Transcriptomic profiling of maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.) seedlings in response to <Emphasis Type="Italic">Pseudomonas putida</Emphasis> stain FBKV2 inoculation under drought stress

Several mechanisms have been proposed for plant growth-promoting rhizobacteria (PGPR)-mediated drought stress tolerance in plants, but little is known about the molecular pathways involved in the drought tolerance promoted by PGPR. We, therefore, aim to study the differential gene response between Pseudomonas putida strain FBKV2 and maize interaction under drought stress using Illumina sequencing. RNA Seq libraries were generated from leaf tissue of maize seedlings with and without strain FBKV2 subjected to drought stress. The libraries were mapped with maize genome database for the identification of differentially expressed genes (DEGs). The expression studies confirmed the downregulation of ethylene biosynthesis (ET), abscisic acid (ABA) and auxin signaling, superoxide dismutase, catalase, and peroxidase in FBKV2-inoculated seedlings. On the other hand, genes involved in β-alanine and choline biosynthesis, heat shock proteins, and late embryogenesis abundant (LEA) proteins were upregulated, which could act as key elements in the drought tolerance conferred by P. putida strain FBKV2. Another remarkable expression was observed in genes encoding benzoxazinoid (BX) biosynthesis which act as the chemoattractant, which was further confirmed by gfp-labeled P. putida strain FBKV2 root colonization studies. Overall, these results indicate that secretion of BXs attracted P. putida strain FBKV2 resulted in root colonization and mediated drought tolerance by modulating metabolic, signaling, and stress-responsive genes.     

<Emphasis Type="Italic">Platycladus orientalis</Emphasis> PoKub3 is involved in abiotic stress responses in transgenic arabidopsis

Ku70-binding proteins associate with Ku70 and their expression levels can affect DSB repair efficiency via the DNA-PK-dependent repair pathway. However, how Ku70-binding proteins in plants exert a regulatory function under abiotic stress is poorly understood. Here, we cloned and characterized a PoKub3 gene from 500-year-old Platycladus orientalis. With increasing age, PoKub3 expression in P. orientalis increased gradually. The PoKub3 expression levels in leaves were upregulated under salt, heat, UV-C and abscisic acid treatments according to qRT-PCR. Moreover, PoKub3 overexpression in Arabidopsis thaliana improved tolerance to salt and drought stress compared with wild-type (WT) and vector control (VC) plants. High RAB18 and DREB2A expression and low JAZ1 and ABI2 expression provided strong evidence that salt tolerance was enhanced in the overexpression plants. Similarly, high RAB18 and DREB2A expression, accompanied by low JAZ1 and LOX1 expression and high DREB1A, CPK10, GSTF6 and APX1 expression, suggested the drought tolerance mechanism was associated with the abscisic acid pathway. In addition, lower malondialdehyde content, electrolyte leakage and stomatal conductance, and higher soluble sugar and relative water contents in PoKub3 overexpression lines than in WT and VC plants demonstrated its role in salt and drought tolerance. Together, these findings show that PoKub3 positively regulates salt and drought tolerance by regulating stress-related genes.     

Potential role of D-<Emphasis Type="Italic">myo</Emphasis>-inositol-3-phosphate synthase and 14-3-3 genes in the crosstalk between <Emphasis Type="Italic">Zea mays</Emphasis> and <Emphasis Type="Italic">Rhizophagus intraradices</Emphasis> under drought stress

Arbuscular mycorrhizal (AM) symbiosis is known to stimulate plant drought tolerance. However, the mechanisms underlying the synergistic responses of the symbiotic partners to drought stress are largely unknown. A split-root experiment was designed to investigate the molecular interactions between a host plant and an AM fungus (AMF) under drought stress. In the two-compartment cultivation system, an entire or only a half root system of a maize plant was inoculated with an AMF, Rhizophagus intraradices, in the presence of localized or systemic drought treatment. Plant physiological parameters including growth, water status, and phosphorus concentration, and the expression of drought tolerance-related genes in both roots and R. intraradices were recorded. Although mycorrhizal inoculation in either one or both compartments systemically decreased abscisic acid (ABA) content in the whole root system subjected to systemic or local drought stress, we observed local and/or systemic AM effects on root physiological traits and the expression of functional genes in both roots and R. intraradices. Interestingly, the simultaneous increase in the expression of plant genes encoding D-myo-inositol-3-phosphate synthase (IPS) and 14-3-3-like protein GF14 (14-3GF), which were responsible for ABA signal transduction, was found to be involved in the activation of 14-3-3 protein and aquaporins (GintAQPF1 and GintAQPF2) in R. intraradices. These findings suggest that coexpression of IPS and 14-3GF is responsible for the crosstalk between maize and R. intraradices under drought stress, and potentially induces the synergistic actions of the symbiotic partners in enhancing plant drought tolerance.     

