SELF PRUNING 3C is a flowering repressor that modulates seed germination, root architecture, and drought responses.

Allelic variation in the CETS (CENTRORADIALIS, TERMINAL FLOWER 1, SELF PRUNING) gene family controls agronomically important traits in many crops. CETS genes encode phosphatidylethanolamine-binding proteins that have a central role in the timing of flowering as florigenic and anti-florigenic signals. The great expansion of CETS genes in many species suggests that the functions of this family go beyond flowering induction and repression. Here, we characterized the tomato SELF PRUNING 3C (SP3C) gene, and show that besides acting as a flowering repressor it also regulates seed germination and modulates root architecture. We show that loss of SP3C function in CRISPR/Cas9-generated mutant lines increases root length and reduces root side branching relative to the wild type. Higher SP3C expression in transgenic lines promotes the opposite effects in roots, represses seed germination, and also improves tolerance to water stress in seedlings. These discoveries provide new insights into the role of SP paralogs in agronomically relevant traits, and support future exploration of the involvement of CETS genes in abiotic stress responses.

Irradiance-regulated biomass allocation in Raphanus sativus plants depends on gibberellin biosynthesis.

Gibberellin has been proposed to increase leaf elongation in radish (Raphanus sativus L.) plants, which is associated with decreased tuber growth. Since light intensity can control growth through interaction with gibberellin, investigation of the effect of gibberellin levels on the growth of radish plants would be a step forward towards unraveling factors that underlie biomass accumulation and allocation in response to irradiance levels. Here, we report that the gibberellin biosynthesis inhibitor paclobutrazol (PAC) decreased petiole elongation, but not lamina growth of radish plants grown under full sunlight. However, shading promoted an increase in shoot elongation, while in plants treated with PAC the petiole and leaf lamina fail to elongate. Plants treated with PAC allocated proportionally more biomass to their tubers and less to shoot compared to control under shade. Moreover, PAC decreased the abundance of transcripts encoding cell wall expansion proteins in leaf lamina and petiole of plants grown under shade, which was positively correlated with sugar consumption by the tuber, thereby increasing the mass fraction and concentrations of minerals for tuber. Thus, allocation of biomass during the growth of radish plants and nutritional quality of tubers depend on gibberellin and light intensity.

Exogenous ethylene reduces growth via alterations in central metabolism and cell wall composition in tomato (Solanum lycopersicum).

Ethylene is a gaseous hormone with a well-established role in the regulation of plant growth and development. However, its role in the modulation of carbon assimilation and central metabolism remains unclear. Here, we investigated the morphophysiological and biochemical responses of tomato plants (Solanum lycopersicum) following the application of ethylene in the form of ethephon (CEPA - 2-chloroethylphosphonic acid), forcing the classical triple response phenotype. CEPA-treated plants were characterized by growth inhibition, as revealed by significant reductions in both shoot and root dry weights, coupled with a reduced number of leaves and lower specific leaf area. Growth inhibition was associated with a reduction in carbon assimilation due to both lower photosynthesis rates and stomatal conductance, coupled with impairments in carbohydrate turnover. Furthermore, exogenous ethylene led to the accumulation of cell wall compounds (i.e., cellulose and lignin) and phenolics, indicating that exposure to exogenous ethylene also led to changes in specialized metabolism. Collectively, our findings demonstrate that exogenous ethylene disrupts plant growth and leaf structure by affecting both central and specialized metabolism, especially that involved in carbohydrate turnover and cell wall biosynthesis, ultimately leading to metabolic responses that mimic stress situations.

The Lanata trichome mutation increases stomatal conductance and reduces leaf temperature in tomato.

