Development of a duplex-PCR for differential diagnosis of Xanthomonas phaseoli pv. manihotis and Xanthomonas cassavae in cassava (Manihot esculenta)

Flores, Carolina; Zarate, Carlos; Triplett, Lindsay R.; Maillot-Lebon, Véronique; Moufid, Yassine; Kante, Moussa; Bragard, Claude; Gagnevin, Lionel; Szurek, Boris; Robène, Isabelle

doi:10.1016/j.pmpp.2018.07.005

Cited by 5 publications

(4 citation statements)

References 29 publications

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“…The second approach was based on a multiplexed nested PCR including a broadly conserved fragment of Xpm transcription activator‐like (TAL) effector genes and a semispecific region of rpoB , resulting in a wider detection potential for Xpm strains (Bernal‐Galeano et al, 2018 ). The third strategy, consisting of a duplex PCR amplifying highly conserved chromosomal regions in Xpm and Xc to detect and distinguish both cassava pathogens, proved to be highly selective and sensitive (Flores et al, 2019 ).…”

Section: Pathogen Descriptionmentioning

confidence: 99%

Cassava diseases caused byXanthomonas phaseolipv.manihotisandXanthomonas cassavae

Zárate-Chaves¹,

Cruz

López

et al. 2021

Molecular Plant Pathology

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Cassava is an important staple crop as a source of food and income for hundreds of millions of people in tropical countries. This major crop is threatened by several pests and pathogens that significantly affect productivity. Among bacterial pathogens, the vascular and systemic Xanthomonas phaseoli pv. manihotis (Xpm) has received considerable attention due to its devastating potential in the tropics and scientific importance worldwide (Mansfield et al., 2012). This pathogen profile describes the main hallmarks of this pathosystem and updates several reviews and original articles on the subject.Although more bacterial pathogens infect cassava, information in the literature is scarce. In this regard, we also present a compilation of information for a second xanthomonad infecting cassava, the nonvascular pathogen X. cassavae. | D IS E A S E DE SCRIP TI ON AND EPIDEMI OLOGY | Cassava bacterial blight symptomsCassava bacterial blight (CBB), which is caused by the bacterial pathogen Xpm, is characterized by a range of symptoms that

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Section: Pathogen Descriptionmentioning

confidence: 99%

Cassava diseases caused byXanthomonas phaseolipv.manihotisandXanthomonas cassavae

Zárate-Chaves¹,

Cruz

López

et al. 2021

Molecular Plant Pathology

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“…10 3 (cPCR) and 10 2 CFU ml -1 (qPCR). For other Xanthomonas spp., pathogenic on cassava, it was shown that a DNA purification step aided the diagnosis by concentrating DNA and eliminating PCR inhibitors, thus decreasing the detection limit (Flores et al 2019). In our hands the highest sensitivity was obtained by combining bacterial population enrichment and real-time PCR.…”

Section: Discussionmentioning

confidence: 73%

Detection of Xanthomonas citri pv. viticola on grapevine by real-time PCR and BIO-PCR using primers designed from pathogenicity and xanthomonadin gene sequences

