Role of Salicylic Acid in Promoting Salt Stress Tolerance and Enhanced Artemisinin Production in Artemisia annua L.

In the present investigation, the role of salicylic acid (SA) in inducing salinity tolerance was studied in Artemisia annua L., which is a major source of the antimalarial drug artemisinin. SA, when applied at 1.00 mM, provided considerable protection against salt stress imposed by adding 50, 100, or 200 mM NaCl to soil. Salt stress negatively affected plant growth as assessed by length and dry weight of shoots and roots. Salinity also reduced the values of photosynthetic attributes and total chlorophyll content and inhibited the activities of nitrate reductase and carbonic anhydrase. Furthermore, salt stress significantly increased electrolyte leakage and proline content. Salt stress also induced oxidative stress as indicated by the elevated levels of lipid peroxidation compared to the control. A foliar spray of SA at 1.00 mM promoted the growth of plants, independent of salinity level. The activity of antioxidant enzymes, namely, catalase, peroxidase, and superoxide dismutase, was upregulated by salt stress and was further enhanced by SA treatment. Artemisinin content increased at 50 and 100 mM NaCl but decreased at 200 mM NaCl. The application of SA further enhanced artemisinin content when applied with 50 and 100 mM NaCl by 18.3 and 52.4%, respectively. These results indicate that moderate saline conditions can be exploited to obtain higher artemisinin content in A. annua plants, whereas the application of SA can be used to protect plant growth and induce its antioxidant defense system under salt stress.     

Effect of Phytohormone Pretreatment on Nitrogen Metabolism in Vigna radiata Under Salt Stress

Application of NaCl (electrical conductivity 4.0 mS cm^–1) resulted in about 52, 50 and 55 % reduction in total nitrogen contents in mung bean [Vigna radiata (L.) Wilczek] leaf, root and nodule, respectively. In nodule, nitrogenase activity was reduced by about 84 % under stress as compared with the control set. Glutamine synthetase activity was reduced by about 31, 16 and 23 %, glutamate oxoglutarate aminotransferase activity was reduced by 78, 57 and 42 % and glutamate dehydrogenase activity was reduced by 9, 8 and 42 % in leaf, root and nodule, respectively, under salt stress. The pretreatment with indole-3-acetic acid, gibberellic acid and kinetin, each ranging from 0.1 to 10 µM, in restoring the metabolic alterations imposed by NaCl salinity was investigated in mung bean. The three phytohormones used were able to overcome to variable extents the adverse effects of stress imposed by NaCl solution.     

Abscisic acid, indole-3-acetic acid and mineral–nutrient changes induced by drought and salinity in longan (Dimocarpus longan Lour.) plants

Longan species (Dimocarpus longan Lour.) exhibit a high agronomic potential in many subtropical regions worldwide; however, little is known about its responses to abiotic stress conditions. Drought and salinity are the most environmental factors inducing negative effects on plant growth and development. In order to elucidate the responses of longan to drought and salinity, seedlings were grown under conditions of drought and salt stresses. Drought was imposed by suspending water supply leading to progressive soil dehydration, and salinity was induced using two concentrations of NaCl, 100 and 150 mM in water solution, for 64 days. Data showed that salt concentrations increased foliar abscisic acid (ABA) and only 150 mM NaCl reduced indole-3-acetic acid (IAA) and increased proline levels. NaCl treatments also increased Na⁺ and Cl^? content in plant organs proportionally to salt concentration. Drought increased leaf ABA but did not change IAA concentrations, and also increased proline synthesis. In addition, drought and salt stresses reduced the photosynthesis performance; however, only drought decreased leaf growth and relative leaf water content. Overall, data indicate that under severe salt stress, high ABA accumulation was accompanied by a reduction of IAA levels; however, drought strongly increased ABA but did not change IAA concentrations. Moreover, drought and high salinity similarly increased (or maintained) ion levels and proline synthesis. Data also suggest that ABA accumulation may mitigate the impact of salt stress through inducing stomatal closure and delaying water loss, but did not mediate the effects of long-term drought conditions probably because leaves reached a strong dehydration and the role of ABA at this stage was not effective to detain leaf injuries.     

