Responses of photosynthesis, chlorophyll fluorescence and ROS-scavenging systems to salt stress during seedling and reproductive stages in rice

BACKGROUND AND AIMS: Salinity is a widespread soil problem limiting productivity of cereal crops worldwide. Rice is particularly sensitive to salt stress during the seedling stage, with consequent poor crop establishment, as well as during reproduction where salinity can severely disrupt grain formation and yield. Tolerance at the seedling stage is weakly associated with tolerance during reproduction. Physiological responses to salinity were evaluated for contrasting genotypes, during the seedling and reproductive stages. METHODS: Three rice genotypes differing in their tolerance of salinity were evaluated in a set of greenhouse experiments under salt stress during both seedling stage and reproduction. KEY RESULTS: Photosynthetic CO2 fixation, stomatal conductance (gs) and transpiration decreased substantially because of salt stress, but with greater reduction in the sensitive cultivar IR29. The tolerant lines IR651 and IR632 had more responsive stomata that tended to close faster during the first few hours of stress, followed by partial recovery after a brief period of acclimation. However, in the sensitive line, gs continued to decrease for longer duration and with no recovery afterward. Chlorophyll fluorescence measurements revealed that non-photochemical quenching increased, whereas the electron transport rate decreased under salt stress. Salt-tolerant cultivars exhibited much lower lipid peroxidation, maintained elevated levels of reduced ascorbic acid and showed increased activities of the enzymes involved in the reactive oxygen scavenging system during both developmental stages. CONCLUSIONS: Upregulation of the anti-oxidant system appears to play a role in salt tolerance of rice, with tolerant genotypes also maintaining relatively higher photosynthetic function; during both the vegetative and reproductive stages.     

Tissue-specific expression and functional role of dehydrins in heat tolerance of sugarcane (Saccharum officinarum)

Studies on the functional roles of dehydrins (DHNs) in heat tolerance of plants are scarce. This study was conducted to immunohistolocalize DHNs in leaves of heat-tolerant (CP-4333) and heat-sensitive (HSF-240) sugarcane (Saccharum officinarum L.) clones at three phenological stages in order to elucidate their putative roles under heat stress. CP-4333 indicated greater amounts of heat-stable proteins than HSF-240 under heat stress. Western blotting revealed the expression of three DHNs in CP-4333 (13- and 15-kDa peptides at 48 h and an additional 18-kDa band at 72 h) and two (13 and 15 kDa at 48 h) in HSF-240 at formative stage; two DHNs in CP-4333 (20 and 25 kDa) and one in HSF-240 (20 kDa) at grand growth stage, while two DHNs in CP-4333 (20 and 22 kDa) and one in HSF-240 (20 kDa) at maturity stage. Tissue-specific immunohistolocalization showed that DHNs were expressed in stele particularly the phloem and the cells intervening bundle sheath and vascular bundles. Furthermore, DHNs were also found scattered along the epidermal and parenchymatous cells. Recovery of sugarcane from heat stress manifested a gradual disappearance of DHNs in both the clones, being quicker in sensitive clone (HSF-240). Results suggested specific implications for DHNs synthesis. Their synthesis in epidermis appears to protect the mesophyll tissues from heat injury. When associated to vascular tissue, they tend to ensure the normal photoassimilate loading into the sieve element–companion cell complex. DHNs diminution during recovery suggested that their expression was transitory. However, prolonged retention of DHNs by tolerant clone appears to be an adaptive advantage of sugarcane to withstand heat stress.     

Does the rate of German chamomile growth and development influence the response of plants to soil drought?

The response of the wild type (WT) and a strain C6/2 of German chamomile to 7-d soil drought and subsequent 7-day rehydration was studied. Shoot and leaf growth, vegetative development, water and protein contents, ascorbate peroxidase activity and gas exchange were compared. At the stress stage, water content of WT plants was slightly influenced and the effect was ceased after rehydration. Also the decrease in gas exchange was temporary. New leaves were formed, although their area was diminished. On the contrary, leaves of C6/2 plants were more desiccated and the durable decrease in water content was accompanied by the impairment in gas exchange also at the recovery stage (20–40% loss when compared to the control). At both stages of the experiment the growth of the long shoots of this genotype was drastically decreased, as well as leaf formation. Ascorbate peroxidase activity was increased by drought in leaves of both genotypes, but the pattern of changes in WT plants reflected the enhancement of metabolism resulting from proper water content and gas exchange at the recovery stage. Different pattern of changes in the protein content during drought was also noticed: a slight increase in WT, while the decrease by ¼ in C6/2 leaves. The response of WT plants to desiccation and rewatering was found to be more elastic than that of C6/2.     