Trichomes are epidermal structures with a large variety of ecological functions and economic applications. Glandular trichomes produce a rich repertoire of secondary metabolites, whereas non-glandular trichomes create a physical barrier on the epidermis: both operate in tandem against biotic and abiotic stressors. A deeper understanding of trichome development and function would enable the breeding of more resilient crops. However, little is known about the impact of altered trichome density on leaf photosynthesis, gas exchange and energy balance. Previous work has compared multiple, closely related species differing in trichome density. Here, we analysed monogenic trichome mutants in the same tomato genetic background (Solanum lycopersicum cv. 'Micro-Tom'). We determined growth parameters, leaf spectral properties, gas exchange and leaf temperature in the hairs absent (h), Lanata (Ln) and Woolly (Wo) trichome mutants. Shoot dry weight, leaf area, leaf spectral properties and cuticular conductance were not affected by the mutations. However, the Ln mutant showed increased net carbon assimilation rate (An), associated with higher stomatal conductance (gs), with no differences in stomatal density or stomatal index between genotypes. Leaf temperature was furthermore reduced in Ln in the hottest, early hours of the afternoon. We show that a single monogenic mutation that modifies trichome density, a desirable trait for crop breeding, concomitantly improves leaf gas exchange and reduces leaf temperature.

Selenium uptake and grain nutritional quality are affected by nitrogen fertilization in rice (Oryza sativa L.).

KEY MESSAGE: The effects of selenium in rice grain composition depend on the soil nitrogen supply. Selenium and nitrogen have the potential to modify rice grain composition; however, it is unclear how the combined effect of Se and nitrogen affects the grain nutritional quality of rice. In our study, grain Se concentration was positively associated with the increased availability of nitrogen in soil. The accumulation of Se in grain of rice plants treated with Se combined with nitrogen was accompanied by an increase in expression of NRT1.1B, a rice nitrate transporter and sensor, in root. Moreover, Se potentiates the response of nitrogen supply in expression of sulfate transporter OsSULTR1.2, phosphate transporter OsPT2 and silicon transporter OsNIP2.1 in root, thereby increasing root Se uptake capacity. The combination of Se with high nitrogen increased the concentrations of protein, carbohydrates, Se, Mo and Mg, but decreased concentrations of Fe, Mn, Cu and Zn in grain. Overall, our results revealed that many of the effects of Se in rice grain composition are due to a shift in the nitrogen status of the plant.

Physiological and metabolic bases of increased growth in the tomato ethylene-insensitive mutant Never ripe: extending ethylene signaling functions.

KEY MESSAGE: The tomato mutant Never ripe (Nr), a loss-of-function for the ethylene receptor SlETR3, shows enhanced growth, associated with increased carbon assimilation and a rewiring of the central metabolism. Compelling evidence has demonstrated the importance of ethylene during tomato fruit development, yet its role on leaf central metabolism and plant growth remains elusive. Here, we performed a detailed characterization of Never ripe (Nr) tomato, a loss-of-function mutant for the ethylene receptor SlETR3, known for its fruits which never ripe. However, besides fruits, the Nr gene is also constitutively expressed in vegetative tissues. Nr mutant showed a growth enhancement during both the vegetative and reproductive stage, without an earlier onset of leaf senescence, with Nr plants exhibiting a higher number of leaves and an increased dry weight of leaves, stems, roots, and fruits. At metabolic level, Nr also plays a significant role with the mutant showing changes in carbon assimilation, carbohydrates turnover, and an exquisite reprogramming of a large number of metabolite levels. Notably, the expression of genes related to ethylene signaling and biosynthesis are not altered in Nr. We assess our results in the context of those previously published for tomato fruits and of current models of ethylene signal transduction, and conclude that ethylene insensitivity mediated by Nr impacts the whole central metabolism at vegetative stage, leading to increased growth rates.

Control of water-use efficiency by florigen.

A major issue in modern agriculture is water loss through stomata during photosynthetic carbon assimilation. In water-limited ecosystems, annual plants have strategies to synchronize their growth and reproduction to the availability of water. Some species or ecotypes of flowers are early to ensure that their life cycles are completed before the onset of late season terminal drought ("drought escape"). This accelerated flowering correlates with low water-use efficiency (WUE). The molecular players and physiological mechanisms involved in this coordination are not fully understood. We analyzed WUE using gravimetry, gas exchange, and carbon isotope discrimination in florigen deficient (sft mutant), wild-type (Micro-Tom), and florigen over-expressing (SFT-ox) tomato lines. Increased florigen expression led to accelerated flowering time and reduced WUE. The low WUE of SFT-ox was driven by higher stomatal conductance and thinner leaf blades. This florigen-driven effect on WUE appears be independent of abscisic acid (ABA). Our results open a new avenue to increase WUE in crops in an ABA-independent manner. Manipulation of florigen levels could allow us to produce crops with a life cycle synchronized to water availability.

