Morpho-physiological responses and growth indices of triticale to drought and salt stresses.

Salinity and drought are two major abiotic stresses challenging global crop production and food security. In this study, the effects of individual and combined effects of drought (at different phenological stages) and salt stresses on growth, morphology, and physiology of triticale were evaluated. For this purpose, a 3 x 4 factorial design in three blocks experiment was conducted. The stress treatments included three levels of salinity (0, 50, and 100 mM NaCl) and four levels of drought (regular irrigation as well as irrigation disruption at heading, flowering, and kernel extension stages). The stresses, individual as well as combined, caused a significant decrease in chlorophyll contents, total dry matter, leaf area index, relative water content, and grain yield of triticale. In this regard, the highest reduction was recorded under combined stresses of 100 mM NaCl and drought stress at flowering. However, an increase in soluble sugars, leaf free proline, carotenoid contents, and electrolyte leakage was noted under stress conditions compared to the control. In this regard, the highest increase in leaf free proline, soluble sugars, and carotenoid contents were noted under the combination of severe salinity and drought stress imposed at the flowering stage. Investigating the growth indices in severe salinity and water deficit stress in different phenological stages shows the predominance of ionic stress over osmotic stress under severe salinity. The highest grain yield was observed under non-saline well-watered conditions whereas the lowest grain yield was recorded under severe salinity and drought stress imposed at the flowering stage. In conclusion, the flowering stage was more sensitive than the heading and kernel extension stages in terms of water deficit. The impact of salinity and water deficit was more pronounced on soluble sugars and leaf free proline; so, these criteria can be used as physiological indicators for drought and salinity tolerance in triticale.

Proteomic, biochemical, and anatomical influences of nanographene oxide on soybean (Glycine max).

Nano-graphene oxide (NGO) is an engineered nanostructure that is used in various fields including biology, chemistry, medicine, and environmental protection. This kind of highly used nanomaterial (NM) is being released and accumulated gradually in nature and can have some adverse influences on living organisms including plants. Soybean as a cultivated plant with a high importance in food industry, but sensitive to stresses, was chosen in the present study to be examined in terms of proteomic, biochemical, and anatomical properties under the NGO stress. Accordingly, a 2-dimensional gel electrophoresis (2-DE) approach was adopted for proteomic analysis of the NGO treated soybean roots, where significant changes were observed in the abundance of 48 proteins. MALDI TOF/TOF analysis revealed the upregulation of the proteins involved in the redox regulation in plants. Furthermore, anatomical examination of soybean roots under light microscopy showed that the NGO could enter into the root epidermis through the apoplastic pathway and accumulated in some parts of the root. With increasing NGO concentration, the diameter of the vascular apertures increased and then decreased at higher concentrations. To evaluate the toxicity of NGO, some of the growth parameters including fresh and dry weight, and height of the shoots, as well as some stress-related biochemical properties such as H2O2 production, antioxidant enzymes activity, and phenolics and flavonoids contents were measured. The results indicated that NGO could cause an oxidative stress, which can be considered a toxic effect evoking antioxidative and detoxification mechanisms in soybean.

Comparative proteomic analysis of salt-responsive proteins in canola roots by 2-DE and MALDI-TOF MS.

Salinity stress is a major abiotic stress that affects plant growth and limits crop production. Roots are the primary site of salinity perception, and salt sensitivity in roots limits the productivity of the entire plant. To better understand salt stress responses in canola, we performed a comparative proteomic analysis of roots from the salt-tolerant genotype Safi-7 and the salt-sensitive genotype Zafar. Plants were exposed to 0, 150, and 300 mM NaCl. Our physiological and morphological observations confirmed that Safi-7 was more salt-tolerant than Zafar. The root proteins were separated by two-dimensional gel electrophoresis and MALDI-TOF mass spectrometry was applied to identify proteins regulated in response to salt stress. We identified 36 and 25 protein spots whose abundance was significantly affected by salt stress in roots of plants from the tolerant and susceptible genotype, respectively. Functional classification analysis revealed that the differentially expressed proteins from the tolerant genotype could be assigned to 14 functional categories, while those from the susceptible genotype could be classified into 9 functional categories. The most significant differences concerned proteins involved in glycolysis (Glyceraldehyde-3-phosphate dehydrogenase, Fructose-bisphosphate aldolase, Phosphoglycerate kinase 3), stress (heat shock proteins), Redox regulation (Glutathione S-transferase DHAR1, L-ascorbate peroxidase), energy metabolism (ATP synthase subunit B), and transport (V-type proton ATPase subunit B1) which were increased only in the tolerant line under salt stress. Our results provide the basis for further elucidating the molecular mechanisms of salt-tolerance and will be helpful for breeding salt-tolerant canola cultivars.

Metabolomics and proteomics reveal drought-stress responses of leaf tissues from spring-wheat.

