Identification of Ditylenchus species associated with Fabaceae seeds based on a specific polymerase chain reaction of ribosomal DNA-ITS regions

Kerkoud, Mohamed; Esquibet, Magali; Plantard, Olivier; Avrillon, Myriam; Guimier, Corinne; Franck, Martine; Lechappé, Joël; Mathis, René

doi:10.1007/s10658-006-9092-6

Cited by 21 publications

(18 citation statements)

References 8 publications

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“…This nematode, formerly considered to be a ‘giant race’ of D. dipsaci ( Ditylenchus sp. B), is now considered to be a species distinct from D. dipsaci on the basis of body size, chromosome number, host‐plant range, biology, RLFP, ALFP and RAPD profiles and rDNA sequence . It has been reported as more damaging to its host than D. dipsaci because of more severe symptoms and more infested seeds .…”

Section: Introductionmentioning

confidence: 99%

A fast and sensitive method for the simultaneous identification of three important nematode species of the genusDitylenchus

Jeszke

Dobosz

Obrępalska‐Stęplowska

2014

Pest. Manag. Sci.

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Section: Introductionmentioning

confidence: 99%

A fast and sensitive method for the simultaneous identification of three important nematode species of the genusDitylenchus

Jeszke

Dobosz

Obrępalska‐Stęplowska

2014

Pest. Manag. Sci.

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“…Considering plant parasitic nematodes ITS sequences were used e.g. for the discrimination of Ditylenchus dipsaci populations (Marek et al 2005;Kerkoud et al 2007).…”

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confidence: 99%

Variability of D2/D3 segment sequences of several populations and pathotypes of potato cyst nematodes (Globodera rostochiensis, Globodera pallida)

Douda¹,

Zouhar²,

Nováková³

et al. 2010

Plant Prot. Sci.

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Douda O., Zouhar M., Nováková E., Mazáková J., Ryšánek P. (2010): Variability of D2/D3 segment sequences of several populations and pathotypes of potato cyst nematodes (Globodera rostochiensis, Globodera pallida). Plant Protect. Sci., 46: 171-180.Potato cyst nematodes (Globodera rostochiensis, Globodera pallida) remain a key pest in the main potato growing regions of the Czech Republic. Due to difficult direct management and presence of diverse pathotypes attacking different potato cultivars the rapid and reliable diagnostics is of crucial importance. Currently, efforts are aimed at a description of different pathotypes based on DNA analysis. The main objective of this study was to evaluate the homogeneity of sequences of D2/D3 segments of the 28S rDNA gene obtained from 3 populations of G. rostochiensis and 5 populations of G. pallida and estimate their value for diagnostic purposes. PCR amplification yielded a single fragment of the length of 700 bp approximately in all populations. The alignment score of the vast majority of all pair comparisons of G. rostochiensis and G. pallida populations varied from 98 to 99. In total 14 point deletions and 3 substitutions were observed. The variability of D2/D3 segments of potato cyst nematodes is rather low and this DNA region can be used for diagnostics on a species level because more differences were found after comparing with G. tabacum and G. millefolii sequences obtained from Gene Bank; however the applicability of D2/D3 sequences to routine diagnostics of potato cyst nematodes could be complicated by its similarity to corresponding sequences of the nematode G. artemisiae.

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“…This PCR method decreases the diagnostic time and costs compared with ITS-PCR-RFLP. Previously, PCR with species-specific primers was developed for differentiation of D. dipsaci and D. gigas using species-specific SCAR or ITS-rRNA primers (Esquibet et al 2003;Marek et al 2005;Subbotin et al 2005;Kerkoud et al 2007;Zouhar et al 2007). The results of the current study allows for additional differentiation of D. weischeri by a PCR species-specific method.…”

Section: Resultsmentioning

confidence: 99%

Diagnostics of stem and bulb nematodes,Ditylenchus weischeriandD. dipsaci(Nematoda: Anguinidae), using PCR with species-specific primers