Selenomethionine induces oxidative stress and modifies growth in rice (Oryza sativa L.) seedlings through effects on hormone biosynthesis and primary metabolism.

Although the chemical characteristics of selenomethionine (SeMet) are similar to those of methionine (Met), the physiological activity of SeMet apparently differs in its ability to stimulate ethylene production in plant tissues. Since selenium alters root architecture of rice seedlings by modifying ethylene production, the investigation of the effect of SeMet and Met on rice growth would be a step forward towards unraveling factors that underlie selenium toxicity. Here, we report that SeMet increased concentrations of reactive oxygen species (ROS), inhibiting auxin and increasing ethylene production in rice seedlings. The effect of SeMet on seedlings was mediated by the inhibition of the abundance of transcripts encoding auxin transport and cell expansion proteins. Moreover, SeMet led to increased seedling respiration, which was positively correlated with organic acids consumption, but negatively with sugars consumption, thereby decreasing seedling growth. In contrast with SeMet treatment, Met did not affect ROS production, hormone biosynthesis and seedling growth, indicating an exclusive selenium effect. The singlet oxygen scavenger, 1,4-diazabicyclooctane, overrode the repressive effect of SeMet in seedling growth. Our results demonstrate a phytotoxic effect of SeMet for rice seedlings and reveal a relationship between reactive oxygen species, hormone homeostasis and carbon availability, which regulates growth responses.

Elevated CO2 induces age-dependent restoration of growth and metabolism in gibberellin-deficient plants.

MAIN CONCLUSION: The effect of elevated [CO2] on the growth of tomato plants with reduced gibberellin content is influenced by developmental stage. The impact of increased atmospheric carbon dioxide (CO2) on plants has aroused interest in the last decades. Signaling molecules known as plant hormones are fundamental controllers of plant growth and development. Elevated CO2 concentration ([CO2]) increases plant growth; however, whether plant hormones act as mediators of this effect is still an open question. Here, we show the response to elevated [CO2] in tomato does not require a functional gibberellin (GA) biosynthesis pathway. We compared growth and primary metabolism between wild-type (WT) and GA-deficient mutant (gib-1) plants transferred from ambient (400 ppm) to elevated (750 ppm) [CO2] at two different growth stages (either 21 or 35 days after germination, DAG). Growth, photosynthetic parameters and primary metabolism in the stunted gib-1 plants were restored when they were transferred to elevated [CO2] at 21 DAG. Elevated [CO2] also stimulated growth and photosynthetic parameters in WT plants at 21 DAG; however, only minor changes were observed in the level of primary metabolites. At 35 DAG, on the other hand, elevated [CO2] did not stimulate growth in WT plants and gib-1 mutants showed their characteristic stunted growth phenotype. Taken together, our results reveal that elevated [CO2] enhances growth only within a narrow developmental window, in which GA biosynthesis is dispensable. This finding could be relevant for breeding crops in the face of the expected increases in atmospheric CO2 over the next century.

Selenium downregulates auxin and ethylene biosynthesis in rice seedlings to modify primary metabolism and root architecture.

MAIN CONCLUSION: Selenium modulates the formation of primary and lateral roots through alterations in auxin and ethylene, leading to new patterns of root architecture in rice seedlings. Selenium (Se) at low concentrations can control root growth through interaction with hormone biosynthesis. Auxin and ethylene have been shown to control the root architecture, with most of the information obtained from the eudicots such Arabidopsis and Nicotiana tabacum. Here, we presented the effects of Se on auxin and ethylene pathways and examined their impact on primary metabolism and root system architecture in rice (Oryza sativa L.) seedlings. Se treatment increased elongation of primary root, but decreased the number and length of lateral roots. Se led to decreased expression of genes associated with the biosynthesis of auxin and ethylene, concomitantly with reduced production of these hormones by the roots. Moreover, Se decreased the abundance of transcripts encoding auxin transport proteins. Indole-3-acetic acid (IAA) treatment overrode the repressive effect of Se on lateral root growth. The ethylene synthesis inhibitor L-α-(2-aminoethoxyvinyl)-glycine (AVG) increased elongation of primary root, whereas the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC) resulted in the opposite effect. Soluble sugars accumulate in roots of rice seedlings under Se treatment. Thus, Se modulates the formation of primary and lateral roots through alterations in auxin and ethylene, leading to new patterns of root architecture in rice seedlings.

