Response of tropical trees to elevated Ozone: a Free Air Ozone Enrichment study.

Tropospheric ozone (O3) has become one of the main urban air pollutants. In the present study, we assessed impact of ambient and future ground-level O3 on nine commonly growing urban tree species under Free Air Ozone Enrichment (FAOE) condition. During the study period, mean ambient and elevated ozone (EO3) concentrations were 48.59 and 69.62 ppb, respectively. Under EO3 treatment, stomatal density (SD) significantly decreased and guard cell length (GCL) increased in Azadirachta indica, Bougainvillea spectabilis, Plumeria rubra, Saraca asoca and Tabernaemontana divaricata, while SD increased and GCL decreased in Ficus benghalensis and Terminalia arjuna. Proline levels increased in all the nine plant species under EO3 condition. EO3 significantly reduced photosynthetic rate, stomatal conductance (gs), and transpiration rates (E). Only A. indica and N. indicum showed higher gs and E under EO3 treatment. Water use efficiency (WUE) significantly increased in F. benghalensis and decreased in A. indica and T. divaricata. Air Pollution Tolerance Index (APTI) significantly increased in Ficus religiosa and S. asoca whereas it decreased in B. spectabilis and A. indica. Of all the plant species B. spectabilis and A. indica were the most sensitive to EO3 (high gs and less ascorbic acid content) while S. asoca and F. religiosa were the most tolerant (lowgs and more ascorbic acid content). The sensitivity of urban tree species to EO3 is a cause of concern and should be considered for future urban forestry programmes. Our study should guide more such studies to identify tolerant trees for urban air pollution abatement.

Impact of chronic elevated ozone exposure on photosynthetic traits and anti-oxidative defense responses of Leucaena leucocephala (Lam.) de wit tree under field conditions.

In this study, the impact of long term exposure of elevated ozone (+20 ppb above ambient) on photosynthetic traits and anti-oxidative defense system of Leucaena leucocephala, a tree of great economic importance, was studied in a Free Air Ozone Concentration Enrichment (O3-FACE) facility at different time intervals (6, 12, 18, and 24 months). Results showed that net photosynthesis, photosynthetic pigments and lipid peroxidation were significantly reduced after 6, 12 and 24 months of exposure to elevated ozone (eO3) whereas stomatal conductance and transpiration rate were significantly decreased after 12 months of exposure to eO3. Antioxidant enzymatic activities (catalase, ascorbate peroxidase and glutathione reductase) were significantly increased after 12 months of exposure to eO3. Ascorbate was increased significantly after 6 and 12 months of exposure to eO3 while reduced glutathione content declined significantly after 6 and 24 months of exposure to eO3. The study showed that there were several negative long lasting physiological and biochemical responses in Leucaena. The results provide evidence that Leucaena exhibited greater sensitivity to O3 during initial exposure (up to 12 months) but showed moderate tolerance by the end of the 2nd year.

Root system architecture, physiological analysis and dynamic transcriptomics unravel the drought-responsive traits in rice genotypes.

Drought is the major abiotic factors that limit crop productivity worldwide. To withstand stress conditions, plants alter numerous mechanisms for adaption and tolerance. Therefore, in the present study, 106 rice varieties were screened for drought tolerance phenotype via exposing different concentrations of polyethylene glycol 6000 (PEG) in the hydroponic nutrient medium at the time interval of 1, 3, and 7 days to evaluate the changes in their root system architecture. Further, based on root phenotype obtained after PEG-induced drought, two contrasting varieties drought-tolerant Heena and -sensitive Kiran were selected to study transcriptional and physiological alterations at the same stress durations. Physiological parameters (photosynthesis rate, stomatal conductance, transpiration), and non-enzymatic antioxidants (carotenoids, anthocyanins, total phenol content) production indicated better performance of Heena than Kiran. Comparatively higher accumulation of carotenoid and anthocyanin content and the increased photosynthetic rate was also observed in Heena. Root morphology (length, numbers of root hairs, seminal roots and adventitious roots) and anatomical data (lignin deposition, xylem area) enable tolerant variety Heena to better maintain membrane integrity and relative water content, which also contribute to comparatively higher biomass accumulation in Heena under drought. In transcriptome profiling, significant drought stress-associated differentially expressed genes (DEGs) were identified in both the varieties. A total of 1033 and 936 uniquely upregulated DEGs were found in Heena and Kiran respectively. The significant modulation of DEGs that were mainly associated with phytohormone signaling, stress-responsive genes (LEA, DREB), transcription factors (TFs) (AP2/ERF, MYB, WRKY, bHLH), and genes involved in photosynthesis and antioxidative mechanisms indicate better adaptive nature of Heena in stress tolerance. Additionally, the QTL-mapping analysis showed a very high number of DEGs associated with drought stress at AQHP069 QTL in Heena in comparison to Kiran which further distinguishes the drought-responsive traits at the chromosomal level in both the contrasting varieties. Overall, results support the higher capability of Heena over Kiran variety to induce numerous genes along with the development of better root architecture to endure drought stress.

