Micro-Tom Plants Research Articles

Transcriptional reprogramming in sound-treated Micro-Tom plants inoculated with Pseudomonas syringae pv. tomato DC3000.

Sound vibrations (SV) are known to influence molecular and physiological processes that can improve crop performance and yield. In this study, the effects of three audible frequencies (100, 500 and 1000 Hz) at constant amplitude (90 dB) on tomato Micro-Tom physiological responses were evaluated 1 and 3 days post-treatment. Moreover, the potential use of SV treatment as priming agent for improved Micro-Tom resistance to Pseudomonas syringae pv. tomato DC3000 was tested by microarray. Results showed that the SV-induced physiological changes were frequency- and time-dependent, with the largest changes registered at 1000 Hz at day 3. SV treatments tended to alter the foliar content of photosynthetic pigments, soluble proteins, sugars, phenolic composition, and the enzymatic activity of polyphenol oxidase, peroxidase, superoxide dismutase and catalase. Microarray data revealed that 1000 Hz treatment is effective in eliciting transcriptional reprogramming in tomato plants grown under normal conditions, but particularly after the infection with Pst DC3000. Broadly, in plants challenged with Pst DC3000, the 1000 Hz pretreatment provoked the up-regulation of unique differentially expressed genes (DEGs) involved in cell wall reinforcement, phenylpropanoid pathway and defensive proteins. In addition, in those plants, DEGs associated with enhancing plant basal immunity, such as proteinase inhibitors, pathogenesis-related proteins, and carbonic anhydrase 3, were notably up-regulated in comparison with non-SV pretreated, infected plants. These findings provide new insights into the modulation of Pst DC3000-tomato interaction by sound and open up prospects for further development of strategies for plant disease management through the reinforcement of defense mechanisms in Micro-Tom plants.

AP3 promotes the synthesis of condensed tannin in fruit by positively regulating ANR expression

Condensed tannins are often found in fruits and nuts and have an astringent flavor. The synthesis pathway of condensed tannins is already clear, but few related regulatory factors have been explored. Previous studies about MADS-box transcription factors have mainly focused on the regulation of floral organ development. Recent studies have shown that MADS-box are also involved in fruit development, maturation, and quality. The fruit of Quercus fabri is rich in starch and nutrients in its kernel but is difficult to eat directly because of its high condensed tannin content. This study identified and functionally characterized the MADS-box transcription factor QfAP3 in Q. fabri. Functional analysis based on overexpression in Micro-Tom showed that QfAP3 promoted condensed tannin synthesis. By analyzing the expression trend of key genes in the condensed tannin synthesis pathway in Micro-Tom plants, we found that the expression trend of ANR was consistent with that of QfAP3, and QfAP3 could bind to the promoter of ANR and positively regulate it. This study has discovered new functions of MADS-box transcription factors in fruit quality formation, developed new regulatory factors for the synthesis pathway of condensed tannin, and provided a biotechnological method that can effectively reduce astringency in fruit.

Effects of gamma radiation on engineered tomato biofortified for space agriculture by morphometry and fluorescence-based indices.

