CRYPTOCHROME 1a-mediated blue light perception regulates tomato seed germination via changes in hormonal balance and endosperm-degrading hydrolase dynamics.

MAIN CONCLUSION: Blue light exposure delays tomato seed germination by decreasing endosperm-degrading hydrolase activities, a process regulated by CRY1a-dependent signaling and the hormonal balance between ABA and GA. The germination of tomato seeds (Solanum lycopersicum L.) is tightly controlled by an internal hormonal balance, which is also influenced by environmental factors such as light. In this study, we investigated the blue light (BL)-mediated impacts on physiological, biochemical, and molecular processes during the germination of the blue light photoreceptor CRYPTOCHROME 1a loss-of-function mutant (cry1a) and of the hormonal tomato mutants notabilis (not, deficient in ABA) and procera (pro, displaying a GA-constitutive response). Seeds were germinated in a controlled chamber in the dark and under different intensities of continuous BL (ranging from 1 to 25 µmol m-2 s-1). In general, exposure to BL delayed tomato seed germination in a fluency rate-dependent way due to negative impacts on the activities of endosperm-degrading hydrolases, such as endo-ß-mannanase, ß-mannosidase, and α-galactosidase. However, not and pro mutants presented higher germination speed index (GSI) compared to WT despite the BL influence, associated with higher hydrolase activities, especially evident in pro, indicating that the ABA/GA hormonal balance is important to diminish BL inhibition over tomato germination. The cry1a germination percentage was higher than in WT in the dark but its GSI was lower under BL exposure, suggesting that functional CRY1a is required for BL-dependent germination. BL inhibits the expression of GA-biosynthetic genes, and induces GA-deactivating and ABA-biosynthetic genes. The magnitude of the BL influence over the hormone-related transcriptional profile is also dependent upon CRY1a, highlighting the complex interplay between light and hormonal pathways. These results contribute to a better understanding of BL-induced events behind the photoregulation of tomato seed germination.

Effect of Indole-3-Acetic Acid on Tomato Plant Growth.

Plant growth-promoting bacteria have several abilities to promote plant growth and development. One of these skills is the synthesis of indole-3-acetic acid (IAA), which mainly promotes root and shoot development. The bacteria Bacillus subtilis and Azospirillum brasilense have been widely used in agriculture with this function. However, little is known about whether the joint inoculation of these bacteria can reduce plant development by the excess of IAA produced as a result of the joint inoculation. The objective of the present study was to verify the effect of IAA on the inoculation of B. subtilis and A. brasilense in three tomato genotypes. The Micro-Tom genotype without mutation for IAA synthesis, Entire, has high sensitivity to IAA, and the diageotropic genotype (dgt) has low sensitivity to IAA. The results show that the plant parameter most sensitive to microbial inoculation is the number of roots. No treatment increased the shoot dry mass parameters for the Micro-Tom genotype and dgt, root dry mass for the Micro-Tom genotype, plant height for the Micro-Tom and Entire genotypes, root area and root volume for the genotype dgt. The Azm treatment reduced plant height compared to the control in the dgt, the BS + Azw and BS + Azm treatments in the Micro-Tom genotype and the Azw + Azm treatment in the dgt genotype reduced the plant diameter compared to the control. BS and BS + Azw reduced the number of roots in the Micro-Tom. The results strongly support that the mixture of B. subtilis and A. brasilense can reduce some parameters of plant development; however, this effect is possibly an interference in the mode of action of growth promotion of each isolate and is not related to an excess of IAA produced by the bacteria.

Tomato phytochromes B1 and B2 are part of the responses to the nutritional stress induced by NPK deficiency.

Phytochromes are red-light photoreceptors that play an important role in regulating many responses of plants, including its nutritional control. Nutrient deficiency in plants has become a constraint for agricultural production; thus, we investigated the role of phytochromes B1 and B2 in the nutritional, physiological, and growth changes of the control genotype (WT) and both phyB1 and phyB2 tomato mutants (deficient in phyB1 and phyB2) under nutritional sufficiency and individual deficiency of N, P, and K. Under complete solution, the plants of phyB1 and phyB2 had a decreased N, P, and K accumulation compared with WT and consequently a reduced content of chlorophyll and carotenoids, and dry weight production. In the condition of N deficiency, phyB1 had decreased N absorption, pigments concentration, and plant dry weight, while increased oxidative stress of membranes (MDA content). Similarly, phyB2 also had reduced N absorption. The deficiency of phyB1 mitigated the effects of P deficiency as phyB1 mutant had improved nutritional and physiological responses, increasing plant dry weight production. In contrast, phyB2 reduced N accumulation, quantum efficiency of photosystem II (Fv/Fm), and the concentration of pigments, while it increased MDA. Under K deficiency, phyB1 displayed a reduced P accumulation, as well as the total concentration of chlorophylls and carotenoids and K use efficiency. An increased concentration of MDA was found in phyB2 plants, as well as a reduction in chlorophylls concentration and in the use efficiency of K. Together, these results indicate a new perspective on the control of phytochromes in the nutrition of tomato plants under nutritional stress.

