Abeer Hashem

Nitric Oxide Mitigates Salt Stress by Regulating Levels of Osmolytes and Antioxidant Enzymes in Chickpea

This work was designed to evaluate whether external application of nitric oxide (NO) in the form of its donor S-nitroso-N-acetylpenicillamine (SNAP) could mitigate the deleterious effects of NaCl stress on chickpea (Cicer arietinum L.) plants. SNAP (50 μM) was applied to chickpea plants grown under non-saline and saline conditions (50 and 100 mM NaCl). Salt stress inhibited growth and biomass yield, leaf relative water content (LRWC) and chlorophyll content of chickpea plants. High salinity increased electrolyte leakage, carotenoid content and the levels of osmolytes (proline, glycine betaine, soluble proteins and soluble sugars), hydrogen peroxide (H2O2) and malondialdehyde (MDA), as well as the activities of antioxidant enzymes, such as superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and glutathione reductase in chickpea plants. Expression of the representative SOD, CAT and APX genes examined was also up-regulated in chickpea plants by salt stress. On the other hand, exogenous application of NO to salinized plants enhanced the growth parameters, LRWC, photosynthetic pigment production and levels of osmolytes, as well as the activities of examined antioxidant enzymes which is correlated with up-regulation of the examined SOD, CAT and APX genes, in comparison with plants treated with NaCl only. Furthermore, electrolyte leakage, H2O2 and MDA contents showed decline in salt-stressed plants supplemented with NO as compared with those in NaCl-treated plants alone. Thus, the exogenous application of NO protected chickpea plants against salt stress-induced oxidative damage by enhancing the biosyntheses of antioxidant enzymes, thereby improving plant growth under saline stress. Taken together, our results demonstrate that NO has capability to mitigate the adverse effects of high salinity on chickpea plants by improving LRWC, photosynthetic pigment biosyntheses, osmolyte accumulation and antioxidative defense system.

Role of Trichoderma harzianum in mitigating NaCl stress in Indian mustard (Brassica juncea L) through antioxidative defense system

Salinity stress affected crop production of more than 20% of irrigated land globally. In the present study the effect of different concentrations of NaCl (0, 100, and 200 mM) on growth, physio-biochemical attributes, antioxidant enzymes, oil content, etc. in Brassica juncea and the protective role of Trichoderma harzianum (TH) was investigated. Salinity stress deteriorates growth, physio-biochemical attributes, that ultimately leads to decreased biomass yield in mustard seedlings. Higher concentration of NaCl (200 mM) decreased the plant height by 33.7%, root length by 29.7% and plant dry weight (DW) by 34.5%. On the other hand, supplementation of TH to NaCl treated mustard seedlings showed elevation by 13.8, 11.8, and 16.7% in shoot, root length and plant DW respectively as compared to plants treated with NaCl (200 mM) alone. Oil content was drastically affected by NaCl treatment; however, TH added plants showed enhanced oil percentage from 19.4 to 23.4% in the present study. NaCl also degenerate the pigment content and the maximum drop of 52.0% was recorded in Chl. ‘a’. Enhanced pigment content was observed by the application of TH to NaCl treated plants. Proline content showed increase by NaCl stress and maximum accumulation of 59.12% was recorded at 200 mM NaCl. Further enhancement to 70.37% in proline content was recorded by supplementation of TH. NaCl stress (200 mM) affirms the increase in H2O2 by 69.5% and MDA by 36.5%, but reduction in the accumulation is recorded by addition of TH to mustard seedlings. 200 mM NaCl elevated SOD, POD, APX, GR, GST, GPX, GSH, and GSSG in the present study. Further enhancement was observed by the application of TH to the NaCl fed seedlings. NaCl stress suppresses the uptake of important elements in both roots and shoots, however, addition of TH restored the elemental uptake in the present study. Mustard seedlings treated with NaCl and TH showed restricted Na uptake as compared to seedlings treated with NaCl alone. In conclusion, TH proved to be very beneficial in imparting resistance to the mustard plants against NaCl stress through improved uptake of essential elements, modulation of osmolytes and antioxidants.

