Pseudomonas aeruginosa and Bacillus cereus Isolated from Brazilian Cerrado Soil Act as Phosphate-Solubilizing Bacteria.

The phosphate-solubilizing microorganism is essential for soil quality and plant development and can serve as an alternative to reduce such Brazilian needs for importing phosphate overseas. Here, we isolated and selected bacteria from Brazilian Cerrado soils capable of solubilize phosphate. We obtained 53 bacteria isolates, of which 23 could solubilize phosphate at a pH of 7.0, 17 could solubilize phosphate at a pH of 6.0, and 8 could solubilize at a pH of 5.5. Using 16S rRNA gene sequences, we identified nine bacteria species clustered in four groups: Bacillus sp., Pseudomonas sp., Priestia sp., and Klebsiella sp. Our results revealed that the UFT01 (P. aeruginosa) and UFT42 (B. cereus) isolates exhibited the best phosphate solubilization performance at all tested pH values. We further recorded higher levels of solubilization and phosphate availability six days after the soil inoculation with P. aeruginosa, and enzymatic analysis of the soil samples revealed that the P. aeruginosa-inoculated samples resulted in four-fold higher enzymatic activities when compared to non-inoculated soils. The B. cereus soil inoculation increased ß-glucosidase activities and resulted in reduced the activities of arylsulfatase. Altogether, our findings demonstrated that P. aeruginosa and B. cereus isolated from Cerrado soils showed high phosphate solubilization potential.

Elevated carbon assimilation and metabolic reprogramming in tomato high pigment mutants support the increased production of pigments.

KEY MESSAGE: High pigment mutants in tomato (Solanum lycopersicum L.), a loss of function in the control of photomorphogenesis, with greater pigment production, show altered growth, greater photosynthesis, and a metabolic reprogramming. High pigment mutations cause plants to be extremely responsive to light and produce excessive pigmentation as well as fruits with high levels of health-beneficial nutrients. However, the association of these traits with changes in the physiology and metabolism of leaves remains poorly understood. Here, we performed a detailed morphophysiological and metabolic characterization of high pigment 1 (hp1) and high pigment 2 (hp2) mutants in tomato (Solanum lycopersicum L. 'Micro-Tom') plants under different sunlight conditions (natural light, 50% shading, and 80% shading). These mutants occur in the DDB1 (hp1) and DET1 (hp2) genes, which are related to the regulation of photomorphogenesis and chloroplast development. Our results demonstrate that these mutations delay plant growth and height, by affecting physiological and metabolic parameters at all stages of plant development. Although the mutants were characterized by higher net CO2 assimilation, lower stomatal limitation, and higher carboxylation rates, with anatomical changes that favour photosynthesis, we found that carbohydrate levels did not increase, indicating a change in the energy flow. Shading minimized the differences between mutants and the wild type or fully reversed them in the phenotype at the metabolic level. Our results indicate that the high levels of pigments in hp1 and hp2 mutants represent an additional energy cost for these plants and that extensive physiological and metabolic reprogramming occurs to support increased pigment biosynthesis.

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.

Beauveria bassiana Enhances the Growth of Cowpea Plants and Increases the Mortality of Cerotoma arcuata.

Cowpea (Vigna unguiculata) crops stand out for their efficient adaptation to edaphoclimatic conditions. Insect pests, such as the leaf beetle Cerotoma arcuata, are among the main factors that limit cowpea yield. Chemical control methods are commonly used to control such pests; however, biological methods are an alternative to reduce the indiscriminate use of conventional pesticides. This study aimed to evaluate the effects of Beauveria bassiana inoculation on the growth and physiological parameters of the cowpea plant and assess the influence of the inoculation on the feeding performance and survival of C. arcuata adults. Colonization by B. bassiana was recorded in the stems (63.89%), roots (45.83%), and leaves (25%) of the cowpea plant. It was found that B. bassiana enhanced the plant height, number of leaves, and the dry mass of the inoculated cowpea plants as compared to the control. The treated plants exhibited higher net carbon dioxide (CO2) assimilation rates in the gas exchange evaluation as well as higher stomatal conductance, evapotranspiration rates, and chlorophyll (a + b) content than the control plants. Moreover, the Kaplan-Meier survival analysis showed that the B. bassiana negatively affected the survival of the insect in the leaf disc assays.

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.

Differential content of leaf and fruit pigment in tomatoes culminate in a complex metabolic reprogramming without growth impacts.