外源水杨酸对NaCl胁迫下大豆种子萌发和幼苗生长生理的影响

以大豆种子、幼苗为试验材料,采用砂培的方法,研究了0.2mmol·L-1外源水杨酸(SA)对100mmol·L-1 NaCl胁迫下大豆种子萌发、幼苗形态及生物量、膜脂过氧化和抗氧化酶活性的影响。结果显示:NaCl胁迫下,大豆种子萌发和幼苗生长受到显著抑制,且随着胁迫时间的延长(0~3d),大豆幼苗相对电解质渗漏率、硫代巴比妥酸活性产物(TBARS)含量显著升高,超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)活性均明显降低。外源SA促进NaCl胁迫下大豆种子萌发和根茎生长,增加幼苗生物量积累,降低幼苗叶片相对电解质渗漏率和TBARS含量,增强其叶片SOD、CAT、APX活性。研究表明,NaCl胁迫能显著抑制大豆种子萌发和幼苗生长,而一定浓度的外源SA能有效提高NaCl胁迫下大豆种子活力及幼苗抗氧化酶活性,减轻膜脂过氧化程度,缓解NaCl胁迫所造成的伤害,提高大豆幼苗抗盐胁迫的能力。     

NaCl pretreatment alleviates salt stress by enhancement of antioxidant defense system and osmolyte accumulation in mungbean (Vigna radiata L. Wilczek)

Enhancement of salt (NaCl) tolerance by pretreatment with sublethal dose (50 mM) of NaCl was investigated in V. radiata seedlings. NaCl stress caused drastic effects on roots compared to shoots. Accompanying reductions in length, number of root hairs and branches, roots became stout, brittle and brown in color. Salt stress caused gradual reduction in chlorophyll, carotenoid pigment contents and chlorophyll fluorescence intensity also. Superoxide dismutase and catechol peroxidase activities increased under stress in both roots and leaves. But catalase activity showed an increase in roots and decrease in leaves. In these seedlings, the oxidative stress has been observed under salinity stress and the level of proline, H2O2 and malondialdehyde content were increased. But pretreatment with sublethal dose of NaCl was able to overcome the adverse effects of stress imposed by NaCl to variable extents by increasing growth and photosynthetic pigments of the seedlings, modifying the activities of antioxidant enzymes, reducing malondialdehyde and H2O2 content and increasing accumulation of osmolytes like proline. Thus, mungbean plants can acclimate to lethal level of salinity by pretreatment with sublethal level of NaCl, improving their health and production under saline condition.     

Effect of NaCl on fatty acids, phenolics and antioxidant activity of Nigella sativa organs

The objective of this study was to investigate the effect of salinity on growth, fatty acid composition, phenol content and antioxidant activity of Nigella sativa organs. Plants were grown hydroponically under NaCl stress (0, 20 40 and 60 mM). The results indicated that salinity affected N. sativa growth. The fatty acid composition of the leaves and the roots was investigated for the first time and major fatty acids were linolenic acid (58.1%) in the leaves and linoleic (43.9%) and palmitic (33.3%) acids and in the roots. Total fatty acid (TFA) content of the leaves decreased at 60 mM NaCl while root TFA increased at 20 and 40 mM NaCl. Moreover, the fatty acid composition was affected by NaCl; in leaves, the double bond index (DBI) decreased accompanied by a decrease of the level of linolenic acid which reached 14% at 60 mM NaCl. However, root DBI degree increased at 40 at 60 mM NaCl provoked mainly by the increase of the amount of linoleic acid by 15 and 8%, respectively, and the decrease of the amount of palmitic acid by 20 and 14%, respectively. Salt stress increased total polyphenol and individual phenolic acid contents in shoots. Moreover, the antiradical activity of the shoots (DPPH) increased at 60 mM NaCl. However, in roots, the total polyphenol content and the antiradical activity decreased sharply with increasing NaCl doses. Data reported here revealed the variation of fatty acids and phenolic compound contents in different organs of N. sativa, and the possible role of theses changes in the plant salt response were discussed.     

