scholarly journals Mating Type Protein Mat1-2 from Asexual Aspergillus fumigatus Drives Sexual Reproduction in Fertile Aspergillus nidulans

2008 ◽  
Vol 7 (6) ◽  
pp. 1029-1040 ◽  
Author(s):  
Wioletta Pyrzak ◽  
Karen Y. Miller ◽  
Bruce L. Miller
Keyword(s):  
Aspergillus Fumigatus ◽  
Aspergillus Nidulans ◽  
Mating Type ◽  
Sexual Development ◽  
Genetic System ◽  
Gene Replacement ◽  
Sexual Cycle ◽  
Homologous Gene ◽  
Gene Exchange ◽  
First Report

ABSTRACT The lack of an experimentally amenable sexual genetic system in Aspergillus fumigatus is a major limitation in the study of the organism's pathogenesis. A recent comparative genome analysis revealed evidence for potential sexuality in A. fumigatus. Homologs of mating type genes as well as other genes of the “sexual machinery” have been identified in anamorphic A. fumigatus. The mat1-2 gene encodes a homolog of MatA, an HMG box mating transcriptional factor (MatHMG) that regulates sexual development in fertile Aspergillus nidulans. In this study, the functionalities of A. fumigatus mat1-2 and the Mat1-2 protein were determined by interspecies gene exchange between sterile A. fumigatus and fertile A. nidulans. Ectopically integrated A. fumigatus mat1-2 (driven by its own promoter) was not functional in a sterile A. nidulans ΔmatA strain, and no sexual development was observed. In contrast, the A. fumigatus mat1-2 open reading frame driven by the A. nidulans matA promoter and integrated by homologous gene replacement at the matA locus was functional and conferred full fertility. This is the first report showing that cross species mating type gene exchange between closely related Ascomycetes did not function in sexual development. This is also the first report demonstrating that a MatHMG protein from an asexual species is fully functional, with viable ascospore differentiation, in a fertile homothallic species. The expression of mat1-2 was assessed in A. fumigatus and A. nidulans. Our data suggest that mat1-2 may not be properly regulated to allow sexuality in A. fumigatus. This study provides new insights about A. fumigatus asexuality and also suggests the possibility for the development of an experimentally amenable sexual cycle.

scholarly journals The Asexual Pathogen Aspergillus fumigatus Expresses Functional Determinants of Aspergillus nidulans Sexual Development

2008 ◽  
Vol 7 (10) ◽  
pp. 1724-1732 ◽  
Author(s):  
Verena Große ◽  
Sven Krappmann
Keyword(s):  
Aspergillus Fumigatus ◽  
Aspergillus Nidulans ◽  
Gene Product ◽  
Mating Type ◽  
Sexual Development ◽  
Fungal Pathogen ◽  
Sexual Cycle ◽  
Genes Encoding ◽  
Type Gene ◽  
High Level

ABSTRACT The major fungal pathogen of humans, Aspergillus fumigatus, lacks a defined sexual cycle, although the presence of genes encoding putative mating type idiomorphs and regulators of Aspergillus sexual development heightens the potential for cryptic sexuality in this deuteromycete. To test the functionality of these genetic determinants, we transferred the alpha box-encoding mat1-1 idiomorph from an A. fumigatus isolate to the homothallic fertile species Aspergillus nidulans. Abundant formation of fruiting bodies (cleistothecia) containing viable ascospores establishes functionality of this mating type gene product in the transgenic strain. Using a similar approach, we also established that the conserved transcriptional regulator from A. fumigatus, the nsdD gene product, can act as a functional, positively acting factor for A. nidulans cleistothecium development; moreover, high-level expression of NsdD in the endogenous host A. fumigatus profoundly alters hyphal development by triggering the formation of coiled hyphae. Our findings demonstrate that the presumably asexual pathogen A. fumigatus encodes functional regulators of mating and sexual development, thereby potentiating the case for cryptic sexuality in this fungal pathogen.