et al. 2019

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Grapevine bacterial canker caused byXanthomonas citri pv. viticola (X. campestris pv. viticola) (Xcv), was detected in Brazil in 1998 and is currently regarded as a quarantine disease with limited distribution in the country. To improve sensitivity and speed in the detection of Xcv in asymptomatic grapevines, two pairs of primers were designed, targeting sequences of a pathogenicity gene (hrpB) and the xanthomonadin coding cluster. Both pairs were tested in conventional PCR (cPCR) and real-time PCR (qPCR) formats. Primers targeting the hrpB gene showed cross reactions with other Xanthomonas spp. but were effective for use in both cPCR and qPCR, whereas primers for the xanthomonadin gene were highly specific for Xcv but showed low efficiency in qPCR. Enrichment of plant extracts in semi-selective medium before qPCR allowed a significant increase in sensitivity when compared to total DNA extraction, making it possible to detect as low as 10 1 CFU ml -1 . Under natural infection conditions, symptomatic and asymptomatic grapevines were tested by qPCR with hrpB primers and cPCR with xanthomonadin primers. In both cases, plant extracts were enriched for 36-72h. Xcv was detected in all symptomatic samples by qPCR and the result was confirmed by cPCR. For the asymptomatic samples, Xcv was detected in 93.4% with qPCR and in 89.5% with cPCR. These two methods offer advantages in terms of sensitivity and specificity, and they could be useful in quarantine programs, certification of grapevine propagating material and detection of inoculum sources in alternative hosts, contributing to the prevention of pathogen spread to disease-free areas.Keywords Grapevine bacterial canker . Vitis vinifera . Xanthomonas campestris pv. viticola. PCR-based diagnosis . qPCR

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“…Thus, it can be used for detection of the CBB pathogens on bean plants and seeds, differentiate the fuscans strains and confirm pathogenicity all in one step. Multiplex PCR has also been used to detect other bacterial pathogen complexes such as those involved in tomato bacterial spot (Araújo et al, 2012), bacterial blight and necrosis of cassava (Flores et al, 2019), citrus canker and citrus bacterial spot (Fonseca et al, 2019), leaf spot and black rot of Brassica (Berg et al, 2006) or for different pathogens associated with tomato seeds (Özdemir, 2009).…”

Section: Discussionmentioning

confidence: 99%

Virulence and type III effector diversities of Xanthomonas citri pv. fuscans and X. phaseoli pv. phaseoli in Brazil

Paiva

Wendland

Rossato

et al. 2021

Journal of Phytopathology

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Common bacterial blight (CBB) is caused by four genetic lineages belonging to two species of Xanthomonas, namely Xanthomonas citri pv. fuscans (includes fuscans, NF2 and NF3 lineages) and X. phaseoli pv. phaseoli (lineage NF1). A collection of 117 strains of Xanthomonas isolated from common bean plants grown in several producing regions of Brazil, between 2007 and 2016 was established. For species and lineage identification, the following tests were performed: multiplex PCR with a set of four specific primer pairs, pathogenicity tests on susceptible cultivar BRS Artico and phylogenetic analysis based on housekeeping gene sequences. The presence of the two species were confirmed among the 117 strains, being 62 non‐fuscans strains (NF1, NF2 and NF3) and 55 fuscans strains of X. citri pv. fuscans. To select a set of representative strains for the virulence assay, a PCR‐based analysis of effector diversity was performed with 42 strains belonging to the two species. PCR with primers for xopL, avrBsT, xopE2 and xopE1 genes were positive for all strains, while for the other six effectors there was variation. Six distinct effector profiles were detected, and one strain representing each type was inoculated in 15 common bean cultivars with varying levels of resistance to CBB. The fuscans strains showed uniformity in their effector profiles and were the most virulent. The phylogenetic analyses of our strain collection revealed that all genetic variants of CBB pathogens (NF1, NF2, NF3 and fuscans) are present in Brazil, with significant variability in virulence to common bean cultivars.

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Development of a duplex-PCR for differential diagnosis of Xanthomonas phaseoli pv. manihotis and Xanthomonas cassavae in cassava (Manihot esculenta)

Cited by 5 publications

References 29 publications

Cassava diseases caused byXanthomonas phaseolipv.manihotisandXanthomonas cassavae

Cassava diseases caused byXanthomonas phaseolipv.manihotisandXanthomonas cassavae

Detection of Xanthomonas citri pv. viticola on grapevine by real-time PCR and BIO-PCR using primers designed from pathogenicity and xanthomonadin gene sequences

Virulence and type III effector diversities of Xanthomonas citri pv. fuscans and X. phaseoli pv. phaseoli in Brazil

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