RAPD-PCR Fingerprinting and Southern Analysis of Xanthomonas axonopodis pv. dieffenbachiae Strains Isolated from Different Aroid Hosts and Locations

Abstract

Anthurium blight, caused by Xanthomonas axonopodis pv. dieffenbachiae, is a systemic disease of Anthurium and other aroids. The aims of this work were to study the genetic diversity among X. axonopodis pv. dieffenbachiae strains and to identify, from the polymerase chain reaction (PCR) profiles, DNA probes that would be specific for the pathovar dieffenbachiae. Twenty-five X. axonopodis pv. dieffenbachiae strains, isolated from different hosts and geographical locations including Mauritius, were fingerprinted using the random amplified polymorphic DNA (RAPD)-PCR technique. The fingerprints were analyzed by the National Taxonomy System Software (NTSYS). The specificity of some of the RAPD fragments selected from PCR profiles was tested by Southern analyses of the PCR products. Ten arbitrary primers were chosen from an initial set of 111 decamers. Two hundred and nine RAPD markers were generated in eight individual DNA profiles. A correlation was found between the serotypes and the RAPD profiles for some groups of isolates. A possible link was also observed between the host range of the isolates tested and their RAPD profiles for strains isolated from Dieffenbachia and Philodendron. These results were confirmed by Southern analysis. Cluster analysis by the unweighted pair group method, arithmetic average (UPGMA) confirmed that the pathovar is genetically diverse with some strains that were clustered together showing similar host preferences. DNA probes with a potential use in molecular diagnostics of Anthurium blight were identified. This preliminary work could be used to develop PCR primers that will enable the sensitive detection of the pathogen in latently infected plants.

Analysis of Molecular Variability Among Isolates of Aspergillus Flavus by PCR-RFLP of the its Regions of rDNA

“…This is why, the ability to distinguish between the various Aspergillus species may have diagnostic value. The analysis of genomic DNA using PCR-based methods has proven to be a fast, sensitive and reliable method for determining genetic relationships among pathogenic microorganisms (Zhang et al 2004;Khoodoo and Jaufeerally-Fakim 2004). Nuclear rDNA, and particularly the internal transcribed spacer (ITS) regions are good targets for the phylogenetic analysis in fungi (Bruns et al 1991) because the ITS regions are often highly variable between isolates of the same species (O'Donnell et al 1998;Salazar et al 1999).…”

Ra Pd-Analys Is of Pseudomonas Syringae, Isolated From Weeds in Agro Phyto Cenos Is of Wheat

“…У більшості досліджень генетичного поліморфізму фітопатогенних бак-терій, ізольованих з різних рослин, штами об'єднувалися у групи зі спільною рослиною хазяїном [7,12]. 2.…”

“…RAPD-ПЛР аналіз успішно використову-ється для генетично-популяційного аналізу широкого кола мікроорганізмів. Переваги RAPD-ПЛР аналізу полягають і у тому, що він дозволяє встановити генетичну варіабельність цілого геному, порівняно з методами гібридизації рибосомальної РНК [7].…”

Sensitive detection of Xanthomonas axonopodis pv. dieffenbachiae on Anthurium andreanum by immunocapture-PCR (IC-PCR) using primers designed from sequence characterized amplified regions (SCAR) of the blight pathogen

“…Cloning, sequencing of fragments, and design of specific primers PCR products amplified by RAPD primers as recently reported (Khoodoo and Jaufeerally-Fakim, 2004) were purified from agarose gels by the Freeze and SqueezeÒ columns (Biorad) and were cloned in a pGEMÒ-T Easy plasmid vector (Promega, Madison, WI, USA). Recombinant E. coli cells were used for plasmid minipreps using the WizardÒPlus SV Miniprep kit (Promega) and sequencing was performed by the Spectrumedix SCE 2410 Genetic Analysis System and the BigDye Version 3.1 kit (ABI, Inqaba Biotechnical Industries, SA).…”

“…dieffenbachiae (Khoodoo and Jaufeerally-Fakim, 2004). This study describes the development of a sensitive PCR tool from the sequences of DNA probes for an early and quick diagnosis of Anthurium blight in plant material.…”