Ribosomal DNA spacer probes for yeast identification: Studies in the genus <i>Metschnikowia</i>

Henriques, Marília; Sá‐Nogueira, Isabel; Giménez-Jurado, Gabriella; Uden, N. van

doi:10.1002/yea.320070210

Cited by 17 publications

(8 citation statements)

References 9 publications

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“…is dramatically different in both length and sequence from the NTS regions of P. marinus and P. atlanticus. In contrast to the conserved SSU, ITS and 5.8S regions, the NTS is not transcribed (Dover and Coen 1981) and therefore has the potential to accumulate a high degree of sequence variability, even between closely related species (Henriques et al 1991; Marsh, Gauthier, and Vasta 1995). In Xenopus laevis and Drosophila melanogaster the NTS regions contain sequence motifs associated with putative RNA polymerase A promoters (Challoner et al 1985; Long, Rebbert, and Dawid 1981) and repeats of unknown function have also been found in Saccharomyces cerevisiae and Saccharomyces carlbergensis (Skryabin et al 1984).…”

Section: Discussionmentioning

confidence: 99%

“…provided the basis for the development of a species‐specific diagnostic PCR assay for P. andrewsi n. sp. The high variability of the NTS sequence made it an ideal target for the detection of subtle differences among intraspecific strains or types and the detection of probably greater differences between species and higher taxonomic categories (Henriques et al 1991; Marsh, Gauthier, and Vasta 1995). The PCR‐based diagnostic assay for P. andrewsi n. sp.…”

Section: Discussionmentioning

confidence: 99%

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Description of Perkinsus andrewsi n. sp. Isolated from the Baltic Clam (Macoma balthica) by Characterization of the Ribosomal RNA Locus, and Development of a Species‐Specific PCR‐Based Diagnostic Assay

Coss¹,

Robledo²,

Ruiz³

et al. 2001

J Eukaryotic Microbiology

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A Perkinsus species was isolated from the baltic clam Macoma balthica and an in vitro culture established under conditions described for P. marinus. As reported previously, morphological features remarkable enough to clearly indicate that this isolate is a distinct Perkinsus species were lacking. In this study, regions of the rRNA locus (NTS, 18S, ITS1, 5.8S, and ITS2) of this isolate were cloned, sequenced, and compared by alignment with those available for other Perkinsus species and isolates. Sequence data from the rRNA locus and species-specific PCR assays indicated not only that Perkinsus sp. from M. balthica was not P. marinus, but it was different from P. atlanticus and P. olseni. The degree of difference was comparable to or greater than differences between accepted Perkinsus species. In particular, NTS sequence and length were dramatically different from that of P. marinus and P. atlanticus. Therefore, we formally propose to designate the Perkinsus sp. from M. balthica as a separate species, P. andrewsi n. sp. Primers based on P. andrewsi NTS sequence were used to develop a PCR-based diagnostic assay that was validated for species-specificity and sensitivity. PCR-based assays specific for either P. andrewsi or P. marinus were used to test for their presence in bivalve species sympatric to M. balthica. Although isolated from M. balthica, P. andrewsi was also detected in the oyster Crassostrea virginica and clams Macoma mitchelli and Mercenaria mercenaria, and could coexist with P. marinus in all four bivalve species tested.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Description of Perkinsus andrewsi n. sp. Isolated from the Baltic Clam (Macoma balthica) by Characterization of the Ribosomal RNA Locus, and Development of a Species‐Specific PCR‐Based Diagnostic Assay

Coss¹,

Robledo²,

Ruiz³

et al. 2001

J Eukaryotic Microbiology

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“…1990) are useful in differentiating wine yeast strains. However, these approaches do not find an easy application in the wine industry because they are complex and laborious techniques when compared to PCR‐typing techniques (Henriques et al . 1991; Baleiras Couto et al .…”

Section: Introductionmentioning

confidence: 99%

Typing of Saccharomyces cerevisiae and Kloeckera apiculata strains from Aglianico wine

