Desenvolvimento de um biossensor portátil de campo como um dispositivo específico e confiável para a deteção precoce da doença do fogo bacteriano

Given the FCUP team's expertise in Erwinia amylovora (Ea) [6-7;14-16] and the current state of the art, which highlights the absence of effective control and preventive measures, there is a risk of significant damage to pear 'Rocha' within a 10-year timeframe [13]. This situation could potentially drive producers to resort to desperate measures, such as the unauthorized application of antibiotics [12], emphasizing the urgent need for the development of a new tool to combat this disease. Thus, Sens4FireBlight main goal is to develop and validate a new biosensor based on the genomic characterization of Ea to obtain a competitive analytical tool, that allows the early detection of the disease in apple and pear cultivars. This will enable us to tackle the constraints associated with the lack of Ea early detection whilst avoiding economic losses for the country. To achieve this, specific aims are to: 1) Identify genomic regions of Ea that can easily be used to separate it from other pathogens that affect pome fruit trees, that are suitable to be used in the detection of early stages of fire blight (Task 1); 2) Develop and validate a reliable, portable, fast, and highly selective biosensor for the early detection of the presence of Ea in plant hosts on-field (Task 2); 3) Evaluate the analytical usefulness of this new biosensor through in vitro and on-field measurements (Task 3). Sens4FireBlight (Fig.1) will be pivotal to disclose Ea genetic determinants behind its possible “evolving clonal population” in Portugal and worldwide, essential to the development of a reliable DNA-based biosensor for early detection that will prevent the severe damage caused by Ea, thus reducing the costs of inefficient cultural practices and risks of agrochemicals application [8;12]. This project presents integrated trans-inter multidisciplinary approaches through multi-omics analysis and biotechnological tools, to tackle a severe problem in pome-fruit trees. This interdisciplinary is based on the collaboration of three Leading Research teams: a) LAQV-REQUIMTE a leading group in development of electrochemical (bio)sensors; b) FCUP which stands at the forefront in Portugal for its studies on Ea ecology, genomic and phenotypic studies, sustainable control measures, and plant pathology, using omics approaches; c) BIOPOLIS currently spearheading, alongside FCUP, the whole genome sequencing (WGS) of the available collection of Ea at FCUP, exploring the genomic and transcriptomics studies of this pathogen. This collaboration will create synergies that will release original contributions to reduce yield losses of important chain-of-values pome fruits in Portugal caused by Ea, especially the important commodity pear ‘Rocha’, with socio-economic impact in Portugal: 1) The first genomic analysis of the Portuguese population of Ea; 2) New knowledge regarding specific genomic targets for Ea identification; 3) Identify key infection targets of Ea in the hosts; 4) The first field-portable electrochemical biosensor(s) for fire blight; 5) Patent a new fast, specific, and reliable Ea identification sensor Sens4FireBlight will be a worldwide reference for early detection of Ea, and this project is easily interchangeable with other plant diseases, allowing faster development of other biosensors related to different plant pathogens. Sens4FireBlight is also aligned with the European Green Deal (Farm to Fork strategy), corresponding to goals 2 and 12 of the 2030 UN Agenda for SD.