Drought stress response in winter wheat varieties – changes in leaf proteins and proteolytic activities

Radiation mutagenesis has been used in sustainable agriculture as a tool for increasing plant variability and providing new lines for selection. This necessitates a comparison, by using suitable stress markers, of the newly created lines with some well-established varieties, which are stress tolerant or susceptible. Drought is one of the most frequently encountered stresses with deleterious effects on plant performance and crop yield. Winter wheat seedlings (soil cultures at 3–4th leaf stage) from one mutant line (M181/1338K), one drought-tolerant (Guinness) and one sensitive variety (Farmer) were subjected to severe drought stress by water withholding, followed by recovery. Changes in leaf protein profiles, the amount of Rubisco large subunit (RLS), some specific chloroplast proteins such as Rubisco binding protein (RPB), Rubisco activase (RA), the chaperone subunit clpA/C of clp protease, as well as the activities of exo- and endo-proteases were analyzed. At the protein level, some differences were found in the drought response of genotypes – stability of RLS and RBP in M181/1338K and Guinness, diminution of RLS and increase in RBP in Farmer. RA presented strong up-regulation at recovery in Guinness but decreased in content under drought in M181/1338K and Farmer. Increase in ClpA/C level was found in all compared varieties under stress. Strong increase in total proteolytic activity was detected under drought only in Farmer. Inhibitory analysis revealed a predominance of cysteine and serine protease types. Aminopeptidase activities remained higher at recovery in M181/1338K and Farmer. Results are discussed in terms of genotype-linked different stress coping strategies

Transcript profiling of serine- and cysteine protease inhibitors in Triticum aestivum varieties with different drought tolerance

A high number of protease inhibitors (PI) have been identified in diverse plant species but information about their role in plant stress responses is still fragmentary. Transcript profiling of six published serine and cysteine protease inhibitor sequences in water-deprived plants from four winter wheat (Triticum aestivum) varieties with varying tolerance was performed in order to outline PIs predominantly accumulating under drought. Expression was analyzed by real time RT-qPCR. Considerable transcript accumulation of Bowman-Birk type PI WALI3 (BBPI) was detected in drought stressed leaves suggesting an important regulatory role of BBPI in adjustment of protein metabolism in leaves under dehydration. Serpin transcripts were less represented in water-deprived plants. Transient accumulation of cystatin transcripts revealed organ-specificity. Under drought cystatin and serpin expression in the leaves of the most drought tolerant variety “Katya” tended to preserve relatively stable levels close to the controls. This preliminary data will serve for future detailed study of regulation of proteolysis in winter wheat subjected to unfavorable environmental factors for development of molecular-based strategies for selection of tolerant varieties

2-DE proteomics analysis of drought treated seedlings of Quercus ilex supports a root active strategy for metabolic adaptation in response to water shortage

Holm oak is a dominant tree in the western Mediterranean region. Despite being well adapted to dry hot climate, drought is the main cause of mortality post-transplanting in reforestation programs. An active response to drought is critical for tree establishment and survival. Applying a gel-based proteomic approach, dynamic changes in root proteins of drought treated Quercus ilex subsp. Ballota [Desf.] Samp. seedlings were followed. Water stress was applied on 20 day-old holm oak plantlets by water limitation for a period of 10 and 20 days, each followed by 10 days of recovery. Stress was monitored by changes in water status, plant growth, and electrolyte leakage. Contrary to leaves, holm oak roots responded readily to water shortage at physiological level by growth inhibition, changes in water status and membrane stability. Root proteins were extracted using trichloroacetate/acetone/phenol protocol and separated by two-dimensional electrophoresis. Coomassie colloidal stained gel images were analyzed and spot intensity data subjected to multivariate statistical analysis. Selected consistent spots in three biological replicas, presenting significant changes under stress, were subjected to MALDI-TOF mass spectrometry (peptide mass fingerprinting and MS/MS). For protein identification, combined search was performed with MASCOT search engine over NCBInr Viridiplantae and Uniprot databases. Data are available via ProteomeXchange with identifier PXD002484. Identified proteins were classified into functional groups: metabolism, protein biosynthesis and proteolysis, defense against biotic stress, cellular protection against abiotic stress, intracellular transport. Several enzymes of the carbohydrate metabolism decreased in abundance in roots under drought stress while some related to ATP synthesis and secondary metabolism increased. Results point at active metabolic adjustment and mobilization of the defense system in roots to actively counteract drought stress

