Association of some plant defense enzyme activities with systemic resistance to early leaf blight and leaf spot induced in tomato plants by azoxystrobin and Pseudomonas fluorescens

Abstract

Azoxystrobin at three different concentrations, namely, 31.25, 62.50 and 125 g a.i. ha^?1 mancozeb (1 kg ha^?1) and Pseudomonas fluorescens (10 kg ha^?1) were evaluated for their efficacy in inducing defense enzymes in tomato against Alternaria solani and Septoria lycopersici. The activity of defense enzymes peroxidase (PO), polyphenol oxidase (PPO), phenylalanine ammonia lyase (PAL), β-1, 3 glucanase, chitinase, catalase and defense-inducing chemicals (total phenols) was found to be increased in azoxystrobin and P. fluorescens-treated tomato plants. The activity of these defense enzymes and chemicals was higher in azoxystrobin (125 g a.i. ha^?1) and P. fluorescens-treated tomato plants challenge inoculated with the pathogens compared to other treatments. Increased expression of specific isoforms of PO and PPO was also observed due to ISR induction.     

Induced systemic protection in plants

Plants, like animals, have inducible disease resistance mechanisms. Induction of systemic protection in plants results in non-specific sensitization to fungal, bacterial and viral agents. The resistance mechanism(s), however, are expressed only after infection. Since, amongst other factors, disease outcome is highly dependent on recognition and activation of defense mechanisms, induced protection may facilitate recognition of the pathogen as non-self and/or provide a faster and more intense host defense response.     

Photosynthetic activities of cadmium-treated tomato plants

Tomato plants (Lycopersicum esculentum Mill. cv. Moneymaker) grown on nutrient medium containing cadmium exhibit reduced net photosynthesis and reduced contents of chlorophyll and accessory pigments. In chloroplasts isolated from cadmiumtreated plants photosystem II activity, as measured by 2,6-dichlorophenolindophenol photoreduction, and photosystem II + I activity (H₂O → methyl viologen) were both inhibited to about 60%. When 1,5-diphenylcarbazide was used as artificial electron donor, no significant cadmium effect was observed. Photosystem I activity was not affected by cadmium. The fine structure of chloroplasts in cadmium-treated plants was degenerated, similarly to senescence response. The principal symptom of cadmium action was the occurrence of large plastoglobules and a disorganization of the lamellar structure, mainly grana stacks. Transfer of cadmium-treated plants into a medium with increased manganese level caused grana stacking and restoration of photosystem II activity.     

Characterization of induced resistance in tomato plants

We have characterized, using several types of bioassays, the resistance induced in young tomato plants by feeding of the corn earworm, Helicoverpa zea. Beet armyworm larvae, Spodoptera exigua, and leafminers, Liriomyza trifolii, were used to assay the induced resistance. In whole-plant experiments, damage localized to a single leaflet of fourleaf tomato plants induced a systemic increase in resistance such that beet armyworm larvae confined to previously damaged (induced) plants grew at a rate about half that of larvae raised on control plants and consumed less leaf tissue from induced plants than from control plants. In experiments using excised leaves, beet armyworm larvae suffered increased mortality when reared on leaves from induced plants. The strength of this induced resistance varied spatially relative to the damaged position; moreover, the spatial distribution of induced resistance changed over a three-week period following damage. Other experiments demonstrated that the mechanisms of induced resistance in tomato foliage involves both a decrease in larval preference for and a decrease in the nutritional value of induced foliage. Induction also retarded the oviposition and/or early development of leafminers. Thus, induced resistance has relatively severe effects on the biology of subsequent herbivores. These data should allow us to begin to elucidate cause-effect relationships between induced resistance and induced chemistry in tomato plants.     

Induction of systemic resistance in plants

When a pathogen is perceived by a host plant, a series of defense responses can be activated. One of these are "local" defenses that occur rapidly at the site of pathogen invasion. Another are "systemic" defenses that are induced in uninoculated parts of the plant. Recently, molecular genetic studies have revealed genes that are signaling components of systemic resistance pathways. Cloning of these genes and characterization of the function of their proteins is now providing insights to processes regulating plant defense against pathogens. Evidence that "systemic" defenses are important for resistance is that when the way is blocked in transgenic plants or in mutants, the plant's defense is compromised. When the pathway is stimulated by exogenous compounds or in mutants, the host resistance is strengthened. A detailed understanding of this pathway is important for both practical and theoretical reasons.     