Root tip structure and volatile organic compound responses to drought stress in Masson pine (<Emphasis Type="Italic">Pinus massoniana</Emphasis> Lamb.)

The Masson pine (Pinus massoniana Lamb.) has a relatively strong tolerance to stressors such as acid rain, drought, and phosphorus deficiency. Previous studies have mainly focused on morphological, physiological, and metabolic responses in the needles of the Masson pine. However, there has been little study of the effects of drought stress on root tip structure or the composition of volatile organic compounds (VOCs) in the root tip. Therefore, this work aimed to evaluate the VOCs and anatomical growth responses of Masson pine root tips to drought stress. The results showed that (1) drought stress inhibited increases in root biomass, root growth, and lateral root number, and damaged and significantly deformed the root apical structure; (2) the relative contents of monoterpenes decreased with the intensity of drought stress, but the relative content of sesquiterpenes increased. (3) The ratio of sesquiterpenes to monoterpenes in the root tip was approximately 1:2 in the absence of stress, and became 3:1 under severe drought stress. The results indicate changes of secondary metabolism in the root tip of the Masson pine in response to drought stress.     

Differentially expressed proteins associated with drought tolerance in bananas (<Emphasis Type="Italic">Musa</Emphasis> spp.)

Bananas are one of the most important fruits in tropical and subtropical regions worldwide. Each year, banana plantations expand, but the areas available are mostly dry lands. The establishment of strategies for obtaining drought tolerant cultivars depends on understanding of biological responses at genetic, molecular and biochemical levels. Proteomics is a powerful tool for functional characterization of the response of plants to abiotic stress and little is known about drought tolerance in Musa spp. Therefore, the aim of this study was to identify proteins related to drought tolerance in two contrasting banana genotypes, Prata Anã and BRS Tropical, susceptible and tolerant to drought, respectively. Proteins were extracted from rhizomes of bananas grown under greenhouse conditions with control, irrigated and water deficit regimes. The differential protein expression pattern was established by two-dimensional (2-D) electrophoresis and spots analyzed in nano Q-Tof Micro UPLC. Twenty-three differentially expressed proteins were found in the tolerant genotype (BRS Tropical) under water deficit, with proteins involved in metabolism, defense and transport. Proteins were classified according to known function and biosynthetic pathways. Signaling proteins in response to water stress, especially for the biological function of growth and development of plants cells, were also encountered, whereas heat shock proteins played a significant role. This is the first report of proteomic analysis for drought tolerance in ‘Pome’ and ‘Silk-type’ bananas containing the ‘B’ genome. Our work provides insights into Musa spp. response to drought and data for further studies regarding molecular mechanisms, which determine how Musa spp. cells better overcome environmental perturbations.     

Responses to drought preconditioning in<Emphasis Type="Italic"> Eucalyptus globulus</Emphasis> Labill. provenances