Response of mycorrhizal and nonmycorrhizal Arachis hypogaea to NaCl and acid stress

 The response of peanut to salt (NaCl) and acid (HCl) stress was studied in association with Glomus caledonium, an arbuscular mycorrhizal (AM) fungus. The plants were exposed to salt stress by irrigation on alternate days with 1% or 5% NaCl solutions, or with 0.1 N HCl to induce acid stress. Plant yield almost tripled in mycorrhizal plants compared with nonmycorrhizal control plants. AM inoculation significantly increased plant yield and biomass at 1% NaCl, while at 5% NaCl AM was less effective in alleviating salt stress. Percentage AM colonization was also lowest at 5% NaCl. AM inoculation was found to promote the establishment of peanut plants under acid stress conditions. Accepted: 2 October 1995     

Sucrose Metabolism in Lupinus albus L. Under Salt Stress

Salt stress (50 and 150 mM NaCl) effects on sucrose metabolism was determined in Lupinus albus L. Sucrose synthase (SS) activity increased under salt stress and sucrose phosphate synthase activity decreased. Acid invertase activity was higher at 50 mM NaCl and decreased to control levels at 150 mM NaCl. Alkaline invertase activity increased with the salt stress. Glucose content decreased with salt stress, sucrose content was almost three times higher in plants treated with 150 mM NaCl and fructose content did not change significantly. The most significant response of lupin plants to NaCl excess is the increase of sucrose content in leaves, which is partially due to SS activity increase under salinity. This revised version was published online in July 2006 with corrections to the Cover Date.     

Comparative evaluation of hydro-, chemo- , and hormonal-priming methods for imparting salt and PEG stress tolerance in Indian mustard (<Emphasis Type="Italic">Brassica</Emphasis><Emphasis Type="Italic">juncea</Emphasis> L.)

The present study was aimed to evaluate the effect of different seed priming methods to enhance the sodium chloride (NaCl) and polyethylene glycol-8000 (PEG-8000) stress tolerance in Indian mustard (Brassica juncea L.). Seeds subjected to different priming treatments such as water (hydro-priming), calcium chloride (CaCl₂) (chemo-priming), and abscisic acid (ABA) (hormonal-priming) showed increased rate of germination as compared to non-primed seeds. The primed and non-primed seeds were grown for 15 days and then the seedlings were independently subjected to iso-osmotic salt (150 mM NaCl) or PEG-8000 (20%) stress. The different biochemical responses were studied 10 days after treatment. Under NaCl and PEG stress, the dry weight and total chlorophyll content were higher in primed sets as compared to non-primed treatment which was also evident by the phenotype of the seedlings. In general, the higher activities of superoxide dismutase and glutathione reductase resulted in lower oxidative damage, in terms of malondialdehyde content, under NaCl and PEG stress in hydro-primed set as compared to non-primed, ABA-, and CaCl₂-primed treatments. Besides, the level of total phenolics and accumulation of osmolytes such as free proline, glycine betaine, and total soluble sugars was also lower in hydro-primed set as compared to other primed and non-primed treatments. The study thus suggests the use of hydro-priming as a simple and cost-effective strategy to alleviate the NaCl and PEG induced stress in B. juncea.     

Investigations on the antioxidative defence responses to NaCl stress in a mangrove, Bruguiera parviflora: Differential regulations of isoforms of some antioxidative enzymes