scholarly journals Deletion of the 389 N-Terminal Residues of the Transcriptional Activator AREA Does Not Result in Nitrogen Metabolite Derepression in Aspergillus nidulans

1998 ◽  
Vol 180 (21) ◽  
pp. 5762-5764 ◽  
Author(s):  
Mark X. Caddick ◽  
Herbert N. Arst
Keyword(s):  
Amino Acids ◽  
Aspergillus Nidulans ◽  
Deletion Mutant ◽  
Transcriptional Activator ◽  
Gene Replacement ◽  
Untranslated Regions ◽  
Homologous Gene ◽  
Heterologous Promoter ◽  
Native Promoter ◽  
N Terminus

ABSTRACT Utilizing a homologous gene replacement in order to retain the native promoter and 5′ and 3′ untranslated messenger regions (and thereby ensure physiological validity), we have shown that deletion of the N-terminal 389 amino acids of the transcriptional activator AREA does not result in nitrogen metabolite derepression inAspergillus nidulans. Our results provide no evidence for a modulating interaction involving the N terminus of AREA and contrast with those of H. K. Lamb, A. L. Dodds, D. R. Swatman, E. Cairns, and A. R. Hawkins (J. Bacteriol. 179:6649–6656, 1997), who used nontargeted ectopic copies of a construct containing a heterologous promoter and untranslated regions. Results obtained with this deletion mutant, nevertheless, provide further evidence for the dispensability of large portions of AREA.

scholarly journals First Report of Homothallic Isolates of Phytophthora infestans in Commercial Potato Crops (Solanum tuberosum) in the Toluca Valley, Mexico

Plant Disease ◽  
2013 ◽  
Vol 97 (8) ◽  
pp. 1112-1112 ◽  
Author(s):  
C. A. López Orona ◽  
A. R. Martínez ◽  
T. T. Arteaga ◽  
H. G. García ◽  
D. Palmero ◽  
...  
Keyword(s):  
Solanum Tuberosum ◽  
Phytophthora Infestans ◽  
Mating Type ◽  
Sexual Cycle ◽  
First Report ◽  
Single Culture ◽  
Ability To Act ◽  
Commercial Potato ◽  
Toluca Valley ◽  
Potato Crops

