Molecular screening of astaxanthin-producing yeasts of birch sap-flows in Alborz Province of Iran

Subject Areas : Environmental Microbiology

Faezeh Ajorloo ¹ , Mohsen Vaez ² , Jafar Hemmat ³

1 - PhD Student, Biotechnology Department, Iranian Research Organization for Science and Technology (IROST)
2 - Assistant Professor, Biotechnology Department, Iranian Research Organization for Science and Technology (IROST)
3 - Assistant Professor, Biotechnology Department, Iranian Research Organization for Science and Technology (IROST)

Received: 2020-10-18 Accepted : 2020-11-15 Published : 2020-12-21

Keywords: Carotenoid, Astaxanthin, Yeast, Multiple polymerase chain reaction (Multiplex-PCR), Birch trees,

Abstract :

Background & Objectives: The carotenoid-producing yeasts have a widespread distribiution  worldwide. However, isolation of carotenoid-producing yeasts with valuable astaxanthin have been only reported from limited areas and specific habitats. The aim of this study was to apply the multiple polymerase chain reaction method for faster molecular screening of isolates for the yeasts in the exudation sap from birch trees (Betula pendula Roth.) in Alborz Province of Iran with astaxanthin‌ production capacity and toevaluate its efficiency.Materials & Methods: This cross-sectional study was carried out by sampling in May 2018 from one of the natural habitats of birch trees Batula Pendula in Alborz Province, Iran. After isolation of yeasts using selected media and purification of yeast colonies with various pink to red colors, yeasts genomic nucleic acids were extracted and molecular screening for astaxanthin-producing detection of isolates was performed by multiple polymerase chain reaction (Multiplex-PCR).Results: Out of 42 pigmented colonies during sampling and isolation in one cross-sectional study, 23 pink to red colonies were evaluated by Multiplex-PCR method. Of these isolates, three were positive and the rest were negative. The results were validated by determining the ribosomal gene sequencing.Conclusion: The present study demonstrates the specificity of the multiple polymerase chain     reaction method in fast and cost-effective molecular screening of astaxanthin-producing yeast   isolates and the applicability of this method to study other natural habitats of these yeasts. 

References:

