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.     

Proteome Dynamics and Physiological Responses to Short-Term Salt Stress in Brassica napus Leaves

Salt stress limits plant growth and crop productivity and is an increasing threat to agriculture worldwide. In this study, proteomic and physiological responses of Brassica napus leaves under salt stress were investigated. Seedlings under salt treatment showed growth inhibition and photosynthesis reduction. A comparative proteomic analysis of seedling leaves exposed to 200 mM NaCl for 24 h, 48 h and 72 h was conducted. Forty-four protein spots were differentially accumulated upon NaCl treatment and 42 of them were identified, including several novel salt-responsive proteins. To determine the functional roles of these proteins in salt adaptation, their dynamic changes in abundance were analyzed. The results suggested that the up-accumulated proteins, which were associated with protein metabolism, damage repair and defense response, might contribute to the alleviation of the deleterious effect of salt stress on chlorophyll biosynthesis, photosynthesis, energy synthesis and respiration in Brassica napus leaves. This study will lead to a better understanding of the molecular basis of salt stress adaptation in Brassica napus and provides a basis for genetic engineering of plants with improved salt tolerance in the future.     

Fruit Spray of 24-Epibrassinolide and Fruit Shade Alter Pericarp Photosynthesis Activity and Seed Lipid Accumulation in <Emphasis Type="Italic">Styrax tonkinensis</Emphasis>

As demand for biofuel feedstock production increases, the substitutable woody oilseed benzoin (Styrax tonkinensis (Pierre) Craib ex Hartwich) has received increased attention among Chinese researchers. However, the mechanism of seed oil regulation has not been revealed in this species. Maternal control of seed lipid content has been observed in many plant species. In this study, in planta treatments were applied to create different pericarp photosynthesis activation levels in benzoin. Compared with fruit shading treatment, superior photosynthetic physiological characteristics were found in the brassinosteroid hormone 24-epibrassinolide-treated pericarps during maturation (56–112 days after flowering); specifically, these superior characteristics were reflected by higher average and peak values of the Chl a/b ratio, NADPH/NADP⁺ ratio, NADP-dependent malate dehydrogenase activity, and ribulose 1,5-bisphosphate carboxylase/oxygenase activity. Next, we found that the pericarp-encapsulated seed yield and oil content were closely connected to the maternal-specific photosynthetic events. The enhanced pericarp photosynthesis activity (treated with 10 μM brassinosteroid) was accompanied by higher fatty acid synthase activity and a faster lipid deposition rate, which was verified by transmission electron microscopy. These changes led to a final increase in the total fatty acid (FA) content of mature seeds of 19%, which could be largely accounted for by the elevated saturated FA (mostly long-chain FAs) percentage in benzoin oil. When benzoin fruits grew in low light (bagged in three layers of black non-woven fabrics), lipid synthesis capability was markedly impaired. The dynamics of cellular storage reserves were further analyzed, suggesting that the pericarp-seed carbohydrate translocation efficiency in fruits and the carbon partitioning among seed reserves were directly responsible for the seed oil diversification under the treatments. Taken together, our results highlighted the maternal control, via pericarp photosynthesis, over the regulation of the seed oil content in benzoin.     

Overexpression of a wheat Na+/H+ antiporter gene (TaNHX2) enhances tolerance to salt stress in transgenic tomato plants (Solanum lycopersicum L.)

Soil salinity is a major environmental stress limiting plant productivity. Vacuole Na⁺/H⁺ antiporters play important roles for the survival of plants under salt stress conditions. We have developed salt stress tolerant transgenic tomato plants (Solanum lycopersicum cv. PED) by overexpression of the wheat Na⁺/H⁺ antiporter gene TaNHX2 using Agrobacterium tumefaciens strain LBA4404 harbouring a binary vector pBin438 that contains the TaNHX2 gene under the control of double CaMV 35S promoter and npt II as a selectable marker. PCR and Southern blot analysis confirmed that TaNHX2 gene has been integrated and expressed in the T₁ generation transgenic tomato plants. When TaNHX2 expressing plants were exposed to 100 or 150 mM NaCl, they were found to be more tolerant to salt stress compared to wild type plants. Biochemical analyses also showed that transgenic plants have substantial amount of relative water content and chlorophyll content under salt stress conditions compared to wild type plants. The relative water content in transgenic and wild type plants ranged from 68 to 75 % and 46–73 % and chlorophyll content fall in between 1.8 to 2.4 mg/g fw and 1.0 to 2.4 mg/g fw, respectively, in all stress conditions. In the present study, we observed a better germination rate of T₁ transgenic seeds under salt stress conditions compared with wild type plants. Our results indicated that TaNHX2-transgenic tomato plants coped better with salt stress than wild type plants.     

