One fungus, which genes? Development and assessment of universal primers for potential secondary fungal DNA barcodes.
نویسندگان
چکیده
The aim of this study was to assess potential candidate gene regions and corresponding universal primer pairs as secondary DNA barcodes for the fungal kingdom, additional to ITS rDNA as primary barcode. Amplification efficiencies of 14 (partially) universal primer pairs targeting eight genetic markers were tested across > 1 500 species (1 931 strains or specimens) and the outcomes of almost twenty thousand (19 577) polymerase chain reactions were evaluated. We tested several well-known primer pairs that amplify: i) sections of the nuclear ribosomal RNA gene large subunit (D1-D2 domains of 26/28S); ii) the complete internal transcribed spacer region (ITS1/2); iii) partial β -tubulin II (TUB2); iv) γ-actin (ACT); v) translation elongation factor 1-α (TEF1α); and vi) the second largest subunit of RNA-polymerase II (partial RPB2, section 5-6). Their PCR efficiencies were compared with novel candidate primers corresponding to: i) the fungal-specific translation elongation factor 3 (TEF3); ii) a small ribosomal protein necessary for t-RNA docking; iii) the 60S L10 (L1) RP; iv) DNA topoisomerase I (TOPI); v) phosphoglycerate kinase (PGK); vi) hypothetical protein LNS2; and vii) alternative sections of TEF1α. Results showed that several gene sections are accessible to universal primers (or primers universal for phyla) yielding a single PCR-product. Barcode gap and multi-dimensional scaling analyses revealed that some of the tested candidate markers have universal properties providing adequate infra- and inter-specific variation that make them attractive barcodes for species identification. Among these gene sections, a novel high fidelity primer pair for TEF1α, already widely used as a phylogenetic marker in mycology, has potential as a supplementary DNA barcode with superior resolution to ITS. Both TOPI and PGK show promise for the Ascomycota, while TOPI and LNS2 are attractive for the Pucciniomycotina, for which universal primers for ribosomal subunits often fail.
منابع مشابه
Phylogenetic Assessment of Some Species of Crocus Genus Using DNA Barcoding
DNA barcoding is a simple method for the identification of any species using a short genetic sequence from a standard genome section. The present study aimed at examining the nuclear and chloroplast diversity as well as the phylogenetic relationships of eight species of saffron including four spring-flowering and five autumn-flowering species from different parts of Iran, using the nuclear barc...
متن کاملMolecular Characterization of a Squalene epoxidase Gene in Dermatophyte Pathogen Trichophyton tonsurans
Background: Trichophyton tonsurans is one of the dermatophyte fungi which invades the skin and hair of human. Several properties of this fungus have been investigated so far. However a few studies were carried out in the field of molecular biology of this fungus. In the present study, we tried to identify the Squalene epoxidase gene which is related to synthesis of ergosterol in this fungus. Me...
متن کاملAssessment of fungal diversity in deep-sea sediments by multiple primer approach.
Increasing evidence of the fungal diversity in deep-sea sediments has come from amplification of environmental DNA with fungal specific or eukaryote primer sets. In order to assess the fungal diversity in deep-sea sediments of the Central Indian Basin (CIB) at ~5,000 m depth, we amplified sediment DNA with four different primer sets. These were fungal-specific primer pair ITS1F/ITS4 (internal t...
متن کاملSpecific PCR-primers for detection of Picoa lefebvrei desert truffle in Carex stenophylla roots
Picoa species are hypogeous desert truffles, which can be found in semi-arid ranges of North Africa, West Asian, South of Europe and Middle East, including Iran (Moreno et al. 2000, Jamali & Banihashemi 2012, 2013). Picoa species form symbiosis mainly on roots of annual and perennial herbaceous plants of the Helianthemum, including H. ledifolium and H. salicifolium var. salicifolium (Jamali & B...
متن کاملDETECTION OF BACTERIA BY AMPLIFYING THE 16S rRNA GENE WITH UNIVERSAL PRIMERS AND RFLP
Background: There is a conserved portion in the 16S rRNA gene of bacteria which can be amplified by the universal PCR method. This fragment is 996 bp in length. In this method, only one set of universal primers is used for the amplification of the conserved region of the 16S rRNA gene, in common bacterial pathogens. Therefore, using the universal PCR method, these bacteria are detectable on...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Persoonia
دوره 35 شماره
صفحات -
تاریخ انتشار 2015