scholarly journals Novel Pathogenic Mucorales Identified Using the Silkworm Infection Model

2021 ◽  
Vol 7 (11) ◽  
pp. 995
Author(s):  
Suresh Panthee ◽  
Hiroshi Hamamoto ◽  
Yayoi Nishiyama ◽  
Atmika Paudel ◽  
Kazuhisa Sekimizu

Mucormycosis, a rare but highly fatal infection, is caused by fungi of the order Mucorales. Due to their ubiquitous nature, reduced susceptibility to antifungals, acid tolerance, and ability to infect immunocompromised patients through rapid dissemination, these fungi have been frequently reported to infect the COVID-19 patients. In order to develop strategies to overcome mucormycosis, it is essential to understand and identify novel Mucorales present in the environment. In this study, we report the identification of four novel pathogenic Mucorales using the silkworm (Bombyx mori) model. The strains’ phylogeny was analyzed using the genome sequence of the large subunit ribosomal ribonucleic acid (LSU rRNA) and the internal transcribed spacer (ITS) region, where strains 1-3, 5-3, and S286-1101 claded with Mucor orantomantidis, and strain 827-14 claded with Backusella lamprospora. All the strains had a cold-sensitive phenotype with their inability to grow prominently at 4 °C. Mucor sp. 1-3 and 5-3 were characterized by their filamentous and yeast-like growth under aerobic and anaerobic conditions, respectively. The yeast colonies of Mucor sp. 5-3 had multipolar budding cells often observed with cleaved cell surfaces under a scanning electron microscope. We further found that these strains were able to kill immunocompromised mice suggesting their pathogenicity to mammals. Our study established an invertebrate model-based screening system to identify novel pathogenic Mucorales from the natural environment and provided a clue towards the rapid increase in COVID-19 related mucormycosis.

2007 ◽  
Vol 57 (2) ◽  
pp. 414-418 ◽  
Author(s):  
Puja Saluja ◽  
G. S. Prasad

Two novel anamorphic yeast strains (S-15LT and 3-C1) were isolated from the inflorescences of plants collected in two different towns in Rajasthan State, India. Sequencing of the D1/D2 domains of the large-subunit (LSU) rDNA and the internal transcribed spacer (ITS) regions suggested they are strains of the same species. Phenotypic characteristics such as the absence of fermentation, the absence of sexual structures and ballistoconidia, the assimilation of myo-inositol and d-glucuronate, and positive Diazonium blue B and urease reactions indicated that these strains belong to the genus Cryptococcus. The novel strains differed from Cryptococcus laurentii in six physiological tests and differed from other related species in more than six tests. A phylogenetic analysis of the sequences of the D1/D2 domains of the LSU rDNA and the ITS regions placed these strains in the Bulleromyces clade within the order Tremellales, with C. laurentii as their closest described relative. The novel strains showed 1.6 and 7.5 % divergence in the D1/D2 domain of the LSU rDNA and ITS regions, respectively, with respect to C. laurentii. The divergence from other species was more than 3 % for the D1/D2 domain and more than 9 % for the ITS region. On the basis of the phenotypic and molecular data, strains S-15LT and 3-C1 represent a novel species within the genus Cryptococcus, for which the name Cryptococcus rajasthanensis sp. nov. is proposed. The type strain is S-15LT (=MTCC 7075T=CBS 10406T).


Plant Disease ◽  
2021 ◽  
Author(s):  
Xiulan Xu ◽  
Si-yi Liu ◽  
Yicong Lv ◽  
Qian Zeng ◽  
Yinggao Liu ◽  
...  