Effect of two systemic fungicides on cellulose decomposing fungi of tomato plants and on some enzymatic activities

Kocide 101 (77% cupric hydroxide) and Ridomil plus (15% metalaxyl and 35% copper oxychloride) were used in the treatment of tomato plants. The two fungicides exerted a depressive effect on the total counts and on the individual cellulose decomposing fungal species associated with the roots and shoots of tomato. When these fungicides were incorporated in the liquid culture medium specified for growth and extracellular enzyme production by some selected fungal species, there was a significant reduction in mycelial growth as well as in amylase, cellulase, lipase and protease production by the fungi tested, particularly at the higher doses (200-400 ppm). Exceptions were observed with lower doses (50 and 100 ppm) especially in case of Aspergillus flavus, Cunninghamella echinulata, Penicillium chrysogenum and Fusarium oxysporum grown for amylase or cellulase production.     

Induced systemic resistance in wounded rice plants

Wounding of one leaf of young rice plants caused a strong and transient accumulation of non-conjugated (–)-JA, followed by induction of a number of pathogenesis-related genes, in the treated leaf. The non-treated leaf of wounded plants that emerged after the treatment was more resistant to challenge infection by the rice blast fungus Magnaporthe grisea (Hebert) Barr. The systemic leaf also showed a transient, but delayed accumulation of jasmonic acid. Unlike the wounded leaf, there was no accumulation of pathogenesis-related mRNAs or proteins in the systemically protected leaf. Local application of jasmonic acid, the putative signal of the wound response, resulted in a similar degree of systemic disease resistance as wounding. The results suggest the operation of systemic, wound-related signalling processes in rice that induce systemic disease resistance.     

茉莉酸介导丛枝菌根真菌诱导番茄抗早疫病的机制

丛枝菌根真菌的定殖可以提高寄主植物的抗病性,但机制并不十分清楚。利用番茄茉莉酸信号转导途径前系统素过表达材料35S::PS、茉莉酸合成突变体spr2、茉莉酸信号识别突变体jai1及其野生型CM 4个不同基因型材料,分别在根系接种丛枝菌根真菌摩西斗管囊霉(Funneliformis mosseae, Fm),待菌根形成后,在叶片外源喷施10 mL 0.5μmol/L茉莉酸甲酯(MeJA)和接种番茄早疫病病原菌(Alternaria solani, As),比较不同基因型抗病防御反应以及对早疫病抗性的差异。结果表明:预先接种菌根真菌的CM和35S::PS番茄,在叶片接种病菌5 d和10 d后,其叶片中过氧化物酶(POD)、多酚氧化酶(PPO)和脂氧合酶(LOX)活性以及丙二烯氧化物环化酶基因(AOC)和茉莉酸信号受体基因(COI1)的转录水平显著高于只接种早疫病菌的处理、只接种菌根菌的处理以及未进行任何处理的健康植株,其早疫病发病率和病情指数也显著降低;外源喷施MeJA可增强预先接种菌根菌的CM和35S::PS番茄植抵抗早疫病的能力。与此同时,对预先接种菌根菌的spr2番茄外源喷施MeJA...     

Autoradiographic and biochemical evidence for the systemic translocation of systemin in tomato plants

The movement of systemin, the 18-amino-acid polypeptide inducer of proteinase inhibitors in tomato (Lycopersicon esculentum L.) plants, was investigated in young tomato plants following the application of [¹⁴C]systemin to wounds on the surface of leaves. Wholeleaf autoradiographic analyses revealed that [¹⁴C]systemin was distributed throughout the wounded leaf within 30 min, and then during the next several hours was transported to the petiole, to the main stem, and to the upper leaves. The movement of [¹⁴C]systemin was similar to the movement of [¹⁴C]sucrose when applied to leaf wounds, except that sucrose was slightly more mobile than systemin. Analyses of the radioactivity in the petiole phloem exudates at intervals over a 5-h period following the application of [¹⁴C]systemin to a wound demonstrated that intact [¹⁴C]systemin was present in the phloem over the entire time, indicating that the polypeptide was either stable for long periods in the phloem or was being continually loaded into the phloem from the source leaf. The translocation pathway of systemin was also investigated at the cellular level, using light microscopy and autoradiography. Within 15 min after application of [³H]systemin to a wound on a terminal leaflet, it was found distributed throughout the wounded leaf and was primarily concentrated in the xylem and phloem tissues within the leaf veins. After 30 min, the radioactivity was found mainly associated with vascular strands of phloem tissue in the petiole and, at 90 min, label was found in the phloem of the main stem. Altogether, these and previous results support a role for systemin as a systemic wound signal in tomato plants.The authors acknowledge the Washington State University Electron Microscope Center and staff for their technical advice and collaboration. We also thank Greg Wichelns for growing our plants and Dr. Steven Doares for providing [³H]systemin. This research was supported in part by the Washington State College of Agriculture and Home Economics Project No. 1791 and National Science Foundation grants IBN 9117795 and IBN 9104542     

