PCR-RFLP ITS-based analysis of wine yeast autochthonous strains isolated from different grape cultivars in Taman subregion

The studies present the results of morphological, cultural and genetic analysis of the ITS1-ITS4 region of the autochthonous yeast strains genome by using the HaeIII restriction enzyme. On the red and white grapes varieties, based on the morphology of the cells, autochthonous strains belonging to the genus Saccharomyces prevail – 83.3%, what is confirmed by genetic analysis of rDNA ITS region. Restriction analysis showed that all strains of the genus Saccharomyces belong to the species Saccharomyces cerevisiae / S. paradoxus. The percentage of Saccharomyces isolated on the Pervenets Magaracha variety is 86.7%, Krasnostop Anapsky - 80%. The non-Saccharomyces yeast had a product size of 750 bp, presumably of the species Hanseniaspora uvarum.

First report of Rhizoctonia solani Kühn AG 2-2 LP in Zoyzia japonica Steud in Brazil

ABSTRACT The use of cultivated grasses in Brazil has grown by 40% between 2010 and 2015, and the species Zoysia japonica Steud, especially the cultivar ‘Esmeralda’, corresponds to 81% of the grass market in the country. The most important disease affecting zoysia grass, known as large patch, is caused by Rhizoctonia solani and occurs in the Brazilian lawns particularly during winter months. The aim of this study was to contribute to the identification and characterization of the anastomosis group of R. solani isolates from lesions typical of large patch collected from ‘Esmeralda’ grass at gardens and golf courses in the states of São Paulo and Bahia, Brazil. All 12 obtained isolates presented dark-brown colonies with aerial mycelial growth, multinucleated hyphae and absence of concentric zonation or sclerotia, and showed their greatest mycelial growth rate at 25°C. In pathogenicity experiments, except three out of R. solani isolates, reduced the growth of zoysia grass. Based on the analysis of sequences of the rDNA-ITS region, the isolates clustered with reference isolates of the anastomosis group AG 2-2 LP. Phylogenetic inference showed that the Brazilian isolates are grouped into two clades that shared the same common ancestral with 96% bootstrap. One of the clades includes only Brazilian isolates while the other one also includes American and Japanese R. solani isolates AG 2-2 LP. This is the first report and characterization of R. solani AG 2-2 LP in zoysiagrass in Brazil.

Phylogenetic Analysis and Molecular Diversity of Capsicum Based on rDNA-ITS Region

The genus Capsicum is comprised of 5 domesticated and more than 30 wild species. The region of nuclear ribosomal DNA internal transcribed spacers (rDNA-ITS) has widely been used for species identification, but has rarely been used in Capsicum. In this study, the evaluation of genetic diversity and a phylogenetic analysis were conducted using rDNA-ITS of 28 Capsicum accessions, including five domesticated and two wild species. We surveyed six conventional keys of domesticated species and another five traits in Capsicum accessions. Specific morphological characteristics were found in C. annuum, C. baccatum, and C.pubescens. Three subclones of each accession were sequenced, and rDNA-ITS polymorphisms were detected in all accessions excluding C. annuum, suggesting that incomplete concerted evolution occurred in rDNA-ITS of Capsicum. The genetic diversity was evaluated using nucleotide polymorphism and diversity. C. annuum had the lowest genetic diversity of all species in this study. The phylogenetic tree formed a species-specific clade for C. annuum, C. baccatum, and C. pubescens. The C. chinense clade existed in the C. frutescens clade, implying that it was a cultivated variant of C. frutescens. C. chacoense likely belonged to the C. baccatum complex according to its morphologic and genetic features. This study indicated that the rDNA-ITS region can be used for simple identification of domesticated Capsicum species.

Notes on Crocus L. Series Flavi Mathew (Iridaceae) and a new species with unique corm structure

Crocus asymmetricus (Iridaceae) is described as a new species endemic to the southern part of the Anatolian diagonal in Turkey. It is phylogenetically related to C. vitellinus and morphologically to C. antalyensis, but differs from these species in showing an asymmetric corm and a single point of root emergence. Both C. antalyensis and C. asymmetricus are illustrated and compared in this paper. A Bayesian phylogenetic tree of the nuclear rDNA ITS region confirms the affiliation of C. asymmetricus to C. ser. Flavi, and its close relationship to C. vitellinus. A new identification key to the species of C. ser. Flavi occurring in Turkey is also presented.

How to resolve cryptic species of polypores: an example in Fomes

Species that cannot be easily distinguished based on morphology, but which form distinct phylogenetic lineages based on molecular markers, are often referred to as cryptic species. They have been proposed in a number of fungal genera, including the basidiomycete genus Fomes. The main aim of this work was to test new methods for species delimitation in cryptic lineages of polypores, and to define useful characters for species identification. A detailed examination of a number of different Fomes strains that had been collected and isolated from different habitats in Italy and Austria confirmed the presence of distinct lineages in the Fomes fomentarius clade. Our zero hypothesis was that the Mediterranean strains growing on Quercus represent a species which can be delimited based on morphological and physiological characters when they are evaluated in statistically relevant numbers. This hypothesis was tested based on phylogenetic analysis of the rDNA ITS region, morphological characters of basidiomes and pure cultures, growth rates and optimum growth temperature experiments, mycelial confrontation tests, enzyme activity tests and volatile organic compound (VOC) production. The Mediterranean lineage can unambiguously be delimited from F. fomentarius. A syntype of an obscure and previously synonymized name, Polyporus inzengae, represents the Mediterranean lineage that we recognize as Fomes inzengae, a distinct species. The rDNA ITS region is useful for delimitation of Fomes species. Moreover, also a variety of morphological characters including hymenophore pore size, basidiospore size, and diameter of skeletal hyphae are useful delimiting characters. The ecology is also very important, because the plant host appears to be a central factor driving speciation. Physiological characters turned also out to be species-specific, e.g. daily mycelial growth rates or the temperature range of pure cultures. The production of VOCs can be considered as a very promising tool for fast and reliable species delimitation in the future.

