Mating patterns of dermatophytes of diverse origin in India

The mating patterns of Trichophyton mentagrophytes var. interdigitale (74 isolates) and the Microsporum gypseum complex (17 isolates) of diverse origin and T. rubrum (25 isolates) and T. tonsurans (10 isolates) of clinical origin were studied. The results of the study showed that the teleomorph of the Indian isolates of T. mentagrophytes belong to Arthroderma vanbreuseghemii, undetermined teleomorphs of T. mentagrophytes var. interdigitale (+) mating types, and undetermined teleomorphs of T. mentagrophytes var. interdigitale indeterminate mating types. All the isolates of T. rubrum and T. tonsurans were found to be of the (-) mating type.     

Serotypes and mating types of clinical strains ofCryptococcus neoformans isolated in Taiwan

Twenty-one strains ofCryptococcus neoformans isolated from patients in Taiwan were characterized for serotypes and mating types. Slide agglutination test was performed with 8 factor-specific sera (Iatron Company, Japan) to determine the serotypes. Wheat bran agar (WBA) and malt extract agar (MEA, Wickerham) media were used for the mating tests. Twenty of the isolates were of serotype A, and one was serotype B. Except for 2 strains of serotype A, all of the serotype A strains mated withFilobasidiella neoformans var.neoformans, mating type a. The only serotype B strain mated withF. neoformans var.bacillispora mating type a in MEA medium. These data revealed the low prevalence (1/21; 4.8%) ofC. neoformans var.gattii in Taiwan, a subtropically located island.     

Phylogeny of the genera Trichophyton using mitochondrial DNA analysis

Diversity of mitochondrial DNA (mtDNA) was investigated in 92 Trichophyton rubrum strains, 2 T. mentagrophytes var. mentagrophytes, 2 T. m. vor. interdigitale, 2 T. m. var. goetzii, 1 T. m. var. erinacei, 2 T. quinckeanum, 2 T. schoenleinii, 1 T. tonsurans, 2 T. verrucosum var. album, 2 T. v. var. discoides, 1 T. violaceum var. violaceum, 1 Arthroderma benhamiae, and 1 A. vanbreuseghemii using endonucleases, Hae III, Msp I, Hind III, Xba I, and Bgl II. Trichophyton species were divided into 7 groups, and a phylogenetic tree was produced based on sequence divergence within mtDNA. The following results were obtained: (1) T. rubrum was divided into 2 groups Type I and Type II, and was suggested to be a complex. (2) A. benhamiae was closely related to T. m. var. erinacei. (3) T. rubrum Type II, T. tonsurans, and A. vanbreuseghemii showed identical restriction profiles, and were suggested to be closely related to each other or identical. (4) T. quinckeanum and T. schoenlenii showed identical restriction profiles, which differed slightly from those of A. vanbreuseghemii. (5) mtDNA analysis was useful in identifying pleomorphic strains.     

Equine ringworm due to Trichophyton mentagrophytes var. quinckeanum

Summary T. mentagrophytes var.quinckeanum was isolated from a case of equine tinea and this finding is thought to be unique. Although it is considered that the fungus is closely related toT. mentagrophytes, the small differences in the relative pathogenicity for animals and man, the unusual clinical pattern which can be seen, and the downy nature of primary isolates on Sabouraud's medium are features not to be expected withT. mentagrophytes infections. The ability of this fungus to produce green fluorescence in the hairs of infected guinea pigs is felt to be its most unique feature and warrants its separate identity asT. mentagrophytes var.quinckeanum.     

Assessment ofGibberella fujikuroi mating type by PCR

Mating type (MAT)-specific fragments of the two idiomorphs ofGibberella fujikuroi (anamorph,Fusarium moniliforme) were obtained by PCR amplification using primers to conserved regions ofMAT homologs from other fungal species and used to assign mating type by molecular criteria rather than the arbitrary historical designation. Mating type—strains of mating populations A-E and a mating type+strain of mating population F carry an α-box motif and should therefore be designatedMAT-1. Mating type+strains of mating populations A-E and a mating type—strain of mating population F carry an HMG-box motif and should be designatedMAT-2. Thus, assessment of mating type ofG. fujikurol strains can be easily achieved usingMAT-specific primers.     