Genetic diversity of sugar beet under heat stress and deficit irrigation

In the light of climate changes and globalwarming, as well as the rapid expansion in sugar beet (Beta vulgaris L.) cultivation in Egypt, the development of sugar beet varieties with improved tolerance to high temperature and deficit irrigation is of great importance. The objective of this studywas to evaluate sugar beet genotypes under high temperatures and deficit irrigation conditions for further identification and selection of heat and drought tolerant genotypes. In the current study, a panel of 18 sugar beet breeding lines produced at the USDA–ARS–NWISRL, Kimberly, ID, and the commercial sugar beet cultivar Kawimera were evaluated for yield and quality under high temperature. Six promising lines in terms of yield and quality were further evaluated under both high temperature and deficit irrigation for two growing seasons. All lines performed differently under deficit irrigation, indicating a high degree of genetic variability in the evaluated lines. Additionally, yield traits showed negative effect due to deficit irrigation. A significant positive correlation was observed between stress tolerance index (STI), and average root and sugar yields under stressed and non-stressed conditions. A linear relationship between STI and average root and sugar yields indicates that STI is a reliable stress index to select high yielding genotypes under both optimum- and deficit-irrigation conditions. USKPS25 and USC944-6-68 breeding lines are most likely adapted to deficit irrigation and high temperature and suitable to be utilized in the proposed sugar beet breeding programs in Egypt

Heat and water stress induce unique transcriptional signatures of heat-shock proteins and transcription factors in grapevine

Grapevine is an extremely important crop worldwide. In southern Europe, post-flowering phases of the growth cycle can occur under high temperatures, excessive light, and drought conditions at soil and/or atmospheric level. In this study, we subjected greenhouse grown grapevine, variety Aragonez, to two individual abiotic stresses, water deficit stress (WDS), and heat stress (HS). The adaptation of plants to stress is a complex response triggered by cascades of molecular networks involved in stress perception, signal transduction, and the expression of specific stress-related genes and metabolites. Approaches such as array-based transcript profiling allow assessing the expression of thousands of genes in control and stress tissues. Using microarrays, we analyzed the leaf transcriptomic profile of the grapevine plants. Photosynthesis measurements verified that the plants were significantly affected by the stresses applied. Leaf gene expression was obtained using a high-throughput transcriptomic grapevine array, the 23K custom-made Affymetrix Vitis GeneChip. We identified 1,594 genes as differentially expressed between control and treatments and grouped them into ten major functional categories using MapMan software. The transcriptome of Aragonez was more significantly affected by HS when compared with WDS. The number of genes coding for heat-shock proteins and transcription factors expressed solely in response to HS suggesting their expression as unique signatures of HS. However, a cross-talk between the response pathways to both stresses was observed at the level of AP2/ERF transcription factors

Holm oak proteomic response to water limitation at seedling establishment stage reveals specific changes in different plant parts as well as interaction between roots and cotyledons

Quercus ilex is a dominant tree species in the Mediterranean region with double economic and ecological importance and increasing use in reforestation. Seedling establishment is extremely vulnerable to environmental stresses, particularly drought. A time course study on physiological and proteomic response of holm oak to water limitation stress and recovery during early heterotrophic growth is reported. Applied stress led to diminution in plant water content and root growth, oxidative stress in roots and some alterations in the anti-oxidative protection. Plant parts differed substantially in soluble sugar and free phenolic content, and in their changes during stress and recovery. Proteomic response in holm oak roots and cotyledons was estimated using combined 1-DE/2-DE approach and protein identification by MALDI TOF-TOF PMF and MS/MS. A total of 127 differentially abundant protein species (DAPs) were identified. DAPs related to starch metabolism, lipid to sugar conversion, reserve proteins and their mobilization were typical for cotyledons. DAPs in roots were involved in sugar utilization, secondary metabolism and defense, including pathogenesis related proteins from PR-5 and PR-10 families. Results emphasize specific proteome signatures of separate plant parts as well as importance of sink-source interaction between root and cotyledon in the time course of stress and in recovery