Future long-term space missions will focus to the solar system exploration, with the Moon and Mars as leading goals. Plant cultivation will provide fresh food as a healthy supplement to astronauts' diet in confined and unhealthy outposts. Ionizing radiation (IR) are a main hazard in outer space for their capacity to generate oxidative stress and DNA damage. IR is a crucial issue not only for human survival, but also for plant development and related value-added fresh food harvest. To this end, efforts to figure out how biofortification of plants with antioxidant metabolites (such as anthocyanins) may contribute to improve their performances in space outposts are needed. MicroTom plants genetically engineered to express the Petunia hybrida PhAN4 gene, restoring the biosynthesis of anthocyanins in tomato, were used. Seeds and plants from wild type and engineered lines AN4-M and AN4-P2 were exposed to IR doses that they may experience during a long-term space mission, simulated through the administration of gamma radiation. Plant response was continuously evaluated along life cycle by a non-disturbing/non-destructive monitoring of biometric and multiparametric fluorescence-based indices at both phenotypic and phenological levels, and indirectly measuring changes occurring at the primary and secondary metabolism level. Responses to gamma radiation were influenced by the phenological stage, dose and genotype. Wild type and engineered plants did not complete a seed-to-seed cycle under the exceptional condition of 30 Gy absorbed dose, but were able to cope with 0.5 and 5 Gy producing fruits and vital seeds. In particular, the AN4-M seeds and plants showed advantages over wild type: negligible variation of fluorimetric parameters related to primary metabolism, no alteration or improvement of yield traits at maturity while maintaining smaller habitus than wild type, biosynthesis of anthocyanins and maintained levels of these compounds compared to non-irradiated controls of the same age. These findings may be useful in understanding phenotypic effects of IR on plant growth in space, and lead to the exploitation of new breeding efforts to optimize plant performances to develop appropriate ideotypes for future long-term space exploration extending the potential of plants to serve as high-value product source.

Functional characterization of transcriptional activator gene SIARRI in tomato reveals its role in fruit growth and ripening.

Auxins regulate several characteristics of plant development and growth. Here, we characterized a new transcriptional activator SIARRI which binds specific DNA sequences and was revealed in Arabidopsis (ARR1). SIARRI acts as a two-component response regulator and its Arabidopsis homologous gene is AT3G16857. It belongs to the subfamily of type-B response regulators in the cytokinin signaling pathway. The study aimed to characterize the transgenic Micro-Tom plants by the overexpression of Solanum lycopersicum two-component response regulator ARR1. Overexpression of SIARRI results in a pleiotropic phenotype during fruit development and ripening. This study indicates that SIARRI is a primary regulator of leaf morphology and fruit development. Moreover, overexpressed plants showed variations in growth related to auxin as well as shorter hypocotyl elongation, enlarged leaf vascularization, and decreased apical dominance. The qRT-PCR investigation revealed that expression was downregulated at the breaker stage and high at Br+6 at various stages of fruit growth and ripening. In contrast to the fruit color, lycopene and β-carotene concentrations in red-yellow overexpression line fruits were reduced significantly, and also slightly reduced in some red fruits. The quantity of β-carotene in the transgenic fruits was lower than that of lycopene. This study showed that this gene might be a new transcriptional activator in fruit development and ripening. Furthermore, this study will provide new insights into tomato fruit ripening.

Tomato SlGSTU38 interacts with the PepMV coat protein and promotes viral infection.

Pepino mosaic virus (PepMV) is pandemic in tomato crops, causing important economic losses world-wide. No PepMV-resistant varieties have been developed yet. Identification of host factors interacting with PepMV proteins is a promising source of genetic targets to develop PepMV-resistant varieties. The interaction between the PepMV coat protein (CP) and the tomato glutathione S-transferase (GST) SlGSTU38 was identified in a yeast two-hybrid (Y2H) screening and validated by directed Y2H and co-immunoprecipitation assays. SlGSTU38-knocked-out Micro-Tom plants (gstu38) generated by the CRISPR/Cas9 technology together with live-cell imaging were used to understand the role of SlGSTU38 during infection. The transcriptomes of healthy and PepMV-infected wild-type (WT) and gstu38 plants were profiled by RNA-seq analysis. SlGSTU38 functions as a PepMV-specific susceptibility factor in a cell-autonomous manner and relocalizes to the virus replication complexes during infection. Besides, knocking out SlGSTU38 triggers reactive oxygen species accumulation in leaves and the deregulation of stress-responsive genes. SlGSTU38 may play a dual role: On the one hand, SlGSTU38 may exert a proviral function depending on its specific interaction with the PepMV CP; and on the other hand, SlGSTU38 may delay PepMV-infection sensing by participating in the redox intracellular homeostasis in a nonspecific manner.

