Salicylic and Jasmonic acids in regulation of the proantioxidant state in wheat leaves infected by <Emphasis Type="Italic">Septoria nodorum</Emphasis> Berk

Influence of mediators of the signal systems of salicylic (SA) and jasmonic (JA) acids and their mixture on reactive oxygen species’ (ROS) (superoxide radical and O₂^·− H₂O₂) generation and activity of oxidoreductases (oxalate oxidase, peroxidase and catalase) in leaves of wheat Triticum aestivum L. infected by Septoria leaf blotch pathogen Septoria nodorum Berk has been studied. Presowing treatment of seeds by SA and JA decreased the development rate of fungus on wheat leaves. SA provided earlier inductive effect on production of O₂^·− and H₂O₂ compared with JA. The protective effect of the salicylic and jasmonic acids against Septoria leaf blotch pathogen was caused by activation of oxalate oxidase, induction of anion and cation peroxidases, and decrease of catalase activity. Ability of compounds to stimulate ROS in the plant tissues can be used as criteria for evaluation of immune-modulating activity of new substances for protection of the plants.     

Effect of jasmonic acid on in vitro explant growth and microtuberization in potato

The shoot fresh mass, root length and root numbers of two potato (Solanum tuberosum L.) cultivars Favorita and Helanwuhua were increased significantly by the application of 0.2 – 2 mg dm⁻³ jasmonic acid (JA) in the Murashige and Skoog medium. However, the growth of potato explants was inhibited by JA at high concentrations (20 – 50 mg dm⁻³). Chlorophyll content in explant leaves decreased with an increase in the concentration of JA. In leaves treated with 0.2 mg dm⁻³ JA acid peroxidase activity increased, while in the leaves treated with more than 2 mg dm⁻³ JA peroxidase activity decreased. Under the dark, the microtuber numbers, fresh mass and percentage of big microtubers of two potato cultivars were not promoted by the application of 0.2 – 50 mg dm⁻³ JA.     

Superoxide anion regulates plant growth and tuber development of potato

A higher concentration of H₂O₂ was detected in the sense transgenic potato plant (SS4) with the lily chCu,ZnSOD sequence, whereas higher levels of O₂⁻ was detected in the antisense transgenic plant (SA1) than the WT plant. The elongation growth in SA1 was significantly inhibited by treatment with diphenyleneiodonium, an inhibitor of O₂⁻ generation, and promoted in the SS4 on treatment with herbicide methyl viologen, a generator of apoplastic O₂⁻. Higher concentrations of GAs were detected during plant growth and the early stage of tuberization in SA1. Complete recovery of the above elongation growth and microtuberization pattern in transgenic plants following treatment of GA₃ or an inhibitor of gibberellin synthesis, paclobutrazol, indicate that these changes were mainly caused by active GA levels. In conclusion, a specific ROS (O₂⁻ ) acts as a signal transducer via GA biosynthetic pathways for the regulation of plant growth and tuber development of potato.     

Accumulation of Four Metals in Tissues of Corchorus olitorius and Possible Mechanisms of Their Tolerance

Corchorus olitorius plants treated by 5 μg cm⁻³ of Cd, Pb, Al or Cu in hydroponic culture accumulated in leaves 190, 150, 350 and 325 μg g⁻¹(d.m.) of these metals, respectively, after 6 d of exposure. Exposure of Corchorus plants to tested metals resulted in a sharp rise in content of amino acids in leaf tissues, however the magnitude of accumulation was different from one metal to another. Presence of sulphur in the growth medium significantly increased uptake of Cd and Pb and cysteine (cyst) was more effective than K₂SO₄. Similarly, addition of salicylic acid (SA) in the growth medium significantly enhanced the ability of Corchorus plants to accumulate all these metals. Growth of Corchorus plants was significantly reduced by treatment with any of the four metals except Cu and added cyst, K₂SO₄ or SA alleviated the growth retarding effect of metals. This revised version was published online in July 2006 with corrections to the Cover Date.     