Leaf heteroblasty in Passiflora edulis as revealed by metabolic profiling and expression analyses of the microRNAs miR156 and miR172.

BACKGROUND AND AIMS: Juvenile-to-adult phase transition is marked by changes in leaf morphology, mostly due to the temporal development of the shoot apical meristem, a phenomenon known as heteroblasty. Sugars and microRNA-controlled modules are components of the heteroblastic process in Arabidopsis thaliana leaves. However, our understanding about their roles during phase-changing in other species, such as Passiflora edulis, remains limited. Unlike Arabidopsis, P. edulis (a semi-woody perennial climbing vine) undergoes remarkable changes in leaf morphology throughout juvenile-to-adult transition. Nonetheless, the underlying molecular mechanisms are unknown. METHODS: Here we evaluated the molecular mechanisms underlying the heteroblastic process by analysing the temporal expression of microRNAs and targets in leaves as well as the leaf metabolome during P. edulis development. KEY RESULTS: Metabolic profiling revealed a unique composition of metabolites associated with leaf heteroblasty. Increasing levels of glucose and α-trehalose were observed during juvenile-to-adult phase transition. Accumulation of microRNA156 (miR156) correlated with juvenile leaf traits, whilst miR172 transcript accumulation was associated with leaf adult traits. Importantly, glucose may mediate adult leaf characteristics during de novo shoot organogenesis by modulating miR156-targeted PeSPL9 expression levels at early stages of shoot development. CONCLUSIONS: Altogether, our results suggest that specific sugars may act as co-regulators, along with two microRNAs, leading to leaf morphological modifications throughout juvenile-to-adult phase transition in P. edulis.

Bundle sheath extensions affect leaf structural and physiological plasticity in response to irradiance.

Coordination between structural and physiological traits is key to plants' responses to environmental fluctuations. In heterobaric leaves, bundle sheath extensions (BSEs) increase photosynthetic performance (light-saturated rates of photosynthesis, Amax ) and water transport capacity (leaf hydraulic conductance, Kleaf ). However, it is not clear how BSEs affect these and other leaf developmental and physiological parameters in response to environmental conditions. The obscuravenosa (obv) mutation, found in many commercial tomato varieties, leads to absence of BSEs. We examined structural and physiological traits of tomato heterobaric and homobaric (obv) near-isogenic lines grown at two different irradiance levels. Kleaf , minor vein density, and stomatal pore area index decreased with shading in heterobaric but not in homobaric leaves, which show similarly lower values in both conditions. Homobaric plants, on the other hand, showed increased Amax , leaf intercellular air spaces, and mesophyll surface area exposed to intercellular airspace (Smes ) in comparison with heterobaric plants when both were grown in the shade. BSEs further affected carbon isotope discrimination, a proxy for long-term water-use efficiency. BSEs confer plasticity in traits related to leaf structure and function in response to irradiance levels and might act as a hub integrating leaf structure, photosynthetic function, and water supply and demand.

Modifications in Organic Acid Profiles During Fruit Development and Ripening: Correlation or Causation?

The pivotal role of phytohormones during fruit development and ripening is considered established knowledge in plant biology. Perhaps less well-known is the growing body of evidence suggesting that organic acids play a key function in plant development and, in particular, in fruit development, maturation and ripening. Here, we critically review the connection between organic acids and the development of both climacteric and non-climacteric fruits. By analyzing the metabolic content of different fruits during their ontogenetic trajectory, we noticed that the content of organic acids in the early stages of fruit development is directly related to the supply of substrates for respiratory processes. Although different organic acid species can be found during fruit development in general, it appears that citrate and malate play major roles in this process, as they accumulate on a broad range of climacteric and non-climacteric fruits. We further highlight the functional significance of changes in organic acid profile in fruits due to either the manipulation of fruit-specific genes or the use of fruit-specific promoters. Despite the complexity behind the fluctuation in organic acid content during fruit development and ripening, we extend our understanding on the importance of organic acids on fruit metabolism and the need to further boost future research. We suggest that engineering organic acid metabolism could improve both qualitative and quantitative traits of crop fruits.