Molecular and physiological strategies to increase aluminum resistance in plants

Aluminum (Al) toxicity is a primary limitation to plant growth on acid soils. Root meristems are the first site for toxic Al accumulation, and therefore inhibition of root elongation is the most evident physiological manifestation of Al toxicity. Plants may resist Al toxicity by avoidance (Al exclusion) and/or tolerance mechanisms (detoxification of Al inside the cells). The Al exclusion involves the exudation of organic acid anions from the root apices, whereas tolerance mechanisms comprise internal Al detoxification by organic acid anions and enhanced scavenging of free oxygen radicals. One of the most important advances in understanding the molecular events associated with the Al exclusion mechanism was the identification of the ALMT1 gene (Al-activated malate transporter) in Triticum aestivum root cells, which codes for a plasma membrane anion channel that allows efflux of organic acid anions, such as malate, citrate or oxalate. On the other hand, the scavenging of free radicals is dependent on the expression of genes involved in antioxidant defenses, such as peroxidases (e.g. in Arabidopsis thaliana and Nicotiana tabacum), catalases (e.g. in Capsicum annuum), and the gene WMnSOD1 from T. aestivum. However, other recent findings show that reactive oxygen species (ROS) induced stress may be due to acidic (low pH) conditions rather than to Al stress. In this review, we summarize recent findings regarding molecular and physiological mechanisms of Al toxicity and resistance in higher plants. Advances have been made in understanding some of the underlying strategies that plants use to cope with Al toxicity. Furthermore, we discuss the physiological and molecular responses to Al toxicity, including genes involved in Al resistance that have been identified and characterized in several plant species. The better understanding of these strategies and mechanisms is essential for improving plant performance in acidic, Al-toxic soils.     

Tocopherol content and enzymatic antioxidant activities in chloroplasts from NaCl-stressed tomato plants

Changes in tocopherol, chlorophyll and TBARS levels and the activities of antioxidant enzymes i.e., GSH-Px, GST, and SOD in chloroplasts of tomato plants subjected to moderate (50 mM) and severe (150 mM) NaCl stress were determined. Increase in tocopherol content around the second day under both stresses did not correlate with the chlorophyll degradation while such correlation was observed from the fifth day of severe stress. The activities of GSH-Px and GST as well as TBARS content showed NaCl-induced enhancement which was dose- and time-dependent. However, chloroplastic SOD was rather not involved in the response of tomato plants to NaCl stress. The obtained results suggest that under the moderate stress similarly as in the early phase of severe stress tocopherol functions as a typical antioxidant, while in the late phase of the latter it may be involved in senescence signaling pathway and enables the recovery and recycling of the compounds significant for a plant organism.     

Penicillium oxalicum directed systemic resistance in tomato against Alternaria alternata

Enhanced yield of tomato has ever been an important issue due to its nutritional value and various dietary consumption forms. But, efficient yield increase can never be achieved without using environmentally safe means e.g. innate resistance. In present study, tomato innate antifungal resistance has been boosted up using Penicillium oxalicum, and then various aspects of resistance modulation have been explored in details. Two tomato varieties of differential antifungal resistance (Dinaar and Red Tara) were treated with six P. oxalicum strains which screened the best inducer strain (Pn 5); which remarkably controlled disease incidence (DI) of Alternaria alternata. Inducer was not only responsible for almost two times production of phenolics, alkaloids and terpenoids in Red Tara, but it also non-significantly triggered same biochemicals in Dinaar. Hemicellulose showed only 40 % increase in variety of least antifungal resistance. During quantification assays of peroxidase (POD), phenyl ammonia lyase and polyphenol oxidase, more or less the same doubling trend was recorded in susceptible variety, while only POD had significant enhancement in resistant variety under the influence of fungal inducer. It was also recorded that inducer not only modulated quantity of enzyme (glucanase), but its isozyme package was also altered. Colorimetric quantifications of lignin, cellulose and pectins proved that biotic inducer strengthened the physical structure of plant cells by increasing these contents from 30 to 120 %. The above investigation collectively comes with the recommendation of an efficient and environmentally safe inducer (P. oxalicum); which, can be used to control fungal pathogens.     

Pre-sowing magnetic treatments of tomato seeds increase the growth and yield of plants