Shoot water relations and morphological responses to drought preconditioning were studied by subjecting 5-month-old seedlings of three provenances of Eucalyptus globulus to different water regimes for 36 days in a greenhouse pot study. Moderately stressed plants were watered every 6 days and severely stressed plants were watered every 9 days. Control plants were watered daily. Drought cycles induced significant changes in morphological and physiological characteristics. Preconditioned seedlings were smaller in size, root collar diameter, height, and leaf area than control seedlings. Shoot/root ratio was not affected by drought. Osmotic potential at full turgor (ψπ_FT) and osmotic potential at turgor loss point (ψπ_TLP) were significantly lower and the magnitude of osmotic adjustment was significantly higher under the severe than under the moderate stress treatment. In severely stressed plants a decrease of turgid mass/dry mass contributed to osmotic adjustment. In a subsequent acclimation test, preconditioned seedlings showed higher values of stomatal conductance, predawn relative water content and water potential and lower mortality than control plants. These variables were significantly related to ψπ_FT. We assume that the reduced leaf area and osmotic adjustment observed in preconditioned seedlings contributed to drought acclimation in the selected E. globulus provenances leading to better rates of gas exchange and improved water status than non-conditioned plants. Provenances exhibited differences in their responses to drought, albeit mainly morphological differences. E. globulus subsp. bicostata from Tumbarumba grew more quickly (larger diameter and height relative growth rate) than the other provenances, implying a greater ability to tolerate water stress. It can be expected that preconditioned seedlings will display greater tolerance of water stress than non-conditioned plants and perform better during early establishment (higher survival and early growth).     

The abundance of certain metabolites responds to drought stress in the highly drought tolerant plant <Emphasis Type="Italic">Caragana korshinskii</Emphasis>

Metabolomics offers opportunities for studying the systematic response of an organism to a genetic and/or an environmental change. Here, the metabolic consequences of drought stress were characterized in the highly drought tolerant plant Caragana korshinskii. The time-of-flight mass spectrometry platform employed identified several hundred metabolites in extracts of the leaf, stem, root collar, and root of plants which had been either subjected to drought stress or were well-watered. Each of the four organs harbored a number of potential metabolite markers for the drought response. An increased abundance of various small carbohydrates and soluble amino acids in each of the four organs was induced by the stress; these compounds may act as compatible solutes or antioxidants. Across the whole plant, there was a fall in the content of several Krebs cycle and glycolysis intermediates, as well as in that of the amino acids glutamic acid and aspartic acid. Pathway analysis suggested that most of the potential metabolite markers were involved in energy metabolism and amino-acid metabolism. The implication was that energy metabolism and photosynthesis are compromised during the adaptation of C. korshinskii to drought stress. Given the different spectrum of metabolites associated with the drought response in the four organs, it was concluded that each organ employs a distinct strategy to cope with drought stress.     

Differential accumulation of proteins in leaves and roots associated with heat tolerance in two Kentucky bluegrass genotypes differing in heat tolerance

To understand the mechanisms of heat stress responses in perennial grasses, differential proteins in leaves and roots of two genotypes of Kentucky bluegrass (Poa pratensis), including heat-tolerant ‘Midnight’ and heat-sensitive ‘Brilliant’, were analyzed with two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS). Plants were exposed to heat stress for 28 days in growth chambers. Under 7–28 days of heat stress, leaf photochemical efficiency declined significantly while electrolyte leakage increased in leaves and roots, and to a lesser extent for heat-tolerant ‘Midnight’ than for heat-sensitive ‘Brilliant’. Compared with leaves, cell membrane damage due to heat stress was more severe in roots. The 2-DE and MS analysis identified 37 heat-responsive proteins in leaves, 28 heat-responsive proteins in roots; 14 proteins in leaves and 9 proteins in roots exhibited differential expression between the two genotypes. The results indicate that proteins involved in metabolism and energy in leaves and those in antioxidant defense in roots are associated with heat tolerance in Kentucky bluegrass. The differential accumulation of these proteins might be the reason for different heat tolerance in two Kentucky bluegrass genotypes in aerial and underground parts.     

Effects of drought stress on growth and chlorophyll fluorescence of Lycium ruthenicum Murr. seedlings

The present study aimed to determine effects of drought stress on Lycium ruthenicum Murr. seedlings. Our results showed that mild drought stress was beneficial to growth of L. ruthenicum seedlings. Their height, basal diameter, crown, leaf number, stem dry mass, leaf and root dry mass increased gradually when the soil water content declined from 34.7 to 21.2%. However, with further decrease of the soil water content, the growth of L. ruthenicum seedlings was limited. After 28 d of treatment, the seedlings were apparently vulnerable to drought stress, which resulted in significant leaf shedding and slow growth. However, growth was restored after rehydration. Drought treatments led to a decrease in contents of chlorophyll (Chl) a, b, and Chl (a+b) and increase in the Chl a/b ratio. After rewatering, the Chl content recovered to the content of the control plants. Under drought stress, minimal fluorescence and nonphotochemical quenching coefficient increased, thereby indicating that L. ruthenicum seedlings could protect PSII reaction centres from damage. Maximum fluorescence, maximum quantum yield, actual quantum yield of PSII photochemistry, and photochemical quenching decreased, which suggested that drought stress impacted the openness of PSII reaction centres. A comparison of these responses might help identify the drought tolerance mechanisms of L. ruthenicum. This could be the reference for the planting location and irrigation arrangements during the growing period of L. ruthenicum.     

