Additive effects of light and branching on fruit size and chemical fruit quality of greenhouse tomatoes.

Introduction: Greenhouse tomato growers face the challenge of balancing fruit size and chemical quality traits. This study focused on elucidating the interplay between plant branching and light management on these traits, while maintaining consistent shoot density. Methods: We evaluated one- and two-shoot plants under varying top light intensities using high-pressure sodium lamps and light-emitting diode (LED) inter-lighting. Results: The reduced yield in the two-shoot plants was mainly due to smaller fruit size, but not due to source strength limitations, as evaluated through leaf weight ratio (LWR), chlorophyll index, specific leaf area (SLA), leaf dry matter percentage, and stem soluble carbohydrate accumulation. Enhanced lighting improved fruit weight and various fruit traits, such as dry matter content, total soluble carbohydrate content, and phenolic content, for both one- and two-shoot plant types. Despite lower mean fruit weight, two-shoot plants exhibited higher values for chemical fruit quality traits, indicating that the fruit growth of two-shoot plants is not limited by the available carbohydrates (source strength), but by the fruit sink strength. Diurnal analysis of fruit growth showed that two-shoot plants had reduced expansion during light transitions. This drop in fruit expansion was not related to changes in root pressure (measured as xylem sap exudation from decapitated plants), but might be related to diminished xylem area in the stem joint of the two-shoot plants. The concentration of several hormones, including cytokinins, was lower in two-shoot plants, suggesting a reduced fruit sink capacity. Discussion: The predominant impact of branching to two-shoot plants on sink capacity suggests that the fruit growth is not limited by available carbohydrates (source strength). Alongside the observation that light supplementation and branching exert independent additive effects on fruit size and chemical traits, this illuminates the potential to independently regulate these aspects in greenhouse tomato production.

Heterogeneous nutrient supply modulates root exudation and accumulation of medicinally valuable compounds in Artemisia annua and Hypericum perforatum.

Plants have evolved complex mechanisms to adapt to nutrient-deficient environments, including stimulating lateral root proliferation into local soil patches with high nutrient content in response to heterogeneous nutrient distribution. Despite the widespread occurrence of this phenomenon in soil, the effect of heterogeneous nutrient distribution on the accumulation of secondary compounds in plant biomass and their exudation by roots remains largely unknown. This study aims to fill this critical knowledge gap by investigating how deficiency and unequal distributions of nitrogen (N), phosphorus (P), and iron (Fe) affect plant growth and accumulation of the antimalarial drug artemisinin (AN) in leaves and roots of Artemisia annua, as well as AN exudation by roots. Heterogeneous N and P supplies strongly increased root exudation of AN in half of a split-root system exposed to nutrient deficiency. By contrast, exposure to a homogeneous nitrate and phosphate deficiency did not modulate root exudation of AN. This indicates that a combination of local and systemic signals, reflecting low and high nutritional statuses, respectively, were required to enhance AN exudation. This exudation response was independent of the regulation of root hair formation, which was predominantly modulated by the local signal. In contrast to the heterogeneous supply of N and P, heterogeneous Fe supply did not modulate AN root exudation but increased AN accumulation in locally Fe-deficient roots. No modulation of nutrient supply significantly changed the accumulation of AN in A. annua leaves. The impact of a heterogeneous nitrate supply on growth and phytochemical composition was also investigated in Hypericum perforatum plants. Unlike in A. annue, the uneven N supply did not significantly influence the exudation of secondary compounds in the roots of H. perforatum. However, it did enhance the accumulation of several biologically active compounds, such as hypericin, catechin, and rutin isomers, in the leaves of H. perforatum. We propose that the capacity of plants to induce the accumulation and/or differential exudation of secondary compounds under heterogeneous nutrient supply is both species- and compound-specific. The ability to differentially exude AN may contribute to A. annua's adaptation to nutrient disturbances and modulate allelopathic and symbiotic interactions in the rhizosphere.

Light exposure of roots in aeroponics enhances the accumulation of phytochemicals in aboveground parts of the medicinal plants Artemisia annua and Hypericum perforatum.