Cryptochrome 1a depends on blue light fluence rate to mediate osmotic stress responses in tomato.

The participation of plant cryptochromes in water deficit response mechanisms has been highlighted in several reports. However, the role of tomato (Solanum lycopersicum L.) cryptochrome 1a (cry1a) in the blue light fluence-dependent modulation of the water deficit response remains largely elusive. The tomato cry1a mutant and its wild-type counterpart were grown in water (no stress) or PEG6000 (osmotic stress) treatments under white light (60 µmol m-2 s-1) or from low to high blue light fluence (1, 5, 10, 15 and 25 µmol m-2 s-1). We first demonstrate that under nonstress conditions cry1a regulates seedling growth by mechanisms that involve pigmentation, lipid peroxidation and osmoprotectant accumulation in a blue light-dependent manner. In addition, we further highlighted under osmotic stress conditions that cry1a increased tomato growth by reduced malondialdehyde (MDA) and proline accumulation. Although blue light is an environmental signal that influences osmotic stress responses mediated by tomato cry1a, specific blue light fluence rates are required during these responses. Here, we show that CRY1a manipulation may be a potential biotechnological target to develop a drought-tolerant tomato variety. Nevertheless, the complete understanding of this phenomenon requires further investigation.

The auxin-resistant dgt tomato mutant grows less than the wild type but is less sensitive to ammonium toxicity and nitrogen deficiency.

The low-auxin-sensitivity tomato mutant, dgt, despite displaying reduced plant growth, has been linked to greater resistance to N deficiency. This led us to test the role of auxin resistance of dgt in NH4+ toxicity and N deficiency, compared to wild type tomato (cv. Micro-Tom, MT), grown in hydroponic media. A completely randomized design with three replications in a 2 × 4 factorial scheme was adopted, corresponding to the two tomato genotypes (MT and dgt), involving four nutritional treatments: NO3- (5 mM); NH4+ (5 mM); NO3- (5 mM) plus exogenous auxin (10 µM IAA); and N omission. The results show that NH4+ was toxic to MT but not to dgt. Under N deficiency, MT displayed a lower shoot NO3- content, a lower photosynthetic rate, and a decrease in both shoot and root dry weight. However, in dgt, no difference was observed in shoot NO3- content and photosynthetic rate between plants grown on NO3- or under N deficiency. In addition, dgt showed an increase in shoot dry weight under N deficiency. We highlight the role of auxin resistance in the adaptation of plants to NH4+ toxicity and N deficiency.

Cryptochrome-Related Abiotic Stress Responses in Plants.

It is well known that light is a crucial environmental factor that has a fundamental role in plant growth and development from seed germination to fruiting. For this process, plants contain versatile and multifaceted photoreceptor systems to sense variations in the light spectrum and to acclimate to a range of ambient conditions. Five main groups of photoreceptors have been found in higher plants, cryptochromes, phototropins, UVR8, zeitlupes, and phytochromes, but the last one red/far red wavelengths photoreceptor is the most characterized. Among the many responses modulated by phytochromes, these molecules play an important role in biotic and abiotic stress responses, which is one of the most active research topics in plant biology, especially their effect on agronomic traits. However, regarding the light spectrum, it is not surprising to consider that other photoreceptors are also part of the stress response modulated by light. In fact, it has become increasingly evident that cryptochromes, which mainly absorb in the blue light region, also act as key regulators of a range of plant stress responses, such as drought, salinity, heat, and high radiation. However, this information is rarely evidenced in photomorphogenetic studies. Therefore, the scope of the present review is to compile and discuss the evidence on the abiotic stress responses in plants that are modulated by cryptochromes.

Constitutive gibberellin response in grafted tomato modulates root-to-shoot signaling under drought stress.