Taxonomic position of a Chinese <Emphasis Type="Italic">Pleurotus</Emphasis> “Bai-Ling-Gu”: it belongs to <Emphasis Type="Italic">Pleurotus eryngii</Emphasis> (DC.: Fr.) Quél. and evolved independently in China

The Chinese Bai-Ling-Gu is a mushroom named Pleurotus eryngii var. tuoliensis C.J. Mou. This species has been identified as P. nebrodensis or P. eryngii var. nebrodensis. We examined its taxonomic position by analysis of mating, cultivation, and rDNA sequences, and concluded as follows. (1) Bai-Ling-Gu mated with P. eryngii var. eryngii, and the F₁ and F₂ formed fruit bodies. (2) Bai-Ling-Gu mated with P. eryngii var. ferulae, and the F₁ formed fruit bodies. (3) In the di-mon mating test, P. eryngii var. nebrodensis from Sicily mated with monokaryons of P. eryngii var. eryngii but mated hardly at all with those of Bai-Ling-Gu and P. eryngii var. ferulae. The di-mon mating pattern of Bai-Ling-Gu resembled those of P. eryngii var. ferulae. (4) The partial sequences of rDNA ITS1 and IGS1 from the epitype of P. nebrodensis were identical with those from P. eryngii var. nebrodensis from Sicily but differed from those from Bai-Ling-Gu. (5) The strains of P. eryngii var. eryngii and P. eryngii var. ferulae were in a group, the strains of P. eryngii var. nebrodensis from Sicily were in another group, and the strains of Bai-Ling-Gu were in the other group in both the phylogenetic trees based on the ITS1 and the IGS1 sequences. These results led to the conclusion that Bai-Ling-Gu is a variety of P. eryngii and evolved independently in China. It is satisfactory to identify Bai-Ling-Gu with P. eryngii var. tuoliensis C.J. Mou.     

Dermatophyte species,microscopic and cultural examination

The dermatophytes Epidermophyton floccosum, Microsporum canis, Trichophyton mentagrophytes var. interdigitale, T. rubrum, and T. verrucosum were compared with respect to the direct microscopic examination of a clinical material and the number of colonies obtained by culture.It was found that the results of microscopy as well as of culture depended to a marked extent upon which species were the cause of the mycosis.The extremes were E. floccosum and T. mentagrophytes var. interdigitale which showed 7.5 % and 32.5 % isolates with negative microscopic findings and 45.5 % and 5.0 % isolates with 10 colonies.     

Comparative Immunology of Selected Wild Types,Varieties and Mutants of Chlamydomonas*

SYNOPSIS. Rabbit antisera to strains of different mating types of Chlamydomonas moewusii and to one strain of C. eugametos were tested against strains of C. reinhardti, C. eugametos, and C. moewussi, and against strains of 2 varieties and 6 mutant types of C. moewusii. Technics of double diffusion, absorption, and immunoelectrophoresis revealed marked serological differences between the sexually incompatible, distinct genetic species C. moewusii and C. reinhardti. Less distinct serological differences were resolved between C. moewusii and the so-called “species”C. eugametos, which is sexually compatible with the former, thus reconfirming the conspecificity between the 2 strains as suggested by Gowans. Marked serological differences were noted between C. moewusii and 2 of its varieties (C. moewusii var. tenuichloris and C. moewusii var. rotunda) which constitute 2 additional genetic species because of sexual incompatibility between themselves and with C. moewusii. Wild types and certain mutants of C. moewusii were compared serologically and could be distinguished on this basis. Strains of different mating types as well as certain mutant strains (e.g., paralyzed flagella, flagella-less, twins and monsters) could be differentiated serologically altho differences were often very subtle. Some antigens were common to organisms tested.     

Multicenter study of dermatophyte distribution in the metropolitan area of Barcelona (Catalonia,Spain)

The results of a multicenter study on the distribution of dermatophyte species in the metropolitan area of Barcelona (Spain) are presented.In the period 1977–1983, 638 dermatophyte strains were isolated from tineas by three different mycological laboratories. Zoophilic strains were prevalent comprising 64.4% of the isolates. T. mentagrophytes var. granulosum was isolated in 51% of the cases.A signigicant drop in the prevalence of T. tonsurans and T. violaceum was noted. Important variations were observed in dermatophyte distribution in relation to other geographical areas of Spain.     