Alleviation of salt-induced adverse impact via mycorrhizal fungi in Ephedra aphylla Forssk

The current investigation was carried out to examine the role of arbuscular mycorrhizal fungi (AMF) in alleviating adverse effects of salt stress in Ephedra aphylla. The plants were exposed to 75 and 150 mM sodium chloride (NaCl) stress with and without application of AMF. Salt stress caused significant decrease in chlorophyll and carotenoid contents; however, the application of AMF restored the pigments content in salt-affected plants. Proline, phenols, and lipid peroxidation were increased with increasing concentration of NaCl, but lower accumulation has been reported in plants treated with AMF. NaCl stress also showed increase in different antioxidant enzymes activities (catalase, ascorbate peroxidase, peroxidase, glutathione reductase, and superoxide dismutase), and further increase was observed in plants treated with AMF. The nutrient uptake, Na+ and Na/K ratio increased and potassium and phosphorus were decreased with increasing concentration of NaCl in the present study. However, the colonization with AMF significantly increased K+ and P and reduced Na+ uptake. It is concluded that presown soil treatment with AMF reduced severity of salt stress in E. aphylla through alterations in physiological parameters, antioxidants and uptake of nutrients.

Salinity Stress and Arbuscular Mycorrhizal Symbiosis in Plants

Soil salinity is one of the main abiotic stresses, which restrict the plant growth and development, and therefore causes major threat to crop productivity. To minimize the crop loss, plant biologists are searching alternatives to develop salt-tolerant crop plants through different means such as plant breeding and genetic engineering. These approaches are successful and presently under use in developed countries but are too costly for the developing countries. In the present scenarios, various methods are utilized all over the globe to mitigate the adverse effect of salinity. One of the commonly used methods is to use beneficial bacteria and fungi that colonize with the plant roots and ultimately alleviate the salinity stress in plants. Among them, application of arbuscular mycorrhizal (AM) fungi has been found to be very effective in mitigating the salinity stress and also improves the crop yield. Various parameters were thoroughly studied and reported in the literature and have demonstrated positive effect in plants subjected to AM fungi under different environmental stress including saline stress. This is attributed to increased antioxidative activities, osmolytes and osmoprotectants in tolerant plants. Deficiency of mineral content in plants under salt stress is compensated by the use of AM fungi as the hyphae of AM fungi acquire minerals in abundance and thereby prevent the plants against deleterious effects of salinity stress. Efficient use of AM fungi can bring the wonders in the field of agriculture with improved yield of crop under salt stress and soil health. Besides this, identification of novel genes that regulate and maintain the biosynthesis of proline and water status in plant cells will help plant researchers to utilize this beneficial interaction in more sustainable ways. The chapter will throw light on the use of AM fungal association in alleviating salt stress in plants and how to exploit this for improved productivity under growth-limited conditions.

The Interaction between Arbuscular Mycorrhizal Fungi and Endophytic Bacteria Enhances Plant Growth of Acacia gerrardii under Salt Stress.

Microbes living symbiotically in plant tissues mutually cooperate with each other by providing nutrients for proliferation of the partner organism and have a beneficial effect on plant growth. However, few studies thus far have examined the interactive effect of endophytic bacteria and arbuscular mycorrhizal fungi (AMF) in hostile conditions and their potential to improve plant stress tolerance. In this study, we investigated how the synergistic interactions of endophytic bacteria and AMF affect plant growth, nodulation, nutrient acquisition and stress tolerance of Acacia gerrardii under salt stress. Plant growth varied between the treatments with both single inoculants and was higher in plants inoculated with the endophytic B. subtilis strain than with AMF. Co-inoculated A. gerrardii had a significantly greater shoot and root dry weight, nodule number, and leghemoglobin content than those inoculated with AMF or B. subtilis alone under salt stress. The endophytic B. subtilis could alleviate the adverse effect of salt on AMF colonization. The differences in nitrate and nitrite reductase and nitrogenase activities between uninoculated plants and those inoculated with AMF and B. subtilis together under stress were significant. Both inoculation treatments, either B. subtilis alone or combined with AMF, enhanced the N, P, K, Mg, and Ca contents and phosphatase activities in salt-stressed A. gerrardii tissues and reduced Na and Cl concentration, thereby protecting salt-stressed plants from ionic and osmotic stress-induced changes. In conclusion, our results indicate that endophytic bacteria and AMF contribute to a tripartite mutualistic symbiosis in A. gerrardii and are coordinately involved in the plant adaptation to salt stress tolerance.