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.     

Improvement of element uptake and antioxidative defense in Brassica napus under lead stress by application of hydrogen sulfide

Heavy metal pollution is one of the major constraints in oilseed rape (Brassica napus L.) production. In this study, protective role of hydrogen sulfide (H₂S) on plant growth under lead (Pb) stress was studied in B. napus. Plants were grown hydroponically in greenhouse conditions under three levels (0, 100, and 400 μM) of Pb and three levels (0, 100 and 200 μM) of H₂S donor sodium hydrosulfide. Outcomes demonstrated that Pb stress significantly reduced the plant biomass, leaf chlorophyll contents, nutrients uptake in the leaves and roots of B. napus plants. Exogenous application of H₂S significantly improved the plant biomass, chlorophyll contents and concentration of macro- and micronutrients in the leaves and roots of B. napus plants under Pb-toxicity conditions. The data indicated that application of Pb alone significantly increased the reactive oxygen species (ROS) as well as malondialdehyde (MDA) in the leaves and roots of plants. Meanwhile, application of H₂S decreased the production of MDA and ROS in the leaves and roots by increasing antioxidant activities under Pb stress. Moreover, this study also revealed that plants treated with H₂S at different concentrations enhanced the contents of total glutathione and glutathione reduced/glutathione oxidized ratio in leaves and roots under different levels of Pb. The results depicted that H₂S improved the plant biomass, uptake of nutrients in the leaves and roots of B. napus plants and enhanced the performance of antioxidant defense system due to its ameliorative potential under Pb stress conditions.     

In vitro Propagation and Isozyme Polymorphism of the Medicinal Plant Hypericum brasiliense

Responses of 20 d-old plants of two Brassica napus L. cultivars Dunkeld and Cyclon to NaCl salinized soil [electrical conductivity 2.4 (control), 4.0, 8.0 or 12.0 dS m⁻¹] were examined. The salt tolerant line Dunkeld had significantly higher fresh and dry masses of shoots, and seed yield than salt sensitive line Cyclon in all salinities. The effect of salt stress on reduction in total leaf soluble sugars was markedly greater in Dunkeld as compared to that in Cyclon. No effect of salt stress was observed on leaf soluble proteins but there was a slight increase in total free amino acids of both cultivars. Leaf proline content increased markedly in both cultivars and Dunkeld had greater proline content than Cyclon at all salinities. Salt stress had no significant effect on seed oil content and erucic acid content of seed oil, however, content of glucosinolates in the seed meal increased and Cyclon had greater content of glucosinolates than Dunkeld. This revised version was published online in July 2006 with corrections to the Cover Date.     

Indigenous salt-tolerant rhizobacterium <Emphasis Type="Italic">Pantoea dispersa</Emphasis> (PSB3) reduces sodium uptake and mitigates the effects of salt stress on growth and yield of chickpea