PGPR-induced OsASR6 improves plant growth and yield by altering root auxin sensitivity and the xylem structure in transgenic Arabidopsis thaliana.

Plant-growth-promoting rhizobacteria (PGPR) improve plant growth by altering the root architecture, although the mechanisms underlying this alteration have yet to be unravelled. Through microarray analysis of PGPR-treated rice roots, a large number of differentially regulated genes were identified. Ectopic expression of one of these genes, OsASR6 (ABA STRESS RIPENING6), had a remarkable effect on plant growth in Arabidopsis. Transgenic lines over-expressing OsASR6 had larger leaves, taller inflorescence bolts and greater numbers of siliques and seeds. The most prominent effect was observed in root growth, with the root biomass increasing four-fold compared with the shoot biomass increase of 1.7-fold. Transgenic OsASR6 over-expressing plants showed higher conductance, transpiration and photosynthesis rates, leading to an ˜30% higher seed yield compared with the control. Interestingly, OsASR6 expression led to alterations in the xylem structure, an increase in the xylem vessel size and altered lignification, which correlated with higher conductance. OsASR6 is activated by auxin and, in turn, increases auxin responses and root auxin sensitivity, as observed by the increased expression of auxin-responsive genes, such as SAUR32 and PINOID, and the key auxin transcription factor, ARF5. Collectively, these phenomena led to an increased root density. The effects of OsASR6 expression largely mimic the beneficial effects of PGPRs in rice, indicating that OsASR6 activation may be a key factor governing PGPR-mediated changes in rice. OsASR6 is a potential candidate for the manipulation of rice for improved productivity.

Sterol glycosyltransferases required for adaptation of Withania somnifera at high temperature.

Heat is a major environmental stress factor that confines growth, productivity, and metabolism of plants. Plants respond to such unfavorable conditions through changes in their physiological, biochemical and developmental processes. Withania somnifera, an important medicinal plant, grows in hot and dry conditions, however, molecular mechanisms related to such adaptive properties are not known. Here, we elucidated that members of the sterol glycosyltransferases (SGT) gene family play important roles in the survival of W. somnifera under adverse conditions through maintaining the integrity of the membrane. SGTs are enzymes involved in sterol modifications and participate in metabolic flexibility during stress. Silencing of WsSGT members, for instance WsSGTL1, WsSGTL2 and WsSGTL4, was inimical for important physiological parameters, such as electron transport rate, photochemical quantum yield, acceptor side limitation, non-photochemical quenching (NPQ), Fv/Fm and net photosynthetic rate, whereas stomatal conductance, transpiration rate and dark respiration rates (Rds) were increased. Decreased NPQ and increased Rds helped to generate significant amount of ROS in the Wsamisgt lines. After heat stress, H2 O2 , lipid peroxidation and nitric oxide production increased in the Wsamisgt lines due to high ROS generation. The expression of HSPs in Wsamisgt lines might be involved in regulation of physiological processes during stress. We have also observed increased proline accumulation which might be involved in restricting water loss in the Wsamisgt lines. Taken together, our observations revealed that SGTL enzyme activity is required to maintain the internal damages of the cell against high temperature by maintaining the sterol vs sterol glycosides ratio in the membranes of W. somnifera.

Physiological performance and differential expression profiling of genes associated with drought tolerance in root tissue of four contrasting varieties of two Gossypium species.