Phytophthora infestans causes severe symptoms of wilt disease on potato crops (Solanum tuberosum) in the Toluca Valley (Mexico) despite the use of fungicides. P. infestans oospores produced by sexual reproduction can survive in the soil for many years, resisting harsh environments. In many agroecosystems, oospores germinate in the beginning of a season, which represents the initial inoculum for epidemics. The sexual cycle of the pathogen allows the generation of recombinant genotypes that can be more pathogenic or even resistant to chemicals. This paper presents a study of 20 isolates of P. infestans collected from potato crops in the Toluca Valley within the municipality of Zinacantepec (Mexico State). Isolates were obtained from potato foliar infected tissues. The pathogen was confirmed as P. infestans on the basis of morphological characters (1). Sporangia were caducous, ovoid, limoniform, semipapillate, and were 28.4 ± 1.3 × 17.6 ± 1.2 μm (height × width). Mycelium was coenocytic with hyphal diameter of 5 to 8 μm. Five isolates were collected in 2011, and 15 in 2012. Isolates were transferred by hyphal tip to culture medium plates with V8 juice agar and incubated at 19°C. All the isolates were mated to determine the mating type with the reference isolates J104 (A1) and J204 (A2), which were provided by the Michoacana University of San Nicolás de Hidalgo (Mx). Isolates that produced oospores with both A1 and A2 testers (J104 and J204) and in a single culture were designated homothallic. Results show that two out of the five isolates collected in 2011 were homothallic and the other three were type A1. Regarding the 15 isolates collected in 2012, six were typed as A1, five as A2, and four were homothallic. The heterothallic isolates only produce oospores when mated with the opposite mating type. The homothallic isolates possessed the ability to act as A1 and A2 during heterothallic mating and were found capable of producing sexual structures (oogonia and amphigynous antheridia) in a single culture, a phenomenon not observed in isolates that are strictly A1 or A2. Oospores formed were aplerotic and measured 32.2 ± 3.3 μm in diameter. Single-sporangium progeny were produced from the six homothallic isolates to be analyzed to confirm the occurrence of the self-fertility. Assessment of 48 single-sporangium progeny from the homothallic isolates resulted in 22 homothallic, 12 A1, 10 A2, and four sterile. These results differ from those found by Grünwald et al. (3), who conducted a study with isolates collected from the Toluca Valley region in 1997 and 1998, finding a 1:1 frequency between compatibility types A1 and A2. Fernandez et al. (2) studied a broad population of 27 isolates from potato crops in the state of Michoacán (Mx), and found two homothallic isolates among heterothallic isolates; the ratio was 1:1. Also, homothallic isolates have been found in Spain and China (4). To our knowledge, this is the first report of the occurrence of homothallic P. infestans isolates in commercial potato crops (S. tuberosum) in the Toluca Valley, Mexico. References: (1) D. C. Erwin and O. K. Ribeiro, Page 346 in: Phytophthora Diseases Worldwide. The American Phytopathological Society. St. Paul, MN, 1996. (2) S. P. Fernández et al. Rev. Mexicana Fitopatol. 23:191, 2005. (3) N. J. Grünwald et al. Phytopathology 91:883, 2001. (4) M. Han et al. J. Eukaryotic Microbiol. 60:79, 2013.

scholarly journals Presence and Functionality of Mating Type Genes in the Supposedly Asexual Filamentous Fungus Aspergillus oryzae

2012 ◽  
Vol 78 (8) ◽  
pp. 2819-2829 ◽  
Author(s):  
Ryuta Wada ◽  
Jun-ichi Maruyama ◽  
Haruka Yamaguchi ◽  
Nanase Yamamoto ◽  
Yutaka Wagu ◽  
...  
Keyword(s):  
Gene Expression ◽  
Aspergillus Oryzae ◽  
Mating Type ◽  
Genetic Manipulation ◽  
Gene Replacement ◽  
Soy Sauce ◽  
Sexual Cycle ◽  
Dependent Manner ◽  
Content Type ◽  
Mating Type Genes

ABSTRACTThe potential for sexual reproduction inAspergillus oryzaewas assessed by investigating the presence and functionality ofMATgenes. Previous genome studies had identified aMAT1-1gene in the reference strain RIB40. We now report the existence of a complementaryMAT1-2gene and the sequencing of an idiomorphic region fromA. oryzaestrain AO6. This allowed the development of a PCR diagnostic assay, which detected isolates of theMAT1-1andMAT1-2genotypes among 180 strains assayed, including industrialtane-kojiisolates. Strains used for sake and miso production showed a near-1:1 ratio of the MAT1-1 and MAT1-2 mating types, whereas strains used for soy sauce production showed a significant bias toward the MAT1-2 mating type. MAT1-1 and MAT1-2 isogenic strains were then created by genetic manipulation of the resident idiomorph, and gene expression was compared by DNA microarray and quantitative real-time PCR (qRT-PCR) methodologies under conditions in whichMATgenes were expressed. Thirty-three genes were found to be upregulated more than 10-fold in either the MAT1-1 host strain or the MAT1-2 gene replacement strain relative to each other, showing that both theMAT1-1andMAT1-2genes functionally regulate gene expression inA. oryzaein a mating type-dependent manner, the first such report for a supposedly asexual fungus.MAT1-1expression specifically upregulated an α-pheromone precursor gene, but the functions of most of the genes affected were unknown. The results are consistent with a heterothallic breeding system inA. oryzae, and prospects for the discovery of a sexual cycle are discussed.