Designing a novel tomato ideotype for future cultivation in space manned missions

Introduction: Methods for production of fresh, health food are needed in view of long-term, deep-space manned missions. To this end, crops tailored for better performance under non-terrestrial conditions may be obtained by the exploitation of biochemical patterns related to specialized metabolites known to confer protection against environmental challenges and to be beneficial to human health.Methods: In this work, for the first time, MicroTom plants have been engineered specifically for agrospace applications to express PhAN4, a MYB-like transcription factor able to regulate the biosynthesis of anthocyanins that influence tomato genes possibly involved in agrospace-relevant functions. Results:PhAN4 engineering underpinned the genetic background of the dwarf tomato MicroTom while maintaining yield and photosynthetic capacity. PhAN4 expression resulted in the accumulation of anthocyanins and polyphenols, a differential carotenoid profile, increased antioxidant scavenging capacities of fruits compared to the original genotype. Improved ability to counteract ROS generation and to preserve plant protein folding after ex-vivo gamma irradiation was observed.Discussion: These results highlights that the manipulation of specific metabolic pathways is a promising approach to design novel candidate varieties for agrospace applications.

Effect of Light Quality on Metabolomic, Ionomic, and Transcriptomic Profiles in Tomato Fruit.

Light quality affects plant growth and the functional component accumulation of fruits. However, there is little knowledge of the effects of light quality based on multiomics profiles. This study combined transcriptomic, ionomic, and metabolomic analyses to elucidate the effects of light quality on metabolism and gene expression in tomato fruit. Micro-Tom plants were grown under blue or red light-emitting diode light for 16 h daily after anthesis. White fluorescent light was used as a reference. The metabolite and element concentrations and the expression of genes markedly changed in response to blue and red light. Based on the metabolomic analysis, amino acid metabolism and secondary metabolite biosynthesis were active in blue light treatment. According to transcriptomic analysis, differentially expressed genes in blue and red light treatments were enriched in the pathways of secondary metabolite biosynthesis, carbon fixation, and glycine, serine, and threonine metabolism, supporting the results of the metabolomic analysis. Ionomic analysis indicated that the element levels in fruits were more susceptible to changes in light quality than in leaves. The concentration of some ions containing Fe in fruits increased under red light compared to under blue light. The altered expression level of genes encoding metal ion-binding proteins, metal tolerance proteins, and metal transporters in response to blue and red light in the transcriptomic analysis contributes to changes in the ionomic profiles of tomato fruit.

Overexpression of the mitochondrial MT-sHSP23.6 gene during antioxidant activity in tomato cv. Micro-Tom subjected to flooding

Plants grown under flooding stress conditions form reactive oxygen species, both during oxygen deficiency and after reoxygenation. The tolerance of plants under these conditions depends on an efficient antioxidant system, which may be influenced by the higher expression of heat shock proteins, such as mitochondrial MT-sHSP23.6. The objective of the present study was to determine whether the overexpression of this gene increases the antioxidant activity in tomato cv. Micro-Tom plants subjected to flooding during the vegetative stage of four fully expanded leaves. The experiment was performed with three genotypes of tomato plants (Solanum lycopersicum Mill. cv. Micro-Tom), one non-transformed denominated “WT” and two modified for high and low expression of the MT-sHSP23.6 gene, subjected to normal irrigation conditions, 72 hours of flooding, and 24 and 192 hours of recovery after the removal of flooding, consisting a 3x4 split-plot factorial design with four replications. Plants were collected at the vegetative stage of 4 fully expanded leaves to determine superoxide dismutase, ascorbate peroxidase and catalase activities, hydrogen peroxide accumulation and lipid peroxidation. Overexpression of the mitochondrial MT-sHSP23.6 gene in the evaluated plants resulted in higher antioxidant system activity. However, it did not necessarily lead to less damage caused by oxidative stress. Low expression of this gene resulted in lower antioxidant system activity and greater damage caused by reactive oxygen species

The auxin-producing Bacillus thuringiensis RZ2MS9 promotes the growth and modifies the root architecture of tomato (Solanum lycopersicum cv. Micro-Tom).

