Brassinolide alleviates the drought-induced adverse effects in barley by modulation of enzymatic antioxidants and ultrastructure

In the present study, possible role of exogenously applied brassinolide (BL) in alleviating the detrimental effects of polyethylene glycol (PEG)-inducted drought stress in barley was evaluated. Barley seedlings were subjected to two PEG levels (0 and 8% w/v) for 2 weeks followed by a foliar spray of BL (0, 0.01, 0.1 and 1 µM) to assess the changes in growth, gas exchange, chlorophyll contents, antioxidant enzyme activities and ultrastructural changes. PEG-induced drought considerably inhibited the barley growth in terms of shoot and root length, shoot, root fresh and dry weights, and chlorophyll contents. Moreover, extensive decrease by PEG-induced drought in gas exchange attributes i.e. net photosynthetic rate, transpiration rate, stomatal conductance and intercellular CO₂ were also recorded. However, exogenous application of BL remarkably improved the gas exchange attributes, chlorophyll contents and plant growth under either drought or control condition. Furthermore, foliar spray of BL decreased the production of malondialdehyde (MDA) and reactive oxygen species (ROS) in the leaves and roots by increasing enzymatic antioxidant activities under drought stress. The microscopic examination indicated that exogenous application of BL improved the cell structure and enabled a clean mesophyll cell having a well-developed chloroplast with thylakoid membranes of the drought stressed plants. On the basis of these findings, it can be concluded that exogenous application of BL could alleviate drought stress to barley by ameliorating photosynthetic parameters, improving antioxidants enzyme activities, reducing ultrastructural changes in leaf mesophyll and finally maintaining the plant growth.     

Exogenously applied polyamines increase drought tolerance of rice by improving leaf water status,photosynthesis and membrane properties

Drought stress hampers rice performance principally by disrupting the plant–water relations and structure of biological membranes. This study appraised the role of polyamines (PAs) in improving drought tolerance in fine grain aromatic rice (Oryza sativa L.). Three PAs [putrescine (Put), spermidine (Spd) and spermine (Spm)] were used each at 10 μM as seed priming (by soaking seeds in solution) and foliar spray. Primed and non-primed seeds were sown in plastic pots with normal irrigation in a phytotron. At four-leaf stage, plants were subjected to drought stress by bringing the soil moisture down to 50% of field capacity by halting water supply. For foliar application, 10 μM solutions each of Put, Spd and Spm were sprayed at five-leaf stage. Results revealed that drought stress severely reduced the rice fresh and dry weights, while PAs application improved net photosynthesis, water use efficiency, leaf water status, production of free proline, anthocyanins and soluble phenolics and improved membrane properties. PAs improved drought tolerance in terms of dry matter yield and net photosynthesis was associated with the maintenance of leaf water status and improved water use efficiency. Among the antioxidants, catalase activity was negatively related to H₂O₂ and membrane permeability, which indicated alleviation of oxidative damage on cellular membranes by PAs application. Foliar application was more effective than the seed priming, and among the PAs, Spm was the most effective in improving drought tolerance.     

Overexpression of a Chloroplast-located Peroxiredoxin Q Gene, SsPrxQ, Increases the Salt and Low-temperature Tolerance of Arabidopsis

Ablotlc stress, such as salt, drought and extreme temperature, can result in enhanced production of reactive oxygen species （ROS）. Plants have developed both enzymatic ROS-scavenging and non-enzymatic ROS-scavenging systems. The major ROS-scavenging enzymes of plants include superoxide dismutase （SOD）, ascorbate peroxldaae （APX）, catalaae （CAT）, glutathione peroxldaae （GPX） and peroxiredoxina （Prxa）. In the present work, we identified a gene encoding chloroplast-located peroxiredoxin Q, SsPrxQ, from Suaeda salsa L. located at chloroplast. Overexpression of SsPrxQ In Arabidopsis leads to an increase In salt and low-temperature tolerance.     

