Alsaheli, Zeinab
(2020)
Investigation and characterization of viruses and phytoplasmas infecting fig trees, [Dissertation thesis], Alma Mater Studiorum Università di Bologna.
Dottorato di ricerca in
Scienze e tecnologie agrarie, ambientali e alimentari, 32 Ciclo. DOI 10.6092/unibo/amsdottorato/9261.
Documenti full-text disponibili:
|
Documento PDF (English)
- Richiede un lettore di PDF come Xpdf o Adobe Acrobat Reader
Disponibile con Licenza: Salvo eventuali più ampie autorizzazioni dell'autore, la tesi può essere liberamente consultata e può essere effettuato il salvataggio e la stampa di una copia per fini strettamente personali di studio, di ricerca e di insegnamento, con espresso divieto di qualunque utilizzo direttamente o indirettamente commerciale. Ogni altro diritto sul materiale è riservato.
Download (2MB)
|
Abstract
Early detection of fig mosaic disease (FMD) is considered as a fundamental step for an appropriate disease management through the production of pathogen-free plant material. Accordingly, singleplex and multiplex-TaqMan RT-PCR assays were developed to detect single and multiple infections of seven fig viruses, i.e. fig leaf mottle-associated virus 1 (FLMaV-1), fig leaf mottle-associated virus 2 (FLMaV-2), fig mild mottle-associated virus (FMMaV), fig mosaic virus (FMV), fig latent virus 1 (FLV-1), fig cryptic virus 1 (FCV-1) and fig fleck-associated virus (FFkaV). The newly designed primers and probes were constructed upon alignment of viral nucleotide sequences of different isolates retrieved from GenBank. The new techniques were validated and found to be more sensitive and reliable than RT-PCR. Multiplex PCR was able to detect with high efficiency up to five fig viruses simultaneously. These two diagnostic techniques can be useful to control and limit fig virus spread and to support quarantine and certification programs. On the other hand, besides to many viral and fungal diseases, nothing was reported before on phytoplasma infections in fig plants. A small-scale survey was therefore conducted in a fig plot for varietal collection (Locorotondo, South of Italy) to investigate the possible presence of phytoplasmas. Forty-three samples were collected and subjected to PCR and nested PCR assays using universal primers that amplify the 16S rDNA gene. Twenty plants were shown positive to phytoplasma. Sequence analysis generated three sequences sharing the highest similarity with ‘Ca. P. asteris’ and ‘Ca. P. solani’, belonging to the 16SrI and 16SrXII ribosomal groups, respectively. The presence of two phylogenetically distinct phytoplasmas was further verified with group specific qPCR assays and RFLP with TruI conducted on 16S rDNA amplified with M1/M2 and fU5/rU3 primers. This is the first record of phytoplasma occurrence in fig plants.
Abstract
Early detection of fig mosaic disease (FMD) is considered as a fundamental step for an appropriate disease management through the production of pathogen-free plant material. Accordingly, singleplex and multiplex-TaqMan RT-PCR assays were developed to detect single and multiple infections of seven fig viruses, i.e. fig leaf mottle-associated virus 1 (FLMaV-1), fig leaf mottle-associated virus 2 (FLMaV-2), fig mild mottle-associated virus (FMMaV), fig mosaic virus (FMV), fig latent virus 1 (FLV-1), fig cryptic virus 1 (FCV-1) and fig fleck-associated virus (FFkaV). The newly designed primers and probes were constructed upon alignment of viral nucleotide sequences of different isolates retrieved from GenBank. The new techniques were validated and found to be more sensitive and reliable than RT-PCR. Multiplex PCR was able to detect with high efficiency up to five fig viruses simultaneously. These two diagnostic techniques can be useful to control and limit fig virus spread and to support quarantine and certification programs. On the other hand, besides to many viral and fungal diseases, nothing was reported before on phytoplasma infections in fig plants. A small-scale survey was therefore conducted in a fig plot for varietal collection (Locorotondo, South of Italy) to investigate the possible presence of phytoplasmas. Forty-three samples were collected and subjected to PCR and nested PCR assays using universal primers that amplify the 16S rDNA gene. Twenty plants were shown positive to phytoplasma. Sequence analysis generated three sequences sharing the highest similarity with ‘Ca. P. asteris’ and ‘Ca. P. solani’, belonging to the 16SrI and 16SrXII ribosomal groups, respectively. The presence of two phylogenetically distinct phytoplasmas was further verified with group specific qPCR assays and RFLP with TruI conducted on 16S rDNA amplified with M1/M2 and fU5/rU3 primers. This is the first record of phytoplasma occurrence in fig plants.
Tipologia del documento
Tesi di dottorato
Autore
Alsaheli, Zeinab
Supervisore
Co-supervisore
Dottorato di ricerca
Ciclo
32
Coordinatore
Settore disciplinare
Settore concorsuale
Parole chiave
Fig, mosaic, viruses, single and multiple detection, phytoplasmas,RT-PCR, PCR, Real-time PCR, RFLP and sequence analysis
URN:NBN
DOI
10.6092/unibo/amsdottorato/9261
Data di discussione
27 Marzo 2020
URI
Altri metadati
Tipologia del documento
Tesi di dottorato
Autore
Alsaheli, Zeinab
Supervisore
Co-supervisore
Dottorato di ricerca
Ciclo
32
Coordinatore
Settore disciplinare
Settore concorsuale
Parole chiave
Fig, mosaic, viruses, single and multiple detection, phytoplasmas,RT-PCR, PCR, Real-time PCR, RFLP and sequence analysis
URN:NBN
DOI
10.6092/unibo/amsdottorato/9261
Data di discussione
27 Marzo 2020
URI
Statistica sui download
Gestione del documento: