Melon and watermelon bacterial fruit blotch, incited by Acidovorax citrulli, is limited to some areas in Brazil but causes important losses, mainly in melon-producing regions. Although genetic diversity has been observed among strains belonging to the species, they are considered a homogeneous group based on the fact that they show only slight physiological or nutritional differences. The objective of this study was to compare Brazilian strains from melon and watermelon by means of biochemical, pathogenicity, serological and molecular assays. Fifteen biochemical tests, cross inoculation between strains and hosts, ELISA and repetitive sequence analysis (rep-PCR) with the primers REP, ERIC and BOX were conducted. No differences were revealed by nutritional characterization or serology, but cross inoculation showed different pathogenicity groups, which could explain high aggressiveness of the bacteria to melon crops in some regions. Molecular analysis by BOX-PCR clustered strains according to their geographical origin, while ERIC- and REP-PCR, analyzed together, indicated genetic diversity, but without geographical or host origin relationships. One test that could be used to verify the pathogenicity of strains by inoculating detached leaf petioles, showing results in 36 h, is proposed here.

O algodoeiro é atacado por Colletotrichum gossypii (CG) e C. gossypii var. cephalosporioides (CGC). Ambos os patógenos são transmitidos pela semente e sua distinção morfológica é extremamente difícil e inconsistente. Tentativas foram feitas no presente trabalho para verificar a variabilidade genética entre CG e CGC através de RAPD-PCR, ERIC- e REP-PCR e PCR-RFLP da região ITS rDNA. Foram utilizados 53 isolados coletados de sementes e folhas de plantas de diferentes cultivares nos estados do Paraná, São Paulo, Mato Grosso, Minas Gerais, e Paraiba, entre 1999 e 2003. Baseado em testes de patogenicidade, vinte e um isolados foram classificados como CG e 32 como CGC. Os resultados obtidos por RAPD-PCR, utilizando-se oito primers, revelaram dois grupos distintos sendo que o primeiro foi formado por 94% dos isolados de sementes e o segundo por 95% dos isolados de folhas. Na análise de ERIC- e REP-PCR, resultados semelhantes a RAPD foram obtidos, sendo que o primeiro grupo foi formado por 93% dos isolados provenientes das sementes e o segundo por 78% dos isolados provenientes das folhas. Quando o produto de amplificação da região ITS rDNA foi digerido com oito enzimas de restrição, um perfil de bandas semelhante para todos os isolados foi obtido. Resultados de RAPD, ERIC- e REP-PCR demonstraram que existem diferenças genéticas entre os isolados provenientes das sementes e aqueles provenientes de parte aérea, e esses dois grupos foram claramente distintos. Estudos futuros devem ser realizados utilizando outras técnicas moleculares para a obtenção de marcadores capazes de distinguir entre isolados de CG e CGC.

Cotton is attacked by Colletotrichum gossypii (CG) and C. gossypii var. cephalosporioides (CGC). Both the pathogens are transmitted by seed and their morphological distinction is extremely difficult and inconsistent. In the present study, attempts were made to verify the genetic variability among 53 isolates of CG and CGC using RAPD-PCR, ERIC- and REP-PCR, as well as PCR-RFLP of the ITS rDNA region. Based on pathogenicity tests, 21 isolates were classified as CG and 32 as CGC. RAPD analysis was performed using eight primers and the results revealed two major groups. The first group contained 94% of the isolates originated from seeds and the second contained 95% of the isolates originated from leaves. Similar results were obtained by ERIC- and REP-PCR, where the first group contained 93% of the isolates originated from seeds and the second contained 78% of the isolates originated from leaves. When the amplified product of the ITS rDNA region was digested with eight restriction enzymes, similar banding patterns were observed for all isolates. Results of RAPD, ERIC- and REP-PCR demonstrated the existence of genetic variability among the seed and leaf isolates and these two groups could be clearly distinguished. Further studies need to be performed using other molecular techniques in order to obtain markers that can distinguish CG and CGC isolates in infected seeds.