Changes in photosynthetic apparatus of mustard (<Emphasis Type="Italic">Sinapis alba</Emphasis>) cotyledons following cycloheximide and kinetin treatments

Biochemical and accompanying structural characteristics of the photosynthetic process were studied in mustard seedlings cultivated on medium with increasing concentrations of cycloheximide alone as well as in combination with various kinetin concentrations. After 7 days of cultivation the contents of total chlorophyll, carotenoids and content of Rubisco in mustard cotyledons were determined. The content of chlorophyll pigments and carotenoids decreased in dependence of cycloheximide concentration. Following antibiotic treatment the content of both Rubisco subunits markedly decreased. In addition cycloheximide caused disturbance in mesophyll organization and chloroplast ultrastructure. Kinetin applied with cycloheximide increased the amount of photosynthetic pigments as well as of Rubisco, compared to the cycloheximide alone. In the seedlings treated with cycloheximide+kinetin the structure of leaf mesophyll and chloroplast membrane system was similar to control. Our results indicate that kinetin diminished the negative effects of cycloheximide on photosynthetic pigments and Rubisco as well as on the structural traits of the cotyledons.     

Combined effect of salicylic acid and potassium mitigates drought stress through the modulation of physio-biochemical attributes and key antioxidants in wheat

Improving physio-biochemical traits in wheat under drought stress conditions has received more research attention in recent years for better adaptability and higher yield. In this study, we explored the potential bio-physiological mechanisms underlying improved plant growth and water use efficiency in wheat following soil application of potassium (0 and 100 kg ha^?1) and seed primed salicylic acid (SA) (150 mg per L) and SA foliar application (100 mg per L) under drought stresses (100%, 60% and 30% FC). Two years' average data revealed that inducing drought stress resulted in a decrease in plant pigments content, growth traits, and plant water status however, the influence was substantially reduced with the combined application of K and SA under drought stress conditions. The SA foliar spray in combination with K had increased chlorophyll a (174% and 83%), chl b (130% and 192%), chl a + b (156% and 120), carotenoid (22% and 11%), proline contents (24% and 29%) leaf relative water content (24% and 29%) while reduced leaf WSD (17% and 20%), WRC (6% and 7%), and WUC (23% and 28%) under mild and severe drought stresses, respectively. The increase in grain yield by 41% and 37% with enhanced water use efficiency was obtained with combined foliar SA and K under mild and severe drought stress, respectively indicating its vital role in overcoming the deleterious effects of drought via regulation of osmotic and metabolic processes and stabilizes cell components. RDA analysis revealed that the studied traits were completely discriminated under severe stress than mild or no drought stress. A positive and significant association was found between plant pigments with seed yield whereas a negative and significant correlation existed between water leaf traits and plant pigments. It was concluded that both foliar SA and seed primed SA with K fertilization combat the adverse effects of drought and improved plant water status as well as growth and bio-physiological traits of wheat under drought stress conditions.     

Cultivar specific variations in morphological and biochemical characteristics of mung bean due to foliar spray of ascorbic acid under elevated ozone

Effect of exogenous application of ascorbic acid (Asc) solution was examined on the growth, photosynthetic pigments, biochemical and yield characteristics of mung bean cultivars against ozone (O₃). Experiment was performed on six mung bean cultivars in open top chambers under field conditions and Asc was applied as foliar spray prior to the exposure of ambient (AO+) and elevated (EO+) levels of O₃. Application of Asc showed increment in growth attributes as compared to plants not provided Asc (AO). However, O₃ induced the production of reactive oxygen species led to membrane damage. Reductions were depicted in lipid peroxidation, solute leakage and foliar injury % in AO+ and EO+ as compared to plants not provided with Asc. Photosynthetic pigments and ascorbic acid content along with activities of anti-oxidative enzymes (APX, CAT, GR and SOD) showed increments in AO+ and EO+ with cultivar-specific variations. Cultivars HUM-1 and HUM-2 restored yield with Asc application while less response was observed in other test cultivars. Quality of the seeds was also affected by Asc treatment in the plants exposed to ambient and elevated levels of O₃. Therefore, exogenous application of Asc promotes plant’s performance by providing protection against O₃ induced oxidative stress and may be used in screening of the mung bean cultivars against O₃ phytotoxicity.     