Light acts as a trigger to enhance the accumulation of secondary compounds in the aboveground part of plants; however, whether a similar triggering effect occurs in roots is unclear. Using an aeroponic setup, we investigated the effect of long-term exposure of roots to LED lighting of different wavelengths on the growth and phytochemical composition of two high-value medicinal plants, Artemisia annua and Hypericum perforatum. In A. annua, root exposure to white, blue, and red light enhanced the accumulation of artemisinin in the shoots by 2.3-, 2.5-, and 1.9-fold, respectively. In H. perforatum, root exposure to white, blue, red, and green light enhanced the accumulation of coumaroylquinic acid in leaves by 89, 65, 84, and 74%, respectively. Root lighting also increased flavonol concentrations. In contrast to its effects in the shoots, root illumination did not change phytochemical composition in the roots or root exudates. Thus, root illumination induces a systemic response, resulting in modulation of the phytochemical composition in distal tissues remote from the light exposure site.

Decoupling of Plant Growth and Accumulation of Biologically Active Compounds in Leaves, Roots, and Root Exudates of Hypericum perforatum L. by the Combination of Jasmonate and Far-Red Lighting.

The plant hormone jasmonic acid (JA) fine tunes the growth-defense dilemma by inhibiting plant growth and stimulating the accumulation of secondary compounds. We investigated the interactions between JA and phytochrome B signaling on growth and the accumulation of selected secondary metabolites in Hypericum perforatum L., a medically important plant, by spraying plants with methyl jasmonate (MeJA) and by adding far-red (FR) lighting. MeJA inhibited plant growth, decreased fructose concentration, and enhanced the accumulation of most secondary metabolites. FR enhanced plant growth and starch accumulation and did not decrease the accumulation of most secondary metabolites. MeJA and FR acted mostly independently with no observable interactions on plant growth or secondary metabolite levels. The accumulation of different compounds (e.g., hypericin, flavonols, flavan-3-ols, and phenolic acid) in shoots, roots, and root exudates showed different responses to the two treatments. These findings indicate that the relationship between growth and secondary compound accumulation is specific and depends on the classes of compounds and/or their organ location. The combined application of MeJA and FR enhanced the accumulation of most secondary compounds without compromising plant growth. Thus, the negative correlations between biomass and the content of secondary compounds predicted by the growth-defense dilemma were overcome.

Organic Waste-Based Fertilizer in Hydroponics Increases Tomato Fruit Size but Reduces Fruit Quality.

In regions with intensive agricultural production, large amounts of organic waste are produced by livestock animals. Liquid digestate from manure-based biogas production could potentially serve as fertilizer if integrated with closed horticultural irrigation systems. The aim of this experiment was to investigate how fertilizer based on liquid biogas by-products of pig manure digestion can affect the growth and production of tomato plants. Integration of a nitrification bioreactor presumes a significantly lower concentration of nutrient solutions and a higher level of oxygenation than classical mineral cultivation. Therefore, additional controls were included. We compared plant growth and fruit quality traits of tomato plants grown in a hydroponic solution with organic fertilizer with two levels of mineral fertilizer. The tomatoes grown with organic waste-based liquid fertilizer showed reduced growth rates but increased mean fruit size, resulting in no significant change in total yield compared with high-mineral cultivation. The growth rate was similarly reduced in plants cultivated with low-mineral fertilizer. Plants cultivated with organic waste-based fertilizer had high Cl- concentration in xylem sap, leaves, and, ultimately, fruits. The leaves of plants cultivated with organic waste-based fertilizer contained higher concentrations of starch and soluble carbohydrate and low concentrations of phosphorous (P) and sulfur (S). The plants grown with organic waste-based or low-mineral medium showed significantly poorer fruit quality than the plants cultivated with the high-mineral solution. The low-mineral treatment increased xylem sap contribution to fruit weight because of higher root power. The organic waste-based fertilization did not change the root power but increased fruit size. In conclusion, organic waste-based cultivation is a possible solution for sustainable plant production in greenhouses. However, additional adjustment of nutrient supply is required to improve fruit quality.

Nitrogen Deficiency and Synergism between Continuous Light and Root Ammonium Supply Modulate Distinct but Overlapping Patterns of Phytohormone Composition in Xylem Sap of Tomato Plants.