Characterization of mat A-2, mat A-3 and ΔmatA Mating-Type Mutants of Neurospora crassa

Genetics ◽  
1998 ◽  
Vol 148 (3) ◽  
pp. 1069-1079 ◽  
Author(s):  
Adlane V-B Ferreira ◽  
Zhiqiang An ◽  
Robert L Metzenberg ◽  
N Louise Glass
Keyword(s):  
Neurospora Crassa ◽  
Mating Type ◽  
Sexual Development ◽  
Vegetative Growth ◽  
Double Mutant ◽  
Sexual Cycle ◽  
Wild Type ◽  
Type Locus ◽  
Isolation And Characterization

AbstractThe mating-type locus of Neurospora crassa regulates mating identity and entry into the sexual cycle. The mat A idiomorph encodes three genes, mat A-1, mat A-2, and mat A-3. Mutations in mat A-1 result in strains that have lost mating identity and vegetative incompatibility with mat a strains. A strain containing mutations in both mat A-2 and mat A-3 is able to mate, but forms few ascospores. In this study, we describe the isolation and characterization of a mutant deleted for mat (ΔmatA), as well as mutants in either mat A-2 or mat A-3. The ΔmatA strain is morphologically wild type during vegetative growth, but it is sterile and heterokaryon compatible with both mat A and mat a strains. The mat A-2 and mat A-3 mutants are also normal during vegetative growth, mate as a mat A strain, and produce abundant biparental asci in crosses with mat a, and are thus indistinguishable from a wild-type mat A strain. These data and the fact that the mat A-2 mat A-3 double mutant makes few asci with ascospores indicate that MAT A-2 and MAT A-3 are redundant and may function in the same pathway. Analysis of the expression of two genes (sdv-1 and sdv-4) in the various mat mutants suggests that the mat A polypeptides function in concert to regulate the expression of some sexual development genes.

scholarly journals The Function of the Coding Sequences for the Putative Pheromone Precursors in Podospora anserina Is Restricted to Fertilization

2005 ◽  
Vol 4 (2) ◽  
pp. 407-420 ◽  
Author(s):  
Evelyne Coppin ◽  
Christelle de Renty ◽  
Robert Debuchy
Keyword(s):  
Amino Acid ◽  
Mating Type ◽  
Male Fertility ◽  
Northern Blot Analysis ◽  
Gene Replacement ◽  
Podospora Anserina ◽  
Sexual Cycle ◽  
Normal Male ◽  
Posttranscriptional Level ◽  
Caax Motif

ABSTRACT We cloned the pheromone precursor genes of Podospora anserina in order to elucidate their role in the biology of this fungus. The mfp gene encodes a 24-amino-acid polypeptide finished by the CAAX motif, characteristic of fungal lipopeptide pheromone precursors similar to the a-factor precursor of Saccharomyces cerevisiae. The mfm gene encodes a 221-amino-acid polypeptide, which is related to the S. cerevisiae α-factor precursor and contains two 13-residue repeats assumed to correspond to the mature pheromone. We deleted the mfp and mfm coding sequence by gene replacement. The mutations specifically affect male fertility, without impairing female fertility and vegetative growth. The male defect is mating type specific: the mat + Δmfp and mat− Δmfm mutants produce male cells inactive in fertilization whereas the mat− Δmfp and mat + Δmfm mutants show normal male fertility. Genetic data indicate that both mfp and mfm are transcribed at a low level in mat + and mat− vegetative hyphae. Northern-blot analysis shows that their transcription is induced by the mating types in microconidia (mfp by mat + and mfm by mat−). We managed to cross Δmfp Δmfm strains of opposite mating type, by complementation and transient expression of the pheromone precursor gene to trigger fertilization. These crosses were fertile, demonstrating that once fertilization occurs, the pheromone precursor genes are unnecessary for the completion of the sexual cycle. Finally, we show that the constitutively transcribed gpd::mfm and gpd::mfp constructs are repressed at a posttranscriptional level by the noncognate mating type.