Does a terminal drought tolerance QTL contribute to differences in ROS scavenging enzymes and photosynthetic pigments in pearl millet exposed to drought?

The control of reactive oxygen species (ROS) and the stability of photosynthetic pigments under stress conditions are hypothesized to contribute to drought tolerance. Here we studied how ascorbic peroxidase (APX), superoxide dismutase (SOD), catalase (CAT) isozyme activities and chlorophyll a, b (Chl a, b) and carotenoids (Car) contents responded to water stress and whether they related to presence of a terminal drought tolerance QTL in pearl millet. We used PRLT2/89-33 (QTL donor), H77/833-2 (sensitive), and near-isogenic lines (QTL-NILs) introgressed with the QTL in H77/833-2 background. Under water stress there was no significant change in the total APX activity; only the proportional APX5 activity increased, with higher band intensity in tolerant genotypes. There were no significant changes in total activities of CAT and SOD under water stress, with similar band intensities in all genotypes, and a new CAT isozyme was induced in all genotypes. The photosynthetic pigment content decreased under water stress, although not differently in any genotype. Under water stress, the activities of most APX, CAT and SOD isozymes were closely related to the total chlorophyll/carotenoids ratio. Overall, besides APX5, water stress did not lead to major changes in the profile of isoenzymes involved in ROS scavenging. Similarly, the pigment content under stress did not discriminate genotypes according to the presence/absence of the QTL. This absence of discrimination for the ROS scavenging enzymes and for the pigment content under stress suggests that these traits may not play a key role in terminal drought tolerance in pearl millet.     

Effects of homobrassinolide application on plant metabolism and grain yield under irrigated and moisture-stress conditions of two wheat varieties

Homobrassinolide (BR) was applied either as a seed treatment or foliar spray to two contrasting wheat varieties, viz. C306 (drought tolerant) and HD2329 (drought susceptible), to examine its effects on plant metabolism and grain yield under irrigated and moisture-stress/rainfed conditions. BR application resulted in increased relative water content, nitrate reductase activity, chlorophyll content and photosynthesis under both conditions. BR application also improved membrane stability (lower injury). These beneficial effects resulted in higher leaf area, biomass production, grain yield and yield related parameters in the treated plants. All the treatments were significantly better than the untreated control. Generally, 0.05 ppm either as a seed treatment or foliar spray was more effective than the 0.01 ppm treatment. The drought-tolerant genotype C306 showed more response to BR application under moisture-stress/rainfed condition than HD 2329. Increased water uptake, membrane stability and higher carbon dioxide and nitrogen assimilation rates under stress seemed to be related to homobrassinolide-induced drought tolerance.     

Effects of salt stress on the structure and function of the photosynthetic apparatus in Cucumis sativus and its protection by exogenous putrescine

With the objective to clarify the physiological significance of polyamines (PAs) in the photosynthetic apparatus, the present study investigated the effects of salt stress with and without foliar application of putrescine (Put) on the structure and function of the photosynthetic apparatus in cucumber. Salt stress at 75 mM NaCl for 7 days resulted in a severe reduction of photosynthesis. The fast chlorophyll afluorescence transient analysis showed that salt stress inhibited the maximum quantum yield of PSII photochemistry (F(v) /F(m) ), mainly due to damage at the receptor side of PSII. In addition, salt stress decreased the density of active reaction centers and the structure performance. The microscopic analysis revealed that salt stress-induced destruction of the chloroplast envelope and increased the number of plastoglobuli along with aberrations in thylakoid membranes. Besides, salt stress caused a decrease in the content of endogenous PAs, conjugated and bound forms of spermidine and spermine in particular, in thylakoid membranes. However, applications of 8 mM Put alleviated the salt stress-mediated decrease in net photosynthetic rates (Pn) and actual efficiency of PSII (Φ(PSII) ). Put increased PAs in thylakoid membranes and overcame the damaging effects of salt stress on the structure and function of the photosynthetic apparatus in salt-stressed plant leaves. Put application to control plants neither increased PAs in thylakoid membranes nor affected photosynthesis. These results indicate that PAs in chloroplasts play crucial roles in protecting the thylakoid membranes against the deleterious influences of salt stress. In addition, the present results point to the probability that the salt-induced dysfunction of photosynthesis is largely attributable to the loss of PAs in the photosynthetic apparatus.     

