Functional dissection of drought-responsive gene expression patterns in Cynodon dactylon L.

Water deficit is one of the main abiotic factors that affect plant productivity in subtropical regions. To identify genes induced during the water stress response in Bermudagrass (Cynodon dactylon), cDNA macroarrays were used. The macroarray analysis identified 189 drought-responsive candidate genes from C. dactylon, of which 120 were up-regulated and 69 were down-regulated. The candidate genes were classified into seven groups by cluster analysis of expression levels across two intensities and three durations of imposed stress. Annotation using BLASTX suggested that up-regulated genes may be involved in proline biosynthesis, signal transduction pathways, protein repair systems, and removal of toxins, while down-regulated genes were mostly related to basic plant metabolism such as photosynthesis and glycolysis. The functional classification of gene ontology (GO) was consistent with the BLASTX results, also suggesting some crosstalk between abiotic and biotic stress. Comparative analysis of cis-regulatory elements from the candidate genes implicated specific elements in drought response in Bermudagrass. Although only a subset of genes was studied, Bermudagrass shared many drought-responsive genes and cis-regulatory elements with other botanical models, supporting a strategy of cross-taxon application of drought-responsive genes, regulatory cues, and physiological-genetic information.     

Different acclimatization mechanisms of two grass pea cultivars to osmotic stress in in vitro culture

Grass pea (Lathyrus sativus L.) is a legume crop known from its tolerance to various abiotic stresses, especially drought. In this study, we investigated: (1) the response of grass pea seedlings to osmotic stress generated in vitro by polyethylene glycol (PEG); (2) potential drought acclimatization mechanisms of two polish grass pea cultivars. Grass pea seeds of two cultivars were sown on media containing different PEG concentrations (0, 5.5, 11.0 mM) and cultivated for 14 days in controlled conditions. Plants’ dry matter increased under osmotic stress (regardless of PEG concentration). In turn, the highest dose of PEG caused a reduction in seedling growth in both cultivars. Furthermore, PEG caused the peroxidase activity increase in whole seedlings and catalase (CAT) activity in roots. However, differences between cultivars were noted in: CAT activity in shoots; while phenols and anthocyanin content as well as electrolyte leakage in shoots and roots. In turn, in both tested genotypes, accumulation of proline increased in shoots under osmotic stress. Obtained results indicate that the examined plants, although belonging to the same species, differ in acclimatization processes leading to elevated tolerance to osmotic stress.     

Thermal stress responses in Antarctic yeast, Glaciozyma antarctica PI12, characterized by real-time quantitative PCR

Living organisms have some common and unique strategies to response to thermal stress. However, the amount of data on thermal stress response of certain organism is still lacking, especially psychrophilic yeast from the extreme habitat. Therefore, it is not known whether psychrophilic yeast shares the common responses of other organisms when exposed to thermal stresses. In this work, the cold shock and heat shock responses in Antarctic psychrophilic yeast Glaciozyma antarctica PI12 which had an optimal growth temperature of 12 °C were determined. The expression levels of 14 thermal stress-related genes were measured using real-time quantitative PCR (qPCR) when the yeast cells were exposed to cold shock (0 °C), mild cold shock (5 °C), and heat shock (22 °C) conditions. The expression profiles of the 14 genes at these three temperatures varied indicating that these genes had their specific roles to ensure the survival of the yeast. Under cold shock condition, the afp4 and fad genes were over-expressed possibly as a way for the G. antarctica PI12 to avoid ice crystallization in the cell and to maintain the membrane fluidity. Under the heat shock condition, hsp70 was significantly up-regulated possibly to ensure the proteins fold properly. Among the six oxidative stress-related genes, MnSOD and prx were up-regulated under cold shock and heat shock, respectively, possibly to reduce the negative effects caused by oxidative stress. Interestingly, it was found that the trehalase gene, nth1 that plays a role in degrading excess trehalose, was down-regulated under the heat shock condition possibly as an alternative way to accumulate trehalose in the cells to protecting them from being damaged.     