The effects of pre-sowing magnetic treatments on growth and yield of tomato (cv Campbell-28) were investigated under field conditions. Tomato seeds were exposed to full-wave rectified sinusoidal non-uniform magnetic fields (MFs) induced by an electromagnet at 100 mT (rms) for 10 min and at 170 mT (rms) for 3 min. Non-treated seeds were considered as controls. Plants were grown in experimental plots (30.2 m(2)) and were cultivated according to standard agricultural practices. During the vegetative and generative growth stages, samples were collected at regular intervals for growth rate analyses, and the resistance of plants to geminivirus and early blight was evaluated. At physiological maturity, the plants were harvested from each plot and the yield and yield parameters were determined. In the vegetative stage, the treatments led to a significant increase in leaf area, leaf dry weight, and specific leaf area (SLA) per plant. Also, the leaf, stem, and root relative growth rates of plants derived from magnetically treated seeds were greater than those shown by the control plants. In the generative stage, leaf area per plant and relative growth rates of fruits from plants from magnetically exposed seeds were greater than those of the control plant fruits. At fruit maturity stage, all magnetic treatments increased significantly (P < .05) the mean fruit weight, the fruit yield per plant, the fruit yield per area, and the equatorial diameter of fruits in comparison with the controls. At the end of the experiment, total dry matter was significantly higher for plants from magnetically treated seeds than that of the controls. A significant delay in the appearance of first symptoms of geminivirus and early blight and a reduced infection rate of early blight were observed in the plants from exposed seeds to MFs. Pre-sowing magnetic treatments would enhance the growth and yield of tomato crop.     

Trichoderma virens 106 inoculation stimulates defence enzyme activities and enhances phenolic levels in tomato plants leading to lowered Rhizoctonia solani infection

Interaction of tomato roots with Trichoderma virens TRS106 provided protection against Rhizoctonia solani-induced disease. In tomato, plants inoculated with R. solani disease symptoms were observed on the roots as brown, necrotic lesions. These symptoms were limited on plants treated with TRS106 and inoculated with R. solani. It was shown that TRS106 did not directly inhibit Rhizoctonia growth in in vitro test. The tested Trichoderma isolate stimulated systemic defence responses in tomato plants, by activating defence enzymes including guaiacol peroxidase (GPX), syringaldazine peroxidase (SPX) and phenylalanine ammonia lyase (PAL). Simultaneously, it enhanced accumulation of phenolics and hydrogen peroxide (H₂O₂) accompanied by decrease in lipid peroxidation in the leaves. HPLC analysis indicated remarkable increases in the concentrations of 22 phenolics in the leaves of Trichoderma-treated tomato, both uninoculated and inoculated with R. solani. Some of the phenolics were present in a free form, the others were accumulated in a bound form as glycosylated conjugates belonging to phenylpropanoids, hydroxybenzoic and cinnamic acid derivatives and flavonoids. Several of the detected phenolics: ferulic and salicylic acids, pyrocatechol and hesperetin were strongly toxic to R. solani in plate tests. The systemic mobilisation of phenolic metabolism might be an element of tomato defence response positively involved in biocontrol of R. solani by TRS106. Based on the results, T. virens TRS106 may have potential to develop a new biofungicide for integrated management of R. solani-induced disease.     

Induced systemic resistance (ISR) in plants: mechanism of action

Plants possess a range of active defense apparatuses that can be actively expressed in response to biotic stresses (pathogens and parasites) of various scales (ranging from microscopic viruses to phytophagous insect). The timing of this defense response is critical and reflects on the difference between coping and succumbing to such biotic challenge of necrotizing pathogens/parasites. If defense mechanisms are triggered by a stimulus prior to infection by a plant pathogen, disease can be reduced. Induced resistance is a state of enhanced defensive capacity developed by a plant when appropriately stimulated. Systemic acquired resistance (SAR) and induced systemic resistance (ISR) are two forms of induced resistance wherein plant defenses are preconditioned by prior infection or treatment that results in resistance against subsequent challenge by a pathogen or parasite. Selected strains of plant growth-promoting rhizobacteria (PGPR) suppress diseases by antagonism between the bacteria and soil-borne pathogens as well as by inducing a systemic resistance in plant against both root and foliar pathogens. Rhizobacteria mediated ISR resembles that of pathogen induced SAR in that both types of induced resistance render uninfected plant parts more resistant towards a broad spectrum of plant pathogens. Several rhizobacteria trigger the salicylic acid (SA)-dependent SAR pathway by producing SA at the root surface whereas other rhizobacteria trigger different signaling pathway independent of SA. The existence of SA-independent ISR pathway has been studied in Arabidopsis thaliana, which is dependent on jasmonic acid (JA) and ethylene signaling. Specific Pseudomonas strains induce systemic resistance in viz., carnation, cucumber, radish, tobacco, and Arabidopsis, as evidenced by an enhanced defensive capacity upon challenge inoculation. Combination of ISR and SAR can increase protection against pathogens that are resisted through both pathways besides extended protection to a broader spectrum of pathogens than ISR/SAR alone. Beside Pseudomonas strains, ISR is conducted by Bacillus spp. wherein published results show that several specific strains of species B. amyloliquifaciens, B. subtilis, B. pasteurii, B. cereus, B. pumilus, B. mycoides, and B.sphaericus elicit significant reduction in the incidence or severity of various diseases on a diversity of hosts.