Treatment with acetyl salicylic acid protects muskmelon seedlings against drought stress

Salicylic acid (SA) is a common plant-produced signal molecule that is responsible for inducing tolerance to a number of biotic and abiotic stresses. An experiment was, therefore, conducted to test whether acetyl salicylic acid (ASA) application at various concentrations through seed soaking or foliar spray would protect muskmelon [Cucumis melo L. (reticulatus group)] seedlings, subjected to drought stress. Twenty-three-day-old plants pre-treated with ASA (0, 0.1, 0.25, 0.50 or 1.0 mM) were subjected to drought stress for 1 week in a greenhouse. ASA applied either through seed soaking or through foliar spray was effective within the range of 0.1–1 mM in providing drought stress protection in muskmelon seedlings; however, there was no difference between application methods indicating that both methods provided similar levels of protection. ASA significantly affected all seedling growth and stress indicator variables measured except leaf number and root dry weight. The best protection appeared to be obtained from seedlings pre-treated with lower concentrations of ASA. Even though both methods provided similar means of protection, due to its simplicity and practicality, soaking muskmelon seeds prior to sowing in up to 0.5 mM ASA would be a more desirable method.     

Ectopic expression of PgRab7 in rice plants (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) results in differential tolerance at the vegetative and seed setting stage during salinity and drought stress

In this work, we have overexpressed a vesicle trafficking protein, Rab7, from a stress-tolerant plant, Pennisetum glaucum, in a high-yielding but stress-sensitive rice variety Pusa Basmati-1 (PB-1). The transgenic rice plants were tested for tolerance against salinity and drought stress. The transgenic plants showed considerable tolerance at the vegetative stage against both salinity (200 mM NaCl) and drought stress (up to 12 days after withdrawing water). The protection against salt and drought stress may be by regulating Na⁺ ion homeostasis, as the transgenic plants showed altered expression of multiple transporter genes, including OsNHX1, OsNHX2, OsSOS1, OsVHA, and OsGLRs. In addition, decreased generation and maintenance of lesser reactive oxygen species (ROS), with maintenance of chloroplast grana and photosynthetic machinery was observed. When evaluated for reproductive growth, 89–96 % of seed setting was maintained in transgenic plants during drought stress; however, under salt stress, a 33–53 % decrease in seed setting was observed. These results indicate that PgRab7 overexpression in rice confers differential tolerance at the seed setting stage during salinity and drought stress and could be a favored target for raising drought-tolerant crops.     

Exogenous application of urea and a urease inhibitor improves drought stress tolerance in maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.)

Drought is believed to cause many metabolic changes which affect plant growth and development. However, it might be mitigated by various inorganic substances, such as nitrogen. Thus, the study was carried out to investigate the effect of foliar-applied urea with or without urease inhibitor N-(n-butyl) thiophosphoric triamide (NBPT) on a maize cultivar under drought stress simulated by 15% (w/v) polyethylene glycol 6000. Foliar-applied urea resulted in a significant increase in plant dry weight, relative water content, and photosynthetic pigments under water stress condition. Furthermore, the activities of superoxide dismutase (SOD), peroxidase (POD), and hydrogen peroxidase (CAT), were enhanced with all spraying treatments under drought stress, which led to decreases in accumulation of hydrogen peroxide (H₂O₂), superoxide anion (\({\text{O}}_2^{ \cdot - }\)) and malondialdehyde (MDA). The contents of soluble protein and soluble sugar accumulated remarkably with urea-applied under drought stress condition. Moreover, a further enhancement in above metabolites was observed by spraying a mixture of urea and urease inhibitor as compared to urea sprayed only. Taken together, our findings show that foliar application of urea and a urease inhibitor could significantly enhance drought tolerance of maize through protecting photosynthetic apparatus, activating antioxidant defense system and improving osmoregulation.     