Strains of Bacillus thuringiensis (Bt) are commonly commercialized as bioinoculants for insect pest control, but their benefits go beyond their insecticidal property: they can act as plant growth-promoters. Auxins play a major role in the plant growth promotion. However, the mechanism of auxin production by the Bacilli group, and more specifically by Bt strains, is unclear. In previous work, the plant growth-promoting rhizobacterium (PGPR) B. thuringiensis strain RZ2MS9 increased the corn roots. This drew our attention to the strain's auxin production trait, earlier detected in vitro. Here, we demonstrate that in its genome, RZ2MS9 harbours the complete set of genes required in two pathways that are used for Indole acetic acid (IAA) production. We also detected that the strain produces almost five times more IAA during the stationary phase. The bacterial application increased the shoot dry weight of the Micro-Tom (MT) tomato by 24%. The application also modified MT root architecture, with an increase of 26% in the average lateral root length and inhibition of the axial root. At the cellular level, RZ2MS9-treated MT plants presented elongated root cortical cells with intensified mitotic activity. Altogether, these are the best characterized auxin-associated phenotypes. Besides that, no growth alteration was detected in the auxin-insensitive diageotropic (dgt) plants either with or without the RZ2MS9 inoculation. Our results suggest that auxins play an important role in the ability of B. thuringiensis RZ2MS9 to promote MT growth and provide a better understanding of the auxin production mechanism by a Bt strain.

Seed priming with ascorbic acid enhances salt tolerance in micro-tom tomato plants by modifying the antioxidant defense system components

Although ascorbic acid (AsA) has been known to improve plant defense system components against salt stressful-conditions, little is known about physiological and biochemical responses of AsA-mediated alleviation of salt stress in tomatoes. The main objective of this work was, therefore, to explore the role of ascorbic acid application via seed priming on antioxidant defense responses and growth and fruit yield regulation in tomato Micro-Tom under salt stress. The experiment was carried out using a 2 × 2 factorial completely randomized design with four replicates. In this way, seed priming with AsA (0 and 100 mM AsA) was used in Micro-Tom seeds to improve the tolerance of subsequent salt-stress exposure (0 and 100 mM NaCl). The AsA priming effect was assessed on subsequent exposure to salt stress conditions with a physiological and biochemical approaches, involving antioxidant responses. Overall results indicated that AsA plays a role in alleviating the negative effects of salt stress by increasing water potential and water use efficiency, reducing Na + content, lipid peroxidation and H 2 O 2 content, and increasing total pigments content, superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), peroxidase (POX), glutathione peroxidase (GPX) and glutathione reductase (GR) activities and GSH content, leading to better plant growth and higher yields. Thus, our results indicated that priming with AsA is an efficient management technique that can be used to alleviate the deleterious effects of salt stress and enhance the salt tolerance of Micro-Tom plants by modulation of antioxidant mechanisms. • Priming with AsA is a management technique to alleviate deleterious effects of salt-stressful. • Conditions. • Seed priming with AsA enhances salt tolerance in Micro-tom tomato plants by modifying the antioxidant defense system. • Salt tolerance conferred by seed priming can be associated with the modulation of ROS scavenging enzymes.

Comparative analysis of two phytochrome mutants of tomato (Micro-Tom cv.) reveals specific physiological, biochemical, and molecular responses under chilling stress