A mancha angular do algodoeiro, causada por Xanthomonas axonopodis pv. malvacearum (Xam), é uma doença de importância econômica e pode causar perdas apreciáveis no rendimento. A doença pode ser controlada por resistência varietal desde que haja conhecimento sobre a variabilidade das populações do patógeno. A variabilidade genética e a estabilidade patogênica entre os isolados deste patógeno não foram suficientemente estudadas, principalmente considerando a introdução de novas cultivares e a expansão da cultura. O objetivo do presente estudo foi avaliar a variabilidade genética entre os 61 isolados de Xam, provenientes de diversas cultivares e regiões do Brasil, através de ensaios moleculares. Análises de ERIC e REP-PCR demonstraram dois grupos distintos de Xam associados a região geográfica de origem. Não foram observadas diferenças nos perfis dos isolados através de PCR-RFLP da região 16S-23S rDNA. A região espaçadora 16S-23S rDNA de três linhagens de Xam foi analisada através de clonagem e sequenciamento e seis diferenças nas seqüências foram encontradas. A técnica de RAPD revelou um maior nível de polimorfismo, distinguindo 6 grupos de Xam a 85% de similaridade. Os resultados indicam a existência de variabilidade muito restrita entre os isolados analisados.

Angular leaf spot or black arm of cotton, caused by Xanthomonas axonopodis pv. malvacearum (Xam), is a disease of economic importance. The disease can be controlled by using resistant cultivars if the knowledge regarding the genetic variability of the pathogen population is available. The genetic variability and the pathogen stability among the isolates of this pathogen have not been sufficiently studied, especially considering the introduction of new cultivars and the expansion of areas under cotton cultivation. The objective of the present study was to identify genetic variability among 61 isolates of Xam collected from different cultivars and geographic regions. Analysis of ERIC and REP-PCR revealed two distinct groups of Xam based on the geographic region of isolation. PCR-RFLP of 16S-23S rDNA did not reveal differences in the restriction profiles of the isolates. The 16S-23S rDNA region of three isolates of Xam was also analysed by cloning and sequencing and 6 differences in the sequences were observed. The RAPD technique revealed a higher level of polymorphism and distinguished 6 groups of Xam. The results indicate a very narrow genetic variability among Xam isolates used in this study.

The Bemisia tabaci complex is formed by approximately 41 biotypes, two of which (B and BR) occur in Brazil. In this work we aimed at obtaining genetic markers to assess the genetic diversity of the different biotypes. In order to do that we analyzed Bemisia tabaci biotypes B, BR, Q and Cassava using molecular techniques including RAPD, PCR-RFLP and sequencing of the ITS1 rDNA region. The analyses revealed a high similarity between the individuals of the B and Q biotypes, which could be distinguished from the BR individuals. A phylogenetic tree based on ITS1 rDNA sequence was constructed. This is the first report of the ITS1 rDNA sequence of Bemisia tuberculata and of the BR biotype of B. tabaci.

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.

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.

Xanthomonas axonopodis pv. citri is a phytopathogenic bacterium responsible for citrus canker, a serious disease which causes severe losses in citriculture around the world. In this study we report the differential expression of X. axonopodis pv. citri in response to specific treatments by using Representational Difference Analysis of cDNA (cDNA RDA). cDNAs from X. axonopodis pv. citri cultured in the presence of leaf extract of the host plant (Citrus sinensis), in vivo, as well as in the complex medium were hybridized against cDNA of the bacterium grown in the minimal medium. Sequencing of the difference products obtained after the second and third hybridizations revealed a total of 37 distinct genes identified by homology searches in the genome of X. axonopodis pv. citri. These genes were distributed in different functional categories, including genes that encode hypothetical proteins, genes involved in metabolism, cellular processes and pathogenicity, and mobile genetic elements. Most of these genes are likely related to growth and/or acquisition of nutrients in specific treatments whereas others might be important for the bacterium pathogenicity.