Immunogold localization of beta-1,3-glucanases in two plants infected by vascular wilt fungi.

An antiserum raised against a purified tobacco beta-1,3-glucanase (PR-N) was used to study the subcellular localization of enzyme in fungus-infected plant tissues by means of post-embedding immunogold labeling. In susceptible tomato plants, the enzyme accumulation was found to occur as a result of successful tissue colonization, whereas it appeared to be an early event associated with limited spread of the fungus in resistant tissues. Although marked differences between susceptible and resistant tomato cultivars were observed in the rate of production of beta-1,3-glucanase, the pattern of enzyme distribution was similar. The enzyme was found to accumulate predominantly in host cell walls and secondary thickenings of xylem vessels. By contrast, a very low amount of enzyme was associated with compound middle lamellae. The occurrence of beta-1,3-glucanase at the cell surface of invading fungi was an indication of their possible antifungal activity. A low enzyme concentration was detected in vacuoles of both healthy and infected tissues. In infected eggplant tissue, the pattern of beta-1,3-glucanase distribution was similar to that observed with tomato. Whether these hydrolases accumulate first in vacuoles and are subsequently conveyed toward the outside to participate in fungal wall lysis remains to be determined.     

Changes in Phenolic Acid Content During Alternaria alternata Infection in Tomato Fruit

Phenolic acids are components of the plant defence system; however few studies had been done with fruits. The objective of this experiment was to elucidate the changes in phenolic acid content in tomato fruits in response to pathogenic attack. Tomato fruit ‘Pinto’ was inoculated with spores of Alternaria alternata and stored for 10 days at 25°C and 90–92% of relative humidity. Sampling of epicarp and mesocarp tissues of control and infected tomato fruits were done every 2 days. Phenolic acids were extracted from each tissue and identified by HPLC coupled to a mass spectrometer. Quantification was done based in standard curves. In vitro evaluation of the phenolic acid effect either one‐by‐one and in a mixture of phenolic acids (MPA) over A. alternata spore germination was done with 25, 50, 100, 200 and 500 mm . It was found caffeic, chlorogenic and vanillic acids in both epicarp and mesocarp tomato tissues before the beginning of the experiment. Infected tomato fruits showed significantly higher concentrations of vanillic acid in the epicarp, only. Chlorogenic acid and the MPA inhibit spore germination by 30%, whereas caffeic acid and vanillic acid inhibit 16% at 500 mm . It was concluded that chlorogenic, vanillic and caffeic acids are phytoanticipins in tomato fruit. The concentration of vanillic acid also increases in tomato fruit epicarp as part of the defence system during a pathogenic attack.     

Production of oxalic acid by Penicillium oxalicum in culture and in infected yam tissue and interaction with macerating enzyme

Penicillium oxalicum produced the greatest amount of oxalic acid in liquid culture and in infected yam tissue within 2 days of inoculation, whereas production of cell wall-degrading enzymes reached a peak eight days after inoculation. Coupled with the production of oxalic acid was the lowering of the culture pH and of infected yam tissue. Oxalic acid was not produced simultaneously with endo-polygalacturonase (the macerating enzyme), but it acted synergistically with the enzyme by sequestering calcium present in the middle lamella of yam cells and conditioning the cell wall for hydrolysis.     

Effects of Pyrenochaeta lycopersici infection on nutrient uptake by tomato plants

The growth of young tomato plants in nutrient solution or in soil and infected with Pyrenochaeta lycopersici Schneider & Gerlach, the cause of tomato brown root rot, was decreased relative to that of uninfected plants. The roots of plants grown in nutrient solution and infected with a mycelial mat of the pathogen contained lower concentrations of potassium and higher concentrations of calcium than roots of uninfected plants. These changes occurred largely in the visibly affected tissue, as opposed to the root system as a whole. The concentrations of magnesium, total nitrogen and phosphorus in the roots of infected plants were not significantly different from those of control plants. Magnesium, nitrogen and phosphorus concentrations in the tops of infected plants were also not significantly different from those of healthy plants, but no consistent changes were found in the concentrations of calcium and potassium. Young tomato plants grown in soil infested with P. lycopersici contained lower concentrations of phosphorus and potassium in the tops than plants grown in sterilized soil. It was not possible to separate intact damaged root systems of infected plants from soil. The changes in composition found in infected plants are discussed in relation to possible methods of manipulating the nutrition of the plant to offset the effects of the disease on crop yield.     

