Estudo molecular e bioquímico de cultivares de algodão em resposta a Colletotrichum gossypii South var.cephalosporioides A.S. Costa

Abstract

Cotton is a crop of considerable importance worldwide, the production of which is affected by diverse biotic and abiotic factors. Stress caused by insect attacks and phytopathogens stand out among the biotic factors, the control of which is significantly hindered due the variability of these organisms and their consequent capacity for adaptation. Fungi are the phytopathogens that most cause damage to cotton plants, especially Colletotrichum gossypii South var. cephalosporioides A.S. Costa, a ramulosis-causing disease characterized by dwarfism and an over-budding of the branches, affecting the development of the buds and number of bolls. The main form of dissemination is through contaminated seeds and the most effect method of control is by means of chemical fungicides or resistant varieties which are obtained after exhaustive tests conducted in fieldwork and greenhouses. The use of biochemical and molecular tools that assist in the identification of resistant genotypes in improvement programs is of considerable importance, as they offer reliability and reduce selection costs in the development process of new varieties. In the present study, a biochemical/molecular study was carried out on cotton varieties submitted toinfection with the ramulosis fungus with the aim of identifying markers associated to the resistance process. In the first assay, biochemical descriptors were assessed in four infected cotton varieties. The plants were inoculated 20 days after cultivation, receiving a fungal concentration of 1 x 106 conidias/mL. Evaluations occurred at 3, 15 and 30 days following inoculation. Proline, peroxidase and catalase levels were quite expressive in the resistant BRS Antares cultivar, exhibiting a rapid response at 3 days after inoculation. Proline and catalase levels are indicated as useful tools for the identification of ramulosis-resistant varieties in cotton crop selection studies. The second assay sought to identify differentially expressed transcripts in cotton plants submitted to infection with the ramulosis fungus. Two varieties with opposing traits regarding resistance to this disease were used: BRS Antares and CNPA Precoce I, respectively resistant and susceptible. Ten RAPD oligonucleotides were used in the RT-PCR reactions at an annealing temperature of 35 ºC. Following an analysis of the band patterns in the control and inoculated plants, 12 down-regulated transcripts, 13 overregulated transcripts and ten activated transcripts were identified. A band ofapproximately 200 bp, obtained with the V10 oligonucleotide, was sequenced and exhibited 86% homology with the GmChl 3 gene, which codifies for thechlorophyllase III enzyme in soybean. This enzyme has considerable importance for vegetal cells submitted to oxidative damage, as it reduces this damage and modulates different defense pathways. The data obtained in the present study are applicable to studies on cotton plant responses to pathogen attacks and offers relevant information for crop improvement programs.