Relationship Between Phenotypic and Genotypic Characteristics of <Emphasis Type="Italic">Trichophyton mentagrophytes</Emphasis> Strains Isolated from Patients with Dermatophytosis

According to epidemiological, clinical and mycological criteria, it has long been admitted that the Trichophyton mentagrophytes species includes two varieties: a zoophilic variety (var. mentagrophytes) and an anthropophilic variety (var. interdigitale) that involve the upper and the lower part of the body, respectively. The further application of molecular techniques to the characterization of dermatophyte strains showed that this classification is unreliable. The aim of our study was to assess the usefulness of PCR–RFLP (restriction fragment length polymorphism) and sequencing in the characterization of T. mentagrophytes strains taken from Tunisian patients. The study was carried out in 2008 in the laboratory of Parasitology–Mycology of Farhat Hached University Hospital, Sousse, Tunisia. A total of 133 strains were isolated from 133 patients addressed to the laboratory for dermatological lesions very evocative of dermatomycosis. Eighty strains were isolated from lesions located on the lower part of the body (onychomycosis, tinea pedis) and 53 strains from the upper part of the body (tinea capitis, tinea corporis). All strains were submitted to mycological examination (direct microscopic examination and culture on Sabouraud medium) and further investigated by using RFLP analysis of the PCR-amplified ITS1-5.8 s-ITS2 region of the ribosomal DNA and the MvaI restriction enzyme. In addition, 62 strains were further submitted to a sequencing of the ITS1-5.8 s-ITS2 region. On the basis of mycological criteria, all strains were diagnosed as T. mentagrophytes. All strains produced the same RFLP pattern and were identified as T. mentagrophytes interdigitale regardless of the location of lesions. Out of the 62 sequenced strains, 16 were found anthropophilic and 46 were zoophilic. In conclusion, all strains provisionally diagnosed as T. mentagrophytes on the basis of mycological criteria were shown to belong to T. interdigitale by using PCR–RFLP and sequencing irrespective of the site of lesions. The predominance of zoophilic strains needs further investigation.     

Clavispora,a new yeast genus of the Saccharomycetales

In Candida lusitaniae van Uden et do Carmo-Sousa (1959), strains of opposite sex have been found. Cells of the opposite mating types conjugate and form asci with one to four clavate spores. These are easily liberated from the ascus. The type strain of Candida obtusa (Dietrichson) van Uden et do Carmo-Sousa ex van Uden et Buckley (1970) also produces ascospores after mating with one of the strains of Candida lusitaniae. As clavate ascospores are unknow in yeasts, a new genus, Clavispora, is proposed with Clavispora lusitaniae spec. nov. as type species.     

Exocellular enzyme activity of dermatophytes and other fungi isolated from ruminants in Southern Iraq

Sixteen fungal species were isolated from 182 specimens collected from four ruminants (buffalo, camel, cattle and sheep) in Southern Iraq. Fungi represented by five species of dermatophytes and eleven species of other fungi were screened for the activity of four enzymes; keratinase, proteinase, lipase and amylase. Keratinase was found to be produced by all of the dermatophytes and non-dermatophytes, except for Paecillomyces variottii and Scytalidium lignicola. However, high keratinase activity was expressed by the dermatophytic species particularly by Trichophyton mentagrophytes var. erinacei and Microsporum gypseum. Three dermatophytes viz. M. gypseum, T. verrucosum and T. mentagrophytes var. nodulare were capable of producing protease, lipase and amylase. Although, T. mentagrophytes var. erinacei showed high protease activity, it did not produce lipase and amylase. On the contrary most of the non-dermatophytic species revealed protease and lipase activities higher than the dermatophytes. The Curvularia spp. isolates showed the highest protease and amylase activity, while Aspergillus parasiticus revealed the highest activity of lipase and amylase. No correlation was observed between enzyme activity and the growth rate of the examined fungi. This revised version was published online in June 2006 with corrections to the Cover Date.     

