Expression of Pinellia ternata leaf agglutinin under rolC promoter confers resistance against a phytophagous sap sucking aphid, Myzus persicae

BACKGROUND: Piercing/sucking insect pests in the order Hemiptera causes substantial crop losses by removing photoassimilates and transmitting viruses to their host plants. Cloning and heterologous expression of plantderived insect resistance genes is a promising approach to control aphids and other sap-sucking insect pests. While expression from the constitutive 35S promoter provides broad protection, the phloem-specific rolC promoter provides better defense against sap sucking insects. The selection of plant-derived insect resistance genes for expression in crop species will minimize bio-safety concerns. RESULTS: Pinellia ternata leaf agglutinin gene (pta), encodes an insecticidal lectin, was isolated and cloned under the 35S and rolC promoters in the pGA482 plant transformation vector for Agrobacterium-mediated tobacco transformation. Integration and expression of the transgene was validated by Southern blotting and qRT-PCR, respectively. Insect bioassays data of transgenic tobacco plants showed that expression of pta under rolC promoter caused 100% aphid mortality and reduced aphid fecundity up to 70% in transgenic tobacco line LRP9. These results highlight the better effectivity of pta under rolC promoter to control phloem feeders, aphids. CONCLUSIONS: These findings suggested the potential of PTA against aphids and other sap sucking insect pests. Evaluation of gene in tobacco under two different promoters; 35S constitutive promoter and rolC phloemspecific promoter could be successfully use for other crop plants particularly in cotton. Development of transgenic cotton plants using plant-derived insecticidal, PTA, would be key step towards commercialization of environmentally safe insect-resistant crops.

Interception of 'Candidatus Phytoplasma solani' in Brazil in apple tree propagative material imported from France / Interceptação de 'Candidatus Phytoplasma solani' no Brasil em material propagativo de macieira importado da França

Apple plants from France introduced in Brazil for research purposes were subjected to a phytosanitary analysis at the Plant Quarantine Laboratory of Embrapa Genetic Resources and Biotechnology (Cenargen). After grafting onto healthy apple rootstock, some plants showed phytoplasma-infection symptoms. Polymerase Chain Reaction (PCR) tests yielded DNA fragments of the expected size for phytoplasmas. DNA sequencing revealed an identity of the 16S rDNA nucleotide sequence of 98-99% with 'Candidatus Phytoplasma solani'. This phytoplasma species is responsible for losses in European apple orchards and has not been reported in Brazil. According to the Federal Legislation on Plant Protection, the plants were incinerated to avoid the introduction of this exotic pest in Brazil.(AU)

Plantas de macieira originárias da França introduzidas no Brasil para fins de pesquisa foram submetidas à análise no Laboratório de Quarentena Vegetal da Embrapa Recursos Genéticos e Biotecnologia (Cenargen). Após realizar a enxertia em porta-enxertos sadios, algumas plantas apresentaram sintomas típicos de infecção por fitoplasmas. Os testes por meio da técnica de reação em cadeia da polimerase (PCR) obtiveram fragmentos de DNA do tamanho esperado para fitoplasmas. O sequenciamento de DNA revelou uma identidade de 98-99% da região 16 rDNA dos fitoplasmas encontrados com o fitoplasma 'Candidatus Phytoplasma solani'. Essa espécie de fitoplasma é responsável por perdas em pomares de maçã europeus e não foi relatada no Brasil. De acordo com a Legislação Federal de Proteção de Plantas, as plantas de macieira foram incineradas para evitar a entrada dessa praga exótica no Brasil.(AU)

Entomopathogenic Fungi as Dual Control Agents against Both the Pest Myzus persicae and Phytopathogen Botrytis cinerea

