Maria das Graças Cardoso

Inheritance of foliar zingiberene contents and their relationship to trichome densities and whitefly resistance in tomatoes

The sesquiterpene zingiberene, present in leaf glandular trichomes, is reportedly responsible for the high level of arthropod resistance found in Lycopersicon hirsutum var. hirsutum. This paper reports on the inheritance of zingiberene contents and of the various types of glandular trichomes in the interspecific cross L. esculentum × Lycopersicon hirsutum var. hirsutum. Plants of L. esculentum ‘TOM-556’ (= P1), L. hirsutum var. hirsutum ‘PI-127826’(= P2), F1 (P1 × P2) and F2 (P1 ×P2) were evaluated for zingiberene contents and densitities of and glandular (types I, IV, VI and VII) trichomes. Broad sense heritabilities were high for all traits studied (0.678, 0.831, 0.996, 0.799 and 0.717 respectively for zingiberene and trichome types I, IV, VI, VII). There were significant positive genetic correlations between zingiberene contents and densities of trichomes types IV, VI and VII. Inheritance of zingiberenecontents can be explained mostly by the action of a single major locus, inwhich the allele from L. hirsutum that conditions high content is incompletely recessive over the allele from L. esculentum. Action of an incompleteley recessive allele in one major locus appears to be evident for densities of trichome types IV, VI and VII, but there is also evidence of the action of other epistatic loci for types IV and VI. F2 genotypes selected for high zingiberene levels showed higher levels of resistance to the silverleaf whitefly Bemisia argentifolii than L. esculentum ‘TOM-556’, levels that were comparable those found in L. hirsutum var. hirsutum ‘PI-127826’ and other whitefly resistant accessions.

Relationships between trichome types and spider mite (Tetranychus evansi) repellence in tomatoes with respect to foliar zingiberene contents

Zingiberene, a sesquiterpene present in glandular trichomes of Lycopersicon hirsutum Dunal var. hirsutum ‘PI-127826’, is responsible for the high level of arthropod resistance in this taxon. The current paper has the following objectives: (a) to quantify zingiberene content in tomato plants obtained from the interspecific cross L. esculentum Mill. × L. hirsutum var hirsutum; (b) to identify, classify and quantify glandular and non-glandular trichome types present in those plants; (c) to assess the level of resistance of those genotypes to spider mites (Tetranychus evansi); (d) to estimate correlations between glandular trichomes, zingiberene contents and mite repellence. Zingiberene content were quantified by a colorimetric method (Freitas, 1999); trichomes were counted from foliar paradermic slide preparations; mite resistance was assessed by a repellence test (Weston & Snyder, 1990). The results indicate that indirect selection for zingiberene content led to correlated increases both in the number of glandular trichomes (particulary type IV) and in the levels of mite repellence. These results were found both in BPX-368 [=F2 (L. esculentum ‘TOM-556’ × L hirsutum var. hirsutum ‘PI-127826’)] and in the subsequent generation BPX-368B (which represents one additional backcross to L.esculentum). Zingiberene appears therefore to be the main factor involved in mite repellence. Density of glandular trichomes in tomato leaflets markedly influences total zingiberene content. Type IV trichome density was the highest, and it was highly correlated to zingiberene content.

Antimicrobial activity of Satureja montana L. essential oil against Clostridium perfringens type A inoculated in mortadella-type sausages formulated with different levels of sodium nitrite

This research evaluated the antimicrobial effect of the winter savory (Satureja montana L.) essential oil (EO) against Clostridium perfringens type A (ATCC 3624) inoculated in mortadella-type sausages formulated with different levels of sodium nitrite (NaNO₂: 0 ppm, 100 ppm and 200 ppm) in addition to EO at concentrations of 0.0%, 0.78%, 1.56% and 3.125% stored at 25°C for 30 days. The EO extracted by hydrodistillation and analyzed by gas chromatography-mass spectrometry (CG-MS) was tested in vitro using an agar well diffusion method for determination of minimum inhibitory concentration (MIC) on C. perfringens. According to compositional analysis of the winter savory EO, 26 chemical compounds were identified, and the major constituents were thymol (28.99%), p-cymene (12.00%), linalool (11.00%) and carvacrol (10.71%). The results obtained showed that EO applied at a concentration of 1.56%, which was defined as the MIC, exhibited antimicrobial activity against C. perfringens in the in vitro assays, and the transmission electron microscopy (TEM) revealed structural damage and cell lysis of C. perfringens caused by EO treatment. A synergistic effect between NaNO₂ and EO was observed. In mortadella-type sausages formulated with 100 ppm of NaNO₂ and EO at all concentrations tested, the population of target microorganisms was reduced (p≤0.05) compared to control samples during all storage period. This data suggests the potential combined use of savory EO and minimal amounts of the synthetic additive, NaNO₂ to control C. perfringens in mortadella, which goes according to current market trends, where consumers are requesting natural products.