Caruso

Capece

Salzano

et al. 2002

Lett Appl Microbiol

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Aims: Kloeckera apiculata and Saccharomyces cerevisiae yeast species are dominant, respectively, at the early and at the following stages of wine fermentation. In the present study, PCR fingerprinting and NTS region amplification and restriction were applied as techniques for monitoring yeast population performing Aglianico of Vulture grape must fermentation. Methods and Results: Thirty S. cerevisiae and 30 K. apiculata strains were typed by PCR fingerprinting with (GAC)5 and (GTG)5 primers and by complete NTS region amplification followed by restriction with HaeIII and MspI enzymes. S. cerevisiae strains generated two patterns with (GAC)5 primer, while (GTG)5 primer yielded a higher genetic polymorphism. Conversely, in K. apiculata Aglianico wine strains (GAC)5 and (GTG)5 primers generated the same profile for all strains. Restriction analysis of the amplified NTS region gave the same profile for all strains within the same species, except for one strain of S. cerevisiae. Conclusions: The PCR fingerprinting technique was useful in discriminating at strain level S. cerevisiae, particularly with the primer (GTG)5. RFLP patterns generated from the NTS region of the two species can be more easily compared than the patterns resulting from PCR fingerprinting, thus RFLP is more suitable for the rapid monitoring of the species involved in different stages of fermentation. Significance and Impact of the Study: The molecular techniques used allow discrimination of S. cerevisiae at strain level and monitoring of the ratio of S. cerevisiae/K. apiculata during the fermentation process. Thus, their application can assure technological adjustments in a suitable time.

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“…Thus, the less conserved rDNA spacer sequences-NTSs and ITSs-should be excellent candidates for the selection of species-specific probes for yeast identification. In a previous report, selected sequences derived from the NTS2 region of the rDNA unit were used, for the first time, as species-specific DNA probes for the identification of the yeast Metschnikowia reukaufii and proved to be very effective (Henriques et al, 1991).…”

Section: Introductionmentioning

confidence: 99%

Specific identification of Candida albicans by hybridization with oligonucleotides derived from ribosomal DNA internal spacers

Botelho¹,

Planta²

1994

Yeast

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In order to develop DNA probes for rapid, sensitive and specific detection of the pathogenic yeast species Candida albicans, we carried out comparative sequence analysis of the two internal transcribed spacer regions (ITS1 and ITS2) of the ribosomal DNA (rDNA) units of C. albicans and the closely related pathogenic species C. tropicalis. While overall sequence similarity between the two species was considerable (65-75%), both ITS1 and ITS2 were found to contain distinct regions with sufficient sequence divergence to make them suitable as specific target sites for the identification of C. albicans. On the basis of these results one ITS1-derived (ANAB1) and two ITS2-derived (ANAB2 and ANAB3) oligonucleotides were selected, chemically synthesized, and used as hybridization probes. Their specificity and reliability were evaluated in dot-blot hybridization experiments with total genomic DNA from 13 strains of medically important Candida species, six strains of other yeast genera associated with man and animals, and ten strains previously identified as C. albicans by phenotypic criteria. Under well-defined hybridization conditions the three probes hybridized exclusively with DNA derived from strains belonging to the species C. albicans, thus demonstrating their potential clinical usefulness. The failure of four of the (presumed) C. albicans strains to show hybridization to the ITS probes sheds doubt upon their taxonomic classification, which is reinforced by other phenotypic aspects of these strains.

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Ribosomal DNA spacer probes for yeast identification: Studies in the genus Metschnikowia

Cited by 17 publications

References 9 publications

Description of Perkinsus andrewsi n. sp. Isolated from the Baltic Clam (Macoma balthica) by Characterization of the Ribosomal RNA Locus, and Development of a Species‐Specific PCR‐Based Diagnostic Assay

Description of Perkinsus andrewsi n. sp. Isolated from the Baltic Clam (Macoma balthica) by Characterization of the Ribosomal RNA Locus, and Development of a Species‐Specific PCR‐Based Diagnostic Assay

Typing of Saccharomyces cerevisiae and Kloeckera apiculata strains from Aglianico wine

Specific identification of Candida albicans by hybridization with oligonucleotides derived from ribosomal DNA internal spacers

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