The green peach aphid (Myzus persicae), a plant pest, and gray mold disease, caused by Botrytis cinerea, affect vegetables and fruit crops all over the world. To control this aphid and mold, farmers typically rely on the use of chemical insecticides or fungicides. However, intensive use of these chemicals over many years has led to the development of resistance. To overcome this problem, there is a need to develop alternative control methods to suppress populations of this plant pest and pathogen. Recently, potential roles have been demonstrated for entomopathogenic fungi in endophytism, phytopathogen antagonism, plant growth promotion, and rhizosphere colonization. Here, the antifungal activities of selected fungi with high virulence against green peach aphids were tested to explore their potential for the dual control of B. cinerea and M. persicae. Antifungal activities against B. cinerea were evaluated by dual culture assays using both aerial conidia and cultural filtrates of entomopathogenic fungi. Two fungal isolates, Beauveria bassiana SD15 and Metarhizium anisopliae SD3, were identified as having both virulence against aphids and antifungal activity. The virulence of these isolates against aphids was further tested using cultural filtrates, blastospores, and aerial conidia. The most virulence was observed in the simultaneous treatment with blastospores and cultural filtrate. These results suggest that the two fungal isolates selected in this study could be used effectively for the dual control of green peach aphids and gray mold for crop protection.

Imidacloprid inhibits IgE-mediated RBL-2H3 cell degranulation and passive cutaneous anaphylaxis

BACKGROUND: Imidacloprid has been commonly used as a pesticide for crop protection and acts as nicotinic acetylcholine receptor agonists. Little information about the relationship between imidacloprid and allergy is available. OBJECTIVE: This study aims to examine the effects of imidacoprid on IgE-mediated mast cell activation. METHODS: The rat basophilic leukemia cell line RBL-2H3 (RBL-2H3 cells) were treated with 10⁻³ – 10⁻¹² mol/L imidacloprid, followed by measuring the mediator production, influx of Ca²⁺ in IgE-activated RBL-2H3 cells, and the possible effects of imidacoprid on anti-dinitrophenyl IgE-induced passive cutaneous anaphylaxis (PCA). RESULTS: It was shown that imidacoprid suppressed the production of histamine, β-hexosaminidase, leukotriene C4, interleukin-6, tumor necrosis factor-α, and Ca²⁺ mobilization in IgE-activated RBL-2H3 cells and decreased vascular extravasation in IgE-induced PCA. CONCLUSION: It is the first time to show that imidacloprid suppressed the activation of RBL-2H3 cells.

O efeito da biodiversidade nativa da Mata Atlântica sobre o metabolismo de amadurecimento da banana (Musa acuminata AAA, Cavendish, cv. Nanicão) / he effect of the Atlantic Coastal Forest´s native biodiversity on the metabolism of banana ripening (Musa acuminata AAA, Cavendish, cv. Nanicão).

A produção de banana no Vale do Ribeira, em São Paulo, está inserida numa região que abriga um fragmento remanescente da Mata Atlântica, considerada um dos principais repositórios de biodiversidade e a floresta mais devastada entre os biomas brasileiros. Medidas sustentáveis, como a adoção de práticas agrícolas alternativas, tem sido o caminho para a conservação da fauna e flora, como também para a redução do uso inapropriado de terras que consequentemente pode aumentar o índice de doenças que atacam as plantas. Apesar, do razoável conhecimento científico sobre a fisiologia pós-colheita da banana, ainda não foram elucidadas quais são as respostas fisiológicas do fruto diante das alterações do meio ambiente decorrentes das práticas agroecológicas. Principalmente quais são os mecanismos de ação de compostos específicos, relacionados com a resistência da planta contra os estresses bióticos e abióticos. Também, não há uma abordagem analítica integrada que identifique os reguladores das vias metabólicas e possibilite um estudo holístico a nível molecular. Neste trabalho, foi avaliado o efeito da proximidade da biodiversidade nativa da Mata Atlântica sobre o perfil de metabólitos da banana (Musa acuminata AAA, Cavendish, cv. Nanicão). Foram comparados os frutos da parcela Biodiversidade, o qual apresenta 60% de seu perímetro limítrofe à floresta com a parcela Controle, inserida em uma área de produção de banana convencional. Neste estudo, foram determinados os perfis de etileno, vida verde, amido, açúcares, textura, cor, metabólitos semi-voláteis e não-voláteis e poliaminas. Como resultado, ambas as parcelas avaliadas apresentaram diferenças significativas no metabolismo primário e secundário, nos frutos verdes e maduros, respectivamente. Os metabólitos com diferença significativa entre as parcelas experimentais apresentaram maior envolvimento no metabolismo primário, sobretudo na via de biossíntese de aminoácidos, compostos precursores de uma ampla faixa de metabólitos secundários. As alterações quanto à abundância destes compostos são uma referência de que as mudanças do meio modificam as respostas da planta a estresses e estímulos