A new species of Bunium (Apiaceae: Apioideae) from Central Anatolia in Turkey: Bunium sivasicum

A new species of Bunium from Central Anatolia (Turkey), Bunium sivasicum (Apiaceae), is described and illustrated. The taxonomic affinities of the new species are determined by detailed morphological investigation and molecular phylogenetic study. Nuclear rDNA-ITS region sequences are used to reveal the phylogenetic relationship of the new species. It belongs to the sect. Elegantia and is related to Bunium simplex, B elegans and B. verruculosum. The differences between the new species and its allies are discussed. Pollen characteristics and fruit surface features of the new species are investigated. The anatomical structure of the fruit is given in detail. In addition, the geographical distributions of the Bunium sivasicum and related taxa in Anatolia are also mapped.

A new species of Psathyrella (Psathyrellaceae, Agaricales) from Italy

Sporophores of a newPsathyrellaspecies have been reported for the first time as growing at the base ofCladiummariscusculms in the Botanical Garden of the University of Calabria, Rende, Cosenza, southern Italy. The fungus was initially identified asP.thujina(=P.almerensis) by means of both ecology and macro- and microscopic characteristics of the basidiomes, then referred toP.cladii-mariscisp. nov. after extraction, amplification, purification and analysis of the rDNA ITS region. We came to this conclusion after comparing our specimen with the descriptions of the taxa available in the literature for the genusPsathyrella.

The genus Coprinellus (Basidiomycota; Agaricales) in Pakistan with the description of four new species

Mushrooms with a thin-fleshed pileus that becomes plicate on opening, deliquescent lamellae and dark brown to blackish basidiospores are commonly called coprinoid mushrooms. The genusCoprinellusis one of the important lineages of coprinoid mushroom in the family Psathyrellaceae. Species-level taxonomy inCoprinellusis based mainly on the presence or absence and the structure of veil and cystidia on the pileus, of cystidia on the lamellae and on basidiospore morphology. In this study, four new species ofCoprinellus(Co.campanulatus,Co.disseminatus-similis,Co.pakistanicusandCo.tenuis) are described from Pakistan. Species descriptions are based on morphological and molecular data. Phylogenetic analyses based on nuc rDNA ITS region show that the new speciesCo.campanulatusandCo.disseminatus-similisare clustered in a clade including members of section Micacei;Co.tenuisfalls in a clade with members of section Domestici; andCo.pakistanicusrecovered in a separate clade adjacent to other recently described clades of genusCoprinellus. Morpho-anatomical descriptions of the new species and comparison with closely allied taxa are provided. With this study, the number of known species ofCoprinellusin Pakistan has reached eight.

Molecular Identification of Endophytic Fungi from Bark of Raru (Cotylelobium melanoxylon) that Produce the Antibacterial Compounds

<p class="IsiAbstrakIndo"><span lang="EN-GB">The Dipterocarpaceae plant, raru (</span><em><span lang="EN-GB">Cotylelobium melanoxylon</span></em><span lang="EN-GB">) is widespread in Southeast Asia. The bark of raru has been used by local communities in North Sumatera as antidiarrheal drugs due to its antibacterial compounds. The antibacterial activity of the </span><em><span lang="EN-GB">raru’s</span></em><span lang="EN-GB"> bark is induced by endophytic fungi that live in the region of the bark. This study aimed to identify the endophytic fungi-producer of antibacterial compounds in the bark of raru (</span><em><span lang="EN-GB">C. melanoxylon</span></em><span lang="EN-GB">) by means of molecular analysis. In general, endophytic fungi have the ability to inhibit the growth of pathogenic bacteria. Thirty-eight isolates of endophytic fungi were isolated from the bark of raru. Selection of isolates for antibacterial activity against </span><em><span lang="EN-GB">Escherichia coli</span></em><span lang="EN-GB"> ATCC 35218 and </span><em><span lang="EN-GB">Staphylococcus aureus</span></em><span lang="EN-GB"> ATCC 25923 used the dual culture assay. Selection using the dual culture assay yielded 6 endophytic fungal isolates that have the ability to inhibit the growth of test bacteria. EF10A sample was the most powerful isolate inhibiting the growth of both bacteria test. Those six bacteria molecularly identified used a sequence generated from ITS rDNA region. Based on rDNA ITS region sequences, isolate, the producers of the antibacterial compound were identified as </span><em><span lang="EN-GB">Talaromyces cellulolyticus, Penicillium purpurogenum, Aspergillus</span></em><span lang="EN-GB"> sp., </span><em><span lang="EN-GB">Trichoderma harzianum</span></em><span lang="EN-GB">, and </span><em><span lang="EN-GB">Aspergillus orizae</span></em><span lang="EN-GB">. The results of this study can be used by researchers to explore more potential endophytic fungi in raru plants (</span><em><span lang="EN-GB">C. melanoxylon</span></em><span lang="EN-GB">) as a source of medicine. The data obtained need to be supported by further research to isolate the bioactive compounds that can inhibit the growth of microbial pathogens.</span></p>