Photinia × fraseri is a well-known green plant mainly distributed in the Yangtze River and Yellow River Basin, east and southwest of China (Guan et al. 2013). In October 2020, typical leaf blight symptoms on roughly 10% leaves in a Photinia × fraseri shrub were observed in the campus of Sichuan Agricultural University (30°42′19″ N, 103°51′29″ E). Initially, chlorotic lesions with brown margins occurred on the leaf margin, then the large patches formed to cause leaves necrotic, finally lesions to dry and acervulus bred in 2–4 months later. Five single conidium isolates were carried out (Chomnunti et al. 2014) cultured on potato dextrose agar (PDA) at 25 ℃. All isolates shared similarly morphological characteristics, which was white and thin, and the reverse were yellowish. Mycelium was hyaline, sparsely septate, measuring 1–4 μm in diam. Conidiogenesis formed after 7 days. Conidiogenous cells were discrete, lageniform, smooth, thin-walled, colorless. Conidia were fusiform, straight to slightly curved, 4-septate, 21–30 × 5–7 μm (x ̅= 27 × 6.0 μm, n=30); basal cells were obconic with truncate base, hyaline, thin- and smooth-walled, 4–7 μm long (x ̅= 5.5 μm, n=30); three median cells were doliiform with thick walls, concolorous, olivaceous, constricted at the septa, and septa and periclinal walls were darker than the rest of the cell, 14–20 μm long (x ̅= 17 μm, n=30); apical cells were hyaline, conic to cylindrical, 3.0–6.5 μm long (x ̅= 4.5 μm, n=30), with 2–4 (mostly 3) tubular apical appendages arising from the upper portion, rarely branched, 7.5–18 μm long (x ̅= 12 μm, n=50); basal appendage was single, unbranched, 3–10 μm long (x ̅= 6.5 μm, n=30). DNA was extracted from the representative strain (SICAUCC 21-0012), and the internal transcribed spacer (ITS) region, the large subunit of the nrDNA (LSU), translation elongation factor 1-alpha (tef1-α), and partial sequences of β-tubulin (tub2) were amplified by polymerase chain reaction and sequenced with primers ITS5/ITS4, LR0R/LR5, 728F/1567R, and Bt2a/Bt2b, respectively (Zhang et al. 2012, Ariyawansa & Hyde 2018). The sequences were deposited in GenBank, viz. MZ453106, MZ453108, MZ467300, MZ467301, respectively. The nucleotide blast showed 99% (ITS, 0 gaps), 100% (tub2, 0 gaps), 100% (tef1-α, 0 gaps) identities with the ex-type Pestalotiopsis trachicarpicola Yan M. Zhang & K. D. Hyde (IFRDCC 2440). The fungus was identified as P. trachicarpicola combined with phylogeny and morphology (Maharachchikumbura et al. 2012, Zhang et al. 2012). To conduct Koch’s postulates, five healthy 6-year-old P. × fraseri were inoculated with 10 µl spore suspension (106 conidia/ml) onto the wounded sites (five leaves per plant, ~1 to 2 years old) via sterile pin, and five healthy plants treated with sterile dH2O as controls (Yang et al. 2021). The plants were placed in a greenhouse at 25°C with relative humidity >80%. After 2 months, leaf blight symptoms gradually emerged on inoculated leaves, and the controls were symptomless. Fungal isolates from symptomatic plants showed similar morphological characteristics as SICAUCC 21-0012, and the pathogen was not isolated from asymptomatic plants. To our knowledge, this is the first report of leaf blight caused by P. trachicarpicola on Photinia × fraseri in China. Disease management should be adopted properly to restore and improve its ornamental value.


2015 ◽  
Vol 65 (Pt_6) ◽  
pp. 1855-1859 ◽  
Author(s):  
Ana Raquel O. Santos ◽  
Elisa S. Faria ◽  
Marc-André Lachance ◽  
Carlos A. Rosa

Five strains of a novel methanol-assimilating yeast species were isolated from mango (Mangifera indica) leaves collected at the campus of the Federal University of Minas Gerais in Brazil. The sequences of the internal transcribed spacer (ITS) region and the D1/D2 domains of the large subunit of the rRNA gene showed that this species belongs to the Ogataea clade and is related to O. allantospora, O. chonburiensis, O. dorogensis, O. kodamae, O. paradorogensis and Candida xyloterini (Ogataea clade). The novel species differs in the D1/D2 domains of the large subunit of the rRNA gene by 12 to 40 substitutions from these Ogataea species. The name Ogataea mangiferae sp. nov. is proposed for this novel species. The type strain of Ogataea mangiferae sp. nov. is UFMG-CM-Y253T ( = CBS 13492T). The Mycobank number is MB 811646.


Phytotaxa ◽  
2019 ◽  
Vol 397 (1) ◽  
pp. 23
Author(s):  
WEI SUN ◽  
BINGJIE LIU ◽  
RONG FU ◽  
XINGZHONG LIU ◽  
MEICHUN XIANG

During survey on rock-inhabiting fungi from karst habitats in Guizhou, China, two new species in Cyphellophora were discovered and identified. Phylogenetic analysis based on combined sequences of the nuclear large subunit (nucLSU) and internal transcribed spacer (ITS) region of ribosomal DNA revealed that the tested isolates, clustered into two clades that well affiliated in the genus of Cyphellophora. Morphological characteristics were also supported the estabolishment of the new species. Herewith Cyphellophora botryose sp. nov. and Cyphellophora guizhouensis sp. nov. were described and their differences from closely related species were discussed.


