Ancient origin of pathogen recognition specificity conferred by the tomato disease resistance gene Pto

We have investigated the origin of the Pto disease resistance (R) gene that was previously identified in the wild tomato species Lycopersicon pimpinellifolium and isolated by map-based cloning. Pto encodes a serine-threonine protein kinase that specifically recognizes strains of Pseudomonas syringae pv. tomato (Pst) that express the avirulence gene avrPto. We examined an accession of the distantly related wild species Lycopersicon hirsutum var. glabratum that exhibits avrPto-specific resistance to Pst. The Pst resistance of L. hirsutum was introgressed into a susceptible Lycopersicon esculentum background to create the near-isogenic line 96T133-3. Resistance to Pst(avrPto) in 96T133-3 was inherited as a single dominant locus and cosegregated with a restriction fragment length polymorphism detected by the Pto gene. This observation suggested that a member of the Pto gene family confers Pst(avrPto) resistance in this L. hirsutum line. Here we report the cloning and characterization of four members of the Pto family from 96T133-3. One gene (LhirPto) is 97% identical to Pto and encodes a catalytically active protein kinase that elicits a hypersensitive response when coexpressed with avrPto in leaves of Nicotiana benthamiana. In common with the Pto kinase, the LhirPto protein physically interacts with AvrPto and downstream members of the Pto signaling pathway. Our studies indicate that R genes of the protein kinase class may not evolve rapidly in response to pathogen pressure and rather that their ability to recognize specific Avr proteins can be highly conserved.

Brassinosteroids interact negatively with jasmonates in the formation of anti-herbivory traits in tomato

Given the susceptibility of tomato plants to pests, the aim of the present study was to understand how hormones are involved in the formation of tomato natural defences against insect herbivory. Tomato hormone mutants, previously introgressed into the same genetic background of reference, were screened for alterations in trichome densities and allelochemical content. Ethylene, gibberellin, and auxin mutants indirectly showed alteration in trichome density, through effects on epidermal cell area. However, brassinosteroids (BRs) and jasmonates (JAs) directly affected trichome density and allelochemical content, and in an opposite fashion. The BR-deficient mutant dpy showed enhanced pubescence, zingiberene biosynthesis, and proteinase inhibitor expression; the opposite was observed for the JA-insensitive jai1-1 mutant. The dpyxjai1-1 double mutant showed that jai1-1 is epistatic to dpy, indicating that BR acts upstream of the JA signalling pathway. Herbivory tests with the poliphagous insect Spodoptera frugiperda and the tomato pest Tuta absoluta clearly confirmed the importance of the JA-BR interaction in defence against herbivory. The study underscores the importance of hormonal interactions on relevant agricultural traits and raises a novel biological mechanism in tomato that may differ from the BR and JA interaction already suggested for Arabidopsis.

Acquired tolerance of tomato (Lycopersicon esculentum cv. Micro-Tom) plants to cadmium-induced stress

The effects of varying concentrations of cadmium (Cd) on the development of Lycopersicon esculentum cv. Micro-Tom (MT) plants were investigated after 40 days (vegetative growth) and 95 days (fruit production), corresponding to 20 days and 75 days of exposure to CdCl(2), respectively. Inhibition of growth was clearly observed in the leaves after 20 days and was greater after 75 days of growth in 1 mM CdCl(2), whereas the fruits exhibited reduced growth following the exposure to a concentration as low as 0.1 mM CdCl(2). Cd was shown to accumulate in the roots after 75 days of growth but was mainly translocated to the upper parts of the plants accumulating to high concentrations in the fruits. Lipid peroxidation was more pronounced in the roots even at 0.05 mM CdCl(2) after 75 days, whereas in leaves, there was a major increase after 20 days of exposure to 1 mM CdCl(2), but the fruit only exhibited a slight significant increase in lipid peroxidation in plants subjected to 1 mM CdCl(2) when compared with the control. Oxidative stress was also investigated by the analysis of four key antioxidant enzymes, which exhibited changes in response to the increasing concentrations of Cd tested. Catalase (EC 1.11.1.6) activity was shown to increase after 75 days of Cd treatment, but the major increases were observed at 0.1 and 0.2 mM CdCl(2), whereas guaiacol peroxidase (EC 1.11.1.7) did not vary significantly from the control in leaves and roots apart from specific changes at 0.5 and 1 mM CdCl(2). The other two enzymes tested, glutathione reductase (EC 1.6.4.2) and superoxide dismutase (SOD, EC 1.15.1.1), did not exhibit any significant changes in activity, apart from a slight decrease in SOD activity at concentrations above 0.2 mM CdCl(2). However, the most striking results were obtained when an extra treatment was used in which a set of plants was subjected to a stepwise increase in CdCl(2) from 0.05 to 1 mM, leading to tolerance of the Cd applied even at the final highest concentration of 1 mM. This apparent adaptation to the toxic effect of Cd was confirmed by biomass values being similar to the control, indicating a tolerance to Cd acquired by the MT plants.

