Arbuscular mycorrhiza influences carbon‐use efficiency and grain yield of wheat grown under pre‐ and post‐anthesis salinity stress

• Soil salinity severely affects and constrains crop production worldwide. Salinity causes osmotic and ionic stress, inhibiting gas exchange and photosynthesis, ultimately impairing plant growth and development. Arbuscular mycorrhiza (AM) have been shown to maintain light and carbon use efficiency under stress, possibly providing a tool to improve salinity tolerance of the host plants. Thus, it was hypothesized that AM will contribute to improved growth and yield under stress conditions.
• Wheat plants (Triticum aestivum L.) were grown with (AMF+) or without (AMF?) arbuscular mycorrhizal fungi (AMF) inoculation. Plants were subjected to salinity stress (200 mm NaCl) either at pre‐ or post‐anthesis or at both stages. Growth and yield components, leaf chlorophyll content as well as gas exchange parameters and AMF colonization were analysed.
• AM plants exhibited a higher rate of net photosynthesis and stomatal conductance and lower intrinsic water use efficiency. Furthermore, AM wheat plants subjected to salinity stress at both pre‐anthesis and post‐anthesis maintained higher grain yield than non‐AM salinity‐stressed plants.
• These results suggest that AMF inoculation mitigates the negative effects of salinity stress by influencing carbon use efficiency and maintaining higher grain yield under stress.

     

Paenibacillus lentimorbus Enhanced Abiotic Stress Tolerance Through Lateral Root Formation and Phytohormone Regulation

Under the stressed conditions plant growth-promoting rhizobacteria (PGPR) are able to stimulate plant growth through several mechanisms, including antioxidants alleviation, regulation of stress responsive genes and phytohormones etc. Present study is conducted to investigate the impact of Paenibacillus lentimorbus B-30488 inoculation on salinity and drought stress mitigation in Arabidopsis thaliana through modulation in defense enzymes, phyto-hormones and root system architecture associated gene expression profiling. In vitro experiments clearly demonstrated the role of B-30488 in stimulating the root length, branches, lateral root formation and biomass under salinity and drought stress. The inoculation of B-30488 modulated the phytohormones levels to protect the plants from salinity and drought stress. Similarly, defence enzymes were also activated under the stressed conditions, but B-30488 inoculation reduced the antioxidants content during salinity and drought stress as compared to their respective controls. Microscopy results showed decrease in lateral roots hair formation under both stresses and B-30488 inoculation not only mitigate but also enhanced the lateral root formation. Gene expression analysis through real time polymerase chain reaction (RT-PCR) showed modulated expression of several genes related to root development, stress and lateral root formation in B-30488 inoculated seedlings. Results based on the present study, B-30488 is also involved in alteration root architecture, its growth regulation via modulation in phytohormones and genes expression and overall significant improvement in plant growth under stress conditions.

     

Plant growth‐promoting endophytic bacteria augment growth and salinity tolerance in rice plants

• Salt stress negatively affects growth and development of plants. However, it is hypothesized that plant growth‐promoting endophytic bacteria can greatly alleviate the adverse effects of salinity and can promote growth and development of plants. In the present research, we aimed to isolate endophytic bacteria from halotolerant plants and evaluate their capacity for promoting crop plant growth.
• The bacterial endophytes were isolated from selected plants inhabiting sand dunes at Pohang beach, screened for plant growth‐promoting traits and applied to rice seedlings under salt stress (NaCl; 150 mm ).
• Out of 59 endophytic bacterial isolates, only six isolates, i.e. Curtobacterium oceanosedimentum SAK1, Curtobacterium luteum SAK2, Enterobacter ludwigii SAK5, Bacillus cereus SA1, Micrococcus yunnanensis SA2, Enterobacter tabaci SA3, resulted in a significant increase in the growth of Waito‐C rice. The cultural filtrates of bacterial endophytes were tested for phytohormones, including indole‐3‐acetic acid, gibberellins and organic acids. Inoculation of the selected strains considerably reduced the amount of endogenous ABA in rice plants under NaCl stress, however, they increased GSH and sugar content. Similarly, these strains augmented the expression of flavin monooxygenase (OsYUCCA1) and auxin efflux carrier (OsPIN1) genes under salt stress.
• In conclusion, the pragmatic application of the above selected bacterial strains alleviated the adverse effects of NaCl stress and enhanced rice growth attributes by producing various phytohormones.

