Alterations in the root phenylpropanoid pathway and root-shoot vessel system as main determinants of the drought tolerance of a soybean genotype.

Climate change increases precipitation variability, particularly in savanna environments. We have used integrative strategies to understand the molecular mechanisms of drought tolerance, which will be crucial for developing improved genotypes. The current study compares the molecular and physiological parameters between the drought-tolerant Embrapa 48 and the sensitive BR16 genotypes. We integrated the root-shoot system's transcriptome, proteome, and metabolome to understand drought tolerance. The results indicated that Embrapa 48 had a greater capacity for water absorption due to alterations in length and volume. Drought tolerance appears to be ABA-independent, and IAA levels in the leaves partially explain the higher root growth. Proteomic profiles revealed up-regulated proteins involved in glutamine biosynthesis and proteolysis, suggesting osmoprotection and explaining the larger root volume. Dysregulated proteins in the roots belong to the phenylpropanoid pathways. Additionally, PR-like proteins involved in the biosynthesis of phenolics may act to prevent oxidative stress and as a substrate for modifying cell walls. Thus, we concluded that alterations in the root-shoot conductive vessel system are critical in promoting drought tolerance. Moreover, photosynthetic parameters from reciprocal grafting experiments indicated that the root system is more essential than the shoots in the drought tolerance mechanism. Finally, we provided a comprehensive overview of the genetic, molecular, and physiological traits involved in drought tolerance mechanisms. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-023-01307-7.

H2O2 priming induces proteomic responses to defense against salt stress in maize.

KEY MESSAGE: H2O2 priming reprograms essential proteins' expression to help plants survive, promoting responsive and unresponsive proteins adjustment to salt stress. ABSTACRT: Priming is a powerful strategy to enhance abiotic stress tolerance in plants. Despite this, there is scarce information about the mechanisms induced by H2O2 priming for salt stress tolerance, particularly on proteome modulation. Improving maize cultivation in areas subjected to salinity is imperative for the local economy and food security. Thereby, this study aimed to investigate physiological changes linked with post-translational protein events induced by foliar H2O2 priming of Zea mays plants under salt stress. As expected, salt treatment promoted a considerable accumulation of Na+ ions, a 12-fold increase. It drastically affected growth parameters and relative water content, as well as promoted adverse alteration in the proteome profile, when compared to the absence of salt conditions. Conversely, H2O2 priming was beneficial via specific proteome reprogramming, which promoted better response to salinity by 16% reduction in Na+ content and shoots growth improvement, increasing 61% in dry mass. The identified proteins were associated with photosynthesis and redox homeostasis, critical metabolic pathways for helping plants survive in saline stress by the protection of chloroplasts organization and carbon fixation, as well as state redox. This research provides new proteomic data to improve understanding and forward identifying biotechnological strategies to promote salt stress tolerance.

Remodeling of the cell wall as a drought-tolerance mechanism of a soybean genotype revealed by global gene expression analysis.

Drought stress is major abiotic stress that affects soybean production. Therefore, it is widely desirable that soybean becomes more tolerant to stress. To provide insights into regulatory mechanisms of the stress response, we compared the global gene expression profiles from leaves of two soybean genotypes that display different responses to water-deficit (BR 16 and Embrapa 48, drought-sensitive and drought-tolerant, respectively). After the RNA-seq analysis, a total of 5335 down-regulated and 3170 up-regulated genes were identified in the BR16. On the other hand, the number of genes differentially expressed was markedly lower in the Embrapa 48, 355 up-regulated and 471 down-regulated genes. However, induction and expression of protein kinases and transcription factors indicated signaling cascades involved in the drought tolerance. Overall, the results suggest that the metabolism of pectin is differently modulated in response to drought stress and may play a role in the soybean defense mechanism against drought. This occurs via an increase of the cell wall plasticity and crosslink, which contributed to a higher hydraulic conductance (K f) and relative water content (RWC%). The drought-tolerance mechanism of the Embrapa 48 genotype involves remodeling of the cell wall and increase of the hydraulic conductance to the maintenance of cell turgor and metabolic processes, resulting in the highest leaf RWC, photosynthetic rate (A), transpiration (E) and carboxylation (A/C i). Thus, we concluded that the cell wall adjustment under drought is important for a more efficient water use which promoted a more active photosynthetic metabolism, maintaining higher plant growth under drought stress. Supplementary Information: The online version contains supplementary material available at 10.1007/s42994-021-00043-4.

