Medium for Isolation and Growth of Bacteria Associated with Plum Leaf Scald and Phony Peach Diseases

Wells, John M.; Raju, B. C.; Nyland, G.; Lowe, S. K.

doi:10.1128/aem.42.2.357-363.1981

Cited by 87 publications

(36 citation statements)

References 22 publications

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“…The 120 other 57 strains were provided by the French Collection of Plant-Associated Bacteria (CIRM-CFBP; 121 https://www6.inra.fr/cirm_eng/CFBP-Plant-Associated-Bacteria). Xf strains were grown on BCYE 122 (Wells et al, 1981) or modified PWG media (agar 12 g.L -1 ; soytone 4 g.L -1 ; bacto tryptone 1 g.L -123 1 ; MgSO4.7H2O 0,4 g.L -1 ; K2HPO4 1.2 g.L -1 ; KH2PO4 1 g.L -1 ; hemin chloride (0.1% in NaOH 0.05 124 M) 10 ml.L -1 ; BSA (7.5%) 24 ml.L -1 ; L-glutamine 4 g.L -1 ) at 28°C for one to two weeks. Other strains 125…”

Section: Bacterial Strains and Growth Conditions 115mentioning

confidence: 99%

Novel tetraplex qPCR assays for simultaneous detection and identification ofXylella fastidiosasubspecies in plant tissues

Dupas

Briand

Jacques

et al. 2019

Preprint

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AbstractXylella fastidiosa is an insect-borne bacterium confined to the xylem vessels of plants. This plant pathogen has a broad host range estimated to 560 plant species. Five subspecies of the pathogen with different but overlapping host ranges have been described, but only three subspecies are widely accepted, namely subspecies fastidiosa, multiplex and pauca. Initially limited to the Americas, Xf has been detected in Europe since 2013. As management of X. fastidiosa outbreaks in Europe depends on the identification of the subspecies, accurate determination of the subspecies in infected plants as early as possible is of major interest. Thus, we developed various tetraplex and triplex qPCR assays for Xylella fastidiosa detection and subspecies identification in planta in a single reaction. We designed primers and probes using SkIf, a bioinformatics tool based on k-mers, to detect specific signatures of the species and subspecies from a dataset of 58 genome sequences representative of X. fastidiosa diversity. We tested the qPCR assays on 39 target and 30 non-target strains, as well as on 13 different plant species spiked with strains of the different subspecies of X. fastidiosa, and on samples from various environmental and inoculated host plants. Sensitivity of simplex assays was equal or slightly better than the reference protocol on purified DNA. Tetraplex qPCR assays had the same sensitivity than the reference protocol and allowed X. fastidiosa detection in all spiked matrices up to 103 cells.mL−1. Moreover, mix infections of two to three subspecies could be detected in the same sample with tetraplex assays. In environmental plant samples, the tetraplex qPCR assays allowed subspecies identification when the current method based on multilocus sequence typing failed. The qPCR assays described here are robust and modular tools that are efficient for differentiating X. fastidiosa subspecies directly in plant samples.

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Section: Bacterial Strains and Growth Conditions 115mentioning

confidence: 99%

Novel tetraplex qPCR assays for simultaneous detection and identification ofXylella fastidiosasubspecies in plant tissues

Dupas

Briand

Jacques

et al. 2019

Preprint

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“…PD2 (Davis et al . ), BCYE (Wells et al ., ) or PWG (modified after Hill & Purcell, ) are widely used for the isolation from different host species. Media are described in Appendix .…”

Section: Detectionmentioning

confidence: 99%

PM 7/24 (2) Xylella fastidiosa

2016

EPPO Bulletin

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“…All the samples showing symptoms similar to those described on olive, almond, oleander and other ornamentals in Southern Italy (Martelli 2013;Regione Puglia 2014) were used in attempts to isolate the bacterium on different media, according to the procedures described by Cariddi et al (2014). Periwinkle wilt gelrite (PWG) (Hill and Purcell 1995), buffered cysteine-yeast extract (BCYE) (Wells et al 1981), Pierce Disease-3 (PD3) (Davis et al 1980) and X. fastidiosa-D1 (Almeida et al 2004) media were used. The plates were incubated at 28°C for 4 weeks and examined with a stereomicroscope at weekly intervals for the presence of Xylella-like colonies (Wells et al 1987).…”

Section: Isolation On Semi-selective Agar Mediamentioning

confidence: 99%

Xylella fastidiosa Does Not Occur in Lebanon

Habib

Nigro

Gerges

et al. 2016

Journal of Phytopathology

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Xylella fastidiosa has been reported as responsible for a devastating disease on olive trees in Apulia region (south-eastern Italy), characterized by a quick decline syndrome. In Lebanon, the pathogen was recently associated with leaf scorch symptoms on oleander, and reports on leaf scorch and dieback of olive trees branches by technicians and farmers have shown an increasing trend in the main agricultural areas. To assess the occurrence and distribution of the pathogen in Lebanon, samples of twigs from olive trees (82), olive seedlings (26), grapevine (30), oleander (32) and ornamentals imported from Italy (48) were analysed by isolation on four agarized media, serological techniques (ELISA and DTBIA) using Xylella fastidiosa-specific antibodies and by PCR, using three specific sets of primers. Results unequivocally demonstrated that all the collected samples were free from the pathogen. As well, both detection protocols and attempts at isolating the pathogen on agarized media demonstrated that oleander samples gathered from American University campus in Beirut, where X. fastidiosa was previously reported, were not infected. Nevertheless, continuous monitoring and rigorous control measures of propagative materials are necessary to prevent the introduction of Xylella fastidiosa in Lebanon.

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Medium for Isolation and Growth of Bacteria Associated with Plum Leaf Scald and Phony Peach Diseases

Cited by 87 publications

References 22 publications

Novel tetraplex qPCR assays for simultaneous detection and identification ofXylella fastidiosasubspecies in plant tissues

Novel tetraplex qPCR assays for simultaneous detection and identification ofXylella fastidiosasubspecies in plant tissues

PM 7/24 (2) Xylella fastidiosa

Xylella fastidiosa Does Not Occur in Lebanon

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