Salt stress has multiple damaging effects on plants including physiological damage, reduced growth, and productivity. Plant growth-promoting rhizobacteria (PGPR) are one of the valuable options to mitigate the negative effects of this stress. In the present study, native bacteria from chickpea’s rhizosphere were isolated, and checked for their salt tolerance and plant growth-promoting attributes (phosphate (P) solubilization, siderophores, indole-3-acetic acid (IAA) production, and 1-aminocyclopropane-1-carboxylate (ACC) deaminase production). One isolate, subsequently identified as Pantoea dispersa, showed appreciable production of IAA (218.3 µg/ml) and siderophores (60.33% SU), P-solubilization (3.64 µg/ml) and ACC deaminase activity (207.45 nmol/mg/h) in the presence of 150 mM NaCl, under laboratory conditions. Salt stress in uninoculated chickpea (GPF2 cultivar) plants induced high accumulation of Na⁺ ions (3.86 mg g^?1 dw) in the leaves, along with significant reduction in K⁺ uptake, membrane integrity, chlorophyll concentration, and leaf water content, thus resulting in impaired growth of the plant and yield (pods and seeds) in a salt concentration-dependent manner. The damage due to salt stress was restored significantly in plants inoculated with P. dispersa. A significant improvement in biomass (32–34%), pods number (31–34.5%), seeds number (32–35.7%), pods weight (30–32.6%), and seeds weight (27–35%) per plant occurred in salt stress-affected plants, which was associated with significant reduction in Na⁺ uptake, reduced membrane damage, significantly improved leaf water content, chlorophyll content, and K⁺ uptake. This study suggests for the first time that native P. dispersa strain PSB3 can be used to alleviate the negative effects of salt stress on chickpea plants and holds the potential to be used as a biofertilizer.     

Seed priming with sorghum extracts and benzyl aminopurine improves the tolerance against salt stress in wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.)

Salt stress impedes the productivity of wheat (Triticum aestivum L.) in many parts of the world. This study evaluated the potential role of benzyl aminopurine (BAP) and sorghum water extract (SWE) in improving the wheat performance under saline conditions. Seeds were primed with BAP (5 mg L^?1), SWE (5% v/v), BAP + SWE, and distilled water (hydropriming). Soil filled pots maintained at the soil salinity levels of 4 and 10 dS m^?1 were used for the sowing of primed and non-primed seeds. Salt stress suppressed the wheat growth; seed priming treatments significantly improved the wheat growth under optimal and suboptimal conditions. Total phenolics, total soluble sugars and proteins, α-amylase activity, chlorophyll contents, and tissue potassium ion (K⁺) contents were increased by seed priming under salt stress; while, tissue sodium ion (Na⁺) contents were decreased. Seed priming with SWE + BAP was the most effective in this regard. Under salt stress, the tissue Na⁺ contents were reduced by 5.78, 28.3, 32.2, 36.7% by hydropriming, seed priming with SWE, seed priming with BAP, and seed priming with SWE + BAP, respectively over the non-primed control. Effectiveness of seed priming techniques followed the order SWE + BAP > BAP > SWE > Hydropriming. In conclusion, seed priming with SWE + BAP may be opted to improve the tolerance against salt stress in wheat.     

Accumulation and distribution of Zn in the shoots and reproductive structures of the halophyte plant species Kosteletzkya virginica as a function of salinity

Kosteletzkya virginica is a wetland halophyte that is a good candidate for rehabilitation of degraded salt marshes and production of oil as biodiesel. Salt marshes are frequently contaminated by heavy metals. The distribution of Zn in vegetative and reproductive organs of adult plants, and the NaCl influence on this distribution remain unknown and were thus explored in the present study. Plants were cultivated in a nutrient film technique system, from seedling stage until seed maturation in a control, Zn (100 μM), NaCl (50 mM) or Zn + NaCl medium. Photosynthesis, ion nutrition, malondialdehyde and non-protein thiol concentrations were quantified. Zinc distribution in reproductive organs was estimated by a laser ablation-inductively coupled plasma-mass spectrometry procedure (LA-ICP-MS). Adult plants accumulated up to 2 mg g^?1 DW Zn in the shoots. Zinc reduced plant growth, inhibited photosynthesis and reduced seed yield. Zinc accumulation in the seeds was only two times higher in Zn-treated plants than in controls. Exogenous NaCl neutralized the damaging action of Zn and modified the Zn distribution through a preferential accumulation of toxic ions in older leaves. Zinc was present in seed testa, endosperm and, to a lower extent, in embryo. Additional NaCl induced a chalazal retention of Zn during seed maturation and reduced final Zn seed content. It is concluded that NaCl 50 mM had a positive impact on the response of K. virginica to Zn toxicity and acts through a modification in Zn distribution rather than a decrease in Zn absorption.     

