DNA probes specific for the yeast species Debaryomyces hansenii: useful tools for rapid identification

Corredor, M

doi:10.1016/s0378-1097(00)00481-x

Cited by 9 publications

(12 citation statements)

References 17 publications

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“…hansenii and D. hansenii var. fabryi, whilst other authors employing different methodologies have not been successful in separating those varieties [23,25,26].…”

Section: Discussionmentioning

confidence: 96%

Differential detection of isolated from intermediate-moisture foods by PCR-RFLP of the IGS region of rDNA

Romero

Patiño

Quirós

et al. 2005

FEMS Yeast Research

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The amplification by PCR of the Intergenic Spacer region (IGS) of rDNA followed by Restriction Fragment Length Polymorphism (RFLP) analysis was evaluated as a potential method for the identification of Debaryomyces hansenii among other yeast species that frequently contaminate Intermediate-Moisture Foods (IMFs). For a first rapid differentiation at the species level, the determination of the IGS-PCR fragment size was found to be a useful approach. The digestion of this region with the enzymes HhaI, HapII and MboI resulted in specific patterns that permit the identification of D. hansenii among other yeast species. This method also permitted the discrimination between the D. hansenii varieties (var. hansenii and var. fabryi) as well as the differentiation of D. hansenii from other species of the genus, such as Debaryomyces pseudopolymorphus or Debaryomyces polymorphus var. polymorphus. The IGS-PCR RFLP method was assayed for the differential detection of D. hansenii in contaminated or spoiled IMF products and compared with traditional identification procedures, resulting in a 100% detection rate for D. hansenii.

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“…hansenii and D. hansenii var. fabryi, whilst other authors employing different methodologies have not been successful in separating those varieties [23,25,26].…”

Section: Discussionmentioning

confidence: 96%

Differential detection of isolated from intermediate-moisture foods by PCR-RFLP of the IGS region of rDNA

Romero

Patiño

Quirós

et al. 2005

FEMS Yeast Research

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“…Several DNA-based methods have been developed for identification of the species D. hansenii (Nishikawa et al 1998;Esteve-Zarzoso et al 1999;Andrighetto et al 2000;Corredor et al 2000;Petersen et al 2001). However, none of these methods are usable for strain typing.…”

Section: Discussionmentioning

confidence: 99%

Genetic diversity of the species Debaryomyces hansenii and the use of chromosome polymorphism for typing of strains isolated from surface-ripened cheeses

Petersen

Jespersen

2004

J Appl Microbiol

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Aims: To investigate the genetic diversity among strains of Debaryomyces hansenii and further to evaluate chromosome polymorphism determined by pulse-field gel electrophoresis (PFGE) as a tool for strain typing. Methods and Results: In total 56 isolates of D. hansenii were analysed by PFGE. The isolates included type strains and other strains obtained from culture collections as well as strains collected during production of Danish surface-ripened cheeses. By use of the PFGE technique the number and size of the chromosomal bands were calculated and the total genome size estimated. The number of chromosomal bands observed was found to vary from five to 10. The most common chromosome number was found to be six and for strains with six chromosomes the total genome size was found to vary from 9AE4 to 12AE6 Mb. The chromosome numbers for the type strain of each variety of D. hansenii (D. hansenii var. hansenii and D. hansenii var. fabryi) appeared to be six and seven respectively. By use of the PFGE technique it was possible to differentiate between all the investigated CBS strains and the vast majority of the dairy isolates. The dairy isolates that were found to have identical profiles (three of 56 isolates) were all isolated during production of one batch of surface-ripened cheeses and are likely to be the same strain isolated several times during cheese production. Further it was shown that the PFGE analysis did not result in a division of the two D. hansenii varieties, i.e. D. hansenii var. fabryi and D. hansenii var. hansenii into separate groups.Conclusion: The present study shows that the chromosomal arrangement of D. hansenii strains is heterogenic and does have a distinct chromosome polymorphism. Further the PFGE technique was proved to have a high discriminative power for strain typing of D. hansenii. Significance and Impact of the Study: The results obtained add to the first knowledge on the genetic diversity of the species D. hansenii. Further the distinct chromosome polymorphism of D. hansenii strains as shown in this study makes the PFGE technique a useful tool for strain typing of D. hansenii, e.g. during cheese production.

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“…NL1 and NL4 primers are described in [21]. Sequences were compiled and analyzed using the Staden package [7] and FASTA [22] in the GCG environment (Genetics Computer Group, Madison, Wisconsin, USA).…”

Section: Sequencingmentioning

confidence: 99%

Use of specific DNA probes for the rapid characterization of yeasts isolated from complex biotopes

Davila¹,

Diez²,

Corredor³

et al. 2001

Genet Sel Evol

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We describe a rapid method developed to identify yeast species commonly found during cheese ripening. It is based on the isolation of species-specific sequences and their hybridization with coarse preparations of genomic DNA. Several strategies were followed for the construction of probes: PCR amplification of sequences available in databases, random cloning of genomic DNA fragments, specific RAPD fragments and PCR-amplified ribosomal DNA fragments. After validation, the probes were applied to the characterization of 400 yeast strains isolated from various French cheeses. Since the strains had been previously identified with classical diagnostic tests, we were able to compare molecular and conventional identification. In addition, the specific probes for one of the species, Debaryomyces hansenii, were used successfully in colony hybridization experiments. The probes developed here proved to be very useful for the screening of large yeast collections and for the assessment of biodiversity within the yeast flora in cheese. yeast / cheese / molecular identification / hybridization / biodiversityResume -Utilisation de sondes ADN specifiques pour la caracterisation rapide de levures isoIees d'ecosystemes complexes. Au cours de ce travail, nous avons developpe une methode a la fois rapide et sensible pour l'identification des levures couramment isolees pendant l'affinage du fromage. Son principe repose sur l'hybridation de sequences specifiques sur les preparations brutes d' ADN genomique des souches a identifier. Pour la construction des sondes, nous avons du suivre plusieurs strategies differentes: amplification de sequences disponibles dans les bases de donnees, clonage au hasard de sequences d' ADN genomique, isolement de sequences specifiques obtenues par amplification RAPD d' ADN genomique ou par amplification d' ADNr. Apres leur validation, les son des ont pu etre mises en ceuvre S354 A.-M. Davila et al.dans differentes applications. Elles ont ete utilisees pour cribler 400 souches de levures isolees de differents from ages fran«ais. Les souches ayant ete prealablement identifiees par des tests classiques, nous avons pu comparer les deux types de methodes d'identification, moleculaire et conventionnelle. La son de specifique de D. hansenii a He testee avec succes dans des essais d'hybridation sur colonie. Au total, ces son des se sont revelees etre de bons outils pour le suivi et l'inventaire de la biodiversite des levures fromageres. levures / fromage / identification moleculaire / hybridation / biodiversite

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DNA probes specific for the yeast species Debaryomyces hansenii: useful tools for rapid identification

Cited by 9 publications

References 17 publications

Differential detection of isolated from intermediate-moisture foods by PCR-RFLP of the IGS region of rDNA

Differential detection of isolated from intermediate-moisture foods by PCR-RFLP of the IGS region of rDNA

Genetic diversity of the species Debaryomyces hansenii and the use of chromosome polymorphism for typing of strains isolated from surface-ripened cheeses

Use of specific DNA probes for the rapid characterization of yeasts isolated from complex biotopes

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