Root growth in drying soil is generally limited by a combination of mechanical impedance and water stress. As the major function of root tissue is water and nutrient uptake, so it imparts an important role in plant growth and stress management. Previously, we have studied physiological performance and expression profiling of gene associated with drought tolerance in leaf tissue of four cotton varieties. Here, we have further continued our studies with the root tissue of these varieties. The Gossypium hirsutum species JKC-770 is drought-tolerant and KC-2 is drought-sensitive, while Gossypium herbaceum species JKC-717 is drought-tolerant and RAHS-187 is drought-sensitive. JKC-770 and JKC-717 the drought-tolerant varieties showed a comparatively high glutathione-S-transferase, superoxide dismutase, proline along with their gene expression, and low malondialdehyde content indicating low membrane damage and better antioxidative defense under drought condition. The expression levels of cellulose synthase, xyloglucan:xyloglucosyl transferase, and glycosyl hydrolases suggest modulation in cell wall structure and partitioning of sugars towards osmoprotectants instead of cell wall biosynthesis in tolerant varieties. Heat shock proteins and serine/threonine protein phosphotases show upregulation under drought condition, which are responsible for temperature tolerance and protein phosphorylation, respectively. These effects many metabolic processes and may be playing a key role in drought tolerance and adaptability of JKC-770 towards drought tolerance. The long-term water use efficiency (WUE) estimated in terms of carbon isotope discrimination (∆(13)C) in the root tissues showed maximum depletion in the ∆(13)C values in JKC-770 variety, while minimum in RAHS-187 under drought stress with reference to their respective control, suggesting a high WUE in JKC-770 variety.

Physiological performance, secondary metabolite and expression profiling of genes associated with drought tolerance in Withania somnifera.

Physiological, biochemical, and gene expression responses under drought stress were studied in Withania somnifera. Photosynthesis rate, stomatal conductance, transpiration rate, relative water content, chlorophyll content, and quantum yield of photosystems I and II (PSI and PSII) decreased in response to drought stress. Comparative expression of genes involved in osmoregulation, detoxification, signal transduction, metabolism, and transcription factor was analyzed through quantitative RT-PCR. The genes encoding 1-pyrroline-5-carboxylate synthetase (P5CS), glutathione S-transferase (GST), superoxide dismutase (SOD), serine threonine-protein kinase (STK), serine threonine protein phosphatase (PSP), aldehyde dehydrogenase (AD), leucoanthocyanidin dioxygenase/anthocyanin synthase (LD/AS), HSP, MYB, and WRKY have shown upregulation in response to drought stress condition in leaf tissues. Enhanced detoxification and osmoregulation along with increased withanolides production were also observed under drought stress. The results of this study will be helpful in developing stress-tolerant and high secondary metabolite yielding genotypes.

Physiological performance and differential expression profiling of genes associated with drought tolerance in contrasting varieties of two Gossypium species.

Cotton is mostly cultivated under rain-fed conditions in India, thus faces frequent drought conditions during its life cycle. Drought being a major stress factor responsible for yield penalty, there has always been a high priority to generate knowledge on adaptation and tolerance of cotton. In the present study, four cotton varieties, JKC-770 and KC-2 (Gossypium hirsutum), and JKC-717 and RAHS-187(Gossypium herbaceum), were imposed to drought. Under drought condition, differential changes in physiological characters like net photosynthesis, transpiration, stomatal conductance, chlorophyll fluorescence, relative water content (RWC), and predawn water potential (ψ 0) showed a change. While proline, malondialdehyde (MDA), and glutathione-S-transferase (GST) content increased along with a concomitant change in the expression of their associated genes. Under moderate stress, tolerant varieties maintain lower ψ 0 probably due to higher proline content as compared to sensitive varieties. Cyclic electron flow (CEF) also plays an important role in tolerance under mild water stress in G. hirsutum varieties. CEF not only activates at high light but also initiates at a very low light intensity. Expression analysis of genes reveals that drought-tolerant varieties showed enhanced detoxifying mechanism by up-regulation of asparagine synthase (AS), glutathione-S-transferase (GST), and methyl glyoxalase (GlyI) genes under drought stress. Up-regulation of Δ(1)-pyrroline-5-carboxylase synthase (Δ(1)P5CS) enhanced accumulation of proline, an osmolyte, under drought in tolerant varieties. While the drought-sensitive varieties showed up-regulation of ethylene responsive factor (ERF) and down-regulation of WRKY70 responsible for senescence of the leaf which correlated well with the high rate of leaf fall in sensitive varieties under water stress.

Characterizing photoinhibition and photosynthesis in juvenile-red versus mature-green leaves of Jatropha curcas L.