BackgroundPhytochromes are plant photoreceptors that have long been associated with photomorphogenesis in plants; however, more recently, their crucial role in the regulation of variety of abiotic stresses has been explored. Chilling stress is one of the abiotic factors that severely affect growth, development, and productivity of crops. In the present work, we have analyzed and compared physiological, biochemical, and molecular responses in two contrasting phytochrome mutants of tomato, namely aurea (aur) and high pigment1 (hp1), along with wild-type cultivar Micro-Tom (MT) under chilling stress. In tomato, aur is phytochrome-deficient mutant while hp1 is a phytochrome-sensitive mutant. The genotype-specific physiological, biochemical, and molecular responses under chilling stress in tomato mutants strongly validated phytochrome-mediated regulation of abiotic stress. ResultsHere, we demonstrate that phytochrome-sensitive mutant hp1 show improved performance compared to phytochrome-deficient mutant aur and wild-type MT plants under chilling stress. Interestingly, we noticed significant increase in several photosynthetic-related parameters in hp1 under chilling stress that include photosynthetic rate, stomatal conductance, stomatal aperture, transpiration rate, chlorophyll a and carotenoids. Whereas most parameters were negatively affected in aur and MT except a slight increase in carotenoids in MT plants under chilling stress. Further, we found that PSII quantum efficiency (Fv/Fm), PSII operating efficiency (Fq′/Fm′), and non-photochemical quenching (NPQ) were all positively regulated in hp1, which demonstrate enhanced photosynthetic performance of hp1 under stress. On the other hand, Fv/Fm and Fq′/Fm′ were decreased significantly in aur and wild-type plants. In addition, NPQ was not affected in MT but declined in aur mutant after chilling stress. Noticeably, the transcript analysis show that PHY genes which were previously reported to act as molecular switches in response to several abiotic stresses were mainly induced in hp1 and repressed in aur and MT in response to stress. As expected, we also found reduced levels of malondialdehyde (MDA), enhanced activities of antioxidant enzymes, and higher accumulation of protecting osmolytes (soluble sugars, proline, glycine betaine) which further elaborate the underlying tolerance mechanism of hp1 genotype under chilling stress. ConclusionOur findings clearly demonstrate that phytochrome-sensitive and phytochrome-deficient tomato mutants respond differently under chilling stress thereby regulating physiological, biochemical, and molecular responses and thus establish a strong link between phytochromes and their role in stress tolerance.

Characterization of the DREBA4-Type Transcription Factor (SlDREBA4), Which Contributes to Heat Tolerance in Tomatoes.

Dehydration-responsive element binding (DREB) transcription factors play crucial regulatory roles in abiotic stress. The only DREB transcription factor in tomato (Solanum lycopersicum), SlDREBA4 (Accession No. MN197531), which was determined to be a DREBA4 subfamily member, was isolated from cv. Microtom using high-temperature-induced digital gene expression (DGE) profiling technology. The constitutive expression of SlDREBA4 was detected in different tissues of Microtom plants. In addition to responding to high temperature, SlDREBA4 was up-regulated after exposure to abscisic acid (ABA), cold, drought and high-salt conditions. Transgenic overexpression and silencing systems revealed that SlDREBA4 could alter the resistance of transgenic Microtom plants to heat stress by altering the content of osmolytes and stress hormones, and the activities of antioxidant enzymes at the physiologic level. Moreover, SlDREBA4 regulated the downstream gene expression of many heat shock proteins (Hsp), as well as calcium-binding protein enriched in the pathways of protein processing in endoplasmic reticulum (ko04141) and plant-pathogen interaction (ko04626) at the molecular level. SlDREBA4 also induces the expression of biosynthesis genes in jasmonic acid (JA), salicylic acid (SA), and ethylene (ETH), and specifically binds to the DRE elements (core sequence, A/GCCGAC) of the Hsp genes downstream from SlDREBA4. This study provides new genetic resources and rationales for tomato heat-tolerance breeding and the heat-related regulatory mechanisms of DREBs.

Salinity impairs photosynthetic capacity and enhances carotenoid-related gene expression and biosynthesis in tomato (Solanum lycopersicum L. cv. Micro-Tom).

