Abstract
BackgroundIn Citrus cultures, three species of Xanthomonas are known to cause distinct diseases. X. citri subsp. citri patothype A, X. fuscans subsp. aurantifolii pathotypes B and C, and X. alfalfae subsp. citrumelonis, are the causative agents of cancrosis A, B, C, and citrus bacterial spots, respectively. Although these species exhibit different levels of virulence and aggressiveness, only limited alternatives are currently available for proper and early detection of these diseases in the fields. The present study aimed to develop a new molecular diagnostic method based on genomic sequences derived from the four species of Xanthomonas.ResultsUsing comparative genomics approaches, primers were synthesized for the identification of the four causative agents of citrus diseases. These primers were validated for their specificity to their target DNA by both conventional and multiplex PCR. Upon evaluation, their sensitivity was found to be 0.02 ng/µl in vitro and 1.5 × 104 CFU ml−1 in infected leaves. Additionally, none of the primers were able to generate amplicons in 19 other genomes of Xanthomonas not associated with Citrus and one species of Xylella, the causal agent of citrus variegated chlorosis (CVC). This denotes strong specificity of the primers for the different species of Xanthomonas investigated in this study.ConclusionsWe demonstrated that these markers can be used as potential candidates for performing in vivo molecular diagnosis exclusively for citrus-associated Xanthomonas. The bioinformatics pipeline developed in this study to design specific genomic regions is capable of generating specific primers. It is freely available and can be utilized for any other model organism.
Highlights
Xanthomonas is a genus of phytopathogenic Gram-negative bacteria that cause a variety of diseases in several agricultural commodities, including citrus (Vauterin et al, 1995)
Since Xanthomonas are quarantine pests in all these pathosystems and controlling them with agrochemicals is very expensive, early detection is of paramount importance for the control and management of the diseases caused by these bacteria
Pathotype C is the cause of cancrosis C, which is characterized by induction of hypersensitivity reaction in its hosts as described in ‘Galego’ acid lime (C. aurantifolia) (Schubert et al, 2001) and ‘Swingle’ citrumelo (Citrus paradisi × Poncirus trifoliata) (Jaciani et al, 2009)
Summary
Xanthomonas is a genus of phytopathogenic Gram-negative bacteria that cause a variety of diseases in several agricultural commodities, including citrus (Vauterin et al, 1995). These diseases cause huge economic losses to the agricultural sector Bacteria from this genus have been targeted as an essential model of study in different pathosystems; for instance, X. oryzae in Oryza spp. Citrumelonis, are the causative agents of cancrosis A, B, C, and citrus bacterial spots, respectively These species exhibit different levels of virulence and aggressiveness, only limited alternatives are currently available for proper and early detection of these diseases in the fields. Primers were synthesized for the identification of the four causative agents of citrus diseases These primers were validated for their specificity to their target DNA by both conventional and multiplex PCR. The bioinformatics pipeline developed in this study to design specific
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