A survey of mixed Begomovirus infection in solanaceae and fabaceae at different altitudes in East Java,Indonesia

A multiplex primer set was developed to detect four Begomoviruses in East Java, Indonesia, i.e. Tomato leaf curl New Delhi virus (ToLCNDV), Tomato yellow leaf curl Kanchanaburi virus (TYLCKaV), Pepper yellow leaf curl Indonesia virus (PepYLCIV) and Mungbean yellow mosaic India virus (MYMIV). Survey at different altitudes found that begomoviruses infecting pepper, tomato and long bean were more variable, while in eggplant and string bean were more uniform. As a single virus, TYLCKaV infected eggplant, and sometimes tomato and pepper; PepYLCIV infected pepper, tomato and long bean; ToLCNDV only infected long bean and tomato at low frequency; and MYMIV infected beans. Mixed infection occurred more frequently in the low altitude areas. Subsequent examination indicated that Cucumber mosaic virus (CMV) and potyviruses were also responsible for diseased fabaceous. Our data suggest a relationship between altitudes and virus species occurrence. However, which viral species infects a crop is mainly influenced by the crop rather than by altitude.     

An untargeted global metabolomic analysis reveals the biochemical changes underlying basal resistance and priming in Solanum lycopersicum,and identifies 1‐methyltryptophan as a metabolite involved in plant responses to Botrytis cinerea and Pseudomonas syringae

In this study, we have used untargeted global metabolomic analysis to determine and compare the chemical nature of the metabolites altered during the infection of tomato plants (cv. Ailsa Craig) with Botrytis cinerea (Bot) or Pseudomonas syringae pv. tomato DC3000 (Pst), pathogens that have different invasion mechanisms and lifestyles. We also obtained the metabolome of tomato plants primed using the natural resistance inducer hexanoic acid and then infected with these pathogens. By contrasting the metabolomic profiles of infected, primed, and primed + infected plants, we determined not only the processes or components related directly to plant defense responses, but also inferred the metabolic mechanisms by which pathogen resistance is primed. The data show that basal resistance and hexanoic acid‐induced resistance to Bot and Pst are associated with a marked metabolic reprogramming. This includes significant changes in amino acids, sugars and free fatty acids, and in primary and secondary metabolism. Comparison of the metabolic profiles of the infections indicated clear differences, reflecting the fact that the plant's chemical responses are highly adapted to specific attackers. The data also indicate involvement of signaling molecules, including pipecolic and azelaic acids, in response to Pst and, interestingly, to Bot. The compound 1‐methyltryptophan was shown to be associated with the tomato–Pst and tomato–Bot interactions as well as with hexanoic acid‐induced resistance. Root application of this Trp‐derived metabolite also demonstrated its ability to protect tomato plants against both pathogens.     

Induced defence responses and protective effects on tomato against Xanthomonas vesicatoria by an aqueous extract from Solanum lycocarpum infected with Crinipellis perniciosa

This study investigated the induced defence responses and protective effects on susceptible tomato (Lycopersicon esculentum Mill.) against Xanthomonas vesicatoria (Doidge) by a heat-treated aqueous extract (VLA) from dry necrotic tissue of ‘Lobeira’ (Solanum lycocarpum St. Hil.) branches infected with the fungus Crinipellis perniciosa (Stahel) compared with acibenzolar-S-methyl (ASM), a commercial inducer of resistance. Plantlets were sprayed with VLA and ASM and challenged 4 days later with a virulent strain of X. vesicatoria, under greenhouse conditions. The disease severity, fresh weight of shoots, the activities of phenol peroxidase (POX), polyphenol oxidase (PPO), chitinase (CHI), phenylalanine ammonia-lyase (PAL), lignin deposition, and soluble phenolic contents were evaluated in the leaf tissues. Reduction of the bacterial spot severity was observed in plantlets treated with VLA which conferred 63% of the ASM protection. This protective effect and lesion reduction promoted by VLA were probably associated particularly with POX and PAL activities, lignin deposition on leaf tissues and, to a less extent, CHI activity.     