2020 ◽  
Vol 8 (5) ◽  
pp. 689 ◽  
Author(s):  
Wenjun Li ◽  
Ming Hu ◽  
Yang Xue ◽  
Zhijun Li ◽  
Yanfei Zhang ◽  
...  

Bayberry (Myrica rubra) is a commercial fruit in China. For the past seven years, twig blight disease has been attacking bayberry plantations in Shantou City, Guangdong Province, China, leading to destructive damage and financial loss. In this study, five fungal species associated with twig dieback and stem blight were identified based on morphological characteristics combined with multilocus sequence analysis (MLSA) on the internal transcribed spacer (ITS) region, partial sequences of β-tubulin (tub2), translation elongation factor 1-α (tef1-α), large subunit ribosomal RNA (LSU) and small subunit ribosomal RNA (SSU) genes, which are Epicoccum sorghinum, Neofusicoccum parvum, Lasiodiplodia theobromae, Nigrospora oryzae and a Pestalotiopsis new species P. myricae. P. myricae is the chief pathogen in fields, based on its high isolation rate and fast disease progression after inoculation. To our knowledge, this is the first study reporting the above five fungi as the pathogens responsible for bayberry twig blight. Indoor screening of fungicides indicates that Prochloraz (copper salt) is the most promising fungicide for field application, followed by Pyraclostrobin, 15% Difenoconazole + 15% Propiconazole, Difenoconazole and Myclobutanil. Additionally, Bacillus velezensis strain 3–10 and zeamines from Dickeya zeae strain EC1 could be used as potential ecofriendly alternatives to control the disease.


2013 ◽  
Vol 63 (Pt_9) ◽  
pp. 3501-3505 ◽  
Author(s):  
Liang Chen ◽  
Lin Zhang ◽  
Zhi-Hui Li ◽  
Feng-Li Hui

Two strains (NYNU 121010T and NYNU 121032) of a novel basidiomycetous yeast species belonging to the genus Sympodiomycopsis were isolated from insect frass collected from trunks of a pagoda tree (Sophora japonica L.) in Yantai, Shandong province, east China. The sequence analyses of the D1/D2 domain of the large subunit (LSU) rRNA gene and the internal transcribed spacer (ITS) region indicated that the closest relatives were Sympodiomycopsis kandeliae FIRDI 007T, Sympodiomycopsis paphiopedili CBS 7429T and Sympodiomycopsis sp. S6A. The D1/D2 sequences of the novel strains differed by 12 nt substitutions (2 %) from the type strain of S. kandeliae, and by 13 nt substitutions (2.2 %) from the type strain of S. paphiopedili and from Sympodiomycopsis sp. S6A. The novel strains differed from closely related species by more than 4.6 % substitutions in the ITS region. The novel strains can also be distinguished from S. kandeliae and S. paphiopedili on the basis of a number of morphological and physiological characteristics and represent a novel species in the genus Sympodiomycopsis, for which the name Sympodiomycopsis yantaiensis sp. nov. is proposed. The type strain is NYNU 121010T ( = CICC 32998T = CBS 12813T). The Mycobank deposit number is MB 804119.


2020 ◽  
Vol 70 (12) ◽  
pp. 6307-6312
Author(s):  
João Drumonde-Neves ◽  
Neža Čadež ◽  
Yazmid Reyes-Domínguez ◽  
Andreas Gallmetzer ◽  
Dorit Schuller# ◽  
...  

During a study of yeast diversity in Azorean vineyards, four strains were isolated which were found to represent a novel yeast species based on the sequences of the internal transcribed spacer (ITS) region (ITS1-5.8S–ITS2) and of the D1/D2 domain of the large subunit (LSU) rRNA gene, together with their physiological characteristics. An additional strain isolated from Drosophila suzukii in Italy had identical D1/D2 sequences and very similar ITS regions (five nucleotide substitutions) to the Azorean strains. Phylogenetic analysis using sequences of the ITS region and D1/D2 domain showed that the five strains are closely related to Clavispora lusitaniae, although with 56 nucleotide differences in the D2 domain. Intraspecies variation revealed between two and five nucleotide differences, considering the five strains of Clavispora santaluciae. Some phenotypic discrepancies support the separation of the new species from their closely related ones, such as the inability to grow at temperatures above 35 °C, to produce acetic acid and the capacity to assimilate starch. Neither conjugations nor ascospore formation were observed in any of the strains. The name Clavispora santaluciae f.a., sp. nov., is proposed to accommodate the above noted five strains (holotype, CBS 16465T; MycoBank no., MB 835794).