Exogenous Salicylic Acid Alleviates Growth Inhibition and Oxidative Stress Induced by Hypoxia Stress in <Emphasis Type="Italic">Malus robusta</Emphasis> Rehd

Salicylic acid (SA) as a signal molecule mediates many biotic and environmental stress-induced physiologic responses in plants. In this study we investigated the role of SA in regulating growth and oxidative stress in Malus robusta Rehd under both normoxic and hypoxic conditions. Hypoxia stress inhibited plant growth and dramatically reduced biomass. Addition of SA significantly alleviated the plant growth inhibition. The amounts of superoxide radicals (O₂ ⁻) and hydrogen peroxide (H₂O₂) significantly increased in leaves of the plants exposed to hypoxia stress and resulted in oxidative stress, which was indicated by accumulated concentration of malondialdehyde (MDA) and electrolyte leakage. Addition of SA significantly decreased the level of O₂ ⁻, electrolyte leakage, and lipid peroxidation and enhanced the activities of superoxide dismutase (SOD), peroxidase (POD), and ascorbate peroxidase (APX) under hypoxia stress. As important antioxidants, ascorbate (AsA) and glutathione (GSH) contents in the plant leaves were slightly increased by SA treatment compared to hypoxia stress treatment alone. It was concluded that SA could alleviate the detrimental effects of hypoxia stress on plant growth and of oxidative stress by enhancing the antioxidant defense system in leaves of M. robusta Rehd.     

Cholic acid changes defense response to oxidative stress in soybean induced by <Emphasis Type="Italic">Aspergillus niger</Emphasis>

The oxidative stress and antioxidant systems in soybean leaves and roots infected with plant pathogen Aspergillus niger were studied following treatment with different concentrations of cholic acid. Several oxidative stress parameters were analyzed: production of superoxide (O₂ ^·−) and hydroxyl radicals (^·OH), lipid peroxidation (LP), and superoxide dismutase (SOD; EC 1.15.1.1) activity, as well as the content of reduced glutathione (GSH). Results showed that inoculation with A. niger led to the increase of O₂ ^·− production and GSH quantities in leaves and ^·OH in roots. The highest activity of SOD occured in infected plants treated with cholic acid in concentrations of 40 and 60 mg L⁻¹ which ultimately led to a decrease in O₂ ^·− production. Inoculation with Aspergillus in combination with elevated cholic acid concentrations also increased ^·OH production which is correlated with increased LP. These results may support the idea of using cholic acid as an elicitor to trigger hypersensitive response in plant cells. Use of cholic acid may also actively contribute to soybean plants defense response against pathogen attack.     

Multiple hormone analysis indicates involvement of jasmonate signalling in the early defence of potato to potato virus YNTN

The involvement of plant hormones in the very early response of plants to virus infection was studied in potato plants (Solanum tuberosum L.) infected with potato virus Y^NTN (PVY^NTN). Endogenous plant hormones and compounds mediating a stress response (JA-jasmonic acid, OPDA-12-oxo phytodienoic acid, SA-salicylic acid, IAA-indole-3-acetic acid, ABA-abscisic acid) were simultaneously quantified in susceptible cv. Désirée and resistant cv. Santé, one and three hours after inoculation. Of the hormones analysed, only the contents of endogenous JA and its precursor OPDA changed in a way that could be clearly connected with the early resistant response. In comparison to susceptible cultivar, a much more pronounced increase of JA was detected in virus-inoculated leaves of resistant cultivar at both time points. The same trend of changes was also observed with OPDA. However, there were no significant changes of JA and its precursor in upper intact systemic leaves and roots, at either time point. These findings implicate the jasmonate signalling pathway in a very early local but not systemic resistant defence of potato to PVY^NTN.     