AM1 is a potential ABA substitute for drought tolerance as revealed by physiological and ultra-structural responses of oilseed rape

Abscisic acid (ABA) is an important signaling molecule for plants under drought tolerance. However, ABA itself has many limitations to be used in agriculture practically. Recently, AM1 (ABA-mimicking ligand) has been found to replace ABA. In this study, we have investigated AM1’s potential role for drought tolerance by growing two contrasting rapeseed (Brassica napus L.) genotypes: Qinyou 8 (drought sensitive) and Q2 (drought resistant) with exogenous ABA or AM1 application under well-watered and drought-stressed conditions. Results demonstrate that drought stress has hampered plant growth (relative height growth rate, plant biomass, leaf area), plant water status (leaf relative water content, root moisture content, leaf water potential), photosynthetic gas exchange attributes like net photosynthesis rate (Pn), stomatal conductance (Gs), intercellular CO₂ concentration (Ci), transpiration rate (E); chlorophyll fluorescence parameters like photosynthetic efficiency (Fv/Fm), effective quantum yield of PSII (Φ _PSII ), photochemical quenching coefficient (qL), electron transport rate (ETR) and chlorophyll content, especially for Qinyou 8 significantly compared to well-watered plants. Whereas increased root/shoot ratio (R/S), water use efficiency (WUE) and non-photochemical quenching (NPQ) was recorded in both genotypes under drought stress. On the other hand, exogenous ABA or AM1 treatment has regulated all the above parameters in a rational way to avoid drought stress. Chloroplast transmission electron microscope images, especially for Qinyou8, have revealed that oxidative stress induced by drought has blurred the grana thylakoids, increased the size or number of plastoglobules due to lipid peroxidation, and the presence of starch granules depict weak capacity to convert them into simple sugars for osmotic adjustment. However, intact grana thylakoid, few plastoglobules with no or very few starch granules were observed in the chloroplast from ABA- or AM1-treated plants under drought. More importantly, AM1-treated plants under drought stress have responded in an extremely similar way like ABA-treated ones. Finally, it is suggested that AM1 is a potential ABA substitute for plant drought tolerance.     

Drought tolerance and proteomics studies of transgenic wheat containing the maize C<Subscript>4</Subscript> phosphoenolpyruvate carboxylase (<Emphasis Type="Italic">PEPC</Emphasis>) gene

Enhancing drought tolerance of crops has been a great challenge in crop improvement. Here, we report the maize phosphoenolpyruvate carboxylase (PEPC) gene was able to confer drought tolerance and increase grain yield in transgenic wheat (Triticum aestivum L.) plants. The improved of drought tolerance was associated with higher levels of proline, soluble sugar, soluble protein, and higher water use efficiency. The transgenic wheat plants had also a more extensive root system as well as increased photosynthetic capacity during stress treatments. The increased grain yield of the transgenic wheat was contributed by improved biomass, larger spike and grain numbers, and heavier 1000-grain weight under drought-stress conditions. Under non-stressed conditions, there were no significant increases in these of the measured traits except for photosynthetic rate when compared with parental wheat. Proteomic research showed that the expression levels of some proteins, including chlorophyll A-B binding protein and pyruvate, phosphate dikinase, which are related to photosynthesis, PAP fibrillin, which is involved in cytoskeleton synthesis, S-adenosylmethionine synthetase, which catalyzes methionine synthesis, were induced in the transgenic wheat under drought stress. Additionally, the expression of glutamine synthetase, which is involved in ammonia assimilation, was induced by drought stress in the wheat. Our study shows that PEPC can improve both stress tolerance and grain yield in wheat, demonstrating the efficacy of PEPC in crop improvement.