Studies on plant growth-regulating substances LV. The Plant growth-promoting properties of some β-aryl-propionic acids

Severe diseases of pepper (Capsicum annuum), tomato (Lycopersicon esculentum), eggplant (Solanum melongena) and tomato eggplant (Solanum integrifolium) in West Africa were induced by pepper veinal mottle virus (PVMV). Five selected virus isolates were serologically similar and readily transmissible by aphids in the non-persistent manner, but they differed in host range and/or symptoms induced in some susceptible species. One isolate from eggplant failed to infect pepper, Chenopodium quinoa and C. amaranticolor, and induced only local infections in tomato. An isolate from tomato failed to infect eggplant, and an isolate from tomato eggplant induced severe stunting in Physalis floridana. The type strain, like the isolate from tomato, failed to infect Nicotiana tabacum systemically, but each caused severe systemic leaf and stem necrosis in tomato. None of the isolates infected S. melongena cv. Long Purple, suggesting that PVMV might be controlled in this and perhaps other crop species by the use of immune or tolerant cultivars. All five isolates were serologically related to potato virus Y and some to six of 12 other potyviruses.     

Olfactory responses of Neoseiulus cucumeris (Acari: Phytoseiidae) to odors of host plants and Frankliniella occidentalis (Thysanoptera: Thripidae)�Cplant complexes

We studied the olfactory responses of Neoseiulus cucumeris (Acari: Phytoseiidae) to odors from eggplant (Solanum melongena L.), pepper (Capsicum aunuum), and tomato (Solanum lycopersicum), three host plants, and their complexes with different treatments (undamaged, mechanically damaged, infested, pre-infested) in an olfactometer. The results showed that N. cucumeris preferred the odors of undamaged eggplant, pepper, and tomato to clean air. N. cucumeris preferred the volatiles from eggplant infested with Frankliniella occidentalis (Thysanoptera: Thripidae) over undamaged, mechanically damaged, and pre-infested with F. occidentalis eggplant volatiles. The same results were observed with pepper and tomato. N. cucumeris adults preferred the odors of undamaged eggplant and tomato to undamaged pepper. Similarly, same results were observed with mechanically damaged eggplant, pepper, tomato, and same plants pre-infested with F. occidentalis. Compared with odors of tomato infested with F. occidentalis, N. cucumeris adults significantly responded to odors of eggplant and pepper infested with F. occidentalis. There was no significant difference of N. cucumeris in making choice between eggplant infested with F. occidentalis and pepper infested with F. occidentalis.     

Phenolic compounds in apple leaves after infection with apple scab

Leaves of the scab-susceptible apple (Malus domestica) cultivar Golden Delicious were harvested from May to August 2008 and 2009. Some leaves were healthy and some infected with fungus Venturia inaequalis. The phenolic compounds were analysed in healthy leaves, infected leaves and in the scab spot tissue. In comparison to healthy leaves, the infected leaves showed higher contents of hydroxycinnamic acid, flavanols and phloridzin, while lower contents on procyanidins, quercetins and phloretin. The total amount of phenolic compounds in the infected tissue was 10 to 20 % higher than in the healthy leaves. Accumulation of phenolic compounds is a post-infection response, and probably their further transformation is a prerequisite for plant resistance.     

Stimulation of Fruit Ethylene Production by Wounding and by Botrytis cinerea and Geotrichum candidum Infection in Normal and Non-Ripening Tomatoes

Inoculations with both Botrytis cinerea and Geotrichum candidum stimulated ethylene evolution in the pre-climacteric normal tomato fruit and the non-ripening nor mutant which did not show any rise in ethylene when uninfected. In the post-climacteric normal fruits, new peaks in ethylene production were formed. The rise in ethylene evolution in all types of infected fruits has already been detected during the incubation period of the disease. Ethylene peaks were detected earlier and were higher in fruits infected with B. cinerea than with G. candidum, coinciding with the faster rate of growth of the former. Mechanical wounding also stimulated ethylene synthesis by the non-ripening fruits, production being directly proportional to wound dimension. Considerably higher rates of ethylene were recorded for infected fruits than for mechanically-injured fruits in which wound dimensions were similar to those of lesion development. Applying aminoxyacetic acid at the site of inoculation inhibited ethylene production by 55–60 % in the normal fruits and by about 80 % in the nor mutant fruits. A similar pathway of ethylene synthesis was suggested for normally ripening tomato fruit and non-ripening infected tissues.     