Effect of salicylic acid on the proton translocation activity of plasmalemma of potato tuber cells

Action of salicylic acid (SA) on the activity of membrane bound H⁺-ATPase and passive proton permeability of plasmalemma membrane vesicles (PMV) from parenchyma cells of potato tubers was detected. A correlation between SA action on germination of tubers and activity of plasmalemma H⁺-ATPase was revealed: the application of growth-stimulating concentrations of SA (10⁻¹⁰–10⁻⁸ M) in the system in vitro resulted in activation of plasmalemma H⁺-ATPase, while the use of growth-inhibiting concentrations (10⁻⁴, 10⁻⁵ M) provoked inhibition of the enzyme activity. Addition of jasmonic acid (JA) to the incubation mix resulted in increase of SA effect on the accumulation of H⁺ in PMV.     

The effects of salicylic acid on pigment contents in ultraviolet radiation stressed pepper plants

Pepper (Capsicum annuum L.) plants were sprayed with salicylic acid (SA) and treated with ultraviolet radiation UV-A (320–390 nm), UV-B (312 nm), and UV-C (254 nm) of 6.1, 5.8, and 5.7 W m⁻², respectively. UV significantly reduced contents of chlorophyll (Chl) a and b, and carotenoids (Car). SA treatment moderated Chl and Car reduction in plants treated with UV-B and UV-C. The quantity of antocyanins, flavonoids, rutin, and UV-absorbing compounds in plants that were treated with UV-B, UV-C, and SA were significantly increased. Foliar spray of SA counteracted the UV effects on pepper.     

Characterization of an ecotype of brake-fern, Pteris vittata, for arsenic tolerance and accumulation in plant biomass

An ecotype of brake fern (Pteris vittata) was assessed for arsenic tolerance and accumulation in its biomass under in vivo and in vitro condition; using soil, and agar-gelled Murashige and Skoog (MS) medium supplemented with different concentrations of arsenic. The plants were raised in soil amended with 100–1000 mg arsenic kg⁻¹ soil, and MS medium was supplemented with 10–300 mg arsenic 1⁻¹ medium using Na₂HAsO₄ · 7H₂O. The spores and haploid gametophytic-prothalli were raised in vitro on MS medium supplemented with arsenic. The field plants showed normal growth and biomass formation in arsenic amended soil, and accumulated 1908–4700 mg arsenic kg⁻¹ dry aerial biomass after 10 weeks of growth. Arsenic toxicity was observed above >200 mg arsenic kg⁻¹ soil. The concentrations of arsenic accumulated in the plant biomass were statistically significant (p < 0.05). Normal plants were developed from spores and gametophyte prothalli on the MS media supplemented with 50–200 mg arsenic 1⁻¹ medium. The in vitro raised plants were tolerant to 300 mg arsenic kg⁻¹ of soil and accumulated up to 3232 mg arsenic kg⁻¹ dry aerial biomass that showed better growth performance, biomass generation and arsenic accumulation in comparison to the field plants. The text was submitted by the authors in English.     

Salicylic and jasmonic acid pathways are necessary for defence against Dickeya solani as revealed by a novel method for Blackleg disease screening of in vitro grown potato

Potato is major crop ensuring food security in Europe, and blackleg disease is increasingly causing losses in yield and during storage. Recently, one blackleg pathogen, Dickeya solani has been shown to be spreading in Northern Europe that causes aggressive disease development. Currently, identification of tolerant commercial potato varieties has been unsuccessful; this is confounded by the complicated etiology of the disease and a strong environmental influence on disease development. There is currently a lack of efficient testing systems. Here, we describe a system for quantification of blackleg symptoms on shoots of sterile in vitro potato plants, which saves time and space compared to greenhouse and existing field assays. We found no evidence for differences in infection between the described in vitro‐based screening method and existing greenhouse assays. This system facilitates efficient screening of blackleg disease response of potato plants independent of other microorganisms and variable environmental conditions. We therefore used the in vitro screening method to increase understanding of plant mechanisms involved in blackleg disease development by analysing disease response of hormone‐ related (salicylic and jasmonic acid) transgenic potato plants. We show that both jasmonic (JA) and salicylic (SA) acid pathways regulate tolerance to blackleg disease in potato, a result unlike previous findings in Arabidopsis defence response to necrotrophic bacteria. We confirm this by showing induction of a SA marker, pathogenesis‐related protein 1 (StPR1), and a JA marker, lipoxygenase (StLOX), in Dickeya solani infected in vitro potato plants. We also observed that tubers of transgenic potato plants were more susceptible to soft rot compared to wild type, suggesting a role for SA and JA pathways in general tolerance to Dickeya.     

