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

2021 ◽  
Vol 285 ◽  
pp. 05020
Author(s):  
Elena Lobodina ◽  
Ivan Suprun ◽  
Natalya Ageeva ◽  
Ekaterina Al-Nakib

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.

Horticulturae ◽  
2020 ◽  
Vol 6 (4) ◽  
pp. 87
Author(s):  
Kumpei Shiragaki ◽  
Shuji Yokoi ◽  
Takahiro Tezuka

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.


2016 ◽  
Vol 10 (1s) ◽  
pp. 35
Author(s):  
Jordan Merkuri ◽  
Stefania Mirela Mang ◽  
Ippolito Camele ◽  
Magdalena Cara ◽  
Gian Luigi Rana

Basidiomata of a wild mushroom macroscopically recognised as <em>Pleurotus ostreatus</em> were observed on an oak trunk in a mixed wood of northern Albania. Pure cultures of the fungus were then obtained on potato-dextrose-agar medium. Molecular analyses of genomic DNA of the fungus confirmed its identification. The rDNA ITS region nucleotide sequence of the studied <em>Pleurotacea</em> matched at 99% those of two <em>P. ostreatus</em> strains already present in NCBI GenBank database. The rDNA ITS nucelotide sequences of two pure cultures of the Albanian <em>P. ostreatus</em> were deposited in EMBL database under the accession numbers LN849458 and LN849459. One of the fungus isolates was subsequently cultivated under protected and semi-natural conditions. Productivity and biological efficiency of the Albanian <em>P. ostreatus</em> ranged from about 10% to 16% and from 33 to 53.33%, respectively. This seems to be the first report on the artificial cultivation of <em>P. ostreatus</em> in Albania and could have, in the next future, a high economic impact on development and diffusion of this important edible mushroom over the country.


Plant Disease ◽  
2010 ◽  
Vol 94 (5) ◽  
pp. 563-569 ◽  
Author(s):  
L. D. Maccarone ◽  
M. J. Barbetti ◽  
K. Sivasithamparam ◽  
R. A. C. Jones

Lettuce plants showing symptoms of lettuce big-vein disease were collected from fields in the Perth Metropolitan region of southwest Australia. When root extracts from each plant were tested by polymerase chain reaction (PCR) using primers specific to the rDNA internal transcribed spacer (ITS) region of Olpidium brassicae and O. virulentus, only O. virulentus was detected in each of them. The nucleotide sequences of the complete rDNA ITS regions of isolates from five of the root samples and 10 isolates of O. virulentus from Europe and Japan showed 97.9 to 100% identities. However, with the six O. brassicae isolates, their identities were only 76.9 to 79.4%. On phylogenetic analysis of the complete rDNA-ITS region sequences of the five Australian isolates and 10 others, the Australian isolates fitted within two clades of O. virulentus (I and II), and within clade I into two of its four subclades (Ia and Id). Japanese isolates had greatest sequence diversity fitting into both clades and into all of clade I subclades except Ib, while European isolates were restricted to subclades Ib and Id. When the partial rDNA-ITS region sequences of two additional southwest Australian isolates, four from Europe, and four from the Americas were included in the analyses, the Australian isolates were within O. virulentus subclades Ia and Id, the European isolates within subclade Ic, and the American isolates within subclades Ia and Ib. These findings suggest that there may have been at least three separate introductions of O. virulentus into the isolated Australian continent since plant cultivation was introduced following its colonization by Europeans. They also have implications regarding numbers of different introductions to other isolated regions. Lettuce big-vein associated virus and Mirafiori lettuce big-vein virus were both detected when symptomatic lettuce leaf tissue samples corresponding to the root samples from southwest Australia were tested using virus-specific primers in reverse transcription–PCR, so presence of both viruses was associated with O. virulentus occurrence.


1996 ◽  
Vol 13 (9) ◽  
pp. 1276-1295 ◽  
Author(s):  
M. A. Hershkovitz ◽  
L. A. Lewis

IMA Fungus ◽  
2019 ◽  
Vol 10 (1) ◽  
Author(s):  
Ursula Peintner ◽  
Regina Kuhnert-Finkernagel ◽  
Viana Wille ◽  
Franco Biasioli ◽  
Anton Shiryaev ◽  
...  

Abstract 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.


Plant Disease ◽  
2006 ◽  
Vol 90 (11) ◽  
pp. 1389-1394 ◽  
Author(s):  
Toshihiro Hayakawa ◽  
Takeshi Toda ◽  
Qu Ping ◽  
Joseph M. Mghalu ◽  
Shigeharu Yaguchi ◽  
...  

Isolates of an unidentified Rhizoctonia sp. (UN isolates) were obtained from Japanese zoysia grass (Zoysia japonica Steud) that exhibited symptoms of a new sheath rot disease. UN isolates were binucleate and showed hyphal fusion with tester isolates of Rhizoctonia anastomosis group (AG)-D. Those isolates were compared with isolates of subgroups I and II of Rhizoctonia AG-D based on cultural morphology, hyphal growth rate at different temperatures, anastomosis frequency, pathogenicity, and sequence analysis of the internal transcribed spacer (ITS) region of ribosomal DNA genes (rDNA-ITS region). The mycelial color of UN isolates was light yellow which differs from AG-D I but is similar to AG-D II. Sclerotia of UN isolates were dark brown in color and larger in size (1 to 3 mm in diameter) than those of AG-D subgroup I (1 mm in diameter), whereas isolates of AG-D II produced white mycelial clamps 4 to 5 mm in size. Hyphal growth rate of UN isolates was slower than that of two AG-D subgroups at several temperatures, especially 25°C. In pathogenicity tests on Japanese zoysia grass, UN isolates showed moderate disease severity and lower pathogenicity than isolates of AG-D subgroups I and II. Sequences of the rDNA-ITS region within UN isolates were almost homologous, but had lower homology with subgroups AG-D I or II. Phylogenetic trees constructed using ITS sequences showed that UN isolates formed an individual cluster that differed from the clusters of the two subgroups. We propose that UN isolates are a new subgroup of Rhizoctonia AG-D, subgroup III, and the name of the disease is “spring-rot” on Japanese zoysia grass.


2000 ◽  
Vol 104 (10) ◽  
pp. 1175-1183 ◽  
Author(s):  
H. Suga ◽  
T. Hasegawa ◽  
H. Mitsui ◽  
K. Kageyama ◽  
M. Hyakumachi

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