Physiological approach to decipher the drought tolerance of a soybean genotype from Brazilian savana.

Drought is one of the major constraints for soybean production in Brazil. In this study we investigated the physiological traits of two soybean parental genotypes under progressive soil drying and rewetting. The plants were evaluated under full irrigation (control) conditions and under water deficit imposed by suspending irrigation until the plants reached predawn leaf water potentials (Ψam) of -1.0 MPa (moderate) and -1.5 MPa (severe). Physiological analyses showed that these genotypes exhibit different responses to water deficit. The Embrapa 48 genotype reached moderate and severe water potential two days after the BR16 genotype and was able to maintain higher levels of A, ETR and ΦPSII even under deficit conditions. This result was not related to changes in gs, 13C isotopic composition and presence of a more efficient antioxidant system. In addition, Fv/Fm values did not decrease in Embrapa 48 genotype in relation to irrigated condition showing that stress was not causing photochemical inhibition of photosynthesis. The greater reduction in the relative growth of the shoots, with concomitant greater growth of the root system under drought, indicates that the tolerant genotype is able to preferentially allocated carbon to the roots, presenting less damage to photosynthesis. Therefore, the physiological responses revealed that the tolerant genotype postponed leaf dehydration by a mechanism involving a more efficient use and translocation of water from root to shoot to maintain cell homeostasis and photosynthetic metabolism under stress.

An integrative overview of the molecular and physiological responses of sugarcane under drought conditions.

Drought is the main abiotic stress constraining sugarcane production. However, our limited understanding of the molecular mechanisms involved in the drought stress responses of sugarcane impairs the development of new technologies to increase sugarcane drought tolerance. Here, an integrated approach was performed to reveal the molecular and physiological changes in two closely related sugarcane cultivars, including the most extensively planted cultivar in Brazil (cv. RB867515), in response to moderate (-0.5 MPa) and severe (-1 MPa) drought stress at the transcriptional, translational, and posttranslational levels. The results show common and cultivar exclusive changes in specific genes related to photosynthesis, carbohydrate, amino acid, and phytohormone metabolism. The novel phosphoproteomics and redox proteomic analysis revealed the importance of posttranslational regulation mechanisms during sugarcane drought stress. The shift to soluble sugar, secondary metabolite production, and activation of ROS eliminating processes in response to drought tolerance were mechanisms exclusive to cv. RB867515, helping to explain the better performance and higher production of this cultivar under these stress conditions.

Integrative Control Between Proton Pumps and SOS1 Antiporters in Roots is Crucial for Maintaining Low Na+ Accumulation and Salt Tolerance in Ammonium-Supplied Sorghum bicolor.

An effective strategy for re-establishing K+ and Na+ homeostasis is a challenge for the improvement of plant performance in saline soil. Specifically, attempts to understand the mechanisms of Na+ extrusion from plant cells, the control of Na+ loading in the xylem and the partitioning of the accumulated Na+ between different plant organs are ongoing. Our goal was to provide insight into how an external nitrogen source affects Na+ accumulation in Sorghum bicolor under saline conditions. The NH4+ supply improved the salt tolerance of the plant by restricting Na+ accumulation and improving the K+/Na+ homeostasis in shoots, which was consistent with the high activity and expression of Na+/H+ antiporters and proton pumps in the plasma membrane and vacuoles in the roots, resulting in low Na+ loading in the xylem. Conversely, although NO3--grown plants had exclusion and sequestration mechanisms for Na+, these responses were not sufficient to reduce Na+ accumulation. In conclusion, NH4+ acts as an efficient signal to activate co-ordinately responses involved in the regulation of Na+ homeostasis in sorghum plants under salt stress, which leads to salt tolerance.