Production of herbicide-resistant transgenic sweet potato plants through <Emphasis Type="Italic">Agrobacterium tumefaciens</Emphasis> method

Transgenic herbicide-resistant sweet potato plants [Ipomoea batatas (L.) Lam.] were produced through Agrobacterium-mediated transformation system. Embryogenic calli derived from shoot apical meristems were infected with Agrobacterium tumefaciens strain EHA105 harboring the pCAMBIA3301 vector containing the bar gene encoding phosphinothricin N-acetyltransferase (PAT) and the gusA gene encoding β-glucuronidase (GUS). The PPT-resistant calli and plants were selected with 5 and 2.5 mg l⁻¹ PPT, respectively. Soil-grown plants were obtained 28–36 weeks after Agrobacterium-mediated transformation. Genetic transformation of the regenerated plants growing under selection was demonstrated by PCR, and Southern blot analysis revealed that one to three copies of the transgene were integrated into the plant genome of each transgenic plant. Expression of the bar gene in transgenic plants was confirmed by RT-PCR and application of herbicide. Transgenic plants sprayed with Basta containing 900 mg l⁻¹ of glufosinate ammonium remained green and healthy. The transformation frequency was 2.8% determined by herbicide application which was high when compared to our previous biolistic method. In addition, possible problems with multiple copies of transgene were also discussed. We therefore report here a successful and reliable Agrobacterium-mediated transformation of the bar gene conferring herbicide-resistance and this method may be useful for routine transformation and has the potential to develop new varieties of sweet potato with several important genes for value-added traits such as enhanced tolerance to the herbicide Basta.     

Photosynthetic Rate, Growth, and Yield of Mustard Plants Sprayed with 28-Homobrassinolide

Thirty-day-old plants of mustard (Brassica juncea L.) were sprayed with 10⁻¹⁰, 10⁻⁸, or 10⁻⁶ M aqueous solution of 28-homobrassinolide (HBR). The HBR-treated plants were healthier than those treated with water and yielded more. Maximum increase over control was found in 60-d-old, 10⁻⁸ M-HBR-treated plants in fresh and dry mass per plant, carbonic anhydrase (CA, E.C. 4.2.1.1) activity, and net photosynthetic rate (P _N), at harvest in number of pods per plant and seed yield per plant (the respective values were 25, 30, 34, 69, 24, and 29 %). A further increase in the concentration of HBR (10⁻⁶ M) did not make any additional impact on the growth and yield. Increased CA activity and P _N were correlated with growth and seed yield. This revised version was published online in August 2006 with corrections to the Cover Date.     

Salicylic acid mitigates salinity stress by improving antioxidant defence system and enhances vincristine and vinblastine alkaloids production in periwinkle [<Emphasis Type="Italic">Catharanthus roseus</Emphasis> (L.) G. Don]

A pot experiment was conducted to find out whether the foliar spray of salicylic acid (SA) could successfully ameliorate the adverse effects of salinity stress on periwinkle. Thirty-day-old plants were supplied with Control; 0 mM NaCl + 10⁻⁵ M SA (T₁); 50 mM NaCl + 0 SA (T₂); 100 mM NaCl + 0 SA (T₃); 150 mM NaCl + 0 SA (T₄); 50 mM NaCl + 10⁻⁵ M SA (T₅); 100 mM NaCl + 10⁻⁵ M SA (T₆); 150 mM NaCl + 10⁻⁵ M SA (T₇). The plants were sampled 90 days after sowing to assess the effect of SA on stressed and unstressed plants. Salt stress significantly reduced the growth attributes including plant height, leaf-area index, shoot and root fresh weights, shoot and root dry weights. Increasing NaCl concentrations led to a gradual decrease in photosynthetic parameters and activities of nitrate reductase and carbonic anhydrase. Ascorbic acid, total alkaloids and antioxidants enzymes superoxide dismutase, catalase and peroxidase also declined in NaCl-treated plants. The plants, undergoing NaCl stress, exhibited a significant increase in electrolyte leakage and proline content. Foliar application of SA (10⁻⁵ M) reduced the damaging effect of salinity on plant growth and accelerated the restoration of growth processes. It not only improved the growth parameters but also reversed the effects of salinity. Total alkaloid content was improved by SA application both in unstressed and stressed plants. The highest level of total alkaloid content recorded in leaves of SA-treated stressed plants was 11.1%. Foliar spray of SA overcame the adverse effect of salinity by improving the content of vincristine (14.0%) and vinblastine (14.6%) in plants treated with 100 M NaCl.     