Differences in Mitochondrial Genome Organization of Cryptococcus Neoformans Strains

The organization of the mitochondrial genomes in two strains belonging in different varieties of Cryptococcus neoformans was analysed. Physical maps of the mtDNA of the IFM5844 (var. neoformans) and IFO410 (var. grubii) strains were constructed by using EcoRI and EcoRV restriction enzymes; functional maps were constructed by hybridization, cloning and sequencing. Most of the genes important in the mitochondrial function (ND1, ND2, ND3, ND4, ND4L, ND5, ND6, ATP6, ATP9, COX1, COX2 and COB) and protein synthesis (SsrRNA and LsrRNA) were localized. We did not find any differences between the strains in the order of these genes. However, they differed significantly in the sizes of the mtDNAs: 32.6 kb for IFM5844, and 24.1 kb for IFO410. This can be attributed to two large regions of the mtDNA. In these regions, differences were found in the numbers of introns in COX1 (no intron in var. grubii, 5 introns in var. neoformans), COB (1 intron in var. grubii, 2 introns in var. neoformans), LsrRNA (no intron in var. grubii, 2 introns in var. neoformans), and ND5 (no intron in var. grubii, 1 intron in var. neoformans) genes. In several introns of the COB and COX1 genes LAGLIDADG motifs were found. Differences were also observed in the nucleotide sequences of some genes and in the sizes and sequences of intergenic regions. The nucleotide sequences of the genes of the IFM and IFO strains were compared with those of the H-99 and JEC 21 strains from the database. Surprisingly high similarities were found between the strains belonging in var. grubii (IFO 410 and H-99) and var. neoformans (IFM 5844 and JEC 21).     

Rapid discrimination of <Emphasis Type="Italic">Porphyra tenera</Emphasis> Kjellman var. <Emphasis Type="Italic">tamatsuensis</Emphasis> Miura by PCR-RFLP

To allow to discriminate rapidly the strains of Porphyra tenera var. tamatsuensis, cultivars of which grow more vigorously than strains of P. tenera var. tenera, strains of both varieties were examined by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analysis using mitochondrial DNA related to the ATP synthase F0 subunit 6 (ATP6) gene. The lengths of all sequences in this region of three strains of each variety were 670 bp and had just a single nucleotide substitution. Digestion with the restriction enzyme of TaaI yielded three visible bands which appeared in the three strains of P. tenera var. tamatsuensis, whereas two bands appeared in the three strains of P. tenera var. tenera. We therefore conclude that PCR-RFLP analysis is a valuable tool for discrimination of P. tenera var. tamatsuensis among the stock of P. tenera strains used for mariculture.     

Repeated Ubiquitin Genes in Trichophyton mentagrophytes

Using Trichophyton mentagrophytes cDNA as a template, we performed PCR amplification with the UB1S and UB1R primers for isolation of a partial fragment of T. mentagrophytes ubiquitin gene. We screened the T. mentagrophytes cDNA library prepared with Uni-ZAP phage vector by hybridization with the T. mentagrophytes ubiquitin fragment, UB1S-UB1R. The nucleotide and deduced amino acid sequences of T. mentagrophytes ubiquitin gene encoded two ubiquitin repeats in 162 bp to 624 bp. The deduced amino acid sequences of ubiquitin from T. mentagrophytes shared about 98% similarity with those of Saccharomyces cerevisiae, Neurospora crassa, and Cryptococcus neoformans. Received: 28 April 1999 / Accepted: 22 June 1999     

A MAT1–2 wild‐type strain from Penicillium chrysogenum: functional mating‐type locus characterization,genome sequencing and mating with an industrial penicillin‐producing strain