1. Yurkov AM. Yeasts of the soil–obscure but precious. Yeast. 2018 May;35(5):369-78.
   2. Stefanini I. Yeast‐insect associations: It takes guts. Yeast. 2018 Apr;35(4):315-30.
   3. Fonseca A, Inácio J. Phylloplane yeasts. InBiodiversity and ecophysiology of yeasts 2006 (pp.
   263-301). Springer, Berlin, Heidelberg.
   4. Phaff HJ. Yeasts associated with plants, insects and soil. Yeasts/edited by Anthony H. Rose, JS
   Harrison. 1987.
   5. Kurtzman C, Fell JW, Boekhout T, editors. The yeasts: a taxonomic study. Elsevier;2011May9.
   6. Boynton PJ. The ecology of killer yeasts: Interference competition in natural habitats. yeast.
   2019 Aug;36(8):473-85.
   7. Pereira da Costa D, Campos Miranda-Filho K. The use of carotenoid pigments as food additives
   for aquatic organisms and their functional roles. Reviews in Aquaculture. 2020 Aug;12
   (3):1567-78.
   8. Schmidt I, Schewe H, Gassel S, Jin C, Buckingham J, Hümbelin M, Sandmann G, Schrader J.
   Biotechnological production of astaxanthin with Phaffia rhodozyma/Xanthophyllomyces
   dendrorhous. Applied microbiology and biotechnology. 2011 Feb 1;89(3):555-71.
   9. Hosseinzadeh CA, Fallah F, Yousefzadeh H. Genetic Diversity and Differentiation of the
   Iranian's Betula pendula Populations by DNA Polymorphisms of Three (CD, DT, K1K2)
   Chloroplast Genome Regions. Iranian Journal of Biology. 2015; 28(2) 191-201.
   10. Weber RW. On the ecology of fungal consortia of spring sap-flows. Mycologist. 2006 Nov
   1;20(4):140-3.
   11. Grabek-Lejko D, Kasprzyk I, Zaguła G, Puchalski C. The bioactive and mineral compounds in
   birch sap collected in different types of habitats. Baltic Forestry. 2017 Jan 1;23(2):394-401.
   12. Lõoke M, Kristjuhan K, Kristjuhan A. Extraction of genomic DNA from yeasts for PCR-based
   applications. Biotechniques. 2011 May;50(5):325-8.
   13. Ajorloo, F., Vaez, M., Hemmat, J. Isolation and molecular identification of the carotenoid
   producing yeasts in the exudation sap from birch trees (Betula pendula Roth.) at Marmisho
   region of Northwest Iran. Journal of Microbial World, 2019; 12(2): 139-149. [In Persian]
   14. Scorzetti G, Fell JW, Fonseca A, Statzell-Tallman A. Systematics of basidiomycetous yeasts: a
   comparison of large subunit D1/D2 and internal transcribed spacer rDNA regions. FEMS yeast
   research. 2002 Dec 1;2(4):495-517.
   15. White TJ, Bruns T, Lee SJ, Taylor JL. Amplification and direct sequencing of fungal
   ribosomal RNA genes for phylogenetics. PCR protocols: a guide to methods and applications.
   1990;18(1):315-22.
   16. Fell JW, Boekhout T, Fonseca A, Scorzetti G, Statzell-Tallman A. Biodiversity and
   systematics of basidiomycetous yeasts as determined by large-subunit rDNA D1/D2 domain
   sequence analysis. International journal of systematic and evolutionary microbiology. 2000
   May 1;50(3):1351-71.
   17. ROSA CA, LACHANCE MA. The yeast genus Starmerella gen. nov. and Starmerella
   bombicola sp. nov., the teleomorph of Candida bombicola (Spencer, Gorin & Tullock) Meyer
   & Yarrow. International Journal of Systematic and Evolutionary Microbiology. 1998 Oct 1;48
   (4):1413-7.
   18. Filatov DA. ProSeq: a software for preparation and evolutionary analysis of DNA sequence
   data sets. Molecular Ecology Notes. 2002 Dec;2(4):621-4.
   19. Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ. Basic local alignment search tool.
   Journal of molecular biology. 1990 Oct 5;215(3):403-10.
   20. Woffelman C. DNAMAN for Windows, Version 5.2. 10. Lynon Biosoft. Institute of
   Molecular Plant Sciences, Netherlands: Leiden University. 2004.
   21. Valduga E, Valério A, Tatsch PO, Treichel H, Furigo A, Luccio M. Assessment of Cell
   Disruption and Carotenoids Extraction from Sporidiobolus salmonicolor (CBS 2636). Food
   and Bioprocess Technology. 2009;2(2):234-8.
   22. Weber RW, Davoli P. Teaching techniques for mycology: 20. Astaxanthin, a carotenoid of
   biotechnological importance from yeast and salmonid fish. Mycologist. 2003 Feb1;17(1):30-4.
   23. Barredo JL, García-Estrada C, Kosalkova K, Barreiro C. Biosynthesis of astaxanthin as a main
   carotenoid in the heterobasidiomycetous yeast Xanthophyllomyces dendrorhous. Journal of
   Fungi. 2017 Sep;3(3):44.
   24. Sorokina M, Steinbeck C. Review on natural products databases: where to find data in 2020.
   Journal of cheminformatics. 2020 Dec;12:1-51.
   25. Schroeder WA, Johnson EA. Carotenoids protect Phaffia rhodozyma against singlet oxygen
   damage. Journal of industrial microbiology. 1995 Jun 1;14(6):502-7.
   26. Golubev WI. Xanthophyllomyces Golubev. InThe Yeasts (Fourth Edition) 1998(pp. 718-719).
   27. Nahvi I, Vaez M, Emtiazi G. Evaluation of carotenoid-producing yeasts associated with birch
   trees (Betula pendula) in the North of Iran-Shahrestanak village. Pajouhesh-va-Sazandegi.
   2000; 13(3):70-74. [In Persian]
   28. Weber RW, Davoli P, Anke H. A microbial consortium involving the astaxanthin producer
   Xanthophyllomyces dendrorhous on freshly cut birch stumps in Germany. Mycologist. 2006
   May 1;20(2):57-61.
   29. Libkind D, Moliné M, de García V, Fontenla S, van Broock M. Characterization of a novel
   South American population of the astaxanthin producing yeast Xanthophyllomyces
   dendrorhous (Phaffia rhodozyma). Journal of industrial microbiology & biotechnology. 2008
   Mar 1;35(3):151-8.
   30. Colabella F, Libkind D. PCR-based method for the rapid identification of
   astaxanthin-accumulating yeasts (Phaffia spp.). Revista Argentina de Microbiología. 2016 Jan
   1;48(1):15-20.
   31. Muir A, Harrison E, Wheals A. A multiplex set of species-specific primers for rapid
   identification of members of the genus Saccharomyces. FEMS yeast research.2011 Nov 1;11
   (7):552-63.
   32. Glushakova AM, Chernov IY. Seasonal dynamics in a yeast population on leaves of the
   common wood sorrel Oxalis acetosella L. Microbiology. 2004 Mar 1;73(2):184-8.
   33. Gilbert DG. Dispersal of yeasts and bacteria by Drosophila in a temperate forest. Oecologia.
   1980 Jul 1;46(1):135-7.
   34. Mittelbach M, Yurkov AM, Nocentini D, Nepi M, Weigend M, Begerow D. Nectar sugars and
   bird visitation define a floral niche for basidiomycetous yeast on the Canary Islands. BMC
   ecology. 2015 Dec;15(1):2.