Continuous light (CL) or a predominant nitrogen supply as ammonium (NH4+) can induce leaf chlorosis and inhibit plant growth. The similarity in injuries caused by CL and NH4+ suggests involvement of overlapping mechanisms in plant responses to these conditions; however, these mechanisms are poorly understood. We addressed this topic by conducting full factorial experiments with tomato plants to investigate the effects of NO3- or NH4+ supply under diurnal light (DL) or CL. We used plants at ages of 26 and 15 days after sowing to initiate the treatments, and we modulated the intensity of the stress induced by CL and an exclusive NH4+ supply from mild to strong. Under DL, we also studied the effect of nitrogen (N) deficiency and mixed application of NO3- and NH4+. Under strong stress, CL and exclusive NH4+ supply synergistically inhibited plant growth and reduced chlorophyll content. Under mild stress, when no synergetic effect between CL and NH4+ was apparent on plant growth and chlorophyll content, we found a synergetic effect of CL and NH4+ on the accumulation of several plant stress hormones, with an especially strong effect for jasmonic acid (JA) and 1-aminocyclopropane-1-carboxylic acid (ACC), the immediate precursor of ethylene, in xylem sap. This modulation of the hormonal composition suggests a potential role for these plant hormones in plant growth responses to the combined application of CL and NH4+. No synergetic effect was observed between CL and NH4+ for the accumulation of soluble carbohydrates or of mineral ions, indicating that these plant traits are less sensitive than the modulation of hormonal composition in xylem sap to the combined CL and NH4+ application. Under diurnal light, NH4+ did not affect the hormonal composition of xylem sap; however, N deficiency strongly increased the concentrations of phaseic acid (PA), JA, and salicylic acid (SA), indicating that decreased N concentration rather than the presence of NO3- or NH4+ in the nutrient solution drives the hormone composition of the xylem sap. In conclusion, N deficiency or a combined application of CL and NH4+ induced the accumulation of JA in xylem sap. This accumulation, in combination with other plant hormones, defines the specific plant response to stress conditions.

Differential Regulation of Kernel Set and Potential Kernel Weight by Nitrogen Supply and Carbohydrate Availability in Maize Genotypes Contrasting in Nitrogen Use Efficiency.

Sub-optimal nitrogen (N) conditions reduce maize yield due to a decrease in two sink components: kernel set and potential kernel weight. Both components are established during the lag phase, suggesting that they could compete for resources during this critical period. However, whether this competition occurs or whether different genotypic strategies exist to optimize photoassimilate use during the lag phase is not clear and requires further investigation. We have addressed this knowledge gap by conducting a nutrient solution culture experiment that allows abrupt changes in N level and light intensity during the lag phase. We investigated plant growth, dry matter partitioning, non-structural carbohydrate concentration, N concentration, and 15N distribution (applied 4 days before silking) in plant organs at the beginning and the end of the lag phase in two maize hybrids that differ in grain yield under N-limited conditions: one is a nitrogen-use-efficient (EFFI) genotype and the other is a control (GREEN) genotype that does not display high N use efficiency. We found that the two genotypes used different mechanisms to regulate kernel set. The GREEN genotype showed a reduction in kernel set associated with reduced dry matter allocation to the ear during the lag phase, indicating that the reduced kernel set under N-limited conditions was related to sink restrictions. This idea was supported by a negative correlation between kernel set and sucrose/total sugar ratios in the kernels, indicating that the capacity for sucrose cleavage might be a key factor defining kernel set in the GREEN genotype. By contrast, the kernel set of the EFFI genotype was not correlated with dry matter allocation to the ear or to a higher capacity for sucrose cleavage; rather, it showed a relationship with the different EFFI ear morphology with bigger kernels at the apex of the ear than in the GREEN genotype. The potential kernel weight was independent of carbohydrate availability but was related to the N flux per kernel in both genotypes. In conclusion, kernel set and potential kernel weight are regulated independently, suggesting the possibility of simultaneously increasing both sink components in maize.

Butylated Hydroxytoluene (BHT) Inhibits PIN1 Exocytosis From BFA Compartments in Arabidopsis Roots.