Dithiothreitol and PEG Induced Combined Stress May Affect the Expressions of ABA Aldehyde Oxidase,Sucrose Synthase and Proline Metabolic Genes in Maize Seedlings

The endoplasmic reticulum (ER) is an organelle in the cell where proteins are created and folded. Folding is a very elaborate process that is often interrupted by various biotic and abiotic stresses, leading to the formation of unfolded and misfolded proteins called ER stress. Dithiothreitol (DTT)-induced unfolded protein response (UPR) in endoplasmic reticulum (ER) has been recently reported in plants. Also, previous studies demonstrated that treatment with polyethylene glycol (PEG₆₀₀₀) could stimulate water deficit in crops. However, further researches should be conducted to elucidate the molecular mechanism of ER stress response and the relationship between water deficiency and ER. In this study, we examined the expressions of sucrose synthase (SuS) gene, proline metabolic genes and abscisic aldehyde oxidase (AAO3) gene in maize seedlings that were subjected to DTT and PEG induced combined stresses by using quantitative real-time RT-PCR. Three weeks old detached maize seedlings were treated with or without DTT and PEG₆₀₀₀ for 12 h. The treatment with DTT increased about 2-fold the expression of gene encoding proline synthesis enzyme, pyrroline-5-carboxylate synthetase (P5CS) but no statistically affected the proline catabolism enzyme, proline dehydrogenase (ProDH) in comparison with un-treated seedlings. PEG treatment was also up-regulated P5CS while it was down-regulated ProDH. The relative expression levels of SuS and AAO3 genes statistically enhanced about 2.5 fold under the DTT-induced ER stress. Likewise, the expression levels of SuS and AAO3 genes were up-regulated in the detached seedlings exposed to PEG-induced water deficit. Conversely, the induced gene expressions were down-regulated under the combined stress, the DTT-induced ER stress and PEG-induced water deficit in comparison with the singular stress responses (DTT or PEG). The results indicated that the expressions of genes, related to the synthesis of some signal osmolyte compounds such as proline and sucrose can be suppressed when ER stress occurred under water deficiency in maize seedlings. The changes in the expressions of genes involved in osmolyte and ABA metabolism can be related to ER stress response as well as variations in water status.     

Characterization of Afp1, an antifreeze protein from the psychrophilic yeast Glaciozyma antarctica PI12

The psychrophilic yeast Glaciozyma antarctica demonstrated high antifreeze activity in its culture filtrate. The culture filtrate exhibited both thermal hysteresis (TH) and ice recrystallization inhibition (RI) properties. The TH of 0.1 °C was comparable to that previously reported for bacteria and fungi. A genome sequence survey of the G. antarctica genome identified a novel antifreeze protein gene. The cDNA encoded a 177 amino acid protein with 30 % similarity to a fungal antifreeze protein from Typhula ishikariensis. The expression levels of AFP1 were quantified via real time-quantitative polymerase chain reaction (RT-qPCR), and the highest expression levels were detected within 6 h of growth at ?12 °C. The cDNA of the antifreeze protein was cloned into an Escherichia coli expression system. Expression of recombinant Afp1 in E. coli resulted in the formation of inclusion bodies that were subsequently denatured by treatment with urea and allowed to refold in vitro. Activity assays of the recombinant Afp1 confirmed the antifreeze protein properties with a high TH value of 0.08 °C.     

Physiological and Biochemical Responses Reveal the Drought Tolerance Efficacy of the Halophyte Salicornia brachiata

The drought tolerance of Salicornia brachiata seedlings was assessed by monitoring growth, nutrient uptake, electrolyte leakage, lipid peroxidation, and biochemical responses under drought conditions simulated with 0, 10, 20, and 30 % polyethylene glycol (PEG 6000). After 7 days of drought induction, plants were harvested for measurement of various parameters. The biomass decreased and the plant height remained unchanged with PEG treatment. The total plant water content (TWC%) decreased by 11 % at the highest concentration of PEG (30 %). The electrolyte leakage and lipid peroxidation of shoots increased by 17 and 5 %, respectively, in 30 % PEG-treated plants. K⁺ and Ca²⁺ contents of shoots increased in a dose-dependent manner. However, in roots K⁺ content decreased and Ca²⁺ content remained unaffected by PEG treatment. Mg²⁺ content increased at high concentrations of PEG (20–30 %) in shoots and decreased at the highest concentration of PEG (30 %) in roots. Total free amino acids, proline, and polyphenol contents increased progressively with increase in severity of the drought stress. Total sugar content and reducing sugar content increased in 10 and 20 % PEG-treated plants and decreased in 30 % PEG-treated plants. Our results suggest that proline and other free amino acids, sugars, and polyphenols are the main compatible solutes in S. brachiata for maintenance of osmotic balance, protection of cellular macromolecules, detoxification of the cells, and scavenging of free radicals under drought stress. A greater accumulation of compatible solutes also facilitates the maintenance of nutrient uptake and adequate tissue water status and protection of membranes under drought conditions in S. brachiata. The results from the present study suggest that S. brachiata can be used for restoration of arid and semiarid lands of coastal ecosystems.