways regulating synthesis of UV-B absorbing compounds such as the Phenylpropanoid pathway, the induction of different antioxidant defense systems and the activation of pathways commonly associated with pathogen defense and abiotic stress responses seem to play critical roles in grapevine responses against UV-B radiation. Furthermore, high fluence rate UV-B seemed to specifically modulate additional pathways and processes in order to protect grapevine plantlets against UV-B-induced oxidative stress, stop the cell cycle progression, and control protein degradation. On the other hand, low fluence rate UV-B regulated the expression of specific responses in the metabolism of auxin and abscisic acid as well as in the modification of cell walls that could be involved in UV-B acclimation-like processes.ConclusionOur results show the UV-B radiation effects on the leaf transcriptome of grapevine (Vitis vinifera cv. Malbec) plantlets. Functional categories commonly modulated under both UV-B treatments as well as transcripts specifically regulated in an UV-B-intensity dependent way were identified. While high fluence rate UV-B had regulatory effects mainly on defense or general multiple-stress responses pathways, low fluence rate UV-B promoted the expression of genes that could be involved in UV-B protection or the amelioration of the UV-B-induced damage. This study also provides an extensive list of genes regulating multiple metabolic pathways involved in the response of grapevine to UV-B that can be used for future researches.

Azospirillum sp. Promotes Root Hair Development in Tomato Plants through a Mechanism that Involves Ethylene

Tomato seeds were inoculated with the plant growth–promoting rhizobacteria Azospirillum brasilense FT326, and changes in parameters associated with plant growth were evaluated 15 days after inoculation. Azospirilla were localized on roots and within xylematic tissue. An increase in shoot and root fresh weight, main root hair length, and root surface indicated that inoculation with A. brasilense FT 326 resulted in plant growth improvement. The levels of indole-3-acetic acid (IAA) and ethylene, two of the phytohormones related to plant growth, were higher in inoculated plants. Exogenously supplied ethylene mimicked the effect of inoculation, and the addition of an inhibitor of its synthesis or of its physiological activity completely blocked A. brasilense growth promotion. Based on our results, we propose that the process of growth promotion triggered by A. brasilense inoculation involves a signaling pathway that has ethylene as a central, positive regulator.

Solar UV-B and ABA Are Involved in Phenol Metabolism of Vitis vinifera L. Increasing Biosynthesis of Berry Skin Polyphenols

It has been previously found that abscisic acid (ABA) participates in the activation of grapevine leaf tissue defense against potentially damaging effects of solar ultraviolet-B radiation (UV-B), apparently by triggering biosynthesis of phenols that filter the harmful radiation and act as antioxidants. The present work studies the effect of solar UV-B and exogenously applied ABA on berry growth, sugar accumulation, and phenol (anthocyanin and nonanthocyanin) profiles across berry development and ripening of Vitis vinifera L. cv. Malbec in a vineyard at 1450 m of altitude. The grapevines were exposed to relatively high UV-B irradiation (normal sunlight; +UV-B) and also to a reduced UV-B treatment (filter exclusion; -UV-B). These two UV-B treatments were combined with weekly spray applications to the leaves and berries of 1 mM ABA (+ABA) or H(2)O (-ABA). Reduction of UV-B delayed berry development and maturation, whereas the +UV-B and +ABA combined treatment hastened berry sugar and phenol accumulation. +UV-B/+ABA treatments also reduced berry growth and decreased sugar per berry without affecting sugar concentration (°Brix) at harvest. Berry skin ABA levels were higher in the +UV-B and +ABA combined treatment, which also hastened the onset of ripening up to 20 days. Berry skin ABA levels then decreased toward harvest, implying a possible role for ABA in the control of ripening in this nonclimacteric fruit. Under both +UV-B and +ABA treatments berry skin phenols were additively increased with a change in anthocyanin and nonanthocyanin profiles and increases in the proportion of phenols with high antioxidant capacity.