The activity of polarly localized PIN-FORMED (PIN) auxin efflux carriers contributes to the formation of auxin gradients which guide plant growth, development, and tropic responses. Both the localization and abundance of PIN proteins in the plasma membrane depend on the regulation of PIN trafficking through endocytosis and exocytosis and are influenced by many external and internal stimuli, such as reactive oxygen species, auxin transport inhibitors, flavonoids and plant hormones. Here, we investigated the regulation of endosomal PIN cycling by using a Brefeldin A (BFA) assay to study the effect of a phenolic antioxidant ionol, butylated hydroxytoluene (BHT), on the endocytosis and exocytosis of PIN1 and PIN2. BHT is one of the most widely used antioxidants in the food and feed industries, and as such is commonly released into the environment; however, the effect of BHT on plants remains poorly characterized. Preincubation of Arabidopsis seedlings with BHT before BFA treatment strongly enhanced the internalization of PIN1 into BFA compartments. After the simultaneous application of BHT and NAA, the NAA effect dominated PIN internalization suggesting the BHT effect occurred downstream to that of NAA. Washing seedlings with BHT after BFA treatment prevented the release of PIN1 from BFA compartments back to the plasma membrane, indicating that BHT application inhibited PIN1 exocytosis. Overall rates of PIN2 internalization were less pronounced than those of PIN1 in seedlings pre-incubated with BHT before BFA treatment, and PIN2 exocytosis was not inhibited by BHT, indicating a specific activity of BHT on PIN1 exocytosis. Comparison of BHT activity with other potential stimuli of PIN1 and PIN2 trafficking [e.g., H2O2 (ROS), salt stress, reduced glutathione (GSH), dithiothreitol (DTT), and flavonoids] showed that BHT has a new activity distinct from the activities of other regulators of PIN trafficking. The findings support BHT as a potentially interesting pharmacological tool for dissecting PIN trafficking and auxin transport.

Natural Auxin Does Not Inhibit Brefeldin A Induced PIN1 and PIN2 Internalization in Root Cells.

The vesicle trafficking inhibitor Brefeldin A (BFA) changes the localization of plasma membrane localized PINs, proteins that function as polar auxin efflux carriers, by inducing their accumulation within cells. Pretreatment with the synthetic auxin 1-NAA reduces this BFA-induced PIN internalization, suggesting that auxinic compounds inhibit the endocytosis of PIN proteins. However, the most important natural auxin, IAA, did not substantially inhibit PIN internalization unless a supplementary antioxidant, butylated hydroxytoluene (BHT), was also included in the incubation medium. We asked whether the relatively small inhibition caused by IAA alone could be explained by its instability in the incubation solution or whether IAA might interact with BHT to inhibit endocytosis. Analysis of the IAA concentration in the incubation solution and of DR5 reporter activity in the roots showed that IAA is both stable and active in the medium. Therefore, IAA degradation was not able to explain the inability of IAA to inhibit endocytosis. Furthermore, when applied in the absence of auxin, BHT caused a strong increase in the rate of PIN1 internalization and a weaker increase in the rate of PIN2 internalization. These increases were unaffected by the simultaneous application of IAA, further indicating that endocytosis is not inhibited by the natural auxin IAA under physiologically relevant conditions. Endocytosis was inhibited at the same rate with 2-NAA, an inactive auxin analog, as was observed with 1-NAA and more strongly than with natural auxins, supporting the idea that this inhibition is not auxin specific.

Supplemental Light-Emitting Diode Inter-Lighting Increases Tomato Fruit Growth Through Enhanced Photosynthetic Light Use Efficiency and Modulated Root Activity.

We investigated the effect of supplemental LED inter-lighting (80% red, 20% blue; 70 W m-2; light period 04:00-22:00) on the productivity and physiological traits of tomato plants (Flavance F1) grown in an industrial greenhouse with high pressure sodium (HPS) lamps (235 W m-2, 420 µmol m-2 s-1 at canopy). Physiological trait measurements included diurnal photosynthesis and fruit relative growth rates, fruit weight at specific positions in the truss, root pressure, xylem sap hormone and ion compositions, and fruit quality. In the control treatment with HPS lamps alone, the ratio of far-red to red light (FR:R) was 1.2 at the top of the canopy and increased to 5.4 at the bottom. The supplemental LED inter-lighting decreased the FR:R ratio at the middle and low positions in the canopy and was associated with greener leaves and higher photosynthetic light use efficiency (PLUE) in the leaves in the lower canopy. The use of LED inter-lighting increased the biomass and yield by increasing the fruit weight and enhancing plant growth. The PLUE of plants receiving supplemental LED light decreased at the end of the light period, indicating that photosynthesis of the supplemented plants at the end of the day might be limited by sink capacity. The supplemental LED lighting increased the size of fruits in the middle and distal positions of the truss, resulting in a more even size for each fruit in the truss. Diurnal analysis of fruit growth showed that fruits grew more quickly during the night on the plants receiving LED light than on unsupplemented control plants. This faster fruit growth during the night was related to an increased root pressure. The LED treatment also increased the xylem levels of the phytohormone jasmonate. Supplemental LED inter-lighting increased tomato fruit weight without affecting the total soluble solid contents in fruits by increasing the total assimilates available for fruit growth and by enhancing root activity through an increase in root pressure and water supply to support fruit growth during the night.