Although significant efforts to produce carotenoid-enriched foods either by biotechnology or traditional breeding strategies have been carried out, our understanding of how changes in the carotenoid biosynthesis might affect overall plant performance remains limited. Here, we investigate how the metabolic machinery of well characterized tomato carotenoid mutant plants [namely crimson (old gold-og), Delta carotene (Del) and tangerine (t)] adjusts itself to varying carotenoid biosynthesis and whether these adjustments are supported by a reprogramming of photosynthetic and central metabolism in the source organs (leaves). We observed that mutations og, Del and t did not greatly affect vegetative growth, leaf anatomy and gas exchange parameters. However, an exquisite metabolic reprogramming was recorded on the leaves, with an increase in levels of amino acids and reduction of organic acids. Taken together, our results show that despite minor impacts on growth and gas exchange, carbon flux is extensively affected, leading to adjustments in tomato leaves metabolism to support changes in carotenoid biosynthesis on fruits (sinks). We discuss these data in the context of our current understanding of metabolic adjustments and carotenoid biosynthesis as well as regarding to improving human nutrition.

Physiological and metabolic changes in response to Boron levels are mediated by ethylene affecting tomato fruit yield.

Boron (B) is an essential nutrient for the plant, and its stress (both deficiency and toxicity) are major problems that affect crop production. Ethylene metabolism (both signaling and production) is important to plants' differently responding to nutrient availability. To better understand the connections between B and ethylene, here we investigate the function of ethylene in the responses of tomato (Solanum lycopersicum) plants to B stress (deficiency, 0 µM and toxicity, 640 µM), using ethylene related mutants, namely nonripening (nor), ripening-inhibitor (rin), never ripe (Nr), and epinastic (Epi). Our results show that B stress does not necessarily inhibit plant growth, but both B stress and ethylene signaling severely affected physiological parameters, such as photosynthesis, stomatal conductance, and chlorophyll a fluorescence. Under B toxicity, visible symptoms of toxicity appeared in the roots and margins of the older leaves through necrosis, caused by the accumulation of B which stimulated ethylene biosynthesis in the shoots. Both nor and rin (ethylene signaling) mutants presented similar responses, being these genotypes more sensitive and displaying several morphophysiological alterations, including fruit productivity reductions, in response to the B toxicity conditions. Therefore, our results suggest that physiological and metabolic changes in response to B fluctuations are likely mediated by ethylene signaling.

Soybean Plants Exposed to Low Concentrations of Potassium Iodide Have Better Tolerance to Water Deficit through the Antioxidant Enzymatic System and Photosynthesis Modulation.

Water deficit inhibits plant growth by affecting several physiological processes, which leads to the overproduction of reactive oxygen species (ROS) that may cause oxidative stress. In this regard, iodine (I) is already known to possibly enhance the antioxidant defense system of plants and promote photosynthetic improvements under adverse conditions. However, its direct effect on water deficit responses has not yet been demonstrated. To verify the efficiency of I concerning plant tolerance to water deficit, we exposed soybean plants to different concentrations of potassium iodide (KI) fed to pots with a nutrient solution and subsequently submitted them to water deficit. A decline in biomass accumulation was observed in plants under water deficit, while exposure to KI (10 and 20 µmol L-1) increased plant biomass by an average of 40%. Furthermore, exposure to KI concentrations of up to 20 µM improved gas exchange (~71%) and reduced lipid peroxidation. This is related to the higher enzymatic antioxidant activities found at 10 and 20 µM KI concentrations. However, when soybean plants were properly irrigated, KI concentrations greater than 10 µM promoted negative changes in photosynthetic efficiency, as well as in biomass accumulation and partition. In sum, exposure of soybean plants to 10 µM KI improved tolerance to water deficit, and up to this concentration, there is no evidence of phytotoxicity in plants grown under adequate irrigation.

Endophytic colonization of five Trichoderma species and their effects on growth of a Eucalyptus hybrid.

The study aimed to evaluate the effectiveness of endophytic colonization via leaf and root inoculation of five Trichoderma species in a Eucalyptus hybrid, as well as the effects of inoculation on plant growth. The experimental design was completely randomized in a 6 × 2 factorial scheme. Plant growth was evaluated during the experimental period at three different times: 20 days after inoculation (d.a.i), 40 d.a.i., and 60 d.a.i. A statistical difference was observed between the inoculation methods during each period and between the Trichoderma species. Plants inoculated with T. asperellum showed the greatest growth among the treatments. Root-inoculated plants produced the greatest growth response. This showed that the presence of Trichoderma in the roots assisted in nutrient assimilation, promoted greater plant growth, when compared with leaf-inoculated plants. Evaluation of the effectiveness of endophytic colonization was performed at each sampling period by collecting leaf samples, and at 60 d.a.i., by collecting leaf, stem, and root samples. T. longibrachiatum and T. harzianum were isolated from leaves at 20 d.a.i., with an increase in the number of colonized plants throughout the evaluation of leaf-inoculated plants. In root-inoculated plants, treatment with T. longibrachiatum, T. harzianum, and T. asperellum presented the highest endophytic colonization in the stem and root samples (at 60 d.a.i.).

KI Increases Tomato Fruit Quality and Water Deficit Tolerance by Improving Antioxidant Enzyme Activity and Amino Acid Accumulation: A Priming Effect or Relief during Stress?