Two-month-old healthy seedlings of a true mangrove, Bruguiera parviflora, raised from propagules in normal nursery conditions were subjected to varying concentrations of NaCl for 45 d under hydroponic culture conditions to investigate the defence potentials of antioxidative enzymes against NaCl stress imposed oxidative stress. Changes in the activities of the antioxidative enzymes catalase (CAT), ascorbate peroxidase (APX), guaiacol peroxidase (POX), glutathione reductase (GR) and superoxide dismutase (SOD) were assayed in leaves to monitor the temporal regulation. Among the oxidative stress triggered chemicals, the level of H₂O₂ was significantly increased while total ascorbate and total glutathione content decreased. The ratio of reduced to oxidized glutathiones, however, increased due to decreased levels of oxidized glutathione in the leaf tissue. Among the five antioxidative enzymes monitored, the APX, POX, GR and SOD specific activities were significantly enhanced at high concentration (400 mM NaCl), while the catalase activities declined, suggesting both up and downregulations of antioxidative enzymes occurred due to NaCl imposed osmotic and ionic stress. Analysis of the stress induced alterations in the isoforms of CAT, APX, POX, GR and SOD revealed differential regulations of the isoforms of these enzymes. In B. parviflora one isoform of each of Mn-SOD and Cu/Zn-SOD while three isoforms of Fe-SOD were observed by activity staining gel. Of these, only Mn-SOD and Fe-SOD2 content was preferentially elevated by NaCl treatment, whereas isoforms of Cu/Zn-SOD, Fe-SOD1 and Fe-SOD3 remained unchanged. Similarly, out of the six isoforms of POX, the POX-1,-2,-3 and -6 were enhanced due to salt stress but the levels of POX-4 and -5 remained same as in control plants suggesting preferential upregulation of selective POX isoforms. Activity staining gel revealed only one prominent band of APX and this band increased with increased salt concentration. Similarly, two isoforms of GR (GR1 and GR2) were visualized on activity staining gel and both these isoforms increased upon salt stress. In this mangrove four CAT-isoforms were identified, among which the prominent CAT-2 isoform level was maximally reduced again suggesting differential downregulation of CAT isoforms by NaCl stress. The results presented in this communication are the first report on the resolutions of isoforms APX, POX and GR out of five antioxidative enzymes studied in the leaf tissue of a true mangrove. The differential changes in the levels of the isoforms due to NaCl stress may be useful as markers for recognizing salt tolerance in mangroves. Further, detailed analysis of the isoforms of these antioxidative enzymes is required for using the various isoforms as salt stress markers. Our results indicate that the overproduction of H₂O₂ by NaCl treatment functions as a signal of salt stress and causes upregulation of APX, POX, GR and deactivations of CAT in B. parviflora. The concentrations of malondialdehyde, a product of lipid peroxidation and lipoxygenase activity remained unchanged in leaves treated with different concentrations of NaCl, which again suggests that the elevated levels of the antioxidant enzymes protect the plants against the activated oxygen species thus avoiding lipid peroxidation during salt stress.     

盐胁迫对竹柳幼苗生理响应及结构解剖的研究

为了解盐胁迫对竹柳(Salix spp.)幼苗生长的影响,采用土培方法,对NaCl胁迫下半年生竹柳扦插苗的成活率、生理响应和根叶部结构进行研究。结果表明,在0.25%Na Cl胁迫(轻度盐胁迫)下竹柳能正常生长,而在0.5%NaCl(中度、重度盐胁迫)下生长受到抑制,推断竹柳的耐盐阈值是0.5%。随着NaCl浓度的增大,叶片相对含水量、叶绿素a含量、叶绿素总含量和叶绿素a/b均呈下降趋势;但叶绿素b含量、脯氨酸含量和MDA含量呈升高趋势。在轻度盐胁迫下,叶片SOD活性和可溶性蛋白含量均升高,在中、重度盐胁迫下显著下降。从根叶解剖结构来看,叶片、角质层、栅栏组织厚度和根部周皮和直径在轻度盐胁迫时最大,但在中度盐胁迫时叶片栅栏组织细胞长度减小且排列越来越疏松,根部输导组织细胞不正常。这表明竹柳在轻度胁迫时具有一定的耐盐性,但在中高度盐胁迫下生长不良。     

Salt stress effects on growth,pigments, proteins and lipid peroxidation in <Emphasis Type="Italic">Salicornia persica</Emphasis> and <Emphasis Type="Italic">S. europaea</Emphasis>

The effects of NaCl stress on growth, water status, contents of protein, proline, malondialdehyde (MDA), various sugars and photosynthetic pigments were investigated in seedlings of Salicornia persica and S. europaea grown in vitro. Seeds were germinated under NaCl (0, 100, 200, 300, 400, 500 and 600 mM) on Murashige and Skoog medium for 45 d. The shoot growth of both species increased under low NaCl concentration (100 mM) and then decreased with increasing NaCl concentrations. In contrast to S. persica, root length in S. europaea reduced steadily with an increase in salinity. Proline content in S. persica was higher than in S. europaea at most NaCl concentrations. Proline, reducing saccharide, oligosaccharide and soluble saccharide contents increased under salinity in both species. In contrast, contents of proteins and polysaccharides reduced in both species under salt stress. MDA content remained close to control at moderate NaCl concentrations (100 and 200 mM) and increased at higher salinities. MDA content in S. europaea was significantly higher than S. persica at higher salinities. Salt treatments decreased K⁺ and P contents in seedlings of both species. Significant reduction in contents of chlorophylls and carotenoids due to NaCl stress was also observed in seedlings of both species. Some differences appeared between S. persica and S. europaea concerning proteins profile. On the basis of the data obtained, S. persica is more salt-tolerant than S. europaea.     