Effect of peel and seed removal on the nutritional value and antioxidant activity of tomato (Lycopersicon esculentum L.) fruits

The effect of peel and seed removal, two commonly practiced procedures either at home or by the processing industry, on the physicochemical properties, bioactive compounds contents and antioxidant capacity of tomato fruits of four typical Portuguese cultivars (cereja, chucha, rama and redondo) were appraised. Both procedures caused significant nutritional and antioxidant activity losses in fruits of every cultivar. In general, peeling was more detrimental, since it caused a higher decrease in lycopene, bcarotene, ascorbic acid and phenolics contents (averages of 71%, 50%, 14%, and 32%, respectively) and significantly lowered the antioxidant capacity of the fruits (8% and 10%, using DPPH. and b-carotene linoleate model assays, correspondingly). Although seeds removal favored the increase of both color and sweetness, some bioactive compounds (11% of carotenoids and 24% of phenolics) as well as antioxidant capacity (5%) were loss. The studied cultivars were differently influenced by these procedures. The fruits most affected by peeling were those from redondo cultivar (-66% lycopene, -44% b-carotene, -26% ascorbic acid and -38% phenolics). Seeds removal, in turn, was more injurious for cereja tomatoes (-10% lycopene, -38% b-carotene, -25% ascorbic acid and -63% phenolics). Comparatively with the remaining ones, the rama fruits were less affected by the trimming procedures.

Pre-meal tomato (Lycopersicon esculentum) intake can have anti-obesity effects in young women?

The effect of pre-meal tomato intake in the anthropometric indices and blood levels of triglycerides, cholesterol, glucose, and uric acid of a young women population (n=35, 19.6 ± 1.3 years) was evaluated. During 4 weeks, daily, participants ingested a raw ripe tomato (~90 g) before lunch. Their anthropometric and biochemical parameters were measured repeatedly during the follow-up time. At the end of the 4 weeks, significant reductions were observed on body weight (-1.09 ± 0.12 kg on average), % fat (-1.54 ± 0.52%), fasting blood glucose (-5.29 ± 0.80 mg/dl), triglycerides (-8.31 ± 1.34 mg/dl), cholesterol (-10.17 ± 1.21 mg/ dl), and uric acid (-0.16 ± 0.04 mg/dl) of the participants. The tomato pre-meal ingestion seemed to interfere positively in body weight, fat percentage, and blood levels of glucose, triglycerides, cholesterol, and uric acid of the young adult women that participated in this study.

Exposure of Lycopersicon Esculentum to Microcystin-LR: Effects in the Leaf Proteome and Toxin Translocation from Water to Leaves and Fruits

Natural toxins such as those produced by freshwater cyanobacteria have been regarded as an emergent environmental threat. However, the impact of these water contaminants in agriculture is not yet fully understood. The aim of this work was to investigate microcystin-LR (MC-LR) toxicity in Lycopersicon esculentum and the toxin accumulation in this horticultural crop. Adult plants (2 month-old) grown in a greenhouse environment were exposed for 2 weeks to either pure MC-LR (100 μg/L) or Microcystis aeruginosa crude extracts containing 100 μg/L MC-LR. Chlorophyll fluorescence was measured, leaf proteome investigated with two-dimensional gel electrophoresis and Matrix Assisted Laser Desorption Ionization Time-of-Flight (MALDI-TOF)/TOF, and toxin bioaccumulation assessed by liquid chromatography-mass spectrometry (LC-MS)/MS. Variations in several protein markers (ATP synthase subunits, Cytochrome b6-f complex iron-sulfur, oxygen-evolving enhancer proteins) highlight the decrease of the capacity of plants to synthesize ATP and to perform photosynthesis, whereas variations in other proteins (ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit and ribose-5-phosphate isomerase) suggest an increase of carbon fixation and decrease of carbohydrate metabolism reactions in plants exposed to pure MC-LR and cyanobacterial extracts, respectively. MC-LR was found in roots (1635.21 μg/kg fw), green tomatoes (5.15–5.41 μg/kg fw), mature tomatoes (10.52–10.83 μg/kg fw), and leaves (12,298.18 μg/kg fw). The results raise concerns relative to food safety and point to the necessity of monitoring the bioaccumulation of water toxins in agricultural systems affected by cyanotoxin contamination.