A water deficit can negatively impact fruit yield and quality, affecting critical physiological processes. Strategies to mitigate water deficits are crucial to global food security. Iodine (I) may increase the efficiency of the antioxidant system of plants, but its role against water deficits is poorly understood. This study aimed to evaluate the effectiveness of I in attenuating water deficits and improving fruit quality, investigating whether metabolic responses are derived from a "priming effect" or stress relief during water deficits. Tomato plants were exposed to different concentrations of potassium iodide (KI) via a nutrient solution and subjected to a water deficit. A water deficit in tomatoes without KI reduced their yield by 98%. However, a concentration of 100 µM of KI increased the yield under a water deficit by 28%. This condition is correlated with increased antioxidant activity, photosynthetic efficiency improvement, and malondialdehyde reduction. In addition, the concentration of 100 µM of KI promoted better fruit quality through antioxidant capacity and a decline in the maturation index. Therefore, KI can be an alternative for attenuating water deficits in tomatoes, inducing positive responses during the water deficit period while at the same time improving fruit quality.

Effects of Trichoderma strigosellum in Eucalyptus urophylla Development and Leaf-Cutting Ant Behavior.

Fungal endophytes can protect plants against herbivory and be used to control leaf-cutting ants. In this study, we aimed to evaluate the potential of endophytic colonization of Eucalyptus urophylla by three filamentous fungal species and their influence on the plant development and foraging behavior of Atta sexdens. The study design was completely randomized and comprised a factorial scheme of 4 × 3, three antagonistic fungal species (Escovopsis sp., Metarhizium anisopliae, and Trichoderma strigosellum) of the leaf-cutting ant, and one control and three inoculation methods (conidial suspension via foliar spray [FS] and soil drench [SD] inoculation, and seedlings inoculated with mycelium [SWM]). The SWM method allowed T. strigosellum to colonize all plant organs, and these plants exhibited higher height, leaf number, shoot dry mass, and total dry mass than the ones subjected to the other inoculation methods. The SWM method increased the plant height than the control plants and those inoculated with Escovopsis sp. and M. anisopliae. Trichoderma strigosellum, previously isolated from soil, colonized E. urophylla plants and positively influenced their development, as demonstrated by the SWM method. Trichoderma strigosellum promoted the increase in E. urophylla height compared with when the FS and SD methods were used (by 19.62% and 18.52%, respectively). Our results reveal that A. sexdens workers preferentially began cutting the leaves from plants not previously colonized by T. strigosellum. This behavior can be explained by modifications in the phenotypic traits of the eucalyptus leaves.

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.

Selenium-Ethylene Interplay in Postharvest Life of Cut Flowers.

Selenium (Se) is considered a beneficial element in higher plants when provided at low concentrations. Recently, studies have unveiled the interactions between Se and ethylene metabolism throughout plant growth and development. However, despite the evidence that Se may provide longer shelf life in ethylene-sensitive flowers, its primary action on ethylene biosynthesis and cause-effect responses are still understated. In the present review, we discuss the likely action of Se on ethylene biosynthesis and its consequence on postharvest physiology of cut flowers. By combining Se chemical properties with a dissection of ethylene metabolism, we further highlighted both the potential use of Se solutions and their downstream responses. We believe that this report will provide the foundation for the hypothesis that Se plays a key role in the postharvest longevity of ethylene-sensitive flowers.

Fumonisin Production by Fusarium verticillioides in Maize Genotypes Cultivated in Different Environments.

Fumonisins are mycotoxins (MTs) produced mainly by the fungus Fusarium verticillioides, the main pathogens of maize which cause ear rot. The aim of this work was to evaluate some factors that may lead to high fumonisin production by F. verticillioides in maize grains, correlating the pathogen inoculation method with different genotypes grown in four Brazilian states. Experiments were conducted in 2015-2016 in maize crops from experimental maize fields located in four distinct states of Brazil. Results showed that contamination by fumonisin mycotoxins occurred even on symptomatic or asymptomatic grains. In all municipalities, the samples showed levels of fumonisin B1 that were higher than would be tolerable for the human consumption of corn products (the current tolerance limit for fumonisin is 1.5 µg g-1). High severity of grains infected with F. verticillioides does not always show high concentrations of fumonisins. Environments with higher temperatures may influence the production of high concentrations of fumonisin in maize hybrids. Spray inoculation methods and inoculation at the center of spikes did not influence fumonisin concentrations. Results showed that the hybrids P3630H, P32R48 and P3250 presented higher disease severity, as well as higher mycotoxin levels in the studied locations with higher temperatures.

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.

Genome Sequence of a New Siphoviridae Phage Found in a Brazilian Bacillus thuringiensis Serovar israelensis Strain.

During the fermentation process, Bacillus thuringiensis (Bt) phages can result in bacterial death and decreased yield. In this work, we describe the genome of a new phage related to the Siphoviridae viral family from a Brazilian strain of Bt which showed high nucleotide sequence identity to the genomes of phages phi4l1 and BtCS33.