Influence of Triadimefon on the Metabolism of NaCl Stressed Radish

The effect of triadimefon (TDM) on various biochemical parameters was studied in NaCl stressed radish (Raphanus sativus L.). Stress imposed by 80 mM NaCl decreased the protein content and proline oxidase activity, and increased the proline and glycine betaine contents, and protease, -glutamyl kinase and ATPase activities. The TDM treatment alleviated the stress by increasing protein, and glycine betaine contents, and by decreasing proline accumulation, and proline oxidase and ATPase activities.     

Involvement of kinetin and spermidine in controlling salinity stress in mulberry (Morus alba L. cv. S1)

In mulberry (Morus alba L.) plants NaCl stress imposed through roots by irrigation during growth period decreased the net photosynthetic rate (NPR), physiological water use efficiency (pWUE), which ultimately reflected on the reduction of growth parameters and leaf yield. Foliar spray of kinetin and spermidine (both at 1 mM) on salinized plants reduced the detrimental effects of saline stress. Kinetin and spermidine sprayed plants increased the total chlorophyll, protein content, as well as leaf yield, but reduced the sugar and proline contents as compared to NaCl treated plants. Kinetin was more effective than spermidine in increasing NPR, pWUE and leaf yield both in nonsalinized and salinized condition.     

Accelerated Reactive Oxygen Scavenging System and Membrane Integrity of Two Panicum Species Varying in Salt Tolerance

Plant exhibits various patterns of survival under salinity and their growth and development depend on their capacity to overcome the stress. Present investigation was focused on the response and regulation of the antioxidant defense system and the level of lipid peroxidation in Panicum miliacium and Panicum sumatrense under salt treatments. NaCl stress was imposed for 20 days after sowing of two Panicum species. The changes in the antioxidant enzyme activity like superoxide dismutase, catalase, peroxidase, and ascorbate peroxidase and the rate of lipid peroxidation level in terms of malondialdehyde (MDA) were recorded in both Panicum species. A great correlation exists between the antioxidant enzymes and lipid peroxidation. The defense mechanism activated in Panicum species studied was confirmed by the increased antioxidant enzyme activities under progressive NaCl stress. MDA content remained close to control at moderate NaCl concentrations and increased at higher salinities. Although lipid peroxidation increased in both Panicum species under salt stress the percent of increase was low in P. sumatrense indicating its salt-tolerant nature. Another possible conclusion is that improved tolerance to salt stress may be accomplished by increased capacity of antioxidative system.     

Regulated alterations in redox and energetic status are the key mediators of salinity tolerance in the halophyte <Emphasis Type="Italic">Sesuvium portulacastrum</Emphasis> (L.) L

The present work addresses the importance of antioxidant, redox and energetic parameters in regulating salt-tolerance in Sesuvium portulacastrum. Experiments were conducted on 45 days old plants subjected to 250 and 1,000 mM NaCl stress for 2–8 days. Plants showed no significant change in growth parameters (shoot length, dry weight, and water content) at 250 mM NaCl as compared to control. However, growth of plants was significantly affected at 1,000 mM NaCl. The differential growth behaviour could be attributed to a greater decline in the energetic parameters (in terms of ratios of NADP/NADPH and ATP/ADP) at 1,000 mM NaCl than at 250 mM NaCl. The osmotic stress imposed to plants at 250 mM NaCl was presumably balanced by the accumulation of sodium ions (Na⁺), an energetically favorable process, and did not require an increased synthesis of proline. In contrast, to counter osmotic stress at 1,000 mM NaCl, plants accumulated Na⁺ as well as proline and were, therefore, energetically stressed. Further, the response of enzymatic and molecular antioxidants at 1,000 mM was either close to or even lower than that at 250 mM, which resulted in oxidative damage at 1,000 mM, particularly on longer durations. In conclusion, it is suggested that altered redox and energetic status of the plants could play a key role in mediating the tolerance of Sesuvium under salinity stress.     