Alleviation of cadmium stress in Solanum lycopersicum L. by arbuscular mycorrhizal fungi via induction of acquired systemic tolerance.

Experiments were conducted to evaluate cadmium (Cd) stress-induced changes in growth, antioxidants and lipid composition of Solanum lycopersicum with and without arbuscular mycorrhizal fungi (AMF). Cadmium stress (50 μM) caused significant changes in the growth and physio-biochemical attributes studied. AMF mitigated the deleterious impact of Cd on the parameters studied. Cadmium stress increased malonaldehyde and hydrogen peroxide production but AMF reduced these parameters by mitigating oxidative stress. The activity of antioxidant enzymes enhanced under Cd treatment and AMF inoculation further enhanced their activity, thus strengthening the plant’s defense system. Proline and phenol content increased in Cd-treated as well as AMF-inoculated plants providing efficient protection against Cd stress. Cadmium treatment resulted in great alterations in the main lipid classes leading to a marked change in their composition. Cadmium stress caused a significant reduction in polyunsaturated fatty acids resulting in enhanced membrane leakage. The present study supports the use of AMF as a biological means to ameliorate Cd stress-induced changes in tomato.

Alleviation of abiotic salt stress in Ochradenus baccatus (Del.) by Trichoderma hamatum (Bonord.) Bainier

The current study was taken up to examine the role of bioagent (Trichoderma hamatum) in mitigating the deleterious effects of NaCl stress in Ochradenus baccatus. Varying concentrations of salt (0, 75, and 150 mM) were used to observe the effect on growth, pigments, some key metabolic attributes, antioxidant enzymes, and elemental accumulation in O. baccatus. The results indicated significant decrease in seed germination, plant growth, pigment content, membrane stability index, tissue water content, and total lipid content with salt stress. Lipid peroxidation increases with the increasing concentration of NaCl. Moreover, salinity stimulated the biosynthesis of phenols, diacylglycerol, sterol esters, nonesterified fatty acids, and enzymatic antioxidants like superoxide dismutase, catalase, peroxidase, ascorbate peroxidase, glutathione reductase. The Na+ content in shoot increases with elevated levels of NaCl concentration, accompanied with significant decreases in K+, Mg2+, and Ca2+. Application of bioagent (T. hamatum) has been observed to alleviate the antagonistic effect of salt stress on plant growth and metabolic processes. In absence and presence of salt stress, the bioagent stimulated the plant growth and alter the plant metabolism through the modification of the above parameters.

Calcium and Potassium Supplementation Enhanced Growth, Osmolyte Secondary Metabolite Production, and Enzymatic Antioxidant Machinery in Cadmium-Exposed Chickpea (Cicer arietinum L.)