The new leaves of Jatropha curcas (L.) appear dark red in colour due to the presence of anthocyanin pigments, these leaves subsequently turn green on maturity. The aim of the study was to characterize the photosynthetic efficiency of the juvenile red and mature green leaves and to understand the possible role of anthocyanin pigment in the juvenile leaves of J. curcas. We studied the localization of anthocyanin pigment, reflectance properties, diurnal gas-exchange performance, carboxylation efficiency and photosynthetic efficiency under different light intensities by investigation of the photochemical and non-photochemical energy dissipation processes related to Photosystem II (PSII) and Photosystem I (PSI), of the juvenile and the mature leaves of J. curcas. The JIP test analysis of chlorophyll a fluorescence transients and the gas-exchange studies revels the low photosynthetic efficiency of red leaves is due to the immaturity of the leaf. The low value of quantum yield of non-photochemical energy dissipation due to acceptor side limitation, Y (NA) under high light in the red leaf, suggests that over-reduction of PSI acceptor side was prevented and it results in the accumulation of oxidized P700, which dissipates excess light energy harmlessly as heat and thereby alleviate photoinhibition of PSI in case of the juvenile red leaves. Further our results of photoinhibition and relaxation on exposure of red and green leaves to monochromatic blue light showed that effective quantum yield of PSII recovers faster and completely under darkness in juvenile red leaves as compared to mature green leaves, supporting the role of anthocyanin pigments in protecting both PSII and PSI in the red leaves.

Reflectance and cyclic electron flow as an indicator of drought stress in cotton (Gossypium hirsutum).

The response and the functioning of the photosynthetic machinery of cotton, Gossypium hirsutum during water stress was studied by leaf optical properties, linear (ETRII) and cyclic electron flow (CEF) and chlorophyll a fluorescence. We observed that in G. hirsutum, during water limitation, Chlorophyll b showed the best correlation with reflectance at 731 nm and is a better indicator of drought. Fv /Fm was observed to be very insensitive to mild water stress. However, during severe water stress the leaves exhibit considerable inhibition in Fv /Fm and an increase in anthocyanin levels by about 20-fold. CEF was very responsive to mild water stress. The mild drought stress caused large decrease in the ability of the leaves to utilize the light energy. Photosystem I and photosystem II is protected from photoinhibition by high CEF and nonphotochemical quenching under mild water stress. While during severe drought stress, linear electron flow showed a sharp decrease in comparison to CEF. CEF play a major role in G. hirsutum leaves during mild as well as under severe water stress condition and is thus a good indicator of water stress.

Photosynthetic characteristics and the response of stomata to environmental determinants and ABA in Selaginella bryopteris, a resurrection spike moss species.

Selaginella bryopteris is a spike-moss lycophyte species with resurrection capability. These plants have small sized stomata that occur in higher density than in other fern species. The diurnal gas-exchange studies under natural conditions showed a bell shaped net photosynthesis curve. The effective quantum yield of PSII (ΔF/F(m')) showed an inverse relationship with light and recovered to its maximum at sunset. This suggests that there was a complete recovery of PSII efficiency during the late evening hours. S. bryopteris displayed broad temperature optima for net photosynthesis from 28 °C to 37 °C. The stomatal sensitivity in response to vapor pressure deficit (VPD), was maximum at 25 °C temperature while at temperatures from 30 to 35 °C it was low. Our study demonstrates that S. bryopteris plants show a very poor mechanism for its stomatal regulation in response to high light, high temperature, high VPD, high CO2 and to ABA treatment. At the same time they show a high stomatal conductance leading to unrestricted rates of transpiration and a lack of capacity to optimize water use efficiency (WUE).

Photosynthetic performance of Jatropha curcas fruits.

Jatropha curcas (L.) trees under north Indian conditions (Lucknow) produce fruits in two major flushes, once during autumn-winter (October-December). The leaves at this time are at the senescence stages and already shedding. The second flush of fruit setting occurs during the summer (April-June) after the leaves have formed during spring (March-April). Photosynthetic performance of detached jatropha fruits was studied at three developmental stages, immature, mature and ripe fruits. Studies were made in both winter and summer fruits in response to light, temperature and vapour pressure deficit (VPD) under controlled conditions to assess the influence of these environmental factors on the photosynthetic performance of jatropha fruits. Immature fruits showed high light saturating point of around 2000 µmol m(-2) s(-1). High VPD did not show an adverse effect on the fruit A. Stomatal conductance (g(s)) showed an inverse behaviour to increasing VPD, however, transpiration (E) was not restricted by the increasing VPD in both seasons. During winter in absence of leaves on the jatropha tree the fruits along with the bark contributes maximum towards photoassimilation. Dark respiration rates (R(d)) monitored in fruit coat and seeds independently, showed maximum R(d) in seeds of mature fruit and these were about five times more than its fruit coat, reflecting the higher energy requirement of the developing fruit during maximum oil synthesis stage. Photosynthesis and fluorescence parameters studied indicate that young jatropha fruits are photosynthetically as efficient as its leaves and play a paramount role in scavenging the high concentration of CO(2) generated by the fruit during respiration.

