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Aureobasidium Mangrovei Sp. Nov., an Ascomycetous Species Recovered from Hara Protected Forests in the Persian Gulf, Iran

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Aureobasidium Mangrovei Sp. Nov., an Ascomycetous Species Recovered from Hara Protected Forests in the Persian Gulf, Iran Antonie van Leeuwenhoek https://doi.org/10.1007/s10482-018-1059-z ORIGINAL PAPER Aureobasidium mangrovei sp. nov., an ascomycetous species recovered from Hara protected forests in the Persian Gulf, Iran Shaghayegh Nasr . Mona Mohammadimehr . Marzieh Geranpayeh Vaghei . Mohammad Ali Amoozegar . Seyed Abolhassan Shahzadeh Fazeli Received: 9 November 2017 / Accepted: 28 February 2018 Ó Springer International Publishing AG, part of Springer Nature 2018 Abstract A new ascomycetous black yeast-like M 30265T and the ex-type culture is deposited in the species was recovered from healthy plant (Avicennia CBS yeast collection of the Westerdijk Fungal Bio- marina) of Hara protected mangrove forests at Qeshm diversity Institute, Utrecht, The Netherlands as CBS Island, Iran. Morphological, physiological analysis as 142205T. The GenBank accession numbers for the well as a molecular analysis of the internal transcribed nucleotide sequences of the large subunit ribosomal spacer (ITS) and partial large ribosomal subunit (D1/ DNA and ITS region are KY089084 and KY089085, D2 domains) confirmed the placement of this strain in respectively. The MycoBank number of the new the genus Aureobasidium and based on considerable species is MB 823444. sequence divergence, distinguishable cardinal growth temperatures and salt tolerance a new species Aure- Keywords Avicennia marina Á Biodiversity Á Black obasidium mangrovei sp. nov. is proposed. However, yeast-like Á Molecular phylogeny Á 1 new taxon. the type strain micro-morphologically is not clearly Qeshm Island distinguishable from other members of the genus. The type strain, Aureobasidium mangrovei was preserved in a metabolically inactive state at the Iranian Biological Resource Centre, Tehran, Iran as IBRC- Introduction The ascomycetous genus Aureobasidium is a member of Electronic supplementary material The online version of the family Saccotheciaceae within the class of the this article (https://doi.org/10.1007/s10482-018-1059-z) con- tains supplementary material, which is available to authorized Dothideomycetes (Thambugala et al. 2014; users. S. Nasr (&) Á M. Mohammadimehr Á M. A. Amoozegar M. Geranpayeh Vaghei Á S. A. Shahzadeh Fazeli Extremophiles Laboratory, Department of Microbiology, Microorganisms Bank, Iranian Biological Resource Faculty of Biology and Center of Excellence in Phylogeny Center (IBRC), ACECR, Tehran, Iran of Living Organisms, College of Science, University of e-mail: [email protected]; Tehran, Tehran, Iran [email protected] S. A. Shahzadeh Fazeli S. Nasr Department of Molecular and Cellular Biology, Faculty of Department of Microbial Biotechnology, Faculty of Basic Basic Sciences and Advanced Technologies in Biology, Sciences and Advanced Technologies in Biology, University of Science and Culture, Tehran, Iran University of Science and Culture, Tehran, Iran 123 Antonie van Leeuwenhoek Wijayawardene et al. 2014; Hymphries et al. 2017). In Hormozgan province, Persian Gulf (Ghasemi et al. recent classification, Aureobasidium has synanamorphs 2012). It is considered a unique ecosystem that plays a named Kabatiella and Selenophoma (Schoch et al. multi-purpose role for biodiversity protection and 2006; Bills et al. 2012; Hymphries et al. 2017). biotechnology by protecting the shore line, providing Phenotypic varieties of the fungus were defined by food sources and safe habitat for fish, wild bird and Hermanides-Nijhof (1977) and then redefined using also domestic animals, participating in the biogeo- molecular methods by Zalar et al. (2008). Later on, the chemical cycle, honey production and production of genome sequencing analysis by Gostincˇar et al. (2014) medicinal materials, such as saponin, flavonoid and revealed that the differences between the four varieties tannins (Ali Zahed et al. 2010; Faridah-Hanum et al. of Aureobasidium pullulans are significant enough to 2014). Unfortunately, such ecosystems have been redefine them as separate species: A. pullulans, Aure- facing destructive threats in Iran and also around the obasidium melanogenum, Aureobasidium subglaciale globe. Over the past 50 years, approximately one-third and Aureobasidium namibiae. Members of this genus of the world’s mangrove forests have been lost. are known from their asexual reproduction and there in Natural and anthropogenic threats including growing no teleomorph linked to this genus (Arzanlou 2014). coastal populations, global climate change, increasing While description of several distinguished clades based pollution and industrialization are considered the main on multilocus DNA analyses was evident, yet nomina- threats (Parvaresh 2011; Faridah-Hanum