Salt stress inhibits germination of Stylosanthes humilis seeds through abscisic acid accumulation and associated changes in ethylene production.

In Stylosanthes humilis, salt stress tolerance is associated with ethylene production by the seeds, however, how salt stress controls seed germination and ethylene production is poorly understood. Here, we studied the hormonal and metabolic changes triggered by salt stress on germination of S. humilis seeds. Salt stress led to decreased seed germination and ethylene production, concomitantly with higher abscisic acid (ABA) production by seeds. Treatment with NaCl and ABA promoted distinct changes in energy metabolism, allowing seeds to adapt to salt stress conditions. Treatment with the ABA biosynthesis inhibitor fluridone or ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC) reversed the effects of salt stress on seed germination and ethylene production. Moreover, ethylene concentration was decreased by increasing the pH of the salt solution. High pH, however, did not influence concentration of ABA in seeds under salt stress. We conclude that biosynthesis of ABA and ethylene in response to salt stress constitutes a point of convergence that provides flexibility to regulate energy metabolism and embryo growth potential of S. humilis seeds within a given pH condition.

Low soil pH modulates ethylene biosynthesis and germination response of Stylosanthes humilis seeds.

The tropical forage legume Stylosanthes humilis is naturally distributed in the acidic soils of the tropics. However, data concerning the role of low soil pH in the control of S. humilis seed germination remains limited. Recently, we have demonstrated that acidic soil triggers increased ethylene production during germination of S. humilis seeds, concomitantly with higher root penetration into the soil. Our finding points an important role of low soil pH as a signal allowing penetration of root in the soil through interaction with the ethylene signalling pathway. Herein, we discuss how low soil pH induces changes on seed hormonal physiology.

Growth inhibition by selenium is associated with changes in primary metabolism and nutrient levels in Arabidopsis thaliana.

Although Selenium (Se) stress is relatively well known for causing growth inhibition, its effects on primary metabolism remain rather unclear. Here, we characterized both the modulation of the expression of specific genes and the metabolic adjustments in Arabidopsis thaliana in response to changes in Se level in the soil. Se treatment culminated with strong inhibition of both shoot and root growth. Notably, growth inhibition in Se-treated plants was associated with an incomplete mobilization of starch during the night. Minor changes in amino acids levels were observed in shoots and roots of plants treated with Se whereas the pool size of tricarboxylic acid (TCA) cycle intermediates in root was not altered in response to Se. By contrast, decreased levels of organic acids involved in the first part of the TCA cycle were observed in shoots of Se-treated plants. Furthermore, decreased expression levels of expansins and endotransglucosylases/endohydrolases (XHTs) genes were observed after Se treatment, coupled with a significant decrease in the levels of essential elements. Collectively, our results revealed an exquisite interaction between energy metabolism and Se-mediated control of growth in Arabidopsis thaliana to coordinate cell wall extension, starch turnover and the levels of a few essential nutrients.

Germination of salt-stressed seeds as related to the ethylene biosynthesis ability in three Stylosanthes species.

Stylosanthes, a genus of tropical forage legume, is known to exhibit good persistence in saline soils, yet mechanisms for regulation of seed germination under salt stress are poorly understood. This study was carried out to evaluate the mode of action of salt stress on seed germination of Stylosanthes. 1-Aminocyclopropane-1-carboxylic acid (ACC) increased ethylene biosynthesis and germination of NaCl-inhibited seeds in a dose-dependent manner. Contents of ACC and germination of Stylosanthes humilis seeds increased following transfer from NaCl solution to deionised water, but not after transfer to l-α-(2-aminoethoxyvinyl)-glycine (AVG) solution, an inhibitor of ethylene biosynthesis. Ethylene biosynthesis was much larger in NaCl-treated seeds of Stylosanthes guianensis than in seeds of S. humilis and Stylosanthes capitata, a fact which was reflected in higher germination rates. S. guianensis seedlings also displayed higher growth and survival rates than S. humilis and S. capitata under salt stress. Moreover, smaller ACC levels, as well as reduced ethylene biosynthesis of S. capitata seeds were accompanied by lower germination under salt stress. In addition, S. capitata seedlings treated with NaCl solutions exhibited relatively lower growth and survival rates in comparison with S. humilis and S. guianensis. Thus, different abilities to synthesize ethylene by S. guianensis, S. humilis and S. capitata seeds explain the differences in tolerance to salt stress of the three species.