High Frequency Recombination During the Sexual Cycle of Dictyostelium discoideum

Genetics ◽  
1998 ◽  
Vol 148 (4) ◽  
pp. 1829-1832 ◽  
Author(s):  
David Francis
Keyword(s):  
Dictyostelium Discoideum ◽  
Genetic Map ◽  
High Frequency ◽  
Cell Biology ◽  
Physical Map ◽  
Genetic System ◽  
Sexual Cycle ◽  
Chromosome 2 ◽  
New Combinations ◽  
Restriction Sites

Abstract Analysis of Dictyostelium development and cell biology has suffered from the lack of an ordinary genetic system whereby genes can be arranged in new combinations. Genetic exchange between two long ignored strains, A2Cycr and WS205 is here reexamined. Alleles which differ in size or restriction sites between these two strains were found for seven genes. Six of these are in two clusters on chromosome 2. Frequencies of recombinant progeny indicate that the genetic map of the two mating strains is colinear with the physical map recently worked out for the standard nonsexual strain, NC4. The rate of recombination is high, about 0.1% per kilobase in three different regions of chromosome 2. This value is comparable to rates found in yeast, and will permit fine dissection of the genome.

scholarly journals THE SEARCH FOR MATING-TYPE-LIMITED GENES IN THE HOMOTHALLIC ALGA CHLAMYDOMONAS MONOICA

Genetics ◽  
1986 ◽  
Vol 113 (3) ◽  
pp. 601-619
Author(s):  
Karen P VanWinkle-Swift ◽  
Jang-Hee Hahn
Keyword(s):  
Mating Type ◽  
Resistance Mutation ◽  
Mendelian Inheritance ◽  
Sexual Cycle ◽  
Uniparental Inheritance ◽  
Cell Type ◽  
Erythromycin Resistance ◽  
Type Locus ◽  
Heterothallic Species ◽  
Chlamydomonas Monoica

ABSTRACT The non-Mendelian erythromycin resistance mutation ery-u1 shows bidirectional uniparental inheritance in crosses between homothallic ery-u1 and ery-u1  + strains of Chlamydomonas monoica. This inheritance pattern supports a general model for homothallism invoking intrastrain differentiation into opposite compatible mating types and, further, suggests that non-Mendelian inheritance is under mating-type (mt) control in C. monoica as in heterothallic species. However, the identification of genes expressed or required by one gametic cell type, but not the other, is essential to verify the existence of a regulatory mating-type locus in C. monoica and to understand its role in cell differentiation and sexual development. By screening for a shift from bidirectional to unidirectional transmission of the non-Mendelian ery-u1 marker, a mutant with an apparent mating-type-limited sexual cycle defect was obtained. The responsible mutation, mtl-1, causes a 1000-fold reduction in zygospore germination in populations homozygous for the mutant allele and, approximately, a 50% reduction in germination for heterozygous (mtl-1/mtl-1  +) zygospores. By next screening for strains unable to yield any viable zygospores in a cross to mtl-1, a second putative mating-type-limited mutant, mtl-2, was obtained. The mtl-2 strain, although self-sterile, mates efficiently with mtl-2  + strains and shows a unidirectional uniparental pattern of inheritance for the ery-u1 cytoplasmic marker, similar to that observed for crosses involving mtl-1. Genetic analysis indicates that mtl-1 and mtl-2 define unique unlinked Mendelian loci and that the sexual cycle defects of reduced germination (mtl-1) or self-sterility (mtl-2) cosegregate with the effect on ery-u1 cytoplasmic gene transmission. By analogy to C. reinhardtii, the mtl-1 and mtl-2 phenotypes can be explained if the expression of these gene loci is limited to the mt  + gametic cell type, or if the wild-type alleles at these loci are required for the normal formation and/or functioning of mt  + gametes only.

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