Carotenoids are essential components of the photosynthetic antenna and reaction center complexes, being also responsible for antioxidant defense, coloration, and many other functions in multiple plant tissues. In tomato, salinity negatively affects the development of vegetative organs and productivity, but according to previous studies it might also increase fruit color and taste, improving its quality, which is a current agricultural challenge. The fruit quality parameters that are increased by salinity are cultivar-specific and include carotenoid, sugar, and organic acid contents. However, the relationship between vegetative and reproductive organs and response to salinity is still poorly understood. Considering this, Solanum lycopersicum cv. Micro-Tom plants were grown in the absence of salt supplementation as well as with increasing concentrations of NaCl for 14 weeks, evaluating plant performance from vegetative to reproductive stages. In response to salinity, plants showed a significant reduction in net photosynthesis, stomatal conductance, PSII quantum yield, and electron transport rate, in addition to an increase in non-photochemical quenching. In line with these responses the number of tomato clusters decreased, and smaller fruits with higher soluble solids content were obtained. Mature-green fruits also displayed a salt-dependent higher induction in the expression of PSY1, PDS, ZDS, and LYCB, key genes of the carotenoid biosynthesis pathway, in correlation with increased lycopene, lutein, β-carotene, and violaxanthin levels. These results suggest a key relationship between photosynthetic plant response and yield, involving impaired photosynthetic capacity, increased carotenoid-related gene expression, and carotenoid biosynthesis.

Mechanisms of cadmium-stress avoidance by selenium in tomato plants.

Cadmium (Cd) is probably the most damaging metal to plant species; with a long biological half-life, it can be taken up by plants, disrupting the cell homeostasis and triggering several metabolic pathways. Selenium (Se) improves plant defence systems against stressful conditions, but the biochemical antioxidant responses to Cd stress in tomato plants is poorly understood. To further address the relationship of Cd-stress responses with Se mineral uptake, Cd and Se concentration, proline content, MDA and H2O2 production, and the activity of SOD, APX, CAT and GR enzymes were analyzed in Micro-Tom (MT) plants submitted to 0.5 mM Cd. The results revealed different responses according to Se combination and Cd application. For instance, roots and leaves of MT plants treated with Se exhibited an increase in dry mass and nutritional status, exhibited lower proline content and higher APX and GR activities when compared with plants with no Se application. Plants submitted to 0.5 mM Cd, irrespective of Se exposure, exhibited lower proline, MDA and H2O2 content and higher SOD, CAT and GR activities. Selenium may improve tolerance against Cd, which allowed MT plants exhibited less oxidative damage to the cell, even under elevated Cd accumulation in their tissues. The results suggest that Se application is an efficient management technique to alleviate the deleterious effects of Cd-stress, enhancing the nutritional value and activity of ROS-scavenging enzymes in tomato plants.

Linking phytohormones with growth, transport activity and metabolic responses to cadmium in tomato

This study investigated the role of phytohormones in the mediating effect of cadmium on growth and metabolism of tomato plants. Micro-Tom plants and its hormone mutants, procera (pro), diageotropica (dgt), epinastic (epi), and notabilis (not) have been used. Tomato seedlings were grown for 5 weeks under controlled hydroponics condition and then treated with CdCl2 concentration ranging from 0 to 100 μM for 2- to 6-week. Lipid peroxidation, leaf chlorophyll, elemental (Cd, Fe, Zn, Ca, Mg) content, and antioxidant enzyme activity were determined. Cd stress considerably altered the transcriptional level of the metal transport-related genes. Cadmium inhibited the uptake of iron (Fe) and zinc (Zn), whereas it has no noticeable effect on magnesium (Mg) and calcium (Ca) content, which might have been induced by the defense against Cd toxicity. The overall results demonstrated that not and epi mutants (with altered levels of ABA and ethylene, respectively) had a Cd-sensitive phenotype while dgt mutant (with reduced sensitivity to auxin) avoided damaging effects of Cd stress. This observation is explained by the lowest Cd accumulation in dgt roots. These results provide valuable information for the relative importance of auxin, ethylene, and abscisic acid in regulating plant responses to heavy metal toxicity.

