Open-access, online, rapid taxonomy
ISSN: 2653-4649 (Online)

Australian Journal of

Taxonomy

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A new species of Genolevuria Xin Zhan Liu, F.Y. Bai, M. Groenew. & Boekhout from Queensland, Australia

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Yu Pei Tan [1*], Thomas S. Marney [1] & Roger G. Shivas [1, 2]

[1] Queensland Plant Pathology Herbarium, Department of Agriculture and Fisheries, Dutton Park 4102, Queensland, Australia
[2] Centre for Crop Health, University of Southern Queensland, Toowoomba 4350, Queensland, Australia

yupei.tan@daf.qld.gov.au

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Abstract

One species of fungus from the phylum Basidiomyctoa is introduced in the genus Genolevuria Xin Zhan Liu, F.Y. Bai, M. Groenew. & Boekhout based on unique molecular traits. The phylogenetic relationships of the new species with other species of Genolevuria are revealed by unique nucleotides in the internal transcribed spacer (ITS) region and large subunit (LSU) gene in the ribosomal DNA.

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Introduction

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Materials & Methods

Isolate

The holotype is deposited in the culture collection at the Queensland Plant Pathology Herbarium (BRIP), and permanently preserved in a metabolically inactive state by cryopreservation in glycerol at -80°C.

DNA isolation & amplification

Conidia were scraped from the ex-type culture grown on malt yeast glucose peptone agar (MYPGA). Genomic DNA was extracted with the ISOLATE II Plant DNA kit (Bioline Meridian Bioscience) according to the manufacturer’s instructions. The primers V9G (de Hoog & Gerrits van den Ende 1998) and ITS4 (White et al. 1990) were used to amplify the ITS region, and primers LR0R and LR5 (Schoch et al. 2009) were used to amplify part of the LSU rDNA, using the Phusion High-Fidelity PCR Master Mix (New England Biolabs). The PCR products were purified with ISOLATE II PCR and Gel kit (Bioline Meridian Bioscience) according to the manufacturer’s instructions, and sequenced by Macrogen Incorporated (Seoul, Korea).

Phylogenetic analysis

All sequences generated were assembled using Geneious Prime 2022 (Biomatters Ltd) and deposited in GenBank. The sequences from this study were aligned with selected sequences obtained from GenBank with the MAFFT alignment algorithm. Maximum likelihood (ML) analysis was run with the RAxML v. 8.2.11 plug-in (Stamatakis 2014) in Geneious starting from a random tree topology. The nucleotide substitution model used was general time-reversible (GTR) with a gamma-distributed rate variation. The Bayesian analysis was run with the MrBayes v.3.2.6 (Huelsenbeck and Ronquist 2001) plug-in in Geneious. To remove the need for a priori model testing, the Markov chain Monte Carlo (MCMC) analysis was set to sample across the entire GTR model space with a gamma-distributed rate variation across the nucleotide sites. Five million random trees were generated using a MCMC simulation with four chains. The sample frequency was set at 1 000 and the temperature of the heated chain was 0.1. “Burn-in” was set at 25%, after which the log-likelihood values were stationary.

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Results

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Discussion

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Figure 1. Phylogenetic tree based on a maximum likelihood analysis of concatenated multilocus alignment (ITS and LSU) from Genolevuria species. Analysis was performed on the Geneious Prime 2022 platform using RAxML v.8.2.11 and MrBayes v. 3.2.6 (Huelsenbeck & Ronquist 2001), both based on the GTR substitution model with gamma-distribution rate variation. Bootstrap values are shown as percentages on the branches of the consensus tree that was inferred from 1 000 replicates. Branch lengths are proportional to substitutions per site. Pseudotremella moriformis ex-type strain CBS 7810 was used as the outgroup. Novel taxon is indicated in bold. Ex-type strains are marked with an asterisk (*).

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Table content

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Key title

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Taxonomy

Bulleraceae, Tremellales, Incertae sedis, Tremellomycetes, Agaricomycotina, Basidiomycota, Fungi

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Acknowledgments

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Supplementary Material

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References

de Hoog GS, Gerrits van den Ende AHG (1998) Molecular diagnostics of clinical strains of filamentous basidiomycetes. Mycoses 41:183–189.

Huelsenbeck JP, Ronquist F. 2001. MRBAYES: Bayesian inference of phylogenetic trees. Bioinformatics 17: 754–755.

Schoch CL, Crous PW, Groenewald JZ, Boehm E, Burgess TI, de Gruyter K, de Hoog G, Dixon L, Grube M, Gueidan C (2009) A class-wide phylogenetic assessment of Dothideomycetes. Studies in Mycology 64:1–15.

Stamatakis A. 2014. RAxML version 8: a tool for phylogenetic analysis and post-analysis of large phylogenies. Bioinformatics 30: 1312–1313.

White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds) PCR protocols: a guide to methods and applications. Academic Press, San Diego, pp 315–322.

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Open-access, online, rapid taxonomy ISSN: 2653-4649 (Online)

Australian Journal of

Taxonomy

Open-access, online, rapid taxonomy
ISSN: 2653-4649 (Online)