Auxin-Induced Plasma Membrane Depolarization Is Regulated by Auxin Transport and Not by AUXIN BINDING PROTEIN1.

Auxin is a molecule, which controls many aspects of plant development through both transcriptional and non-transcriptional signaling responses. AUXIN BINDING PROTEIN1 (ABP1) is a putative receptor for rapid non-transcriptional auxin-induced changes in plasma membrane depolarization and endocytosis rates. However, the mechanism of ABP1-mediated signaling is poorly understood. Here we show that membrane depolarization and endocytosis inhibition are ABP1-independent responses and that auxin-induced plasma membrane depolarization is instead dependent on the auxin influx carrier AUX1. AUX1 was itself not involved in the regulation of endocytosis. Auxin-dependent depolarization of the plasma membrane was also modulated by the auxin efflux carrier PIN2. These data establish a new connection between auxin transport and non-transcriptional auxin signaling.

The evolution of nuclear auxin signalling.

BACKGROUND: The plant hormone auxin directs many aspects of plant growth and development. To understand the evolution of auxin signalling, we compared the genes encoding two families of crucial transcriptional regulators, AUXIN RESPONSE FACTOR (ARF) and AUXIN/INDOLE-3-ACETIC ACID (Aux/IAA), among flowering plants and two non-seed plants, Physcomitrella patens and Selaginella moellendorffii. RESULTS: Comparative analysis of the P. patens, S. moellendorffii and Arabidopsis thaliana genomes suggests that the well-established rapid transcriptional response to auxin of flowering plants, evolved in vascular plants after their divergence from the last common ancestor shared with mosses. An N-terminally truncated ARF transcriptional activator is encoded by the genomes of P. patens and S. moellendorffii, and suggests a supplementary mechanism of nuclear auxin signalling, absent in flowering plants. Site-specific analyses of positive Darwinian selection revealed relatively high rates of synonymous substitution in the A. thaliana ARFs of classes IIa (and their closest orthologous genes in poplar) and Ib, suggesting that neofunctionalization in important functional regions has driven the evolution of auxin signalling in flowering plants. Primary auxin responsive gene families (GH3, SAUR, LBD) show different phylogenetic profiles in P. patens, S. moellendorffii and flowering plants, highlighting genes for further study. CONCLUSION: The genome of P. patens encodes all of the basic components necessary for a rapid auxin response. The spatial separation of the Q-rich activator domain and DNA-binding domain suggests an alternative mechanism of transcriptional control in P. patens distinct from the mechanism seen in flowering plants. Significantly, the genome of S. moellendorffii is predicted to encode proteins suitable for both methods of regulation.

Comprehensive transcriptome analysis of auxin responses in Arabidopsis.

In plants, the hormone auxin shapes gene expression to regulate growth and development. Despite the detailed characterization of auxin-inducible genes, a comprehensive overview of the temporal and spatial dynamics of auxin-regulated gene expression is lacking. Here, we analyze transcriptome data from many publicly available Arabidopsis profiling experiments and assess tissue-specific gene expression both in response to auxin concentration and exposure time and in relation to other plant growth regulators. Our analysis shows that the primary response to auxin over a wide range of auxin application conditions and in specific tissues comprises almost exclusively the up-regulation of genes and identifies the most robust auxin marker genes. Tissue-specific auxin responses correlate with differential expression of Aux/IAA genes and the subsequent regulation of context- and sequence-specific patterns of gene expression. Changes in transcript levels were consistent with a distinct sequence of conjugation, increased transport capacity and down-regulation of biosynthesis in the temperance of high cellular auxin concentrations. Our data show that auxin regulates genes associated with the biosynthesis, catabolism and signaling pathways of other phytohormones. We present a transcriptional overview of the auxin response. Specific interactions between auxin and other phytohormones are highlighted, particularly the regulation of their metabolism. Our analysis provides a roadmap for auxin-dependent processes that underpins the concept of an 'auxin code'--a tissue-specific fingerprint of gene expression that initiates specific developmental processes.