Plants are sessile organisms that must perceive and respond to various environmental constraints throughout their life cycle. Among these constraints, drought stress has become the main limiting factor to crop production around the world. Water deprivation is perceived primarily by the roots, which efficiently signal the shoot to trigger drought responses in order to maximize a plant's ability to survive. In this study, the tomato (Solanum lycopersicum L.) mutant procera (pro), with a constitutive response to gibberellin (GA), and its near isogenic line cv. Micro-Tom (MT), were used in reciprocal grafting under well-watered and water stress conditions to evaluate the role of GA signaling in root-to-shoot communication during drought stress. Growth, oxidative stress, gene expression, water relations and hormonal content were measured in order to provide insights into GA-mediated adjustments to water stress. All graft combinations with pro (i.e. pro/pro, MT/pro and pro/MT) prevented the reduction of growth under stress conditions without a reduction in oxidative stress. The increase of oxidative stress was followed by upregulation of SlDREB2, a drought-tolerance related gene, in all drought-stressed plants. Scions harboring the pro mutation tended to increase the abscisic acid (ABA) content, independent of the rootstock. Moreover, the GA sensitivity of the rootstock modulated stomatal conductance and water use efficiency under drought stress, indicating GA and ABA crosstalk in the adjustment of growth and water economy.

Cadmium stress antioxidant responses and root-to-shoot communication in grafted tomato plants.

Many aspects related to ROS modulation of signaling networks and biological processes that control stress responses still remain unanswered. For this purpose, the grafting technique may be a powerful tool to investigate stress signaling and specific responses between plant organs during stress. In order to gain new insights on the modulation of antioxidant stress responses mechanisms, gas-exchange measurements, lipid peroxidation, H2O2 content, proline, superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), ascorbate peroxidase (APX) and guaiacol peroxidase (GPOX) were analyzed in Micro-Tom grafted plants submitted to cadmium (Cd). The results observed revealed that higher amounts of Cd accumulated mainly in the roots and rootstocks when compared to leaves and scions. Macronutrients uptake (Ca, S, P and Mg) decreased in non-grafted plants, but differed among plant parts in all grafted plants. The results showed that the accumulation of proline observed in scions of grafted plants could be associated to the lower MDA contents in the scions of grafted plants. In the presence of Cd, non-grafted plants displayed increased CAT, GR, GPOX and APX activities for both tissues, whilst grafted plants revealed distinct trends that clearly indicate signaling responses from the rootstocks, allowing sufficient time to activate defense mechanisms in shoot. The information available concerning plants subjected to grafting can provide a better understanding of the mechanisms of Cd detoxification involving root-to-shoot signaling, opening new possibilities on strategies which can be used to manipulate heavy metal tolerance, since antioxidant systems are directly involved in such mechanism.

Compostos fenólicos e capacidade antioxidante em frutos de tomateiros mutantes fotomorfogenéticos / Phenolic compounds and antioxidant capacity in fruits of photomorphogenic mutant tomato

Compostos fenólicos e capacidade antioxidante são mecanismos de defesa das plantas aos danos do estresse oxidativo. Os compostos fenólicos são sintetizados pela via dos fenilpropanoides, cuja enzima chave, fenilalanina amônia liase, é influenciada pela luz e ação de fotorreceptores, como o fitocromo. O objetivo do presente trabalho é avaliar a concentração de compostos fenólicos e a capacidade antioxidante de frutos de microtomateiro selvagem, cultivar "Micro-Tom" (MT), e seus mutantes fotomorfogenéticos high pigment 1 (hp1), super-responsivo a eventos mediados por luz e aurea (au), deficiente quantitativo em fitocromos. Vinte frutos maduros de cada genótipo (MT, hp1 e au) foram utilizados para as análises, realizadas em triplicata. Para quantificação dos compostos fenólicos totais, foi utilizado o método de Folin-Ciocalteu e a capacidade antioxidante foi realizada pelos métodos Ferric Reducing Antioxidant Power (FRAP) e 2,2-diphenyl-1-picrylhydrazyl (DPPH). Os frutos do mutante hp1 apresentaram maiores conteúdos de compostos fenólicos totais e também maior capacidade antioxidante em relação à cultivar selvagem ("MT") e ao mutante au, o qual não diferiu significativamente da cultivar "MT".