In heterothallic ascomycetes, mating is controlled by two nonallelic idiomorphs that determine the ‘sex’ of the corresponding strains. We recently discovered mating‐type loci and a sexual life cycle in the penicillin‐producing fungus, Penicillium chrysogenum. All industrial penicillin production strains worldwide are derived from a MAT1‐1 isolate. No MAT1‐2 strain has been investigated in detail until now. Here, we provide the first functional analysis of a MAT1‐2 locus from a wild‐type strain. Similar to MAT1‐1, the MAT1‐2 locus has functions beyond sexual development. Unlike MAT1‐1, the MAT1‐2 locus affects germination and surface properties of conidiospores and controls light‐dependent asexual sporulation. Mating of the MAT1‐2 wild type with a MAT1‐1 high penicillin producer generated sexual spores. We determined the genomic sequences of parental and progeny strains using next‐generation sequencing and found evidence for genome‐wide recombination. SNP calling showed that derived industrial strains had an uneven distribution of point mutations compared with the wild type. We found evidence for meiotic recombination in all chromosomes. Our results point to a strategy combining the use of mating‐type genes, genetics, and next‐generation sequencing to optimize conventional strain improvement methods.     

Variability of biochemical characteristics in strains of Trichophyton mentagrophytes

The characteristics of 23 strains of Trichophyton mentagrophytes isolated from clinical samples, were studied. Sabouraud agar was the most adequate medium to inhance at their utmost the different taxonomical features. Variability of enzymatic reactions in strains of T. mentagrophytes was observed.     

Mating Type Inheritance in Glaucoma*

SYNOPSIS. Mating was observed in collections of Glaucoma from 2 localities in Illinois. The collections could be divided into 2 syngens on the basis of mating type reactions. Corticotype analysis showed that syngen 1 was intermediate in meridian number between Glaucoma chattoni and Glaucoma scintillans, while syngen 2 had the same range as Glaucoma scintillans (Lee, unpublished). The major features of mating type inheritance in syngen 1 are that exconjugant clones usually have identical mating types and that mating types I and II appear in an approximately 1:1 ratio in successive generations. This is the result expected in genic mating type inheritance if one of the original parents was a homozygote and the other a heterozygote at the mating type locus. No significant immaturity period was found in these strains. Only 2 viable pairs were obtained from syngen 2 crosses, but results from these suggest that mating type inheritance may be epigenetic and a longer immaturity period may characterize this syngen.     

风毛菊属6种植物的核型分析

采用常规压片技术对分布于横断山区菊科(Compositae)风毛菊属(Saussurea DC.)的6种植物进行染色体数目和核型分析。结果表明:尖苞雪莲（S.polycolea var.acutisquama）核型公式为:2n=2x=32=20m+12sm,属2B型;球花雪莲核（S.globosa）型公式为:2n=2x=34=16m+18sm,属2B型;重齿风毛菊(S.katochaete)核型公式为:2n=2x=32=8m+18sm+6st,属3B型;柱茎风毛菊(S.columnaris)核型公式为:2n=2x=32=24m+8sm,属2B型;禾叶风毛菊(S.graminea)核型公式为:2n=2x=28=8m+18sm+2st,属3B型;长毛风毛菊(S.hieracioides)核型公式为:2n=2x=32=12m+16sm+4st,属2B型。6个种染色体中均未发现随体。其中尖苞雪莲和柱茎风毛菊染色体为首次报道。     

Mating behaviour of Nannizzia otae (=Microsporum canis)

One hundred and ninety-eight isolates of Microsporum canis, obtained from humans and animals in 12 countries, were paired with the two Japanese tester strains of Nannizzia otae (= M. canis), VUT 74037 (CDC B-2094+) and VUT 74039 (CDC B-2095–). One hundred and forty-one (72%) produced either gymnothecia or pseudogymnothecia in crosses with VUT 74037. Fifty-seven (28%) were nonreactive. None of the paired isolates reacted with VUT 74039. The number of nonreactive isolates decreased to 17% when 104 of the 198 isolates were paired with one additional tester strain of each mating type. All sexually reacting strains, however, belonged to the (–) mating type. Crosses between nonreactive isolates did not result in ascocarp formation.The F₁ generations from three different strongly reactive crosses were all characterized by poor ascospore germination. Most of the monoascospore progeny that germinated to form mature colonies were nonreactive in crosses to determine their mating type. Others reacted predominantly as the (+) mating type, thereby precluding the likelihood of an associated lethal factor accounting for the lack of this mating type in our clinical isolates. Several explanations for this phenomenon are presented.