Salicylic acid-induced salinity tolerance in corn grown under NaCl stress

Effects of salicylic acid on some physiological and biochemical characteristics of maize ( Zea mays L.) seedlings under NaCl stress were studied. Pre-soaking treatments of NaCl (0, 50, 100 and 200 mM) were given to maize seeds in the presence as well as in the absence of 0.5 mM salicylic acid. Two-week-old maize seedlings exhibited significant decrease in dry weight, root length, shoot length and leaf area on 6 h exposure of 100 and 200 mM NaCl stress. Photosynthetic pigments and NR activity in leaves decreased sharply with increasing stress levels. Both proline content and lipid peroxidation (measured in terms of MDA) levels increased significantly under saline conditions. However, seedlings pretreated with 0.5 mM salicylic acid along with the salinity levels showed enhancement in growth parameters, photosynthetic pigments, NR activity while, free proline and MDA levels decreased. The results showed that salt-induced deleterious effects in maize seedlings were significantly encountered by the pretreatment of salicylic acid. It is concluded that 0.5 mM salicylic acid improves the adaptabilities of maize plants to NaCl stress.     

Caffeic acid decreases salinity-induced root nodule superoxide radical accumulation and limits salinity-induced biomass reduction in soybean

Caffeic acid (3,4-dihydroxycinnamic acid; CA) is a cinnamic acid occurring naturally in a variety of plant species. In this study, the effects of caffeic acid (100 μM caffeic acid) on soybean root nodule superoxide content, cell viability and superoxide dismutase (SOD; EC 1.15.1.1) activity were evaluated in the presence and absence of salinity stress (imposed by application of 70 mM NaCl), along with the effects of CA on growth of soybean in the presence or absence of salinity. Treatment with CA caused a decrease in superoxide content, enhanced cell viability and SOD activity, with changes in SOD activity accounted for by increased activity of two manganese SOD isoforms and one copper/zinc SOD isoform. Furthermore, CA improved soybean growth under salinity but reduced soybean biomass in the absence of salinity. We suggest that CA improves soybean salinity stress tolerance, possibly via signals that regulate accumulation of reactive oxygen species (ROS) during salinity stress.     

In vitro Growth and Leaf Composition of Grapevine Cultivars as Affected by Sodium Chloride

In vitro proliferated shoot culture of six grape genotypes (Vitis vinifera L.) were screened for tolerance to NaCl (0 to 200 mM). The cv. Perlette was found to tolerate 175 mM NaCl followed by cvs. Pusa Seedless and Beauty Seedless 150 mM NaCl. Na, K, Cl, Ca and Mg content increased upto 100 mM NaCl in most of the genotypes. Total sugar and proline content of stem tissue gradually increased under NaCl stress while leaf chlorophyll a+b content declined. Studies suggest that the in vitro screening procedure can be used for ranking the grape genotypes for salinity tolerance.     

Provenance variation in tolerance of Melaleuca cajuputi trees to interactive effects of aluminum and salt

A future rise in sea level will expand areas of salt-affected acid sulfate soil, calling for studies on plant tolerance to combined aluminum (Al) and salt (NaCl) stress. We investigated random amplified polymorphic DNA (RAPD) profiles and tolerance to Al and NaCl alone and in combination in 14 Melaleuca cajuputi Powell provenances. Two-month-old seedlings were grown with or without 10 mM Al and/or 50 mM NaCl at pH 3.8 for 3 months. Plant growth was reduced mostly by combined Al and NaCl stress and then by NaCl and least by Al. Moreover, Al enhanced the effect of NaCl on growth and vice versa. There were significant differences in plant growth among provenances under all treatments; however, positive relationships were found among Al tolerance, NaCl tolerance, and combined Al and NaCl tolerance. Provenance variation in stress tolerance increases with the increasing levels of stress effect. Furthermore, NaCl tolerance tended to have a positive relationship with osmotic potential. Leaf sap K concentration was decreased by NaCl and increased by Al; however, provenances that were more tolerant to NaCl tended to have lower K concentrations. RAPD analysis also revealed genetic variation among provenances. These results suggest that the low tolerance to combined Al and NaCl stress in M. cajuputi is largely due to low tolerance to NaCl and the effect of interaction between Al and NaCl. Provenance variation in stress tolerance was significant and could be partly explained by the variation in genetic material and the ability of plants to reduce ion excess stress in their shoots.