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Istituto Agronomico Mediterraneo di Bari (IAMB) 2019

Development of advanced and innovative diagnostic assays for the detection of Xylella fastidiosa in plants and insect-vector

Dakroub, Hiba

Titre : Development of advanced and innovative diagnostic assays for the detection of Xylella fastidiosa in plants and insect-vector

Auteur : Dakroub, Hiba

Etablissement de soutenance : Istituto Agronomico Mediterraneo di Bari (IAMB) - Centre international de hautes études agronomiques méditerranéennes (CIHEAM)

Grade : Master of science : Integrated pest management of Mediterranean fruit crops ; 2019

Résumé
Although DNA- and protein-based methods have revolutionized Xylella fastidiosa diagnosis, in some cases they are not very reliable due, on one hand, to the peculiarities of this pathogen, i.e. (i) irregular distribution inside the plant, (ii) xylem-limited localization making its extraction difficult, (iii) latency in some species that can affect accurate sampling ; (iv) low concentration in some hosts ; (v) genomic variance between subspecies ; and, on the other hand, the poor performance of some serological and molecular techniques to sensitively and differentially detect and discriminate between subspecies and strains of Xylella fastidiosa. Accordingly, the aim of this study was to develop advanced serological (Nanobodies) and molecular (Real-time TaqMan PCR, LAMP and FTPS-LAMP) assays able to detect Xylella fastidiosa, especially the CoDiRO strain present in Apulia, in an easy, sensitive, accurate and discriminating manner . The hemagglutinin gene, found solely in the CoDiRO genome, together with two universal genes of Xylella fastidiosa, were used for the construction of protein- and DNA-based techniques, respectively. These three recombinant proteins were bacterially expressed, purified and quantified for future injections in rabbit and Llama for the production of polyclonal and monoclonal antibodies against Xylella fastidiosa. Real-time TaqMan PCR, LAMP and FTPS-LAMP assays here developed in singleplex and multiplex approaches were found highly sensitive, specific and discriminant to detect Xylella fastidiosa in infected plants and insect-vectors.

Mots clés : CODIRO ; LAMP ; NANOBODIES ; REAL-TIME TAQMAN PCR ; XYLELLA FASTIDIOSA

Présentation (IAMB Library)

Page publiée le 14 mai 2020