Evaluation of canopy transpiration rate by applying a plant hormone “abscisic acid”

A method for evaluation of temporal changes in canopy transpiration rate and stomatal conductance in crop fields by using a plant hormone abscisic acid (ABA) has recently been developed. The method was applied to a corn canopy at different growth stages in the upper Yellow River basin, China. Diurnal changes in the canopy transpiration rate and stomatal conductance were evaluated at the initial stage with a leaf area index (LAI) of 0.37 on June 7 and the crop development stage with an LAI of 4.39 on July 15, 2005. The proportions of the accumulated transpiration rate during daytime to the accumulated evapotranspiration were 24% and 74% at the initial and crop development stages, respectively. Stomatal conductance varied in parallel with transpiration rate in the initial stage of the crop. However, in the crop development stage with low soil water content, stomatal conductance reached the maximum value at 10:00 a.m. and thereafter decreased rapidly at around noon with high evaporative demand to corn canopy. This shows the midday stomatal closure was caused by excessive water stress to corn canopy in the crop development stage. Thus, the proposed method with ABA application is useful for evaluation of temporal changes in transpiration rate and stomatal conductance, and hence, can detect the plant water stress.     

Assessment of climatic indices limiting rainfed wheat yield

In this study variation of six climatic indices including accumulated precipitation (P), accumulated potential evapotranspiration (PET), accumulated actual evapotranspiration (AET), accumulated crop evapotranspiration (ETC), accumulated water stress (S) and climatic water deficit (D), was investigated. Climatic indices and their variation were calculated during seven growth stages of wheat in five locations in the northeast of Iran from 1983 to 2008. Principal component analysis (PCA) technique was applied to explore major modes of variation in the regional climatic indices during different crop growth stages. The principle component obtained for each region was correlated to the regional winter wheat yield. Finally the regional amount of water and precipitation use efficiency (WUE and PUE) were analyzed in order to assess any possible association with wheat yield. The results showed that the highest precipitation occurred during the tillering stage and spatially decreased from north (Bojnord) to south (Birjand) and from east (Mashhad) to west (Sabzevar). The difference between the highest and the lowest precipitation across all locations was 2.5 of standard value. The variation pattern of AET, compared to other indices, showed more similarity to variation of precipitation at different growth stages and the highest AET (more than 2 of standard value in all locations) occurred during the tillering stage. The PCA indicated that effective components varied in different locations. The most positive and effective components were types of evapotranspiration that are associated with crop (ETC and AET) and precipitation. However none of these effective PCs showed a significant correlation with final yield. The PUE and WUE analysis indicated that PUE provides more information to interpret the relationship between total amounts of precipitation and the final yield.     

Effects of different levels of water stress on leaf photosynthetic characteristics and antioxidant enzyme activities of greenhouse tomato

Two greenhouse experiments were conducted in order to investigate the effects of different levels of water stress on gas exchange, chlorophyll fluorescence, chlorophyll content, antioxidant enzyme activities, lipid peroxidation, and yield of tomato plants (Solanum lycopersicum cv. Jinfen 2). Four levels of soil water content were used: control (75 to 80% of field water capacity), mild water stress (55 to 60%), moderate water stress (45 to 50%), and severe water stress (35 to 40%). The controlled irrigation was initiated from the third leaf stage until maturity. The results of two-year trials indicated that the stomatal conductance, net photosynthetic rate, light-saturated photosynthetic rate, and saturation radiation decreased generally under all levels of water stress during all developmental stages, while compensation radiation and dark respiration rate increased generally. Water stress also declined maximum quantum yield of PSII photochemistry, electron transfer rate, and effective quantum yield of PSII photochemistry, while nonphotochemical quenching increased in all developmental stages. All levels of water stress also caused a marked reduction of chlorophyll a, chlorophyll b, and total chlorophyll content in all developmental stages, while activities of antioxidant enzymes, such as superoxide dismutase, peroxidase, and catalase, and lipid peroxidation increased.     