MycoKeys ◽  
2019 ◽  
Vol 56 ◽  
pp. 33-47 ◽  
Author(s):  
Arooj Naseer ◽  
Abdul Nasir Khalid ◽  
Rosanne Healy ◽  
Matthew E. Smith

The genus Hygrophorus is poorly studied from Asia. From Pakistan, only one species has been reported so far. Two new species in the genus have been collected from Himalayan oak forests of Pakistan. Hygrophorusalboflavescens (section Pudorini, subgenus Colorati) is characterised by its pure white, centrally depressed pileus, occurrence of white stipe with yellow patches at lower half and broader (4.98 μm) basidiospores. Hygrophorusscabrellus (section Hygrophorus, subgenus Hygrophorus) is characterised by its yellowish-green stipe with white apex that has fine scales on the entire stipe, an off-white pileus with dark green and greyish fibrils, ovoid to ellipsoid basidiospores and clavate 4-spored basidia. Macro- and micromorphological descriptions have revealed that both these taxa are not yet described. Phylogenetic estimation based on DNA sequences from the internal transcribed spacer (ITS) region and large subunit (LSU) of the nuclear ribosomal DNA (rDNA) genes, is congruent with the morphological characters that help to delimit these as new species of Hygrophorus. Allied taxa are also compared.


F1000Research ◽  
2018 ◽  
Vol 7 ◽  
pp. 1418 ◽  
Author(s):  
Adam R. Rivers ◽  
Kyle C. Weber ◽  
Terrence G. Gardner ◽  
Shuang Liu ◽  
Shalamar D. Armstrong

The internally transcribed spacer (ITS) region between the small subunit ribosomal RNA gene and large subunit ribosomal RNA gene is a widely used phylogenetic marker for fungi and other taxa. The eukaryotic ITS contains the conserved 5.8S rRNA and is divided into the ITS1 and ITS2 hypervariable regions. These regions are variable in length and are amplified using primers complementary to the conserved regions of their flanking genes. Previous work has shown that removing the conserved regions results in more accurate taxonomic classification. An existing software program, ITSx, is capable of trimming FASTA sequences by matching hidden Markov model profiles to the ends of the conserved genes using the software suite HMMER. ITSxpress was developed to extend this technique from marker gene studies using Operational Taxonomic Units (OTU’s) to studies using exact sequence variants; a method used by the software packages Dada2, Deblur, QIIME 2, and Unoise. The sequence variant approach uses the quality scores of each read to identify sequences that are statistically likely to represent real sequences. ITSxpress enables this by processing FASTQ rather than FASTA files. The software also speeds up the trimming of reads by a factor of 14-23 times on a 4-core computer by temporarily clustering highly similar sequences that are common in amplicon data and utilizing optimized parameters for Hmmsearch. ITSxpress is available as a QIIME 2 plugin and a stand-alone application installable from the Python package index, Bioconda, and Github.


2000 ◽  
Vol 38 (5) ◽  
pp. 1869-1875 ◽  
Author(s):  
Aditya K. Gupta ◽  
Yatika Kohli ◽  
Richard C. Summerbell

A system based on PCR and restriction endonuclease analysis was developed to distinguish the seven currently recognizedMalassezia species. Seventy-eight strains, including authentic culture collection strains and routine clinical isolates, were investigated for variation in the ribosomal DNA repeat units. Two genomic regions, namely, the large subunit of the ribosomal gene and the internal transcribed spacer (ITS) region, were amplified by PCR, and products were digested with restriction endonucleases. The patterns generated were useful in identification of five out of sevenMalassezia species. M. sympodialis was readily distinguishable in that its ITS region yielded a 700-bp amplified fragment, whereas the other six species yielded an 800-bp fragment.M. globosa and M. restricta were very similar in the regions studied and could be distinguished only by performing a hot start-touchdown PCR on primers for the β-tubulin gene. Primers based on the conserved areas of the Candida cylindracealipase gene, which were used in an attempt to amplifyMalassezia lipases, yielded an amplification product after annealing at 55°C only with M. pachydermatis. This specific amplification may facilitate the rapid identification of this organism.


Sign in / Sign up

Export Citation Format

Share Document