Anti-proliferative effect of the essential oil of Cymbopogon citratus (DC) Stapf (lemongrass) on intracellular amastigotes, bloodstream trypomastigotes and culture epimastigotes of Trypanosoma cruzi (Protozoa: Kinetoplastida)

This study analyses the anti-proliferative effect of lemongrass essential oil and its main constituent (citral) on all 3 evolutive forms of Trypanosoma cruzi. Steam distillation was used to obtain lemongrass essential oil, with chemical composition determined by gas chromatography (GC) and GC coupled to mass spectrometry (GC-MS). The IC50/24 h (concentration that reduced the parasite population by 50%) of the oil and of citral upon T. cruzi was determined by cell counting in a Neubauer chamber, while morphological alterations were visualized by scanning and transmission electron microscopy. Treatment with the essential oil resulted in epimastigote growth inhibition with IC50=126.5 microg/ml, while the IC50 for trypomastigote lysis was 15.5 microg/ml. The IC50/48 h for the Association Index (% macrophage infection x number of amastigotes per cell) was 5.1 microg/ml, with a strong inhibition of intracellular amastigote proliferation. Ultrastructural analysis demonstrated cytoplasmic and nuclear extraction, while the plasma membrane remained morphologically preserved. Our data show that lemongrass essential oil is effective against T. cruzi trypomastigotes and amastigotes, and that its main component, citral, is responsible for the trypanocidal activity. These results indicate that essential oils can be promising anti-parasitic agents, opening perspectives to the discovery of more effective drugs of vegetal origin for treatment of parasitic diseases. However, additional cytotoxicity experiments on different cell lines and tests in a T. cruzi-mouse model are needed to support these data.

Caracterização química e efeito inibitório de óleos essenciais sobre o crescimento de Staphylococcus aureus e Escherichia coli

Objetivou-se avaliar o efeito inibitorio dos oleos essenciais de Cymbopogon citratus (capim-limao), Origanum vulgare (oregano) e Syzygium aromaticum (cravo-da-india); os experimentos foram realizados com as bacterias Staphylococcus aureus e Escherichia coli, importantes patogenos causadores de contaminacoes em queijos e outros alimentos. Para quantificacao e identificacao dos constituintes quimicos dos oleos, utilizou-se um cromatografo gasoso acoplado a um espectrometro de massa. Os resultados dos testes in vitro, os oleos essenciais de C. citratus, O. vulgare e S. aromaticum promoveram efeito inibitorio sobre as bacterias S. aureus e E. coli, porem o S. aromaticum apresentou melhor formacao de halo de inibicao nas menores concentracoes. Para o efeito sinergistico dos oleos sobre as bacterias nao foram observadas diferencas quando comparados com o efeito individual dos mesmos. Foi possivel verificar que os oleos possuem efeito inibitorio sobre os microrganismos estudados, sendo, portanto uma alternativa no controle microbiologico de alimentos.

A convenient method for the determination of moisture in aromatic plants

A method is proposed for the determination of the moisture content of aromatic plants. This method is based on the co-distillation of the starting material in a modified Clevenger apparatus with four organic solvents (toluene, cyclohexane, dichloromethane and carbon tetrachloride). The results were compared with those obtained by oven drying at 105 oC and steam distillation of the essential oil. The efficiencies of the methods were shown to be equivalent. The solvent distillation method was more practical, especially with respect to operating time (2 h).

Influência da luz e da temperatura sobre a oxidação do óleo essencial de capim-limão (Cymbopogon citratus (D.C.) Stapf)

The identification of the chemical compounds of the essential oil was performed with a gas chromatograph coupled to a mass spectrometer. The oil was left in the presence and absence of light and submitted to different temperatures to evaluate its stability. The yields of the major compounds were evaluated every fifteen days. Citral and myrcene, the major compounds of the essential oil, were degraded over time in both the presence and absence of light, but temperature only influenced the degradation of myrcene.