Salt tolerance conferred by expression of a global regulator IrrE from <Emphasis Type="Italic">Deinococcus radiodurans</Emphasis> in oilseed rape

As salinity is a major threat to sustainable agriculture worldwide, cultivation of salt-tolerant crops becomes increasingly important. IrrE acts as a global regulator and a general switch for stress resistance in Deinococcus radiodurans. In this study, to determine whether the irrE gene can improve the salt tolerance of Brassica napus, we introduced the irrE gene into B. napus by the Agrobacterium tumefaciens-mediated transformation method. Forty-two independent transgenic plants were regenerated. Polymerase chain reaction (PCR) analyses confirmed that the irrE gene had integrated into the plant genome. Northern as well as Western blot analyses revealed that the transgene was expressed at various levels in transgenic plants. Analysis for the T₁ progenies derived from four independent transformants showed that irrE had enhanced the salt tolerance of T₁ in the presence of 350 mM NaCl. Furthermore, under salt stress, transgenic plants accumulated more compatible solutes (proline) and a lower level of malondialdehyde (MDA), and they had higher activities of catalase (CAT), peroxidase (POD) and superoxide dismutase (SOD). However, agronomic traits were not affected by irrE gene overexpression in the transgenic B. napus plants. This study indicates that the irrE gene can improve the salt tolerance of B. napus and represents a promising candidate for the development of crops with enhanced salt tolerance by genetic engineering.     

Brassinosteroid seed priming with nitrogen supplementation improves salt tolerance in soybean

This study was conducted to evaluate the influence of brassinosteroid (24-epibrassinolide, EBL) seed priming and optimal nitrogen (N) supply in improving salt tolerance in soybean. The experimental treatments were (a) control (nutrient solution without N and without EBL priming), (b) nutrient solution without N and EBL seed priming, (c) N supplemented nutrient solution without EBL priming and (d) EBL seed priming + N supplemented nutrient solution under optimal (0 mM NaCl) and salt stress (0 mM NaCl) conditions. Salt stress caused significant reduction in growth and biomass accumulation of soybean. However, EBL seed priming and application of N improved the soybean performance under optimal and salt stress conditions. In this regard, treatments receiving both EBL and N were more effective. EBL priming and N, alone and in combination, triggered the accumulation of osmolytes including proline, glycine betaine and sugars resulting in better photo-protection through maintenance of tissue water content. Antioxidant activity and osmolyte accumulation significantly increased due to combined treatment of N and EBL under normal as well as salt stress conditions. In conclusion, salt stress caused reduction in growth and biomass soybean due to oxidative damage and osmotic stresses. However, soybean performance was improved by seed priming with EBL. Supplementation of N further improved the effectiveness of EBL treatment in improving salt tolerance in soybean.     

Silicon Application to Rice Root Zone Influenced the Phytohormonal and Antioxidant Responses Under Salinity Stress

Silicon (Si) application shows beneficial effects on plant growth; however, its effects on the phytohormone and enzymatic antioxidant regulation have not been fully understood. We studied the effects of short-term (6, 12, and 24 h) silicon (0.5, 1.0, and 2.0 mM) application on salinity (NaCl)-induced phytohormonal [abscisic acid (ABA), jasmonic acid (JA), and salicylic acid (SA)] and antioxidant regulation in Oryza sativa. The results showed that Si treatments significantly increased rice plant growth compared to controls under salinity stress. Si treatments reduced the sodium accumulation resulting in low electrolytic leakage and lipid peroxidation compared to control plants under salinity stress. Enzymatic antioxidant (catalase, peroxidase and polyphenol oxidase) responses were more pronounced in control plants than in Si-treated plants under salinity stress. Stress- and defense-related phytohormones like JA were significantly downregulated and SA was irregular after short-term Si applications under salinity stress compared to control. Conversely, ABA was significantly higher after 6 and 12 h but insignificant after 24 h in Si-treated plants under salinity stress. After 6 and 12 h, Si and salinity stress resulted in upregulation of zeaxanthin epoxidase and 9-cis-epoxycarotenoid dioxygenase 1 and 4 (NCED1 and 4), whereas 24-h treatments significantly downregulated the expressions of these genes compared to those in the control. NCED3 expression increased after 6 and 24 h but it was insignificant after 12 h of Si application compared to control. The current findings indicate that increasing the Si concentrations for longer periods of time can regulate the salinity-induced stress by modulating phytohormonal and enzymatic antioxidants’ responses.     

