Comparison of predicted binders in Rhipicephalus (Boophilus) microplus intestine protein variants BM86 Campo Grande strain, BM86 and BM95 / Comparação da previsão de ligação das variantes de proteína de intestino de Rhipicephalus (Boophilus) microplus Bm86 cepa Campo Grande, Bm86 e Bm95

This paper reports the sequence analysis of Bm86 Campo Grande strain comparing it with Bm86 and Bm95 antigens from the preparations TickGardPLUS and GavacTM, respectively. The PCR product was cloned into pMOSBlue and sequenced. The secondary structure prediction tool PSIPRED was used to calculate alpha helices and beta strand contents of the predicted polypeptide. The hydrophobicity profile was calculated using the algorithms from the Hopp and Woods method, in addition to identification of potential MHC class-I binding regions in the antigens. Pair-wise alignment revealed that the similarity between Bm86 Campo Grande strain and Bm86 is 0.2 percent higher than that between Bm86 Campo Grande strain and Bm95 antigens. The identities were 96.5 percent and 96.3 percent respectively. Major suggestive differences in hydrophobicity were predicted among the sequences in two specific regions.

O objetivo deste estudo foi analisar a seqüência de Bm86 cepa Campo Grande comparando-a com os antígenos Bm86 e Bm95 das preparações TickGardPLUS e GavacTM, respectivamente. O produto de PCR foi clonado em PMOSBlue e seqüenciado. Para calcular os conteúdos de alfa-hélice e fita beta do polipeptídio previsto, foi utilizada a ferramenta de prognóstico de estrutura secundária PSIPRED. O perfil de hidrofobicidade foi calculado usando os algoritmos de Hopp e Woods, além da identificação das possíveis regiões de ligação com MHC classe I nos antígenos. O alinhamento "pair-wise" revelou que a similaridade entre Bm86 cepa Campo Grande e Bm86 é 0,2 por cento maior que aquela entre Bm86 cepa Campo Grande e Bm95. As identidades foram de 96,5 por cento e 96,3 por cento, respectivamente. Com relação à hidrofobicidade, os resultados sugerem que a maior diferença entre as seqüências está localizada em duas regiões específicas.

Signaling pathways in a Citrus EST database

Citrus spp. are economically important crops, which in Brazil are grown mainly in the State of São Paulo. Citrus cultures are attacked by several pathogens, causing severe yield losses. In order to better understand this culture, the Millenium Project (IAC Cordeirópolis) was launched in order to sequence Citrus ESTs (expressed sequence tags) from different tissues, including leaf, bark, fruit, root and flower. Plants were submitted to biotic and abiotic stresses and investigated under different development stages (adult vs. juvenile). Several cDNA libraries were constructed and the sequences obtained formed the Citrus ESTs database with almost 200,000 sequences. Searches were performed in the Citrus database to investigate the presence of different signaling pathway components. Several of the genes involved in the signaling of sugar, calcium, cytokinin, plant hormones, inositol phosphate, MAPKinase and COP9 were found in the citrus genome and are discussed in this paper. The results obtained may indicate that similar mechanisms described in other plants, such as Arabidopsis, occur in citrus. Further experimental studies must be conducted in order to understand the different signaling pathways present.

Genes associated with hypersensitive response (HR) in the citrus EST database (CitEST)

Plants are continuously exposed to pathogen attack, but successful infection is rare because they protect themselves against pathogens using a wide range of response mechanisms. One of them is the hypersensitive response (HR), which is a form of cell death often associated with plant resistance to pathogen infection to prevent the spreadsebpg@cnpq.br sebpg@cnpq.br of the potential pathogen from infected to uninfected tissues. Cell death is activated by recognition of pathogen-derived molecules by the resistance (R) gene products, and is associated with the massive accumulation of reactive oxygen species (ROS), salicylic acid (SA), and other pro-death signals such as nitric oxide (NO). The analysis of the citrus EST (CitEST) database revealed the presence of putative genes likely to be involved in HR through their products, like metacaspases, lipoxygenases, phospholipases, pathogenesis-related proteins, glutathione transferases/peroxidases, enzymes involved in the phenylpropanoid pathway and in the formation and detoxification of ROS, as well as those involved in the formation and regulation of ion channels, SA and NO. By analysis of the EST database of Citrus, it was possible to identify several putative genes that code for key enzymes involved in HR triggering and also in plant defense against biotic and abiotic stress.