Bacterial community structure associated with white band disease in the elkhorn coral Acropora palmata determined using culture-independent 16S rRNA techniques

Culture-independent molecular (16S ribosomal RNA) techniques showed distinct differences in bacterial communities associated with white band disease (WBD) Type I and healthy elkhorn coral Acropora palmata. Differences were apparent at all levels, with a greater diversity present in tissues of diseased colonies. The bacterial community associated with remote, non-diseased coral was distinct from the apparently healthy tissues of infected corals several cm from the disease lesion. This demonstrates a whole-organism effect from what appears to be a localised disease lesion, an effect that has also been recently demonstrated in white plague-like disease in star coral Montastraea annularis. The pattern of bacterial community structure changes was similar to that recently demonstrated for white plague-like disease and black band disease. Some of the changes are likely to be explained by the colonisation of dead and degrading tissues by a micro-heterotroph community adapted to the decomposition of coral tissues. However, specific ribosomal types that are absent from healthy tissues appear consistently in all samples of each of the diseases. These ribotypes are closely related members of a group of alpha-proteobacteria that cause disease, notably juvenile oyster disease, in other marine organisms. It is clearly important that members of this group are isolated for challenge experiments to determine their role in the diseases.     

ACC Synthesis as the Activated Step Responsible for the Rise of Ethylene Production Accompanying Citrus Exocortis Viroid Infection in Tomato Plants

Ethylene production was stimulated during the period when systemic symptoms appeared in tomato plants infected with citrus exocortis viroid (CEV). Neither methionine nor S-adenosylmethionine increased ethylene production in leaf discs. In contrast, 1-aminocyclopropane-l-carboxylic acid (ACC) stimulated ethylene production notably. Whether viroid infection acted upon ACC production, its conversion to ethylene, or both, was studied by determining the time course of the concentration of ACC and its in vivo production and conversion rates. During early symptoms, ACC synthesis increased and then remained steady during the development of symptoms, but no difference in the capacity of conversion of ACC to ethylene between healthy and CEV-infected tissues was observed. This indicates that ethylene production in tomato leaves showing systemic symptoms to CEV is activated at the level of ACC production.     

Life table and biological characteristics of Macrolophus pygmaeus when feeding on Myzus persicae and Trialeurodes vaporariorum

The life table and biological characteristics of the predatory bug Macrolophus pygmaeus Rambur (Hemiptera: Miridae) were studied when the bugs were fed with Myzus persicae (Sulzer) (Homoptera: Aphididae) feeding on eggplant and with Trialeurodes vaporariorum Westwood (Homoptera: Aleyrodidae) feeding on tomato plants. The tests were done at five temperatures between 15 and 30 °C, using a L16:D8 photoperiod and 65 ± 5% r.h. Most eggs (range 85 to 90%) hatched at 15 and 20 °C. Incubation period was shortest at 27.5 °C (8.45 and 8.38 days on eggplant and tomato, respectively). Preoviposition was also shortest at 27.5 °C (5.10 and 4.75 days on eggplant and tomato, respectively) whereas fecundity was highest at 20 °C (213.90 and 228.25 eggs on eggplant and tomato, respectively). Maximum longevity of females was at 15 °C (122.40 and 129.35 days on eggplant and tomato, respectively). Mean generation time was longest at 15 °C on both host plants (122.75 and 124.64 days, respectively). The intrinsic rate of increase of M. pygmaeus was highest at 27.5 °C with similar values on eggplant (0.0981 day^–1) and tomato (0.1040 day^–1). Doubling time was shortest at 27.5 °C (7.06 and 6.67 days on eggplant and tomato, respectively) and, also, finite rate of increase was highest at 27.5 °C (1.1031 and 1.1096 on eggplant and tomato, respectively). The results show that the predator M. pygmaeus develops well on the aphid M. persicae or on the whitefly T. vaporariorum, both of which are important pests of vegetable crops. This predator is also well adapted to the temperatures that occur both in greenhouses and in the open field in the Mediterranean region. Compared to other natural enemies of whiteflies, such as Encarsia formosa Gahan (Hymenoptera: Aphelinidae), Macrolophus pygmaeus can increase at relatively low temperatures.     