et al. 2014). tion of separate taxa has not been performed (Man- During a research project aimed at identifying itchotpisit et al. 2009). The total number of classified biodiversity of yeasts species related to plant flora of Aureobasidium species currently varies in different Hara forests at Qeshm Island, Iran, two conspecific official nomenclatural repositories and it is close to 38 unknown ascomycetous black yeast-like strains were species. The number of Aureobasidium species is isolated. We used a combination of morphological and increasing and the two most recent novel species (A. physiological characteristics, and DNA sequence iranianum and A. thailandense) have been isolated from analyses to characterize the two strains and describe Bamboo stems, leaves and wooden surfaces (Arzanlou them as novel species of the genus Aureobasidium. and Khodaei 2012; Peterson et al. 2013). Species in the genus Aureobasidium have been recovered from plant residues, soil, air, water, glacial ice and even the clinical Materials and methods laboratory as a contaminant or opportunistic infection (Loncaric et al. 2007; Zalar et al. 2008;Moraisetal. Sample collection and fungal isolation 2011; Arfi et al. 2012; Santo et al. 2012). Aureobasidium spp. exhibit diverse life styles such as saprophytes, plant Healthy leaves were collected from hara mangrove associated endophytes or opportunistic human patho- trees (Avicennia marina) growing in the marsh zone gens (Arzanlou 2014). Occurrence, substrates and (approximately coordinates: 26°470N, 55°450E), cut distribution of Aureobasidium spp. in Iran was inves- into small pieces (2 9 2 cm) aseptically and macer- tigated by Arzanlou (2014). Species of the genus ated in 90 ml saline. Serial dilutions in saline (0.9% Aureobasidium show significant biotechnological and NaCl) from healthy plant leaves were directly plated commercial potentials due to their ability to produce on Rose Bengal agar (Himedia) supplemented with extracellular polysaccharide (EPS), a variety of hydro- 0.1 g chloramphenicol l-1 and incubated at 28 °C for lytic enzymes and a possible role as biocontrol agent 48 h. against phytopathogenic fungi in fire blight disease (Manitchotpisit et al. 2009; Martini et al. 2009; Cheng Phenotypical and physiological characterization et al. 2011;Richetal.2011). Furthermore, biofinishing, biosurfactant and single cell oil production recently has Developing colonies were grouped into morphotypes been introduced as a novel biotechnological application based on their cultural characteristics and representa- of some members of this genus (Wang et al. 2014;Kim tives of each colony type per plate purified by et al. 2015; Nieuwenhuijzen et al. 2016). streaking on yeast peptone glucose (YPG) agar Hara Protected Forests is the common name for medium at 28 °C. Morphological characters of the mangrove forests on the southern coast of Iran, in the isolates were studied on potato dextrose agar (PDA, 123 Antonie van Leeuwenhoek Merck), malt extract (MEA) agar (MEA: 20 g malt and yeast isolates were found: Sympodiomycopsis extract (Merck), 1 g peptone (Merck), 20 g glucose kandeliae (two), Quambalaria simpsonii (one), Hor- (Merck) and 20 g agar (Merck), 1000 ml distilled taea werneckii (two), Rhodotorula mucilaginosa water, pH 5.5), Harrold’s M40Y and M60Y media as (one); Aureobasidium pullulans, Exophiala spp. described (Raper and Fennell 1965; Klich 2002). (three) and a newly described species (Nasr et al. Colony diameters and appearance were recorded. 2017) named as Jaminaea pallidilutea (two). In Photographs were taken from 7 days old culture plates addition, we isolated two unknown and potentially incubated at 28 °C unless otherwise mentioned. For conspecific ascomycetous black yeast-like strains micromorphological observations, temporary mounts (IBRC-M 30265T and IBRC-M 30266). We success- were made in lactic acid. The standard methods of fully PCR amplified and sequenced the D1/D2 identification described by Barnett et al. (2000) and domains of the LSU and ITS rDNA region. An initial the key of Kurtzman et al. (2011) were used to explore megablast similarity search in the NCBI nr database the physiological and biochemical characteristics of using the LSU sequence showed the highest similarity the isolates. to Aureobasidium sp. strain RBF-17A2 (FN665420; 98% identity). Megablast similarity search using the DNA extraction and phylogenetic analysis ITS sequence showed the highest similarity to Ka- of marker genes batiella sp. strain SQU-MA24 (KU945924; 99%) and uncultured fungus clone CMH306 (KF800397; 97%). For DNA isolation, the strains were grown on YPG Phylogenetic analyses of LSU and ITS rRNA regions broth at 28 °C for 24 h. The DNA was extracted based on both the neighbor-joining (NJ) and the according to Hanna and Xiao (2006). The internal maximum-likelihood (ML) method were performed to transcribed spacer (ITS) region of ribosomal DNA was assess
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