A new inclusive MLVA assay to investigate genetic variability of Xylella fastidiosa with a specific focus on the Apulian outbreak in Italy

Abstract

The Olive Quick Decline Syndrome by Xylella fastidiosa subspecies pauca is among the most severe phytopathological emergencies nowadays. In few years, the outbreak devastated olive groves in Apulia (Italy), potentially endangering the entire Mediterranean basin. This research aimed to develop a multiple locus VNTR analysis assay, a molecular tool to differentiate between populations of the pathogen. It has already been successfully applied to different X. fastidiosa subspecies from various plant hosts. The previously published TR loci, together with a set of new design, have been tested in silico on the genome of the Apulian De Donno strain. The resulting selection of 37 TR loci was amplified on the genomic DNAs of the Apulian strains and from representatives of X. fastidiosa subspecies, and directly on DNA extracted from infected plants. The assay clearly discerned among subspecies or even sequence types (ST), but also pointed out variants within the same ST so as to provide more detailed information on the dynamics and pathogen diffusion pathways. Its effective application even on total DNAs extracted from infected tissues of different host plants makes it particularly useful for large-scale screening of infection and for the strengthening of containment measures.

Suspicions of two bridgehead invasions ofXylella fastidiosasubsp.multiplexin France

“…Another strength of our MLVA scheme is that it has been developed and optimized to be directly usable on genomic DNA extracted from fresh or frozen infected plant material, as it has already shown to be possible for Italian samples infected with X. fastidiosa subsp. pauca ST53 ( 47 ). Our study further confirms the interest of such direct genotyping techniques.…”

Enfermedades emergentes y reemergentes de plantas en Latinoamérica: una revisión.

“…Xylella fastidiosa es una especie de bacteria Gram negativa, la cual fue reportada inicialmente en Estados Unidos, causa la enfermedad de Pierce (PD) en la vid y es transmitida por vectores de la subfamilia Cicadellinae, superfamilia Cercopoidea y potencialmente de la superamilia Cicadoidea (Mazzaglia et al, 2020) que se alimentan del xilema (Almeida & Nunney, 2015;Chen et al, 2007). X. fastidiosa presenta seis subespecies, cada una con un rango y distribución geográfica específicos: X. fastidiosa subsp.…”

“…tashke, causando la quemadura de la hoja de chitalpa en EE. UU (Mazzaglia et al, 2020). La distribución geográfica del género Xylella sugiere que las diferentes especies evolucionaron separadamente, aumentando su variabilidad genética por introducción de cepas de X. fastidiosa a nuevas áreas geográficas y recombinación con cepas endémicas (Mazzaglia et al, 2020;Nunney et al, 2014).…”

“…Por otra parte, debe mencionarse el caso contrario. Algunas especies patógenas se especializan en determinado número de especies de plantas e incluso se encuentran circunscritas dentro de determinada región geográfica dando lugar a nuevos grupos, como es el caso de Xylella fastidiosa y sus subespecies (Mazzaglia et al, 2020).…”

Rapid Discrimination Among Methicillin Resistant Staphylococcus Aureus Isolates Using Variable Number Tandem Repeat Analysis in a Sample of Iraqi Patients And

“…Pulse Field Gel Electrophoresis (PFGE) (16), Multilocus Sequence Typing (MLST) (5) and spa-sequence typing (10) are the most used typing techniques worldwide. Recently a technique known MLVA which is multiplelocus variable-number tandem repeat analysis that knowing by its ability to typing bacterial pathogens in large number (4,7,8,11,19,20,21). Epidemiological studies has been done through creation a DNA profiles by using the differences in the sequence numbers of short tandem repeat that knowing as VNTR.…”