To reveal the integrative biochemical networks of wheat leaves in response to water deficient conditions, proteomics and metabolomics were applied to two spring-wheat cultivars (Bahar, drought-susceptible; Kavir, drought-tolerant). Drought stress induced detrimental effects on Bahar leaf proteome, resulting in a severe decrease of total protein content, with impairments mainly in photosynthetic proteins and in enzymes involved in sugar and nitrogen metabolism, as well as in the capacity of detoxifying harmful molecules. On the contrary, only minor perturbations were observed at the protein level in Kavir stressed leaves. Metabolome analysis indicated amino acids, organic acids, and sugars as the main metabolites changed in abundance upon water deficiency. In particular, Bahar cv showed increased levels in proline, methionine, arginine, lysine, aromatic and branched chain amino acids. Tryptophan accumulation via shikimate pathway seems to sustain auxin production (indoleacrylic acid), whereas glutamate reduction is reasonably linked to polyamine (spermine) synthesis. Kavir metabolome was affected by drought stress to a less extent with only two pathways significantly changed, one of them being purine metabolism. These results comprehensively provide a framework for better understanding the mechanisms that govern plant cell response to drought stress, with insights into molecules that can be used for crop improvement projects.

Knowledge and Attitude of Iranian Red Crescent Society Volunteers in Dealing with Bioterrorist attacks.

INTRODUCTION: Bioterrorism is a worldwide problem and has been the focus of attention during recent decades. There is no precise information on the knowledge, attitude, and preparedness of Iranian Red Crescent volunteers in dealing with bioterrorism. Therefore, the present study aimed to evaluate the above-mentioned parameters in Mahabad Red Crescent Society volunteers. METHODS: In this prospective cross-sectional study, the knowledge of 120 volunteers was evaluated and rated as poor, moderate, and good. In addition, attitude of the volunteers and preparedness of Mahabad Red Crescent Society was rated as inappropriate and appropriate using a questionnaire. RESULTS: The mean age of volunteers was 32.0 ± 8.2 years (62.5% male). 2 (1.7%) volunteers had good knowledge while 94 (78.3%) had no knowledge regarding bioterrorist attack management. Only 1 (0.8%) volunteer had appropriate attitude and 6 (5.0%) stated their preparedness for being sent out to the crisis zone. 116 volunteers (96.7%) indicated that Mahabad Red Crescent Society has an inappropriate level of preparedness to encounter bioterrorist attacks. CONCLUSION: The findings of the present study showed poor knowledge and inappropriate attitude of Mahabad Red Crescent Society volunteers in encountering probable bioterrorist attacks. Furthermore, the Red Crescent Society of this town had an inappropriate level of preparedness in the field of bioterrorism from the viewpoint of the studied volunteers.

Differential response of root proteome to drought stress in drought sensitive and tolerant sunflower inbred lines.

Productivity of sunflower (Helianthus annuus L.), the fourth most important oilseed crop, is strongly dependent on water availability. To search for genetic variation in the ability of roots to grow into drying soil, 16 sunflower lines were screened in 2 years field experiments by imposing drought stress at flowering stage. The results differentiated RGK 21 and BGK 329 as the most sensitive and tolerant lines respectively. The time course physiological assay of these lines at seedling stage revealed roots as the most affected organ 6 days after imposing drought stress. A proteomics approach was adapted for investigating of differential changes in roots proteome under contrasting moisture regimes. Protein spots with significant changes in protein abundance were identified by nano LC-MS/MS. The results indicated that under drought stress relative abundance of metabolism related proteins were decreased in both sensitive and tolerant lines. Abundance of energy and disease/defence related proteins were decreased in the sensitive but increased in the tolerant line. The results indicate that changes in energy usage, water transport and ROS scavenging are important mechanisms for maintaining root growth as the soil dries.

Proteome analysis of gut and salivary gland proteins of fifth-instar nymph and adults of the sunn pest, Eurygaster integriceps.

In the digestive system of the sunn pest, Eurygaster integriceps Puton (Hemiptera: Scutelleridae), the salivary gland has a key role in extra oral digestion and the gut is the main site for digestion of food. In this study, proteomics was used to study the role of proteins involved in digestion. The amount of feeding on wheat grain by adult insects increased by comparison to fifth-instar nymphs. Proteins of the gut and salivary gland in adults and fifth-instar nymphs were analyzed 1 day after feeding. The proteins related to digestion, metabolism, and defense against toxins were accumulated in the gut of adult insects. Three plant proteins including serpin, dehydroascorbate reductase, and ß-amylase were accumulated in guts of adults. In the salivary gland, phospholipase A2 and arginine kinase were increased in adults. Heat shock protein 70 increased in the gut of fifth-instar nymphs. Proteomic analysis revealed that most of changed proteins in digestive system of sunn pest were increased in adults. This study provided more targets derived from gut and salivary gland for pest management.

Comparative proteomic analysis of canola leaves under salinity stress.