Expression pattern of drought stress marker genes in soybean roots under two water deficit systems.

The study of tolerance mechanisms for drought stress in soybean is fundamental to the understanding and development of tolerant varieties. Using in silico analysis, four marker genes involved in the classical ABA-dependent and ABA-independent pathways of drought response were identified in the Glycine max genome in the present work. The expression profiles of the marker genes ERD1-like, GmaxRD20A-like, GmaxRD22-like and GmaxRD29B-like were investigated by qPCR in root samples of drought sensitive and tolerant soybean cultivars (BR 16 and Embrapa 48, respectively), submitted to water deficit conditions in hydroponic and pot-based systems. Among the four putative soybean homologs to Arabidopsis genes investigated herein, only GmaxRD29B-like was not regulated by water deficit stress. Distinct expression profiles and different induction levels were observed among the genes, as well as between the two drought-inducing systems. Our results showed contrasting gene expression responses for the GmaxRD20A-like and GmaxRD22-like genes. GmaxRD20A-like was highly induced by continuous drought acclimating conditions, whereas GmaxRD22-like responses decreased after abrupt water deprivation. GmaxERD1-like showed a different expression profile for the cultivars in each system. Conversely, GmaxRD20A-like and GmaxRD22-like genes exhibited similar expression levels in tolerant plants in both systems.

Expression analysis in response to drought stress in soybean: Shedding light on the regulation of metabolic pathway genes.

Metabolomics analysis of wild type Arabidopsis thaliana plants, under control and drought stress conditions revealed several metabolic pathways that are induced under water deficit. The metabolic response to drought stress is also associated with ABA dependent and independent pathways, allowing a better understanding of the molecular mechanisms in this model plant. Through combining an in silico approach and gene expression analysis by quantitative real-time PCR, the present work aims at identifying genes of soybean metabolic pathways potentially associated with water deficit. Digital expression patterns of Arabidopsis genes, which were selected based on the basis of literature reports, were evaluated under drought stress condition by Genevestigator. Genes that showed strong induction under drought stress were selected and used as bait to identify orthologs in the soybean genome. This allowed us to select 354 genes of putative soybean orthologs of 79 Arabidopsis genes belonging to 38 distinct metabolic pathways. The expression pattern of the selected genes was verified in the subtractive libraries available in the GENOSOJA project. Subsequently, 13 genes from different metabolic pathways were selected for validation by qPCR experiments. The expression of six genes was validated in plants undergoing drought stress in both pot-based and hydroponic cultivation systems. The results suggest that the metabolic response to drought stress is conserved in Arabidopsis and soybean plants.

Method optimization for proteomic analysis of soybean leaf: Improvements in identification of new and low-abundance proteins.

The most critical step in any proteomic study is protein extraction and sample preparation. Better solubilization increases the separation and resolution of gels, allowing identification of a higher number of proteins and more accurate quantitation of differences in gene expression. Despite the existence of published results for the optimization of proteomic analyses of soybean seeds, no comparable data are available for proteomic studies of soybean leaf tissue. In this work we have tested the effects of modification of a TCA-acetone method on the resolution of 2-DE gels of leaves and roots of soybean. Better focusing was obtained when both mercaptoethanol and dithiothreitol were used in the extraction buffer simultaneously. Increasing the number of washes of TCA precipitated protein with acetone, using a final wash with 80% ethanol and using sonication to ressuspend the pellet increased the number of detected proteins as well the resolution of the 2-DE gels. Using this approach we have constructed a soybean protein map. The major group of identified proteins corresponded to genes of unknown function. The second and third most abundant groups of proteins were composed of photosynthesis and metabolism related genes. The resulting protocol improved protein solubility and gel resolution allowing the identification of 122 soybean leaf proteins, 72 of which were not detected in other published soybean leaf 2-DE gel datasets, including a transcription factor and several signaling proteins.