This work examined the role of exogenously applied calcium (Ca; 50 mM) and potassium (K; 10 mM) (alone and in combination) in alleviating the negative effects of cadmium (Cd; 200 μM) on growth, biochemical attributes, secondary metabolites and yield of chickpea (Cicer arietinum L.). Cd stress significantly decreased the length and weight (fresh and dry) of shoot and root and yield attributes in terms of number of pods and seed yield (vs. control). Exhibition of decreases in chlorophyll (Chl) a, Chl b, and total Chl was also observed with Cd-exposure when compared to control. However, Cd-exposure led to an increase in the content of carotenoids. In contrast, the exogenous application of Ca and K individually as well as in combination minimized the extent of Cd-impact on previous traits. C. arietinum seedlings subjected to Cd treatment exhibited increased contents of organic solute (proline, Pro) and total protein; whereas, Ca and K-supplementation further enhanced the Pro and total protein content. Additionally, compared to control, Cd-exposure also caused elevation in the contents of oxidative stress markers (hydrogen peroxidase, H2O2; malondialdehyde, MDA) and in the activity of antioxidant defense enzymes (superoxide dismutase, SOD; catalase, CAT; ascorbate peroxidase, APX; glutathione reductase, GR). Ca, K, and Ca + K supplementation caused further enhancements in the activity of these enzymes but significantly decreased contents of H2O2 and MDA, also that of Cd accumulation in shoot and root. The contents of total phenol, flavonoid and mineral elements (S, Mn, Mg, Ca and K) that were also suppressed in Cd stressed plants in both shoot and root were restored to appreciable levels with Ca- and K-supplementation. However, the combination of Ca + K supplementation was more effective in bringing the positive response as compared to individual effect of Ca and K on Cd-exposed C. arietinum. Overall, this investigation suggests that application of Ca and/or K can efficiently minimize Cd-toxicity and eventually improve health and yield in C. arietinum by the cumulative outcome of the enhanced contents of organic solute, secondary metabolites, mineral elements, and activity of antioxidant defense enzymes.

Enhancing growth performance and systemic acquired resistance of medicinal plant Sesbania sesban (L.) Merr using arbuscular mycorrhizal fungi under salt stress

Pot experiments were conducted to evaluate the damaging effects of salinity on Sesbania sesban plants in the presence and absence of arbuscular mycorrhizal fungi (AMF). The selected morphological, physiological and biochemical parameters of S. sesban were measured. Salinity reduced growth and chlorophyll content drastically while as AMF inoculated plants improved growth. A decrease in the number of nodules, nodule weight and nitrogenase activity was also evident due to salinity stress causing reduction in nitrogen fixation and assimilation potential. AMF inoculation increased these parameters and also ameliorated the salinity stress to some extent. Antioxidant enzymes like superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and glutathione reductase (GR) as well as non enzymatic antioxidants (ascorbic acid and glutathione) also exhibited great variation with salinity treatment. Salinity caused great alterations in the endogenous levels of growth hormones with abscisic acid showing increment. AMF inoculated plants maintained higher levels of growth hormones and also allayed the negative impact of salinity.

Exogenous Application of Selenium Mitigates Cadmium Toxicity in Brassica juncea L. (Czern & Cross) by Up-Regulating Antioxidative System and Secondary Metabolites

The main aim of the present study was to examine the role of selenium (Se) in ameliorating the toxic effect of cadmium (Cd) in mustard (Brassica juncea) plants. The plants exposed to elevated levels of Cd exhibited reduced biomass, pigment content, and relative water content (RWC). However, supplementation of Se restores the negative effect of Cd and increases biomass, pigment content, and RWC. Osmolyte (proline and glycine betaine) and sugar content were increased under Cd stress and further increase was observed with addition of Se. Cd decreased protein content and supplementation of Se increases it to appreciable levels. Cd also increased production of H2O2 and lipid peroxidation, electrolyte leakage, and the activities of antioxidant enzymes such as superoxide dismutase, ascorbate peroxidase, and glutathione reductase. Supplementation of Se decreased accumulation of H2O2 and lipid peroxidation, increased the activities of antioxidant enzymes to greater levels, and regulates Cd accumulation in roots and shoots. Ascorbic acid (AsA) and flavonoids decreased with elevated concentrations of Cd; however, tocopherol and total phenols were increased with the same concentrations of Cd. Se application maintains AsA and flavonoid content, and further increase in tocopherol and total phenols were observed with Se in the present study. Overall the results confirm that exogenous application of Se mitigates the negative effects of Cd stress in mustard plants through the regulation of osmoprotectants, antioxidant enzymes, and secondary metabolites.