Exogenous Abscisic Acid Increases Carbohydrate Accumulation and Redistribution to the Grains in Wheat Grown Under Field Conditions of Soil Water Restriction

This work investigates the effects of abscisic acid (ABA) on physiologic parameters related to yield in wheat (Triticum aestivum) grown under field conditions with water restriction ranging between 45.7% and 49.5% of field capacity during anthesis and postanthesis. ABA (300 mg L−1) was sprayed onto the plants at the beginning of shoot lengthening which significantly promoted leaf area and higher concentrations of chlorophylls and carotenoids in flag leaf at anthesis. ABA also increased soluble carbohydrates in shoots at anthesis, which were then re-exported to the grains at maturity. This correlated with a yield increase that was achieved by a higher number and weight of grains per spike, but protein content was not significantly affected.

Bacteria isolated from roots and rhizosphere of Vitis vinifera retard water losses, induce abscisic acid accumulation and synthesis of defense-related terpenes in in vitro cultured grapevine.

Eleven bacterial strains were isolated at different soil depths from roots and rhizosphere of grapevines from a commercial vineyard. By 16S rRNA gene sequencing 10 different genera and 8 possible at species level were identified. From them, Bacillus licheniformis Rt4M10 and Pseudomonas fluorescens Rt6M10 were selected according to their characteristics as plant growth promoting rhizobacteria (PGPR). Both produced abscisic acid (ABA), indole-3-acetic acid (IAA) and the gibberellins A1 and A3 in chemically-defined medium. They also colonized roots of in vitro grown Vitis vinifera cv. Malbec plants. As result of bacterization ABA levels in 45 days-old in vitro plants were increased 76-fold by B. licheniformis and 40-fold by P. fluorescens as compared to controls. Both bacteria diminished plant water loss rate in correlation with increments of ABA. Twenty and 30 days post bacterization the plants incremented terpenes. The monoterpenes α-pinene, terpinolene, 4-carene, limonene, eucalyptol and lilac aldehyde A, and the sesquiterpenes α-bergamotene, α-farnesene, nerolidol and farnesol were assessed by gas chromatography-electron impact mass spectrometry analysis. α-Pinene and nerolidol were the most abundant (µg per g of tissue in plants bacterized with P. fluorescens). Only α-pinene, eucalyptol and farnesol were identified at low concentration in non-bacterized plants treated with ABA, while no terpenes were detected in controls. The results obtained along with others from literature suggest that B. licheniformis and P. fluorescens act as stress alleviators by inducing ABA synthesis so diminishing water losses. These bacteria also elicit synthesis of compounds of plant defense via an ABA independent mechanism.

Azospirillum brasilense ameliorates the response of Arabidopsis thaliana to drought mainly via enhancement of ABA levels

Production of phytohormones is one of the main mechanisms to explain the beneficial effects of plant growth-promoting rhizobacteria (PGPR) such as Azospirillum sp. The PGPRs induce plant growth and development, and reduce stress susceptibility. However, little is known regarding the stress-related phytohormone abscisic acid (ABA) produced by bacteria. We investigated the effects of Azospirillum brasilense Sp 245 strain on Arabidopsis thaliana Col-0 and aba2-1 mutant plants, evaluating the morphophysiological and biochemical responses when watered and in drought. We used an in vitro-grown system to study changes in the root volume and architecture after inoculation with Azospirillum in Arabidopsis wild-type Col-0 and on the mutant aba2-1, during early growth. To examine Arabidopsis development and reproductive success as affected by the bacteria, ABA and drought, a pot experiment using Arabidopsis Col-0 plants was also carried out. Azospirillum brasilense augmented plant biomass, altered root architecture by increasing lateral roots number, stimulated photosynthetic and photoprotective pigments and retarded water loss in correlation with incremented ABA levels. As well, inoculation improved plants seed yield, plants survival, proline levels and relative leaf water content; it also decreased stomatal conductance, malondialdehyde and relative soil water content in plants submitted to drought. Arabidopsis inoculation with A. brasilense improved plants performance, especially in drought.