     

Comparison of CAM expression,photochemistry and antioxidant responses in Sedum album and Portulaca oleracea under combined stress

Previous studies of crassulacean acid metabolism (CAM) pathway during stress have been directed at individual drought and salinity stress, here, we studied the effects of a combination of drought and salt on CAM expression, chlorophyll fluorescence and antioxidant parameters in the C₃-CAM facultative Sedum album and C₄-CAM facultative Portulaca oleracea plants. While salinity alone was not able to induce functional CAM expression in P. oleracea leaves, we showed that salinity induced low level of nocturnal acid accumulation in S. album species. After 20 d of exposure to the combination of simultaneous salt and drought stress, P. oleracea plants exhibited more resistance to photoinhibition as compared to S. album plants. The decrease of maximum quantum yield (F_v/F_m) in S. album leaves under combined stress was in parallel with the largest suppression of CAM expression of >50%, probably displaying the withdrawal of functional CAM back to C₃ pathway. However, under drought treatment alone, S. album plants exhibited higher photosynthetic flexibility, which was associated with the up-regulation of antioxidant enzymes activities and maintenance of glutathione (GSH) pool, and consequently higher photochemical functioning. The levels of nitric oxide (NO) correlated well with CAM expression, which was observed only in S. album, suggesting that NO acts in a different way in C₃ and C₄ species during CAM induction. Additionally, in both species, over the course of CAM induction, the changes in CAM expression parameters exhibited a similar pattern to that of antioxidant capacity and photochemical functioning parameters.     

Effect of ellagic acid on growth and physiology of canola (Brassica napus L.) under saline conditions

Salinity stress is limiting growth and productivity of plants in many areas of the world. Plants adopted different strategies to minimize the effect of salt stress. A pot experiment was conducted to investigate the morphological and physiological changes produced in Canola (Brassica napus) by exogenous application of ellagic acid (EA) under saline conditions. EA is an antioxidant, expected to reduce the effect of salinity stress. The seeds of two canola cultivars, Rainbow and Oscar, were soaked for 6?h with different concentrations of EA (0, 55 and 110?µg/ml). The soaked seeds were sown in small pots. Salt stress was imposed on the plants by applying NaCl solutions of different concentrations (0, 60 and 120?mM) and the duration of stress was for four weeks. Salinity stress reduced seed germination and disturbed the morphological and physiological attributes of B. napus. Application of EA as seed soaking reduced the effect of salinity and enhanced the growth of plants. Overall, we could confirm a significant role of EA by inducing salinity tolerance in B. napus.     

The Possible Roles of Priming with ZnO Nanoparticles in Mitigation of Salinity Stress in Lupine (Lupinus termis) Plants

To our knowledge, little attention has been paid to evaluating ZnO nanoparticles (ZNPs) roles in plants grown under salinity stress. In this study, seeds of lupine (Lupinus termis) plants were grown in plastic pots and exposed to 0 (control) and 150 (S) mM NaCl with or without priming with different concentrations of ZnO [20 mg L^?1 (ZNPs1), 40 mg L^?1 (ZNPs2), and 60 mg L^?1 (ZNPs3)] for 20 days. Salinized plants showed a reduction in plant growth parameters (root length, shoot length, fresh weight, and dry weight) and in the contents of photosynthetic pigments (chlorophyll a and b, and carotenoids) and Zn, as well as in the activity of catalase (CAT) against control plants. On the other side, salinity stress boosted the contents of organic solutes (soluble sugar, soluble protein, total free amino acids, and proline), total phenols, malondialdehyde (MDA), ascorbic acid and Na, as well as the activities of superoxide dismutase (SOD), peroxidase (POD), and ascorbate peroxidase (APX) in stressed plants over control plants. However, seed-priming with ZNPs mostly stimulated growth of stressed plants, which was accompanied by reinforcement in the levels of photosynthetic pigments, organic solutes, total phenols, ascorbic acid and Zn, as well as in the activities of SOD, CAT, POD, and APX enzymes over stressed plants alone. On the contrary, priming with ZNPs caused a decrement in the contents of MDA and Na in stressed plants relative to salinized plants alone. It is worthy to mention that, this improvement in salt tolerance of plants primed with ZNPs was more obvious in plants primed with ZNPs3 and grown both in unstressed and stressed regimes. Thus, our findings suggest that seed-priming with ZNPs, especially 60 mg L^?1 ZnO is an effective strategy that can be used to enhance salt tolerance of lupine plants.