Modulation in growth,photosynthetic efficiency,activity of antioxidants and mineral ions by foliar application of glycinebetaine on pea (Pisum sativum L.) under salt stress

A pot experiment was carried out to explore the role of glycinebetaine (GB) as foliar spray foliar on two pea (Pisum sativum L.) varieties (Pea 09 and Meteor Fsd) under saline and non-saline conditions. Thirty-two-day-old plants were subjected to two levels 0 and 150 mM of NaCl stress. Salt treatment was applied in full strength Hoagland’s nutrient solution. Three levels 0, 5 and 10 mM of GB were applied as foliar treatment on 34-day-old pea plants. After 2 weeks of foliar treatment with GB data for various growth and physiochemical attributes were recorded. Rooting-medium applied salt (150 mM NaCl) stress decreased growth, photosynthesis, chlorophyll, chlorophyll fluorescence and soluble protein contents, while increasing the activities of enzymatic (POD and CAT) and non-enzymatic (ascorbic acid and total phenolics) antioxidant enzymes. Foliar application of GB decreased root and shoot Na⁺ under saline conditions, while increasing shoot dry matter, root length, root fresh weight, stomatal conductance (g _s), contents of seed ascorbic acid, leaf phenolics, and root and shoot Ca²⁺ contents. Of three GB (0, 5, 10 mM) levels, 10 mM proved to be more effective in mitigating the adverse effects of salinity stress. Overall, variety Pea 09 showed better performance in comparison to those of var. Meteor Fsd under both normal and salinity stress conditions. GB-induced modulation of seed ascorbic acid, leaf phenolics, g _s, and root Ca²⁺ values might have contributed to the increased plant biomass, reduction of oxidative stress, increased osmotic adjustment and better photosynthetic performance of pea plants under salt stress.     

Use of Transgenic Plants with Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase Antisense DNA to Evaluate the Rate Limitation of Photosynthesis under Water Stress

The biochemical lesion that causes impaired chloroplast metabolism (and, hence, photosynthetic capacity) in plants exposed to water deficits is still a subject of controversy. In this study we used tobacco (Nicotiana tabacum L.) transformed with "antisense" ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) DNA sequences to evaluate whether Rubisco or some other enzymic step in the photosynthetic carbon reduction cycle pathway rate limits photosynthesis at low leaf water potential ([psi]w). These transformants, along with the wild-type material, provided a novel model system allowing for an evaluation of photosynthetic response to water stress in near-isogenic plants with widely varying levels of functional Rubisco. It was determined that impaired chloroplast metabolism (rather than decreased leaf conductance to CO2) was the major cause of photosynthetic inhibition as leaf [psi]w declined. Significantly, the extent of photosynthetic inhibition at low [psi]w was identical in wild-type and transformed plants. Decreasing Rubisco activity by 68% did not sensitize photosynthetic capacity to water stress. It was hypothesized that, if water stress effects on Rubisco caused photosynthetic inhibition under stress, an increase in the steady-state level of the substrate for this enzyme, ribulose 1,5-bisphosphate (RuBP), would be associated with stress-induced photosynthetic inhibition. Steady-state levels of RuBP were reduced as leaf [psi]w declined, even in transformed plants with low levels of Rubisco. Based on the similarity in photosynthetic response to water stress in wild-type and transformed plants, the reduction in RuBP as stress developed, and studies that demonstrated that ATP supply did not rate limit photosynthesis under stress, we concluded that stress effects on an enzymic step involved in RuBP regeneration caused impaired chloroplast metabolism and photosynthetic inhibition in plants exposed to water deficits.     