Promotion of growth by elevated carbon dioxide is coordinated through a flexible transcriptional network in Arabidopsis.

Although gibberellins (GAs) promote many developmental responses in plants, little is known about how the hormone interacts with environmental signals at the molecular level for regulating plant growth. Recently, we have demonstrated that inhibition of growth by the GA biosynthesis inhibitor paclobutrazol (PAC) at ambient [CO2] (350 µmol CO2 mol(-1)) is reverted by elevated [CO2] (750 µmol CO2 mol(-1)). Our finding points to an important role of elevated [CO2] as a signal allowing higher growth rates of low-GA plants. GA promotes plant growth via a complex transcriptional network that integrates multiple signaling pathways. Herein, we discuss how elevated [CO2] stimulates biomass accumulation in a GA-independent manner by regulating the expression of growth-related genes.

Action of gibberellins on growth and metabolism of Arabidopsis plants associated with high concentration of carbon dioxide.

Although the positive effect of elevated CO(2) concentration [CO(2)] on plant growth is well known, it remains unclear whether global climate change will positively or negatively affect crop yields. In particular, relatively little is known about the role of hormone pathways in controlling the growth responses to elevated [CO(2)]. Here, we studied the impact of elevated [CO(2)] on plant biomass and metabolism in Arabidopsis (Arabidopsis thaliana) in relation to the availability of gibberellins (GAs). Inhibition of growth by the GA biosynthesis inhibitor paclobutrazol (PAC) at ambient [CO(2)] (350 µmol CO(2) mol(-1)) was reverted by elevated [CO(2)] (750 µmol CO(2) mol(-1)). Thus, we investigated the metabolic adjustment and modulation of gene expression in response to changes in growth of plants imposed by varying the GA regime in ambient and elevated [CO(2)]. In the presence of PAC (low-GA regime), the activities of enzymes involved in photosynthesis and inorganic nitrogen assimilation were markedly increased at elevated [CO(2)], whereas the activities of enzymes of organic acid metabolism were decreased. Under ambient [CO(2)], nitrate, amino acids, and protein accumulated upon PAC treatment; however, this was not the case when plants were grown at elevated [CO(2)]. These results suggest that only under ambient [CO(2)] is GA required for the integration of carbohydrate and nitrogen metabolism underlying optimal biomass determination. Our results have implications concerning the action of the Green Revolution genes in future environmental conditions.

Translatome and metabolome effects triggered by gibberellins during rosette growth in Arabidopsis.

Although gibberellins (GAs) are well known for their growth control function, little is known about their effects on primary metabolism. Here the modulation of gene expression and metabolic adjustment in response to changes in plant (Arabidopsis thaliana) growth imposed on varying the gibberellin regime were evaluated. Polysomal mRNA populations were profiled following treatment of plants with paclobutrazol (PAC), an inhibitor of GA biosynthesis, and gibberellic acid (GA(3)) to monitor translational regulation of mRNAs globally. Gibberellin levels did not affect levels of carbohydrates in plants treated with PAC and/or GA(3). However, the tricarboxylic acid cycle intermediates malate and fumarate, two alternative carbon storage molecules, accumulated upon PAC treatment. Moreover, an increase in nitrate and in the levels of the amino acids was observed in plants grown under a low GA regime. Only minor changes in amino acid levels were detected in plants treated with GA(3) alone, or PAC plus GA(3). Comparison of the molecular changes at the transcript and metabolite levels demonstrated that a low GA level mainly affects growth by uncoupling growth from carbon availability. These observations, together with the translatome changes, reveal an interaction between energy metabolism and GA-mediated control of growth to coordinate cell wall extension, secondary metabolism, and lipid metabolism.