Response of pigeon pea cultivars to water stress

Decrease in soil water potential during vegetative and flowering stages of two cultivars of pipeon pea (Cajanus cajari) caused higher decrease in relative water content in cv. ICPL-151 than in cv. H-77-216. Both cultivars showed partial recovery during rehydration. Cv. H-77-216 also accumulated more proline and carbohydrates during stress and showed better drought tolerance than cv. ICPL-151.     

Dehydrin stress proteins in <Emphasis Type="Italic">Pinus sylvestris</Emphasis> L. needles under conditions of extreme climate of Yakutia

This is the first study to investigate stress proteins dehydrins with the use of specific antibodies in the Scots pine (Pinus sylvestris L.) needles and their changes in the annual cycle under extreme climate of Yakutia. No pronounced polymorphism of major dehydrins (14–15 and 66 kDa) has been found during the winter dormancy period of P. sylvestris. A clear correlation between the seasonal variations in dehydrins and changes in the water content in needles was revealed. Consistently high levels of dehydrins was retained throughout the period of low negative temperatures. It is assumed that dehydrins can participate in the formation of P. sylvestris L. resistance to the permafrost conditions.     

Effects of drought stress on phosphorus and potassium uptake dynamics in summer maize (Zea mays) throughout the growth cycle

The effects of drought stress on the phosphorus (P) and potassium (K) uptake dynamics of summer maize (Zea mays L.) throughout the growth cycle were studied. Field trials were conducted under a completely randomized design with three field water capacity (FC) regimes: 75?% FC was well watered and considered to be the control, 55?% FC represented moderate stress (MS), and 35?% FC represented severe stress (SS). The water regimes were applied from the third leaf stage until maturity. Drought stress induced sharp decreases in total K and P uptake of maize organs at different developmental stages and, in particular, detrimentally affected the nutrient uptake capability of roots. SS caused more deleterious effect than MS on both total K and P uptake by plant organs. The results suggested maize plants differ in their ability to maintain nutrient uptake under drought stress, and it is highly dependent on the intensity and duration of drought stress and the developmental stage. The decrease in total K and P uptake caused by both MS and SS was accompanied by reduction in biomass production in drought-stressed tissues. The biomass allocation patterns in response to drought stress fluctuated strong mostly because of competitive changes in the shoot and roots at different stages, thus the root:shoot ratio increased at some stages and decreased at other stages. SS induced a dramatic reduction in the harvest index (HI), whereas MS slightly decreased HI. Thus, water limitation caused lower K and P uptake and HI.     

Glycine betaine application modifies biochemical attributes of osmotic adjustment in drought stressed wheat

Nineteen wheat genotypes were used to examine the effects of foliar applied glycine betaine (GB, 100 mM) on concentration of various osmolytes (such as proline, choline, GB and sucrose) under drought stress conditions. Drought stress caused a significant increase in proline content and GB content of wheat genotypes, both at maximum tillering and anthesis stages. Choline and sucrose were accumulated significantly at higher levels under stress conditions at both the stages. GB application increased the proline content and endogenous levels of GB in comparison to their stressed counterparts both at maximum tillering and anthesis stages but this increase was observed to be genotype specific. Furthermore, significant decrease in choline levels and sucrose contents of GB treated plants at anthesis stage and enhanced levels of proline questioned about involvement of GB in production of other osmolytes as well as stage specific response of wheat genotypes to GB spray. But these changes in osmolyte accumulation (OA) were not correlated with relative water content and stress tolerance index observed, under both GB sprayed and non-sprayed drought stressed conditions. So OA could not be considered as a selection criteria for drought tolerance in wheat.     