     

Variability in response to salinity stress in natural Tunisian populations of Hordeum marinum subsp. marinum

• Soil salinity is one of the most serious environmental factors affecting crop productivity around the world.
• In this study, we analysed morpho‐physiological variation in responses to salt stress in Tunisian populations of Hordeum marinum subsp. marinum. The plants were grown under two treatments (0 and 200 mm NaCl) until maturity. A total of 19 quantitative traits were measured before and during the harvest.
• It was observed that most studied traits are influenced by the increasing salinity. High to moderate broad‐sense heritability (H²) were noted for most of parameters under control and salt treatment, implying that salt tolerance is moderately heritable and environmental variation plays an equally important role. The majority of correlations between measured traits under the two treatments are positive, where the strongest correlations were between spike number (SN) and weight (SW). Based on the salt response index (SRI) values, SN and SW are the most affected by salinity. The 150 studied lines formed three groups according to the SRI values of the 19 quantitative parameters, of which 101 were moderately sensitive, 27 tolerant and 22 highly tolerant.
• Overall genetic variation of H. marinum in response to salt stress may provide novel insight to identify genes responsible for salt tolerance.

     

The Irreversible C3 to CAM Shift in Well-watered and Salt-stressed Plants of Mesembryanthemum crystallinum is under Strict Ontogenetic Control

The shift from C₃ to CAM was investigated as a function of both leaf and plant age in well-watered and salt-stressed (300 mM NaCl solution) plants of Mesembryanthemum crystallinum. Initiation of a night-time accumulation of malic acid, the decisive criterion of CAM, was followed in plants that were continuously stressed at different points in their life cycle. The deinducibility of CAM was examined after the release from stress by extensively rinsing the potting soil with de-mineralized water. Our results show that in M. crystallinum CAM is under strict developmental control, since CAM appeared only when a certain stage of development of the whole plant was reached. CAM was not present in any plant before this threshold, which was the same in salt-stressed as well as in well-watered plants. The metabolic shift coincided with the change from the seedling to the juvenile growth phase, and not with that from vegetative to reproductive growth, represented by the start of branching. The latter is timed to the end of extension growth. In well-watered plants, after this decisive point in development, a weak nighttime accumulation of malic acid could be measured (? 0.05 mol kg_DW^?1) in the oldest, mature leaves but not in young, developing ones. This “CAM capacity” gradually increased up to 0.2 mol kg_DE^?1 with further plant ageing. Leaf senescence, characterized by wilting and yellowing, diminished the CAM activity. In mature leaves salt stress drastically enhanced the magnitude of diurnal fluctuation in malic acid content. Removal of salt stress did not deinduce CAM activity, but diminished the amplitude of malic acid oscillations to some extent in those plants which had been stressed from early in their life cycle. In these plants, salt stress delayed plant development and growth thus retarding the life cycle. Well-watered plants, for example, branched about three weeks earlier than those that had been stressed continuously from one week after germination. After removal of stress a quasi-preserved earlier developmental stage in relation to the control plants determined the weaker CAM expression.     

Increased expression of phospholipase Dα1 in guard cells decreases water loss with improved seed production under drought in Brassica napus

The activation of phospholipase Dα1 (PLDα1) produces lipid messenger phosphatidic acid and promotes stomatal closure in Arabidopsis. To explore the use of the PLDα1‐mediated signalling towards decreasing water loss in crop plants, we introduced Arabidopsis PLDα1 under the control of a guard cell–specific promoter AtKatI_pro into two canola (Brassica napus) cultivars. Multiple AtKatI_pro::PLDα1 lines in each cultivar displayed decreased water loss and improved biomass accumulation under hyperosmotic stress conditions, including drought and high salinity. Moreover, AtKatI_pro::PLDα1 plants produced more seeds than did WT plants in fields under drought. The results indicate that the guard cell–specific expression of PLDα1 has the potential to improve crop yield by enhancing drought tolerance.     