AVALIAÇÃO DE PROGÊNIES DE TOMATE (LYCOPERSICON ESCULENTUM) PARA CULTIVO SOB TEMPERATURAS ELEVADAS

Foram avaliadas três progênies F7 de tomate άο cruzamento HT-16, do programa de melhoramento genético do Departamento de Ciências Agronômicas do INPA, para o caráter pegamento de frutos. Os ensaios foram realizados em casa de vegetação com cobertura de plástico transparente e por um período de 37 dias, que cobriu toda a fase de florescimento e frutificação do primeiro ao oitavo racimo; tomou-se dados sobre a temperatura do ar no interior da casa de vegetação, por meio de um termohigrógrafo. A amplitude de variação da temperatura foi de 18° C (mínimo) até 43° C (máximo). Os resultados mostraram a ocorrência de variabilidade genética entre e dentro das progênies avaliadas para o caráter percentagem de pega mento de frutos, sendo possível sua exploração em processos seletivos visando obter linhagens com alta capacidade de pegamento de frutos sob condição de cultivo em temperaturas elevadas.

Mechanism of extracellular silver nanoparticles synthesis by Stereum hirsutum and Fusarium oxysporum

The increasing interest for greener and biological methods of synthesis has led to the development of non-toxic and comparatively more bioactive nanoparticles. Unlike physical and chemical methods of nanoparticle synthesis, microbial synthesis in general and mycosynthesis in particular is cost-effective and environment-friendly. However, different aspects, such as the rate of synthesis, monodispersity and downstream processing, need to be improved. Many fungal-based mechanisms have been proposed for the formation of silver nanoparticles (AgNPs), mainly those involving the presence of nitrate reductase, which has been detected in filtered fungus cell used for AgNPs production. There is a general acceptance that nitrate reductase is the main responsible for the reduction of Ag ions for the formation of AgNPs. However, this generally accepted mechanism for fungal AgNPs production is not totally understood. In order to elucidate the molecules participating in the mechanistic formation of metal nanoparticles, the current study is focused on the enzymes and other organic compounds involved in the biosynthesis of AgNPs. The use of each free fungal mycelium of both Stereum hirsutum and Fusarium oxysporum will be assessed. In order to identify defective mutants on the nitrate reductase structural gene niaD, fungal cultures of S.hirsutum and F.oxysporum will be selected by chlorate resistance. In addition, in order to verify if each compound identified as key-molecule influenced on the production of nanoparticles, an in vitro assay using different nitrogen sources will be developed. Lately, fungal extracellular enzymes will be measured and an in vitro assay will be done. Finally, The nanoparticle formation and its characterization will be evaluated by UV-visible spectroscopy, electron microscopy (TEM), X-ray diffraction analysis (XRD), Fourier transforms infrared spectroscopy (FTIR), and LC-MS/MS.

Contribuição ao estudo da aplicação do método da diagnose foliar ao algodoeiro (Grossypium hirsutum L., var. I. A. C. 817) II

This paper deal with one experiment carried out in order to study the correlation between petioles analysis and seed cotton yield. A 3X3X3 factorial with respect to N, P2 0(5) and K2 O was installed in a sandy soil with low potash content and medium amounts of total N and easily extractable P. Two kinds of petioles, newly mature were collected for analysis: those attached to fruit hearing branches, and petioles located on the stem; the first group is conventionally named "productive petioles"; The second one is called "not productive petioles". Petioles' sampling was done when the first blossoms appeared. Yield date showed a marked response to potash, both nitrogen and phosphorus having no effect. Very good correlation was found between petioles potash and yield. Both types of petioles samples were equally good indicators of the potash status of the plants. By mathematical treatment of the date it followes that the highed yield which was possible under experimental conditions, 1.562 kg of seed cotton per hectare would be reacher by using 128 kg of K2O per hectare. With this amount of potash supplied to the plants the following K levels would be expected in the petioles: "productive petioles" "not productive petioles" 1,93 % K 1,85 % K

Variação da composição de aminoácidos em caules e raízes de algodoeiro (Gossypium hirsutum L.) de acôrdo com a suscetibilidade aFusarium oxysporwn f. vasinfectum (Atk.) SNYD e HANS