Physiological and proteomic responses of cotton (Gossypium herbaceum L.) to drought stress.

Cotton genotype RAHS 187 was analyzed for changes in physiology, biochemistry and proteome due to drought stress. The deleterious effect of drought in cotton plants was mainly targeted towards photosynthesis. The gas-exchange parameters of net photosynthesis (A), stomatal conductance (g(s)) and transpiration (E) showed a decreasing trend as the drought intensity increased. The fluorescence parameters of, effective quantum yield of PSII (Φ(PSII)), and electron transport rates (ETR), also showed a declining trend. As the intensity of drought increased, both H(2)O(2) and MDA levels increased indicating oxidative stress. Anthocyanin levels were increased by more than four folds in the droughted plants. Two-dimensional gel electrophoresis detected more than 550 protein spots. Significantly expressed proteins were analyzed by peptide mass fingerprinting (PMF) using MALDI-TOF-TOF. The number of up-regulated spots was found to be 16 while 6 spots were down-regulated. The reasonable implications in drought response of the identified proteins vis-à-vis physiological changes are discussed. Results provide some additional information that can lead to a better understanding of the molecular basis of drought-sensitivity in cotton plants.

Desiccation-induced physiological and biochemical changes in resurrection plant, Selaginella bryopteris.

Selaginella bryopteris is a lycophyte resurrection plant, which incurves during desiccation and recovers on availability of moisture. The aim of the study was to test and understand the various physiological and biochemical changes the fronds undergo during desiccation and rehydration, to get an insight as to how this plant adapts and survives through the dry phase. Upon desiccation, S. bryopteris fronds showed drastic inhibition in net photosynthesis (A) and maximal photochemical efficiency of PSII (F(v)/F(m)) however, chlorophyll content did not show much variation. Dark respiration (R(d)) continued even at 10% relative water content (RWC), and showed a burst after rehydration, which is proposed to be crucial to establish protection mechanisms. Desiccation caused an enhanced production of reactive oxygen species (ROS) and increased lipid peroxidation. Proline accumulation increased substantially by 11-fold. Sucrose and starch contents decreased upon desiccation as compared to control. The antioxidative enzymes viz. superoxide dismutase (SOD), ascorbate peroxidase (APX) and catalase (CAT) along with soluble acid invertase increased during desiccation. S. bryopteris shows mechanical as well as physiological mechanisms for tolerance to extreme levels of desiccation stress. The rapid and almost complete recovery of F(v)/F(m) after rehydration clearly indicates the absence of marked photoinhibitory or thermal injury to PSII during desiccation. This along with the homoiochlorophyllous characteristics enables S. bryopteris to recover its A. The antioxidant metabolism further plays an important role in the desiccation tolerance of S. bryopteris.

Influence of leaf-to-air vapour pressure deficit (VPD) on the biochemistry and physiology of photosynthesis in Prosopis juliflora.

The effect of leaf-to-air vapour pressure deficit (VPD) was studied in well-watered, potted, 1-2-year-old plants of the leguminous tree P. juliflora grown outside in northern India. The long-term responses to VPD were analysed from diurnal and seasonal variations in gas exchange parameters measured in two cohorts of leaves produced in February and July, respectively. In general, inhibitory effects of high VPD were visible only when the VPD level exceeded a threshold of >3 kPa. There was a substantial decline in net photosynthesis rate and stomatal conductance at high VPD >4 kPa and transpiration showed a decrease in steady-state rate or feedforward response to VPD. The feedforward responses were visible in all seasons, although the plants were exposed to a wide range of VPD during the year and leaf relative water content was constant. The maximum quantum efficiency of PSII measured predawn was constant (around 0.8) in all seasons except summer. Short-term experiments showed that, although gas exchange was severely affected by high VPD in the leaves of both cohorts, the plant maintained a constant, water use efficiency in different seasons. High VPD also caused reductions in Rubisco activity, affecting carboxylation efficiency, and reductions in sucrose and starch content due to a decrease in the activity of sucrose-phosphate synthase. However, the relative quantum yield of PSII and electron transport rates measured at 1500 micromol m(-2) s(-1) were unaffected by increasing VPD, indicating the presence of a large alternative sink possibly, photorespiration. The overall results showed that P. juliflora can withstand high VPD by reducing metabolic activity and by effective adjustments in the partitioning of electron flow between assimilation and non-assimilation processes, which, in turn, imposed a strong limitation on the potential carbon gain.