Coral disease physiology: the impact of Acroporid white syndrome on <Emphasis Type="Italic">Symbiodinium</Emphasis>

Acroporid white syndrome, a disease-like syndrome from the Great Barrier Reef, results from degenerative host tissue at lesion borders. Tissue preceding lesion borders appears visually healthy, but it is currently unclear whether the endosymbiotic zooxanthellae (Symbiodinium) are physiologically impacted. Compared to healthy colonies, this study found no significant differences in symbiont density, mitotic index or chlorophyll a content in tissue bordering (0 cm), and 8 cm away from white syndrome lesions. Using chlorophyll a fluorescence techniques, the border tissue did not appear to be photosynthetically compromised, and Symbiodinium extracted from this area were photosynthetically competent. Transmission electron microscopy revealed extensive degeneration of host tissues surrounding symbionts in affected areas, however, Symbiodinium cells were structurally intact with no sign of in situ degradation. Collectively, these results suggest that Symbiodinium at white syndrome lesion borders exist in a dynamic intra-cellular state during active host tissue loss, yet remain physiologically uncompromised.     

The identification of mild and severe strains of tobacco mosaic virus in doubly inoculated tomato plants

Using specific antisera, it was possible to identify mild (MII-16) and the 0 and 1 strains of tobacco mosaic virus (TMV) in tomato plants infected with the mild strain following challenge inoculation with the wild strains. In addition, with single lesion isolates of the MII-16 and o strains it has been shown that both strains can be assayed simultaneously in mixed isolates on Nicotiana glutinosa based on differences in local lesion size. The accuracy of this differential local lesion assay was c. 90% as determined by back inoculation of single lesion isolates into susceptible tomato cultivars.     

Occurrence of Peroxidases in Sweet Potato Infected by the Black Rot Fungus

Changes in the peroxidase activity and the patterns in sweet potato tissue infected by Ceratocystis fimbriata were investigated, by the method of starch-gel electrophoresis, DEAE-cellulose column chromatography and immunochemical analysis, compared with that in cut and healthy tissue. Time-course analysis of the increment of the total peroxidase activity also showed noticeable differences between diseased and cut tissues. The peroxidases in diseased and cut tissues were composed of four major and several minor components. Though electrophoretic analysis did not show so striking differences in the patterns of peroxidases between diseased and cut tissues, there were distinct differences in the ratio of activities of major peroxidase components between both tissues. Immunochemical works indicated that peroxidase A which showed the most prominent increase in diseased tissue was found to be formed in cut tissue though the amounts were appreciably small. The activity of peroxidase C in diseased tissue was not so high as seen in cut tissue.     

Functional response of Nesidiocoris tenuis (Hemiptera: Miridae) to Trialeurodes vaporariorum (Hemiptera: Aleyrodidae): effect of different host plants

The influence of three host plants, namely cucumber, tomato and eggplant, on functional response of male, virgin and mated female predatory bug Nesidiocoris tenuis was investigated on different densities of Trialeurodes vaporariorum nymphs. The 24-h experiment conducted at laboratory conditions revealed that N. tenuis exhibited a type II functional response to T. vaporariorum on host plants. There were no significant differences between attack rates, as well as handling times estimated for each adult stage of the predator between host plants. However, on each host plant, the handling time estimated for the mated female in comparison with two other adult stages had lower values (0.7952, 0.6827 and 0.8884?h^?1 on cucumber, tomato and eggplant, respectively). Handling time estimated for the mated female on cucumber was significantly lower than that estimated for the male predator. The highest maximum handling rate (T/T_h) was estimated for the mated female followed by the virgin female and male on all host plants. For three adult stages of the predator, the highest value of this parameter was determined on tomato followed by cucumber and eggplant. Unlike virgin and mated females, the host plant significantly affected prey consumption by the male. Prey consumed by mated females was higher than those obtained for two other adult stages of the predator on each host plant. The difference in trichome density between three host plants may be responsible for the obtained results. These results revealed that N. tenuis is more effective in the biological control of T. vaporariorum on tomato in comparison with cucumber and eggplant.     

A Comparison of Pectinases from Ditylenchus dipsaci and Allium cepa Callus Tissue

Ground and whole Ditylenchus dipsaci maintained on onion callus contain no culturable micro-organisms when tested with five check media. Healthy onion callus does not produce pectolytic enzymes. Pectolytic enzymes are present in infected callus. These enzymes are, however, associated with resident nematodes and not host tissues. These results suggest that D. dipsaci is the actual source of the endo-polygalacturonase and endo-pectinmethyltrans-eliminase extracted from them.