Os resultados do estudo da variação na composição dos aminoácidos em caules e raízes de plantas de algodoeiro resistentes e suscetíveis a Fusarium oxysporum f. vasinfectum, revelaram maiores diferenças nos caules, quando comparados com aquêles obtidos para raízes. No estágio cotiledonar, tanto para plantas inoculadas como n]ao inoculadas, os caules de plantas da variedade suscetível apresentaram maior teor de aminoácidos, quando comparados com aquêles de plantas resistentes. Os caules das plantas suscetíveis em estágio de 3a 4 fôlhas verdadeiras, quando inoculadas, apresentaram maior teor de aminoácidos, quando comparadas com aquêles de plantas resistentes inoculadas com a mesma idade. Porém, para o caso de plantas não inoculadas, o teor de aminoácidos das plantas suscetíveis era ligeiramente inferior que o observado para as plantas resistentes.

Adubação foliar do algodoeiro (Gossypium hirsutum, L., var. I.A.C.12), com nitrogênio, fósforo e potássio avaliada pela produção e diagnose foliar

Aplicações de adubos por via foliar no algodoeiro tem sido feitas em outros países; porém, no Brasil, tal prática encontra-se no inicio. Com o presente trabalho, foi estudado o efeito da adubação foliar nos aumentos de produção e foi estabelecido correlações entre a produção e as concentrações de nitrogênio, fósforo e potássio, nos pecíolos das folhas de ramos produtivos e não produtivos.

Efeitos do ácido 2-cloroetilfosfônico na maturação de frutos do tomateiro (Lycopersicon esculentum Mill. cv. 'São Sebastião')

Estudou-se a influência do ácido 2-cloroetilfosfônico (ethephon), aplicado em pré-maturação, na frutificação do tomateiro de crescimento indeterminado, cultivar 'São Sebastião'; em condições de casa de vegetação. Pela análise da produtividade verificou-se que o regulador de crescimento acelerou significativamente a maturação dos frutos, incrementando o peso e o número total de frutos por planta obtidos nas primeiras colheitas, sendo isto compensado por diminuições significativas em colheitas posteriores. Não observou-se porém, diferenças significativas no peso e número de frutos por planta, considerando a totalidade das colheitas, entre o controle e as plantas tratadas com 1000, 2000 e 4000 ppm de ethephon.

Avaliação do comportamento de sementes de algodoeiro (Gossypium hirsutum L.) durante o armazenamento

Sementes de algodoeiro, variedade IAC 13-1, foram armazenadas em câmara seca e em condições normais de ambiente, durante oito meses. Periodicamente, com intervalos bimestrais, foi avaliado o comportamento das sementes das duas porções, através de testes de germinação e de vigor (envelhecimento rápido e irradiação de sementes), no Laboratório de Sementes do Departamento de Agricultura e Horticultura, ESALQ, USP. As análises dos dados e a interpretação dos resultados permitiram concluir que: a) As sementes conservaram-se melhor em câmara seca. b) Há possibilidade do emprego de irradiação para avaliar o vigor de sementes, obtendo-se resultados comparáveis aos do envelhecimento rápido, dependendo da dosagem utilizada.

Influência da adubação foliar com NPK, na cultura algodoeira (G. hirsutum L. var. IAC-12), sobre a qualidade da fibra e da semente

Aplicações de fertilizantes por via foliar e ao solo foram comparados na cultura algodoeira, utilizando-se a variedade IAC-12, com a finalidade de se estudar uma influência sobre alguns caracteres físicos da fibra e da semente. Tanto na adubação ao solo, como na foliar, foram utilizados como fonte de nitrogênio, a uréia (46%N), como fonte de fósforo, o superfosfato concentrado (45% P2O2) e como fonte de potássio, o cloreto de potássio (60% K2O). Os resultados obtidos mostram que entre as características físicas da análise da fibra, encontrou-se um efeito positivo no comprimento da fibra, resistência da fibra, resistência do fio e maior peso de semente (índice de semente) quando o cloreto de potássio foi aplicado por via foliar. Quanto à uniformidade da fibra e ao índice de finura, não houve influência da adubação foliar com nenhum dos três elementos. Nestes casos, a adubação ao solo, proporcionou melhores resultados. Deve-se ressaltar que com a dose mais alta de fósforo (60 kg de P2O5/ha), o valor do índice de finura foi maior. O adubo nitrogenado na dose de 40 kg de N/ha, influenciou na resistência da fibra (Pressley).