Effects of Exogenously Applied Jasmonates on Growth and Intracellular pH in Maize Coleoptile Segments

The effects of jasmonic acid (JA) on elongation growth of coleoptile segments from etiolated maize (Zea mays L.) were investigated in the presence and absence of auxin. When supplied alone, at physiological concentrations (10⁻⁹, 10⁻⁸, and 10⁻⁵ m), JA (or methyl-JA) inhibited growth. JA at a similar range of concentrations also inhibited auxin-induced elongation growth. To determine whether this effect on growth depended on endogenous abscisic acid (ABA), we grew maize coleoptiles in the presence of norflurazon (an inhibitor of carotenoid biosynthesis) that results in reduced endogenous ABA levels. Growth of etiolated coleoptile segments from these plants was inhibited by JA (or methyl-JA) in both the absence and presence of auxin. Previously, we have observed a correlation between elongation growth and cytosolic pH (pH_i), in which auxin lowers pH_i, and growth inhibitors such as ABA raise pH_i. We examined the effect of low concentrations of methyl-JA on pH_i with dual emission dye, carboxy seminaphthorhodafluor-1, and confocal microscopy. To confirm these studies, we also used in vivo ³¹P NMR spectrometry to ascertain the changes in pH_i after addition of jasmonate to maize coleoptiles. Coleoptiles grown in either the absence or presence of norflurazon responded to methyl-JA or JA by increases in pH_i of approximately 0.2 pH unit. This response occurs over a period of 15–20 min and appears to be independent of endogenous ABA. This alkalization induced by JA is likely to form a permissive environment for JA signal transduction pathway(s). Received February 5, 1999; accepted August 25, 1999     

Jasmonic acid and salicylic acid play minor roles in stomatal regulation by CO2, abscisic acid,darkness, vapor pressure deficit and ozone

Jasmonic acid (JA) and salicylic acid (SA) regulate stomatal closure, preventing pathogen invasion into plants. However, to what extent abscisic acid (ABA), SA and JA interact, and what the roles of SA and JA are in stomatal responses to environmental cues, remains unclear. Here, by using intact plant gas-exchange measurements in JA and SA single and double mutants, we show that stomatal responsiveness to CO₂, light intensity, ABA, high vapor pressure deficit and ozone either did not or, for some stimuli only, very slightly depended upon JA and SA biosynthesis and signaling mutants, including dde2, sid2, coi1, jai1, myc2 and npr1 alleles. Although the stomata in the mutants studied clearly responded to ABA, CO₂, light and ozone, ABA-triggered stomatal closure in npr1-1 was slightly accelerated compared with the wild type. Stomatal reopening after ozone pulses was quicker in the coi1-16 mutant than in the wild type. In intact Arabidopsis plants, spraying with methyl-JA led to only a modest reduction in stomatal conductance 80 min after treatment, whereas ABA and CO₂ induced pronounced stomatal closure within minutes. We could not document a reduction of stomatal conductance after spraying with SA. Coronatine-induced stomatal opening was initiated slowly after 1.5–2.0 h, and reached a maximum by 3 h after spraying intact plants. Our results suggest that ABA, CO₂ and light are major regulators of rapid guard cell signaling, whereas JA and SA could play only minor roles in the whole-plant stomatal response to environmental cues in Arabidopsis and Solanum lycopersicum (tomato).