Cross-talk between salicylic acid and NaCl-generated reactive oxygen species and nitric oxide in tomato during acclimation to high salinity

Hydrogen peroxide (H₂O₂) and nitric oxide (NO) generated by salicylic acid (SA) are considered to be functional links of cross‐tolerance to various stressors. SA‐stimulated pre‐adaptation state was beneficial in the acclimation to subsequent salt stress in tomato (Solanum lycopersicum cv. Rio Fuego). At the whole‐plant level, SA‐induced massive H₂O₂ accumulation only at high concentrations (10^?3–10^?2M), which later caused the death of plants. The excess accumulation of H₂O₂ as compared with plants exposed to 100 mM NaCl was not associated with salt stress response after SA pre‐treatments. In the root tips, 10^?3–10^?2M SA triggered the production of reactive oxygen species (ROS) and NO with a concomitant decline in the cell viability. Sublethal concentrations of SA, however, decreased the effect of salt stress on ROS and NO production in the root apex. The attenuation of oxidative stress because of high salinity occurred not only in pre‐adapted plants but also at cell level. When protoplasts prepared from control leaves were exposed to SA in the presence of 100 mM NaCl, the production of NO and ROS was much lower and the viability of the cells was higher than in salt‐treated samples. This suggests that, the cross‐talk of signalling pathways induced by SA and high salinity may occur at the level of ROS and NO production. Abscisic acid (ABA), polyamines and 1‐aminocyclopropane‐1‐carboxylic acid, the compounds accumulating in pre‐treated plants, enhanced the diphenylene iodonium‐sensitive ROS and NO levels, but, in contrast to others, ABA and putrescine preserved the viability of protoplasts.     

Endophytic bacterium Bacillus subtilis (BERA 71) improves salt tolerance in chickpea plants by regulating the plant defense mechanisms

Plant growth-promoting endophytic bacteria can stimulate the growth, nutrient acquisition, symbiotic performance and stress tolerance of chickpea plants under saline soil conditions. The aim of this study was to investigate the stress-adaptive mechanisms of chickpea plants mediated by Bacillus subtilis (BERA 71) under saline conditions. Inoculation with BERA 71 enhanced plant biomass and the synthesis of photosynthetic pigments and reduced the levels of reactive oxygen species (ROS) and lipid peroxidation in plants under conditions of stress. Furthermore, the activities of ROS-scavenging antioxidant enzymes (superoxide dismutase, peroxidase, catalase and glutathione reductase), the levels of non-enzymatic antioxidants (ascorbic acid and glutathione) and the total phenol content were increased in stressed plants during bacterial association. The bacteria decreased sodium accumulation and enhanced the nitrogen, potassium, calcium and magnesium content in the plants. The suppression of ROS generation and of lipid peroxidation and the accumulation of proline in BERA-71-inoculated plants enhanced the membrane stability under salinity stress and non-stress conditions.     

Metabolic and Physiological Changes Induced by Nitric Oxide and Its Impact on Drought Tolerance in Soybean

Nitric oxide (NO) is an important signaling molecule that plays a pivotal role in stress tolerance. To study the role of NO in drought tolerance and elucidate the underlying mechanisms, NO (0 and 100 μM) was applied to drought-treated soybean plants. Drought stress was imposed by PEG (5% (W/V) of PEG 6000. Nitric oxide improved growth of soybean plants under drought as evidenced by enhanced dry weight (30%). Nitric oxide caused a remarkable increase in activities of catalase and superoxide dismutase (SOD) and SOD expression (14.8-fold), which led to a significant decline in malondealdehyde content under drought conditions. Nitric oxide induced proline biosynthesis due to enhancing pyrroline-5- carboxylate synthetase (P5CS) expression (43.66-fold). The growth-promoting effect of NO application in soybean plants was concomitant with change in metabolic profile (phenolic acid and flavonoid compounds). Nitric oxide up-regulated of phenylalanine ammonia-lyase (PAL) expression in drought-treated plants and may influence on the phenylpropanoid production. Nitric oxide increased salicylic acid (SA) content in soybean plants under stress. So, NO and SA are jointly responsible for boosted tolerance to drought stress in soybean plants. The decrease in unsaturated fatty acid through NO application might reflect a reduction in oxidative damage. These results propose a multifaceted contribution of NO through regulation of physiological and metabolic processes in response to drought stress.