Madani

Tenuta

Chizhov

et al. 2015

Canadian Journal of Plant Pathology

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The stem and bulb nematodes of the Ditylenchus dipsaci (Kühn) Filipjev species complex are obligate endoparasites of various agricultural plants, causing stunting and swelling and resulting in significant economic losses. Recently, a new closely related Ditylenchus species, D. weischeri Chizhov, Borisov and Subbotin, a parasite of the cosmopolitan herbaceous perennial weed, Cirsium arvense (L.) Scop., was described. Many countries impose quarantine restrictions for the presence of D. dipsaci in imported plant and soil materials. In the current study, we developed PCR with species-specific primers for the rapid and reliable separation of D. weischeri and D. dipsaci using gel electrophoresis and melting curve analysis. Species-specific primer sets were designed based on the nucleotide sequence of the heat shock protein (hsp90) gene for both nematode species. The PCR protocol was verified using samples of D. weischeri, D. dipsaci and the closely related species, D. gigas, which parasitizes broadbean (Vicia faba L.). The species-specific primer sets were able to detect D. weischeri and D. dipsaci from samples containing mixtures of Ditylenchus species. The PCR species-specific protocol should allow for more rapid identification of Ditylenchus species recovered from plant materials than previously possible.Résumé: Les nématodes de la tige et du bulbe du complexe d'espèces Ditylenchus dipsaci (Kühn) Filipjev sont des endoparasites obligatoires qui s'attaquent à différentes cultures et qui causent du rabougrissement et du renflement, ce qui occasionne d'importantes pertes économiques. Récemment, une nouvelle espèce de Ditylenchus étroitement apparentée, D. weischeri Chizhov, Borisov and Subbotin, un parasite de l'adventice cosmopolite vivace Cirsium arvense (L.) Scop., a été décrite. Plusieurs pays imposent des restrictions phytosanitaires relativement à D. dipsaci quant aux importations de végétaux et de matériaux constitutifs du sol. Dans cette étude, nous avons développé une PCR avec amorces spécifiques de l'espèce pour différencier rapidement et de manière fiable D. weischeri et D. dipsaci, et ce, à l'aide d'une électrophorèse sur gel et de l'analyse de la courbe de fusion. Les jeux d'amorces spécifiques de l'espèce ont été conçus, pour les deux espèces de nématodes, en fonction de la séquence de nucléotides du gène de la protéine de choc thermique (hsp90). Le protocole de la PCR a été vérifié avec des échantillons de D. weischeri, D. dipsaci et de l'espèce étroitement apparentée, D. gigas, qui parasite la féverole à gros grains (Vicia faba L.). Les jeux d'amorces spécifiques de l'espèce ont pu détecter D. weischeri et D. dipsaci dans des échantillons contenant un mélange d'espèces de Ditylenchus. Le protocole spécifique de l'espèce de la PCR devrait permettre d'identifier plus rapidement que cela était possible auparavant les espèces de Ditylenchus extraites des végétaux.Mots clés: analyse de la courbe de fusion, Cirsium arvense, diagnostics, Ditylenchus, D. dipsaci, D. weischeri, gène de la protéine de choc thermi...

show abstract

Identification of Ditylenchus species associated with Fabaceae seeds based on a specific polymerase chain reaction of ribosomal DNA-ITS regions

Cited by 21 publications

References 8 publications

A fast and sensitive method for the simultaneous identification of three important nematode species of the genusDitylenchus

A fast and sensitive method for the simultaneous identification of three important nematode species of the genusDitylenchus

Variability of D2/D3 segment sequences of several populations and pathotypes of potato cyst nematodes (Globodera rostochiensis, Globodera pallida)

Diagnostics of stem and bulb nematodes,Ditylenchus weischeriandD. dipsaci(Nematoda: Anguinidae), using PCR with species-specific primers

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