Phenolic compounds and antioxidant capacity are defense mechanisms of plants against the oxidative stress damage. Phenolic compounds are synthesized through the phenylpropanoid pathway, where the enzyme phenylalanine-ammonia-lyase plays a key role and it is influenced by light and photoreceptors such as phytochromes. The present research aims to evaluate the phenolic compounds content and antioxidant capacity of the wild "Micro-Tom" (MT) cultivar tomato fruits and its photomorphogenic mutant tomato plants high pigment 1 (hp1), super responsive to events mediated by light, and aurea (au), quantitative phytochrome deficient. Twenty mature fruits of each genotype ("MT", hp1, au) were used in triplicate for analyses. To quantify the total phenolic compounds the Folin-Ciocalteu method was used and the antioxidant capacity was analyzed by Ferric Reducing Antioxidant Power (FRAP) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) methods. The hp1 mutant presented the highest total phenolic compounds content and higher antioxidant capacity than wild cultivar ("MT") and au mutant, which did not differ significantly from "MT" cultivar.

Photosynthate partitioning and morphoanatomical aspects of photomorphogenic mutants of tomato / Partição de fotoassimilados e aspectos morfoanatômicos em tomateiros mutantes fotomorfogenéticos

The aim of this study was to analyze photosynthate partitioning in tomato photomorphogenic mutants at the ends of the vegetative (40 days after emergence [DAE]) and reproductive (69 DAE) stages and to determine its interaction with morphoanatomical aspects. The mutants aurea (au), phytochrome-deficient, high pigment-1 (hp1), light-exaggerated response, were studied along with the non-mutant Micro-Tom (MT) cultivar. The plants were analyzed at 40 and 68 DAE to identify photosynthate source organs and tissues as well as the target organs of remobilized photosynthate during the reproductive stage. The plants were evaluated for their internal and external morphology as well as the percentage of dry mass of their organs. Photosynthate allocation in the hp1 mutant occurred primarily in the roots and leaves, and allocation in the au mutant occurred primarily in fruits. The au mutant showed a high capacity for photosynthate remobilization to fruit during the reproductive stage, and the predominant sources of these remobilized photosynthates were the leaf spongy parenchyma, the root vascular cylinder and the marrow stem.

O objetivo deste estudo foi analisar a partição de fotoassimilados em tomateiros mutantes fotomorfogenéticos ao final da fase vegetativa, aos 40 dias após a emergência (DAE), e ao final da fase reprodutiva, aos 69 DAE, e sua interação com aspectos morfoanatômicos. Foram estudados os mutantes aurea (au), deficiente em fitocromo, e hp1, o qual expressa resposta exagerada à luz, e o tomateiro selvagem cultivar Micro-Tom (MT). As plantas foram analisadas 40 dias após a emergência (DAE) e 68 DAE, tentando identificar os órgãos e tecidos dos fotoassimilados remobilizados e seus órgãos de destino durante o estádio reprodutivo. As plantas foram avaliadas quanto à sua morfologia interna e externa e percentagem de massa seca entre os órgãos. A alocação de fotoassimilados no mutante hp1 ocorreu prioritariamente em raízes e folhas comparativamente aos demais órgãos, e no mutante au ocorreu prioritariamente em frutos comparativamente aos demais órgãos. O mutante au deteve alta capacidade de remobilização de fotoassimilados durante sua fase reprodutiva para os frutos e os fotoassimilados remobilizados tiveram origem preponderante do parênquima lacunoso foliar, do cilindro vascular radicular e da medula caulinar.

The role of phytochrome in stress tolerance.

It is well-documented that phytochromes can control plant growth and development from germination to flowering. Additionally, these photoreceptors have been shown to modulate both biotic and abiotic stress. This has led to a series of studies exploring the molecular and biochemical basis by which phytochromes modulate stresses, such as salinity, drought, high light or herbivory. Evidence for a role of phytrochromes in plant stress tolerance is explored and reviewed.

Small and remarkable: The Micro-Tom model system as a tool to discover novel hormonal functions and interactions.

Hormones are molecules involved in virtually every step of plant development and studies in this field have been shaping plant physiology for more than a century. The model plant Arabidopsis thaliana, long used as a tool to study plant hormones, lacks significant important developmental traits, such as fleshy climacteric fruit, compound leaf and multicellular trichomes, suggesting the necessity for alternative plant models. An attractive option often used is tomato, a species also of major economic importance, being ideal to bring together basic and applied plant sciences. The tomato Micro-Tom (MT) cultivar makes it possible to combine the direct benefits of studying a crop species with the fast life cycle and small size required for a suitable biological model. However, few obscure questions are constantly addressed to MT, creating a process herein called "MT mystification". In this work we present evidence clarifying these questions and show the potential of MT, aiming to demystify it. To corroborate our ideas we showed that, by making use of MT, our laboratory demonstrated straightforwardly new hormonal functions and also characterized a novel antagonistic hormonal interaction between jasmonates and brassinosteroids in the formation of anti-herbivory traits in tomato.