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Identification of species in the genus Pichia by restriction of the internal transcribed spacers (ITS1 and ITS2) and the 5.8S ribosomal DNA gene

Identification of Pichia species by 5.8S-ITS rDNA RFLP

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Abstract

In this study, the variability within the ribosomal DNA region spanning the internal transcribed spacers ITS1 and ITS2 and the 5.8S gene (5.8S-ITS rDNA) was used to differentiate species in the genus Pichia. The 5.8S-ITS rDNA region was PCR-amplified and the PCR product digested with the enzymes CfoI, HinfI, and HaeIII. The variability in the size of the amplified product and in the restriction patterns enabled differentiation between species in the genus Pichia, and between Pichia species and yeast species from other genera in the Yeast-id database (http://www.yeast-id.com/). Moreover, the restriction fragment length polymorphism (RFLP) patterns of the 5.8S-ITS enabled misidentified strains to be detected and revealed genetic heterogeneity between strains within the Pichia membranifaciens and Pichia nakazawae species. Ultimately, the RFLP patterns of the 5.8S-ITS rDNA failed to differentiate between some Pichia and Candida species that could be distinguished on the basis of the sequence of the 5.8S-ITS rRNA region or the sequence of the D1/D2 domain of the 26S rDNA gene.

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Acknowledgements

This work was funded by CICYT Grant AGL2000-1492 to AQ and CICYT Grant AEOO-0337-01 to AQ and MVC. CB acknowledges the Ministerio de Educacion, Ciencia y Deportes and CSIC for postdoctoral fellowships.

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Authors and Affiliations

  1. Departamento de Biotecnología, Instituto de Agroquímica y Tecnología de los Alimentos, CSIC, P.O. Box 73, 46100, Burjassot, Valencia, Spain

    Mercedes Villa-Carvajal, Amparo Querol & Carmela Belloch

  2. Centro Tecnológico AINIA, Parque Tecnológico de Valencia, 46980 , Paterna, Valencia, Spain

    Mercedes Villa-Carvajal

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  1. Mercedes Villa-Carvajal
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  2. Amparo Querol
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  3. Carmela Belloch
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Correspondence to Carmela Belloch.

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Villa-Carvajal, M., Querol, A. & Belloch, C. Identification of species in the genus Pichia by restriction of the internal transcribed spacers (ITS1 and ITS2) and the 5.8S ribosomal DNA gene. Antonie Van Leeuwenhoek 90, 171–181 (2006). https://doi.org/10.1007/s10482-006-9071-0

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  • DOI: https://doi.org/10.1007/s10482-006-9071-0

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