Selenium improves photosynthesis and induces ultrastructural changes but does not alleviate cadmium-stress damages in tomato plants.

The application of Se to plants growing under Cd contamination may become an alternative strategy to minimize Cd damage. However, there is no specific information available regarding whether Se can affect the anatomical structure and photosynthetic rates of plants under Cd stress. To address questions related to Se-protective responses under Cd stress, we evaluated the structural and ultrastructural aspects, photosynthetic rates and growth of tomato cv. Micro-Tom plants. Plants were exposed to 0.5mM CdCl2 and further supplemented with 1.0μM of selenite or selenate. The overall results revealed different trends according to the Se source and Cd application. Both Se sources improved growth, photosynthesis, leaf characteristics and middle lamella thickness between mesophyll cells. In contrast, Cd caused decreases in photosynthesis and growth and damage to the ultrastructure of the chloroplast. The number of mitochondria, peroxisomes, starch grains and plastogloboli and the disorganization of the thylakoids and the middle lamella in plants increased in the presence of Cd or Cd + Se. Se plays an important role in plant cultivation under normal conditions. This finding was corroborated by the identification of specific structural changes in Se-treated plants, which could benefit plant development. However, a reversal of Cd stress effects was not observed in the presence of Se.

New insights into cadmium stressful-conditions: Role of ethylene on selenium-mediated antioxidant enzymes

Cadmium (Cd) contamination has generated an environmental problem worldwide, leading to harmful effects on human health and damages to plant metabolism. Selenium (Se) is non essential for plants, however it can improve plant growth and reduce the adverse effects of abiotic stress. In addition, ethylene may interplay the positive effects of Se in plants. In order to investigate the role of ethylene in Se-modulation of antioxidant defence system in response to Cd-stress, we tested the hormonal mutant Epinastic (epi) with a subset of constitutive activation of the ethylene response and Micro-Tom (MT) plants. For this purpose, Se mineral uptake, Cd and Se concentrations, pigments, malondialdeyde (MDA) and hydrogen peroxide (H2O2) contents, ethylene production, glutathione (GSH) compound, and superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT), glutathione reductase (GR) and glutathione peroxidase (GSH-Px) activities were analysed in MT and epi plants submitted to 0.5 mM CdCl2 and 1 μM of selenate or selenite. MT plants treated with both Se forms increased growth in the presence or not of 0.5 mM CdCl2, but not change epi growth. Both Se forms reduced Cd uptake in MT plants and cause reverse effect in epi plants. P, Mg, S, K and Zn uptake increased in epi plants with Se application, irrespective to Cd exposure. Chlorophylls and carotenoids contents decreased in both genotypes under Cd exposure, in contrast to what was observed in epi leaves in the presence of Se. When antioxidant enzymes activities were concerned, Se application increased Mn-SOD, Fe-SOD and APX activities. In the presence of Cd, MT and epi plants exhibited decreased SOD activity and increased CAT, APX and GR activities. MT and epi plants with Se supply exhibited increased APX and GR activities in the presence of Cd. Overall, these results suggest that ethylene may be involved in Se induced-defence responses, that triggers a positive response of the antioxidant system and improve growth under Cd stress. These results showed integrative roles of ethylene and Se in regulating the cell responses to stressful-conditions and, the cross-tolerance to stress could be used to manipulate ethylene regulated gene expression to induce heavy metal tolerance.