     

Day-night cycles of net CO2 exchange in crassulacean acid metabolism as related to day-night changes of abscisic-acid levels

Eight to nine months old seedlings of the Cactaceae Gereus validus HAWORTH grown in soil culture were chosen to study day-night cycles of net CO₂ exchange, indicating the stomatal rhythm of crassulacean acid metabolism, CAM, in relation to day-night changes of abscisic-acid levels. Drought stress was imposed by repotting the seedlings in dry sand and keeping them without watering for up to 37–39 days while control plants were watered regularly. Abscisicacid levels were higher in the stressed plants than in the controls and higher in the light period aa compared to the dark period. In the stressed plants abscisic-acid levels increased throughout the light period. Abscisic acid reached particularly high levels in the late afternoon in plants stressed for 37 days. It is conceivable that stomatal closure during the first part of the light period of CAM is elicited by high internal partial pressures of CO₂ built up by decarboxylation of nocturnally stored malic acid. However, the elimination of late-afternoon stomatal opening and the reduction of stomatal opening during the dark period of CAM observed under conditions of drought stress must have other reasons. The analyses of abscisic acid presented allow the conclusion that this stress hormone is involved in stomatal regulation of CAM under such conditions.     

An osmotin from the resurrection plant Tripogon loliiformis (TlOsm) confers tolerance to multiple abiotic stresses in transgenic rice

Osmotin is a key protein associated with abiotic and biotic stress response in plants. In this study, an osmotin from the resurrection plant Tripogon loliiformis (TlOsm) was characterized and functionally analyzed under abiotic stress conditions in T. loliiformis as well as in transgenic Nicotiana tabacum (tobacco) and Oryza sativa (rice) plants. Real‐time PCR analysis on mixed elicitor cDNA libraries from T. loliiformis showed that TlOsm was upregulated a 1000‐fold during the early stages of osmotic stresses (cold, drought, and salinity) in both shoots and roots but downregulated in shoots during heat stress. There was no change in TlOsm gene expression in roots of heat‐stressed plants and during plant development. The plasma membrane localization of TlOsm was showed in fluorescent‐tagged TlOsm tobacco plants using confocal laser scanning microscopic analysis. Transgenic rice plants expressing TlOsm were assessed for enhanced tolerance to salinity, drought and cold stresses. Constitutively expressed TlOsm in transgenic rice plants showed increased tolerance to cold, drought and salinity stress when compared with the wild‐type and vector control counterparts. This was evidenced by maintained growth, retained higher water content and membrane integrity, and improved survival rate of TlOsm‐expressing plants. The results thus indicate the involvement of TlOsm in plant response to multiple abiotic stresses, possibly through the signaling pathway, and highlight its potential applications for engineering crops with improved tolerance to cold, drought and salinity stress.     

Isolation of a cDNA clone (PcSrp) encoding serine-rich-protein from Porteresia coarctata T. and its expression in yeast and finger millet (Eleusine coracana L.) affording salt tolerance

A 1.4 Kb cDNA clone encoding a serine-rich protein has been isolated from the cDNA library of salt stressed roots of Porteresia coarctata, and designated as P. coarctata serine-rich-protein (PcSrp) encoding gene. Northern analysis and in situ mRNA hybridization revealed the expression of PcSrp in the salt stressed roots and rhizome of P. coarctata. However, no such expression was seen in the salt stressed leaves and in the unstressed tissues of root, rhizome and leaf, indicating that PcSrp is under the control of a salt-inducible tissue-specific promoter. In yeast, the PcSrp conferred increased NaCl tolerance, implicating its role in salinity tolerance at cellular level. Further, PcSrp was cloned downstream to rice Actin-1 promoter and introduced into finger millet through particle-inflow-gun method. Transgenic plants expressing PcSrp were able to grow to maturity and set seed under 250 mM NaCl stress. The untransformed control plants by contrast failed to survive under similar salt stress. The stressed roots of transgenic plants invariably accumulated higher Na⁺ and K⁺ ion contents compared to roots of untransformed plants; whereas, shoots of transgenics accumulated lower levels of both the ions than that of untransformed plants under identical stress, clearly suggesting the involvement of PcSrp in ion homeostasis contributing to salt tolerance.