Abstract Background Amaranthus spp. are problematic weeds and competitors for nutrients in several crops, especially in soybean and corn. Resistance to glyphosate has been detected in several weed species, and a triple mutation in its EPSPS target gene was detected recently in Amaranthus hybridus. Objective The aim of this work was to develop a simple polymerase chain reaction (PCR) method to detect the EPSPS triple mutation in A. hybridus. Methods Two pairs of primers were designed for PCR-based detection of the EPSPS TAP-IVS triple mutation, which confers resistance to glyphosate, in A. hybridus. Results These sets of allele-specific primers were tested on five Amaranthus species and in 65 different field accessions. The PCR reaction using one set of the primers amplifies the wildtype (TAP) allele while the PCR reaction using the other pair of primers amplifies the triple mutation (IVS) allele. The presence of PCR products in both sets of primers identifies the heterozygous resistant individuals, and PCR product amplified only with the triple mutation set of primers identifies the homozygous resistant individuals. A DNA concentration test was performed and the recommend DNA amount to be used is 100 ng. Conclusions We developed and tested two sets of primers to detect the EPSPS TAP-IVS triple mutation and the results showed a 100% genotypic to phenotypic association. The triple mutation detection assay is easy to use and can be applied in a molecular laboratory with basic equipments. Early detection of resistance helps to better manage and control its spreading.

Poucos estudos foram desenvolvidos no Brasil sobre a doença hérnia das crucíferas, causada por Plasmodiophora brassicae. Realizou-se testes de severidade da doença causada por diferentes populações do patógeno em espécies de brássicas (couve-flor, couve-chinesa suscetível, couve-chinesa resistente, brócolis e repolho). As populações de P. brassicae foram obtidas de raízes de brássicas infectadas originárias de algumas regiões produtoras no Brasil. Os testes de severidade foram realizados em casa de vegetação (25±2ºC) mediante inoculações, no colo de cada planta com 2 mL da suspensão de esporos do patógeno, na concentração de 107 esporos mL-1. As avaliações ocorreram 35 dias após a inoculação. Em laboratório foi realizada a extração de DNA dessas populações e análises de PCR-RAPD para a comparação das características genéticas. As populações obtidas nas regiões de Carandaí MG e Colombo PR mostraram-se mais agressivas, manifestando patogenicidade até mesmo em cultivar considerada resistente. Entretanto, não foi observado um padrão genético específico quanto ao local de origem das populações avaliadas, sugerindo que mesmo em locais distantes e com diferenças quanto à severidade, tais populações são geneticamente semelhantes.

Few studies have been conducted with clubroot disease in Brazil, caused by Plasmodiophora brassicae. Tests on severity of disease using different populations were carried out in brassica species (cauliflower, susceptible Chinese cabbage, resistant Chinese cabbage, broccoli, and cabbage). P. brassicae populations were obtained from infected roots in several commercial production regions in Brazil. Severity tests were conducted under greenhouse conditions (25±2ºC) with inoculations (2 mL) of spore suspension at concentration of 10(7) spores. mL-1, applied in the collar root of each plant. Disease evaluations were performed 35 days after inoculation. DNA extraction and PCR-RAPD analyses for these populations were conducted to compare genetic traits. The populations obtained from the regions of Carandaí MG and Colombo PR was more aggressive, and cause symptoms even in cultivars considered resistant. However, no specific genetic pattern was observed as to the place of origin or aggressiveness.

Recentemente, observaram-se plantas de erva-baleeira (Cordia verbenácea Al. DC) sendo atacadas pelo percevejo Dictyla monotropidia Stål. As colônias foram encontradas na superfície abaxial das folhas, onde ninfas e adultos sugavam a seiva do floema, causando encarquilhamento, seguido de amarelecimento e queda de folhas. D. monotropidia já foi relatada como praga de outras espécies de Cordia em países das Américas Central e do Sul, entretanto, este é o primeiro relato do inseto atacando plantas de C. verbenacea no Brasil.

Recently, Dictyla monotropidia Stål was observed feeding on plants of black sage (Cordia verbenacea Al. DC). The colonies of this insect were observed on abaxial surface of leaves, with nymphs and adults sucking the phloem sieve, causing spot, yellow aspect and leaf fall. D. monotropidia was already related as pest in other Cordia species in countries of Central and South America. Although, this is the first report of this insect attacking plants of C. verbenacea in Brazil.

In silico expression profiles, of the discovered 3,103 citrus ESTs putatively encoding for PR protein families (PR-1 to PR-17), were evaluated using the Brazil citrus genome EST CitEST/database. Hierarchical clustering was displayed to identify similarities in expression patterns among citrus PR-like gene families (PRlgf) in 33 selected cDNA libraries. In this way, PRlgf preferentially expressed by organ and citrus species, and library conditions were highlighted. Changes in expression profiles of clusters for each of the 17 PRlgf expressed in organs infected by pathogens or drought-stressed citrus species were displayed for relative suppression or induction gene expression in relation to the counterpart control. Overall, few PRlgf showed expression 2-fold higher in pathogen-infected than in uninfected organs, even though the differential expression profiles displayed have been quite diverse among studied species and organs. Furthermore, an insight into some contigs from four PRlgf pointed out putative members of multigene families. They appear to be evolutionarily conserved within citrus species and/or organ- or stress-specifically expressed. Our results represent a starting point regarding the extent of expression pattern differences underlying PRlgf expression and reveal genes that may prove to be useful in studies regarding biotechnological approaches or citrus resistance markers.

Due to the economic importance of gummosis disease for the citriculture, studies on P. parasitica-Citrus interaction comprise a significant part in the Brazilian Citrus genome data bank (CitEST). Among them, two cDNA libraries constructed from two different growth conditions of the P. parasitica pathogen are included which has generated the PP/CitEST database (CitEST - Center APTA Citros Sylvio Moreira/IAC- Millennium Institute). Through this genomic approach and clustering analyses the following has been observed: out of a total of 13,285 available in the Phytophthora parasitica database, a group of 4,567 clusters was formed, comprising 2,649 singlets and 1,918 contigs. Out of a total of 4,567 possible genes, only 2,651 clusters were categorized; among them, only 4.3% shared sequence similarities with pathogenicity factors and defense. Some of these possible genes (103) corresponding to 421 ESTs, were characterized by phylogenetic analysis and discussed. A comparison made with the COGEME database has shown homology which may be part of an evolutionary pathogenicity pathway present in Phytophthora and also in other fungi. Many of the genes which were identified here, which may encode proteins associated to mechanisms of citrus gummosis pathogenicity, represent only one facet of the pathogen-host Phytophthora - Citrus interaction.