Identification of different ABA biosynthesis sites at seedling and fruiting stages in Arachis hypogaea L. following water stress

Abscisic acid (ABA) is one of the most important phytohormones involved in abiotic stress responses. ABA transport in plants is important in determining endogenous ABA levels and their resulting physiological responses. However, the regulation of ABA transport remains unclear. In this study, we compared the ABA concentrations and AhNCED1 levels at seedling and fruiting stages in peanut (Arachis hypogaea L.), in response to water stress. At the seedling stage, ABA initially accumulated in roots (1 h), followed by the lower stem (2 h) and finally in the upper stem (4 h). The expression/activity of an ABA biosynthesis rate-limiting enzyme, AhNCED1, showed the same accumulation patterns. In contrast, during the fruiting stage, ABA and AhNCED1 increases were initially detected in the first apical leaf of main stem, followed by the stem, and finally in the root. These results imply that biosynthesis of ABA in peanut plants subject to water deficiency could be dependent on developmental stage with the roots being the initial site of ABA biosynthesis during the seedling stage, whereas during the fruiting stage ABA biosynthesis occurs initially in the leaf. The distribution patterns of ABA in seedling stage peanuts in response to water stress were: root-stem-leaf, while in fruiting stage peanuts the distribution patterns of ABA were: leaf-stem-root. These findings will help to understand plant regulatory water deficit resistance mechanisms at seedling and fruiting stages and to advance our total understanding of the regulation of ABA transport.     

Detection of dehydrin-like proteins in embryos and endosperm of mature Euterpe edulis seeds

Summary. Euterpe edulis Martius, a tropical palm species characterized as highly recalcitrant, accumulated dehydrin proteins in both the endosperm and the embryo of the mature seed, as detected by Western blot analysis and immunogold electron microscopy. Three major bands at molecular masses of approximately 16, 18, and 24 kDa were identified in both samples analysed. Immunogold electron microscopy studies detected the presence of dehydrins in the embryo and endosperm. In both cases, dehydrins were immunolocalized in cytoplasm and chromatin. No labelling associated with either membranes or organelles was detected. It is known that dehydrins are produced as part of the developmental program of orthodox seeds and are also present in some recalcitrant seeds of temperate regions. The constitutive presence of dehydrins in embryos of extremely recalcitrant species of tropical origin has not been previously reported. Correspondence and reprints: Departamento de Biodiversidad y Biología Experimental, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Pabellón 2, Ciudad Universitaria, C1428EHA, Ciudad Autónoma de Buenos Aires, Argentina.     

Stage-specific protein regulation during somatic embryo development of Carica papaya L. ‘Golden’

Somatic embryogenesis is an important biotechnological technique for large-scale propagation of elite genotypes. Identifying stage-specific compounds associated with somatic embryo development can help elucidate the ontogenesis of Carica papaya L. somatic embryos and improve tissue culture protocols. To identify the stage-specific proteins that are present during the differentiation of C. papaya somatic embryos, proteomic analyses of embryos at the globular, heart, torpedo and cotyledonary developmental stages were performed. Mass spectrometry data have been deposited in the ProteomeXchange with the dataset identifier PXD021107. Comparative proteomic analyses revealed a total of 801 proteins, with 392 classified as differentially accumulated proteins in at least one of the developmental stages. The globular-staged presented a higher number of unique proteins (16), and 7 were isoforms of 60S ribosomal proteins, suggesting high translational activity at the beginning of somatic embryogenesis. Proteins related to mitochondrial metabolism accumulated to a high degree at the early developmental stages and then decreased with increasing development, and they contributed to cell homeostasis in early somatic embryos. A progressive increase in the accumulation of vicilin, late embryogenesis abundant proteins and chloroplastic proteins that lead to somatic embryo maturation was also observed. The differential accumulation of acetylornithine deacetylase and S-adenosylmethionine synthase 2 proteins was correlated with increases in putrescine and spermidine contents, which suggests that both polyamines should be tested to determine whether they increase the conversion rates of globular- to cotyledonary-staged somatic embryos. Taken together, the results showed that somatic embryo development in C. papaya is regulated by the differential accumulation of proteins, with ribosomal and mitochondrial proteins more abundant during the early somatic embryo stages and seed maturation proteins more abundant during the late stages.     