Salt-induced abnormalities on root tip mitotic cells of Allium cepa: prevention by inositol pretreatment

Salt-induced growth reduction of plants is a well-known phenomenon which poses major problem in crop productivity in places where vast majority of land plants are affected by salt. In this report, studies were carried out to reveal the effect of salt injury on the cell division pattern in roots and the role of myo-inositol in preventing the salt-induced ion disequilibrium on the chromosome and DNA degradation in roots. Present study revealed induction of various chromosomal abnormalities on the root tip mitotic cells of Allium cepa by treatment with different concentrations of NaCl (0–500 mM) for 24 h as also the amelioration of such effect by prior treatment of the roots with different concentration of myo-inositol (0–300 mM). Results showed that a narrow albeit definite range of extracellular myo-inositol (100–150 mM) is effective in preventing internucleosomal fragmentation which is the early response in roots under salt stress. Transgenic tobacco plants overexpressing Oryza (OsINO1) as well as Porteresia (PcINO1) cytosolic l-myo-inositol-1-phosphate synthase coding genes can withstand and retain their chromosomal and DNA integrity in 100 mM NaCl solution and can subsequently prevent DNA fragmentation, caused by intracellular endonuclease activity at this salt concentration.     

Germination and ROS detoxification in bell pepper (Capsicum annuum L.) under NaCl stress and treatment with microalgae extracts

We evaluated the salt tolerance of hybrids of pepper (Capsicum annuum L.) during germination. Treatments were applied at 0, 25, and 50 mM NaCl with preparations of supplemental extracts of the microalgae Dunaliella salina and Phaeodactylum tricornutum to determine the percentage germination rate as well as measured indicators of oxidative stress caused by the salt treatments during seed germination. We found that root growth was favorably influenced by the microalgae leading to increased germination rate. Tissues were analyzed in terms of superoxide radical production, lipid peroxidation, and activity of antioxidant enzymes viz. superoxide dismutase, catalase, and glutathione peroxidase. Our results suggest that application of microalgae extracts significantly reduced (p?<?0.05) superoxide radical production, as well as lower lipid peroxidation in comparison to plants without extracts of microalgae. The antioxidant enzymes increased in the presence of microalgae showing a significant difference (p?<?0.05). The results suggest differences in oxidative metabolism in response to the magnitude of salt stress and concentrations of microalgae help mitigate salt stress in plants during the germination process.     

Plant growth promoting rhizobacteria alleviate salinity induced negative effects on growth, oil content and physiological status in Mentha arvensis

This study was aimed to investigate the effect of inoculation on three salt-tolerant, plant-growth-promoting rhizobacteria (PGPR) STR2 (Bacillus pumilus), STR8 (Halomonas desiderata) and STR36 (Exiguobacterium oxidotolerans), for their growth-promoting potential and efficacy in augmenting salt tolerance in Mentha arvensis, an essential oil-bearing crop and natural source of l-menthol, under varying levels of NaCl stress (0, 100, 300 and 500 mM) imposed through irrigating water. Increase in the levels of salt concentration led to a decrease in the growth, fresh weight, leaf–stem ratio, oil content and yield. However, the negative effects of salinity were observed to be convalesced in the PGPR inoculated plants. At salinity levels of 100 and 300 mM NaCl, H. desiderata inoculated plants recorded the highest herb yield, whereas at 500 mM NaCl, the plants inoculated with E. oxidotolerans yielded maximum herb. The oil content in non-inoculated, salt-stressed plants was observed to be 0.46, 0.42 and 0.35 % at 100, 300 and 500 mM NaCl, respectively, whereas the plants inoculated with H. desiderata recorded the oil content of 0.71 and 0.60 and 0.48 % at similar levels of NaCl stress, respectively. The halotolerant PGPR minimized the deleterious effects of salt toxicity producing at par or higher yields at lower and medium salinity levels (100, 300 mM NaCl) than the un-inoculated non-salt-stressed plants through improved foliar nutrient uptake and enhanced antioxidant machinery. Based on the results of the experiments reported herein, the use of salt-tolerant, plant-growth-promoting bacteria may provide an effective means of facilitating M. arvensis growth in salt-stressed environments.