Resistance of tomato genotypes with high level of acylsugars to Tetranychus evansi Baker & Pritchard

O acesso de Lycopersicon pennellii LA716 tem sido utilizado em programas de melhoramento do tomateiro devido a sua resistencia a artropodos-pragas, mediada pela presenca de acilacucares nos foliolos. Foi verificada a correlacao entre o teor de acilacucares em foliolos de tomateiros e a repelencia ao acaro Tetranychus evansi Baker & Pritchard. Foram selecionadas plantas contrastantes quanto aos niveis de acilacucares nos foliolos de populacoes F2 e F2RC1 (= F2 do primeiro retrocruzamento para Lycopersicon esculentum) do cruzamento interespecifico de L. esculentum TOM 584 (baixo teor de acilacucares) e L. pennellii LA 716 (alto teor). Foi realizado um teste de repelencia ao acaro T. evansi, tomando por base a distância percorrida pelos acaros nos foliolos. Os genotipos selecionados para alto teor de acilacucares, em media, foram responsaveis por reducoes significativas nas distâncias percorridas pelos acaros sobre a superficie do foliolo. A repelencia ao acaro T. evansi exercida pelas plantas selecionadas para alto teor de acilacucares deu-se de forma semelhante aquela conferida pelo genitor LA-716. As magnitudes das correlacoes encontradas confirmam a associacao entre altos teores do aleloquimico e a repelencia ao acaro.

Relação entre zingibereno, tricomas foliares e repelência de tomateiros a Tetranychus evansi

The objective of this work was to quantify zingiberene contents in a segregating population (F2) of the second backcross generation to Lycopersicon esculentum after the initial cross with the wild species L. hirsutum var. hirsutum, and to analyse the relationship between zingiberene and both glandular trichome density on tomato leaves and levels of repellence to spider mites Tetranychus evansi. A spectrophotometric assay was used to quantify zingiberene in leaf disks. Trichome counting was made in paradermic preparations of leaf and mite repellence was accessed by a quick thumbtack assay. Indirect selection for high zingiberene contents increased both glandular trichome density and the level of mite repellence. Zingiberene in glandular trichomes therefore mediates mite repellence in the tomato population. Glandular trichome densities, especially that of type VI trichomes, were highly and positively correlated with zingiberene contents. High zingiberene levels and their relationship with both glandular trichome densities and resistance (repellence) to spider mites remained significant after two backcross generations, indicating an indirect selection for high zingiberene as an effective way of selecting mite resistant tomato genotypes.

Chemical composition and fumigant effect of essentialoil of Lippia sidoides Cham. and monoterpenes against Tenebrio molitor (L.) (coleoptera: tenebrionidae)

A composicao quimica do oleo essencial de Lippia sidoides obtido por hidrodestilacao foi caracterizada e quantificada por GC/MS, bem como sua atividade inseticida por teste de fumigacao foi avaliada sobre Tenebrio molitor. Alem disso, a toxicidade dos monoterpenos carvacrol, 1,8-cineol e timol, tambem foi avaliada quando esses compostos foram aplicados isoladamente, ou em misturas binarias (1:1), ou terciarias (1:1:1). O oleo essencial de L. sidoides tem como principais constituintes o carvacrol (31,68%), ρ-cimeno (19,58%), 1,8-cineol (9,26%) e ϒ-terpineno (9,21%), em 21 compostos identificados, sendo 92,53% do total. Ambos os compostos possuem atividade inseticida contra T. molitor, seguindo a ordem de toxicidade: carvacrol > 1,8-cineol > oleo essencial de L. sidoides > timol; sendo suas respectivas CL50 a 24 h de 5,53; 5,71; 8,04 e 14,71 µL/L ar. Quando as diferentes misturas de carvacrol, 1,8-cineol e timol foram avaliadas contra T. molitor, verificou-se o efeito sinergico. Para a mistura de carvacrol:1,8-cineol a CL50 foi de 5,34 µL/L ar; carvacrol:timol de 7,67 µL/L ar; 1,8-cineol:timol de 7,51 µL/L ar e carvacrol:1,8-cineol:timol de 6,34 µL/L ar. Principalmente o monoterpeno timol teve efeito sinergico, aumentando a toxicidade de carvacrol e 1,8-cineol quando em misturas binarias, tais como carvacrol:timol e 1,8-cineol:timol.