Banana production in the Ribeira Valley, in São Paulo, is set in a region that embraces a remaining fragment of the Atlantic Coastal Forest (Mata Atlântica), considered one of the main biodiversity repositories and the most devastated forest among the Brazilians´ biomes. Sustainable measures such as the adoption of alternative farming practices has been the way for the conservation of fauna and flora, but also to reduce the inappropriate use of land which in turn may increase the disease index that attack plants. Although, reasonable scientific knowledge about banana postharvest physiology is available, the fruit´s physiologic response given the environment changes as a result agroecological practices has not been elucidated yet. Primarily what are the mechanisms of action of specific compounds, related to plant resistance against biotic and abiotic stresses. In addition, there is not an integrated analytical approach to identify regulators of metabolic pathways and enable a holistic study at the molecular level. In this study, we evaluated the effect of proximity to the native biodiversity of the Atlantic Forest on the banana metabolites profile (Musa acuminata AAA, Cavendish, cv. Nanicão). The fruits of Biodiversity group were compared, which has 60% of its perimeter adjacent to the forest with the Control group set in a conventional banana production area. In this study, we determined the ethylene profiles, green life, starch, sugars, texture, colour, semi- volatile and non- volatile metabolites and polyamines. As a result, both evaluated groups showed significant differences in primary and secondary metabolism, green and ripen fruits respectively. The metabolites with significant differences between treatments showed greater involvement in primary metabolism, especially in amino acid biosynthesis, precursors of secondary metabolites. Changes in the abundance of these compounds are indicators that alterations in the environment modify the plant responses to stresses and stimuli

Produção de óleo essencial de Melissa officinalis L. em diferentes épocas, sistemas de cultivo e adubações / Essential oil of Melissa officinalis L. at differents seasons, systems of planting and fertilizations

Os objetivos deste trabalho foram avaliar a biomassa, o rendimento e a composição de óleo essencial de Melissa officinalis L. submetida a diferentes épocas de plantio, sistemas de cultivo, e adubações. O delineamento experimental foi o de blocos casualizados (DBC), em esquema fatorial 2 x 2 x 2, sendo duas épocas de plantio (verão e inverno), dois sistemas de cultivo (estufa e campo), e dois tipos de adubação (mineral e orgânica). A colheita consistiu no corte de todas as plantas da parcela útil. Após a colheita, foi avaliada a massa fresca foliar, a altura de plantas, o comprimento e largura das folhas. A extração de óleo essencial foi realizada por hidrodestilação e a composição química foi analisada em cromatógrafo gasoso acoplado a espectrômetro de massas. Das substâncias encontradas, prevaleceram como constituintes majoritários os monoterpenos geranial e neral na colheita de verão, e geranial, neral e citronelal na colheita de inverno. As condições do ambiente protegido e do verão propiciaram melhor biomassa de melissa, maior rendimento de óleo essencial, e maior taxa do constituinte citral.

The objectives of this study were to evaluate the biomass, yield and composition of essential oil of Melissa officinalis L. under differents seasons, systems of planting and fertilizations. The experimental design was a randomized block design (RBD), in a factorial 2 x 2 x 2 with two seasons of harvest (summer and winter), two systems of planting (greenhouse and field) and two types of fertilizer (mineral and organic). The harvest consisted of cutting all plants of central line. After harvest, it were evaluated the leaf fresh weight, height of the plant, length and width of the leaves. The essential oil extraction was performed by hydrodistillation and the chemical composition was performed by gas chromatography coupled by mass spectrometry. The major constituents were geranial and neral in summer harvest and geranial, neral and citronellal in winter harvest. The conditions of the greenhouse and the summer presented the better biomass, higher yield of oil and higher rate of citral.