Amplification of gibberellins response in tomato modulates calcium metabolism and blossom end rot occurrence

Blossom end rot is one of the main physiological disorders of the tomato crop. Usually, the blossom end rot (BER) occurrence is associated with fruit calcium (Ca2+) deficiency; however, increasing evidence of Ca2+ disorders and hormonal regulation has emerged. Therefore, the present work was undertaken to investigate the role of gibberellin (GA) signaling on the control of tomato plant responses to Ca2+ deficiency. In this study, we used the tomato mutant procera (pro), which has a constitutive response to GA, and its isogenic line cv. Micro-Tom (MT) grown in a hydroponic system with or without Ca2+. Subsequently, the analysis of the growth of the shoot and the root system was performed, and the contents of photosynthetic pigments and oxidative stress were also analyzed. Furthermore, the fruit production, BER incidence, accumulation and metabolism of Ca2+ were also analyzed. The deficiency of Ca2+ reduced both shoot and root growth in both genotypes. Likewise, photosynthetic pigment degradation and oxidative stress were induced by Ca2+ deficiency. However, only pro plants exhibited the inhibition of fruit dry mass, as well as a higher incidence of BER, when exposed to Ca2+ deficiency compared to MT plants. In accordance with the greater pro susceptibility to Ca2+ deficiency, the efficiency of the transport, use and accumulation of Ca2+ in fruits was lower in pro when compared to the MT. Thus, we conclude that the higher sensitivity to GA in pro plants impairs Ca2+ metabolism and favors the occurrence of BER in tomato fruits, opening new approaches for the GA signaling on BER response.

Genome-wide transcriptomic analysis of BR-deficient Micro-Tom reveals correlations between drought stress tolerance and brassinosteroid signaling in tomato

Brassinosteroids (BRs) are plant steroid hormones that play crucial roles in a range of growth and developmental processes. Although BR signal transduction and biosynthetic pathways have been well characterized in model plants, their biological roles in an important crop, tomato (Solanum lycopersicum), remain unknown. Here, cultivated tomato (WT) and a BR synthesis mutant, Micro-Tom (MT), were compared using physiological and transcriptomic approaches. The cultivated tomato showed higher tolerance to drought and osmotic stresses than the MT tomato. However, BR-defective phenotypes of MT, including plant growth and stomatal closure defects, were completely recovered by application of exogenous BR or complementation with a SlDWARF gene. Using genome-wide transcriptome analysis, 619 significantly differentially expressed genes (DEGs) were identified between WT and MT plants. Several DEGs were linked to known signaling networks, including those related to biotic/abiotic stress responses, lignification, cell wall development, and hormone responses. Consistent with the higher susceptibility of MT to drought stress, several gene sets involved in responses to drought and osmotic stress were differentially regulated between the WT and MT tomato plants. Our data suggest that BR signaling pathways are involved in mediating the response to abiotic stress via fine-tuning of abiotic stress-related gene networks in tomato plants.

MicroRNA159-targeted SlGAMYB transcription factors are required for fruit set in tomato.

The transition from flowering to fruit production, namely fruit set, is crucial to ensure successful sexual plant reproduction. Although studies have described the importance of hormones (i.e. auxin and gibberellins) in controlling fruit set after pollination and fertilization, the role of microRNA-based regulation during ovary development and fruit set is still poorly understood. Here we show that the microRNA159/GAMYB1 and -2 pathway (the miR159/GAMYB1/2 module) is crucial for tomato ovule development and fruit set. MiR159 and SlGAMYBs were expressed in preanthesis ovaries, mainly in meristematic tissues, including developing ovules. SlMIR159-overexpressing tomato cv. Micro-Tom plants exhibited precocious fruit initiation and obligatory parthenocarpy, without modifying fruit shape. Histological analysis showed abnormal ovule development in such plants, which led to the formation of seedless fruits. SlGAMYB1/2 silencing in SlMIR159-overexpressing plants resulted in misregulation of pathways associated with ovule and female gametophyte development and auxin signalling, including AINTEGUMENTA-like genes and the miR167/SlARF8a module. Similarly to SlMIR159-overexpressing plants, SlGAMYB1 was downregulated in ovaries of parthenocarpic mutants with altered responses to gibberellins and auxin. SlGAMYBs likely contribute to fruit initiation by modulating auxin and gibberellin responses, rather than their levels, during ovule and ovary development. Altogether, our results unveil a novel function for the miR159-targeted SlGAMYBs in regulating an agronomically important trait, namely fruit set.