Nitrogen distribution index ofCajanus cajan L. during drought and rehydration

Relative competition among various plant parts for N during water stress,i.e. nitrogen distribution index (NDI) was determined in relation to specific nitrogenase activity (SNA) and nodule and soil nitrogen in both indeterminate (H-77-216) and determinate (ICPL-151) types of pigeonpea (Cajanus cajan L.) under greenhouse conditions. Two levels of water stress,i.e. moderate (soil Ψ_w) -0.77 MPa) and severe (soilΨ_w -1.34 MPa) were created by witholding the irrigation at vegetative (40 DAS) and flowering (70 DAS) stages. At vegetative stage under moderate stress the highest NDI was in nodules of cv. H-77-216 and in leaf of cv. ICPL-151. Under severe stress both the cultivars showed negative values of NDI, with maximum loss of N from root and nodules. Cultivar ICPL-151 behaved differently at flowering and vegetative stages. Very high loss of N from different plant parts was seen at flowering under severe stress. All the plant parts showed gain in N during rehydration. Loss and gain in N at both the stages under stress and rehydration respectively, correlated with available N in soil. Specific nitrogenase activity (SNA) and nodule N were maximum at moderate stress and related with NDI values of leaf and nodules.     

Seasonal Changes in the Composition and Content of Dehydrins in Winter Wheat Plants

Seasonal changes in the pattern and content of dehydrins in winter wheat (Triticum aestivum) plants grown under field and laboratory conditions were studied by one-dimensional PAGE and immunochemical methods. During hardening, plants accumulated dehydrin-like polypeptides with mol wts of 209, 196, 66, 50, and 41 kD. In winter, low-molecular-weight dehydrins with mol wts of 24, 22, 17, 15, and 12 kD were synthesized and accumulated as well. Their content dropped sharply in spring when plants became unhardened. Accumulation/disappearance of these proteins corresponded to the fluctuations in wintering plant frost tolerance before winter and in spring. It is assumed that both high- and medium-molecular-weight dehydrins are involved in plant stress responses and adaptation, whereas low-molecular-weight dehydrins are evidently involved only in the process of low-temperature adaptation.     

Effect of high temperature and water stress on pollen germination and spikelet fertility in rice

In future climates, rice could more frequently be subjected to simultaneous high temperature and water stress during sensitive developmental stages such as flowering. In this study, five rice genotypes were exposed to high temperature, water stress and combined high temperature and water stress during flowering to quantify their response through spikelet fertility. Microscopic analyses revealed significant differences in anther dehiscence between treatments and genotypes, with a moderately high association with the number of germinated pollen grains on the stigma. There was a strong relationship between spikelet fertility and the number of germinated pollen on stigmas. Although, all three stress treatments resulted in spikelet sterility, high-temperature stress caused the highest sterility in all five genotypes. A cumulative linear decline in spikelet fertility with increasing duration of independent high-temperature stress and in combination with water stress was quantified. Better anther dehiscence, higher in vivo pollen germination, and higher spikelet fertility were observed in both the N22 accessions compared with IR64, Apo and Moroberekan under high temperature, water stress and combined stress, indicating its ability to tolerate multiple abiotic stresses.     

Global patterns of gene regulation associated with the development of ectomycorrhiza between birch (Betula pendula Roth.) and Paxillus involutus (Batsch) Fr

The formation of ectomycorrhizal (ECM) root tissue is characterized by distinct morphological and developmental stages, such as preinfection and adhesion, mantle, and Hartig net formation. The global pattern of gene expression during these stages in the birch (Betula pendula)-Paxillus involutus ECM association was analyzed using cDNA microarrays. In comparison with nonsymbiotic conditions, 251 fungal (from a total of 1,075) and 138 plant (1,074 in total) genes were found to be differentially regulated during the ECM development. For instance, during mantle and Hartig net development, there were several plant genes upregulated that are normally involved in defense responses during pathogenic fungal challenges. These responses were, at later stages of ECM development, found to be repressed. Other birch genes that showed differential regulation involved several homologs that usually are implicated in water permeability (aquaporins) and water stress tolerance (dehydrins). Among fungal genes differentially upregulated during stages of mantle and Hartig net formation were homologs putatively involved in mitochondrial respiration. In fully developed ECM tissue, there was an upregulation of fungal genes related to protein synthesis and the cytoskeleton assembly machinery. This study highlights complex molecular interactions between two symbionts during the development of an ECM association.