Although canola is a moderately salt-tolerant species, its growth, seed yield, and oil production are markedly reduced under salt stress, particularly during the early vegetative growth stage. To identify the mechanisms of salt responsiveness in canola, the proteins expressed in the second and third newly developed leaves of salt-tolerant, Hyola 308, and salt-sensitive, Sarigol, cultivars were analyzed. Plants were exposed to 0, 175, and 350 mM NaCl during the vegetative stage. An increase in the Na content and a reduction in growth were observed in the third leaves compared to the second leaves. The accumulation of Na was more pronounced in the salt-sensitive compared with the salt-tolerant genotype. Out of 900 protein spots detected on 2-DE gels, 44 and 31 proteins were differentially expressed in the tolerant and susceptible genotypes, respectively. Cluster analysis based on the expression level of total and responsive proteins indicated that the second leaves had a discriminator role between the two genotypes at both salinity levels. Using MS analysis, 46 proteins could be identified including proteins involved in responses to oxidative stress, energy production, electron transport, translation, and photosynthesis. Our results suggest that these proteins might play roles in canola adaptation to salt stress.

Proteomics approach for identifying osmotic-stress-related proteins in soybean roots.

Osmotic stress can endanger the survival of plants. To investigate the mechanisms by which plants respond to osmotic stress, protein profiles from soybean plants treated with polyethylene glycol (PEG) were monitored by a proteomics approach. Treatment with 10% aqueous PEG reduced the lengths of roots and hypocotyls of soybean seedlings. Proteins from soybean roots were separated by two-dimensional polyacrylamide gel electrophoresis, and 415 proteins were detected by Coomassie brilliant blue staining. Thirty-seven proteins changed by PEG treatment were analyzed using Edman sequencing and peptide-mass fingerprinting method and this group included proteins involved in disease/defense. Seven proteins were selected for further experiments using the results of cluster analysis and statistical analysis of the abundance change. A comparison with the effects of other abiotic stresses showed that caffeoyl-CoA-O-methyltransferase and 20S proteasome alpha subunit A were decreased and increased by abiotic stresses, respectively. Expression analyses of these transcripts were also changed by PEG treatment. Caffeoyl-CoA-O-methyltransferase and 20S proteasome alpha subunit A may control the sensitivity of several regulatory genes specific to short exposure to osmotic stress.

Proteome analysis of rice root plasma membrane and detection of cold stress responsive proteins.

To investigate the function of plant plasma membrane, proteins of rice plasma membrane were analyzed and the proteins changed by cold stress were identified. Plasma membrane proteins were purified with an aqueous two-phase partitioning method from root of rice seedlings, and activity of specific H(+)-ATPase localized in plasma membranes was measured. The plasma membrane proteins were separated by SDS-PAGE or 2D-PAGE, and analyzed with nano LC-MS/MS. The number of transmembrane helices was predicted from the amino acid sequence of annotated proteins. Functional categorization revealed that the most of proteins were associated with energy production, signal transduction, protein synthesis, cell growth/division and defense. In addition, 12 cold stress responsive proteins were identified from the plasma membrane using 2D-PAGE based proteomics method. Out of them, cold shock protein-1 was significantly decreased in plasma membrane of rice under cold stress.

Acoustic technology for high-performance disruption and extraction of plant proteins.

Acoustic technology shows the capability of protein pellet homogenization from different tissue samples of soybean and rice in a manner comparable to the ordinary mortar/pestle method and far better than the vortex/ultrasonic method with respect to the resolution of the protein pattern through two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). With acoustic technology, noncontact tissue disruption and protein pellet homogenization can be carried out in a computer-controlled manner, which ultimately increases the efficiency of the process for a large number of samples. A lysis buffer termed the T-buffer containing TBP, thiourea, and CHAPS yields an excellent result for the 2D-PAGE separation of soybean plasma membrane proteins followed by the 2D-PAGE separation of crude protein of soybean and rice tissues. For this technology, the T-buffer is preferred because protein quantification is possible by eliminating the interfering compound 2-mercaptoethanol and because of the high reproducibility of 2D-PAGE separation.

Tagging QTLs for maximum root length in rainfed lowland rice (Oryza sativa L.) using molecular markers.

A number of morphological, physiological and phenological traits are known to improve the performance of rice challenged by drought. Root morphological traits and stress-induced response form important components of drought tolerance. Enhancing grain yield remains the principal objective of most breeding programs. Interaction between primary traits poses a formidable challenge while dealing with grain yield under stress. The evaluation of root morphology at three different growth stages and grain yield along with related characteristics under contrasting moisture regimes was made using nine backcrosses along with their parent and standard checks. The backcrosses invoved transgressant double haploid lines derived from IR64 and Azucena with IR64. Marked genotypic differences were observed for all root morphology as well as grain yield related characteristics across the sampling dates as revealed by individual and combined ANOVA. Among the nine backcrosses studied in this experiment, the BC1F2 population of P124 x IR64 were evaluated for forwarding based on their performance with respect to maximum root length and grain yield under both well-watered and low-moisture stress conditions. Sixty-nine plants - ten percent of the backcross population - were selectively genotyped using RAPD primers. Under well-watered conditions two RAPD markers showed strong linkage to QTLs for maximum root length evaluated under ww conditions. Two other markers could explain the considerable amount of variation in MRL under LMS. One of the markers identified under low-moisture stress conditions was also able to explain variability in maximum root length in the mean environment.