干旱胁迫下发菜光合作用相关基因差异表达分析

该研究以不同失水处理的发菜为研究材料,以充分吸水状态的发菜为对照,利用高通量测序技术和qRT PCR技术检测了干旱胁迫下发菜光合作用相关基因差异表达规律,并对光合色素和酶活在干旱胁迫下的变化进行了研究。结果表明：(1)发菜在不同程度干旱胁迫下有113个光合作用相关基因差异表达,其中失水30%、75%和100%的发菜分别有44个、74个和91个光合作用相关基因差异表达。(2)随着干旱胁迫程度的加深藻胆素、叶绿素a和类胡萝卜素含量逐渐降低,Rubisco活性随着干旱胁迫程度的增强先上升后下降,GAPDH活性随着干旱胁迫的增强呈现下降的趋势。研究表明,发菜通过光合作用相关基因的差异表达调控光合作用以适应干旱胁迫。该研究结果对进一步研究发菜干旱胁迫响应机制及其耐旱光合机理奠定了基础。     

Proteins and Metabolites Regulated by Trinexapac-ethyl in Relation to Drought Tolerance in Kentucky Bluegrass

Plants have developed various mechanisms in adaptation to water deficit stress, including growth retardant to reduce water loss. Previous studies reported that plants treated with a growth inhibitor, trinexapac-ethyl (TE), had improved drought tolerance. The objective of this study was to determine alterations in proteins and metabolite accumulation associated with drought tolerance improvement in a perennial grass species, Kentucky bluegrass (Poa pratensis), induced by TE application. Plants were treated with TE [1.95 ml l⁻¹ (v:v); a.i. TE = 0.113%] through foliar spray for 14 days, and then subjected to drought stress by withholding irrigation for 15 days in growth chambers. TE-treated plants exhibited significantly higher relative water content and photosynthetic capacity and lower membrane leakage than nontreated plants under drought stress, suggesting TE-enhanced drought tolerance in Kentucky bluegrass. Physiological improvement in drought tolerance through TE application was associated with the increased accumulation of various proteins and metabolites, including ferritin, catalase, glutathione-S-transferase, Rubisco, heat shock protein 70, and chaperonin 81, as well as fatty acids (palmitic acid, α-linolenic acid, linoleic acid, and octadecanoic acid). Our results suggest that TE may regulate metabolic processes for antioxidant defense, protective protein synthesis, photorespiration, and fatty acid synthesis, and thereby contribute to better drought tolerance in Kentucky bluegrass.     

<Emphasis Type="Bold">Role of microRNAs and their target genes in salinity response in plant</Emphasis>s

We examined the effects of 2,4-epibrassinolide (EBR) application on photosynthesis, antioxidant enzyme activity, and Rubisco activase (RCA) gene expression in wheat (Triticum aestivum L.) seedlings under a combination of drought and heat stress. The net photosynthetic rates (P_n) of wheat seedlings decreased significantly, the photosynthetic capability was inhibited, and the activities of superoxide (SOD), peroxidase (POD), catalase (CAT), and RCA as well as the initial and total activity of Rubisco declined under the combined stress. These decreases and inhibitory effects were significantly ameliorated by exogenous EBR application. Three subunits (45–46, 41–42, and 38–39 kDa) of RCA were observed in wheat seedlings. The abundances of the 38–39 kDa and 41–42 kDa subunits were significantly lower in plants subjected to stressful conditions than in unstressed plants. Interestingly, a marked increase in 45–46 kDa RCA was observed under heat or heat combined with drought stress. The abundance of 38–39 kDa RCA in seedlings exposed to heat, drought, or their combination was significantly enhanced by EBR pretreatment, which paralleled the changes in initial Rubisco activity and P_n, but was not consistent with observed mRNA abundance. These results indicated that the larger subunit of RCA (45–46 kDa), which is more thermostable and increased in response to moderate heat stress, and the smaller isoform (38–39 kDa) of RCA may play important roles in maintaining the photosynthetic capability by EBR under stress conditions.