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T S Chandra - One of the best experts on this subject based on the ideXlab platform.
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antagonistic effect of myo inositol on riboflavin production in two riboflavinogenic fungi ashbya gossypii and Eremothecium ashbyi
Mycoscience, 2013Co-Authors: Sudeshna Sengupta, S Kiruthiga, T S ChandraAbstract:Abstract Ashbya gossypii and Eremothecium ashbyi are riboflavin over producing fungi. Guanosine tri-phosphate (GTP) cyclohydrolase II, encoded by RIB1 gene, catalyses the rate limiting step of the riboflavin biosynthetic pathway. We report here the antagonistic effect of myo-inositol on riboflavin production in these two fungi by adapting them in Yeast Malt Agar (YMA) media containing 0.1% inositol. The adapted E. ashbyi showed 92% reduction and the adapted Ashbya showed 39% increase in the total riboflavin production, when compared to that produced by E. ashbyi and Ashbya respectively. Reverse transcription-polymerase chain reaction (RT-PCR) study indicated that myo-inositol reduced RIB1 gene expression in adapted E. ashbyi but enhanced the same in adapted Ashbya .
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development of fluorescent reporter tagged rib gene cassettes for replicative transformation early expression and enhanced riboflavin production in Eremothecium ashbyi
Fungal Biology, 2012Co-Authors: Sudeshna Sengupta, Andreas Kaufmann, T S ChandraAbstract:Abstract Eremothecium ashbyi is a riboflavin overproducing filamentous fungus in which the metabolic pathways have not been genetically characterized. Two genes of the riboflavin biosynthetic (RIB) pathway, RIB1 and RIB3 , which encode GTP-cyclohydrolase II (GCH II) and 3,4-dihydroxy-2-butanone 4-phosphate (DHBP) synthase respectively, were selected for the present study. The two RIB genes under their native promoters were obtained from Ashbya gossypii genomic library. Yeast enhanced green fluorescent protein ( yEGFP ) and mCherry genes were tagged to the C-terminal ends of RIB1 and RIB3 genes to analyse the functionality of the RIB transgenes in E. ashbyi . Shuttle vectors with the reporter tagged RIB genes contained the Escherichia coli kan R gene and Saccharomyces cerevisiae ARS element. On transformation with these plasmids, the ARS element was found to be functional in E. ashbyi . The E. ashbyi transcription factors could recognize the Ashbya RIB gene promoters and express the reporter tagged RIB genes as cytoplasmic proteins, in early cell development. Replicative transformants carrying RIB1-mCherry plasmids showed 2.95 times more GCH II activity and 2.44 times more riboflavin production when compared to untransformed. This is the first report of genetic transformation of E. ashbyi and is of significance as the first step towards genetic engineering of this genus.
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Sequence Analysis and Structural Characterization of a Glyceraldehyde-3-Phosphate Dehydrogenase Gene from the Phytopathogenic Fungus Eremothecium ashbyi
Mycopathologia, 2011Co-Authors: Sudeshna Sengupta, T S ChandraAbstract:Eremothecium ashbyi is a phytopathogenic fungus infesting cotton, soybeans and several other plants. This highly flavinogenic fungus has been phylogenetically characterized, but the genetic aspects of its central metabolic and riboflavin biosynthetic pathways are unknown. An ORF of 996 bp was obtained from E. ashbyi by using degenerate primers for glyceraldehyde-3-phosphate dehydrogenase (GPD) through reverse transcriptase polymerase chain reaction (RT–PCR) and 5′-3′ rapid amplification of cDNA ends (RACE-PCR). This nucleotide sequence had a high similarity of 88% with GPD sequence of Ashbya gossypii . The putative GPD peptide of 331-aa had a high similarity of 85% with the GPD sequence from other ascomycetes. The ORF had an unusually strong codon bias with 5 amino acids showing strict preference of a single codon. The theoretical molecular weight for the putative peptide was 35.58 kDa with an estimated pI of 5.7. A neighbor-joining tree showed that the putative peptide from E. ashbyi displayed the highest similarity to GPD of A. gossypii . The gene sequence is available at the GenBank, accession number EU717696 . Homology modeling done with Kluyveromyces marxianus GPD (PDB: 2I5P) as template indicated high structural similarity.
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molecular and structural characterization of gtp cyclohydrolase ii in Eremothecium ashbyi nrrl y 1363 cdna cloning comparative sequence analysis and molecular modeling
Fungal Biology, 2010Co-Authors: Sudeshna Sengupta, T S ChandraAbstract:Abstract GTP-cyclohydrolase II (GCH II) encoded by RIB1 gene catalyzes the first committed step in the riboflavin biosynthetic pathway. We report here the cloning and characterization of the entire RIB1 ORF ( EaRIB1 ) of 942 bp by reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE-PCR) in Eremothecium ashbyi where it was found to be present as a single-copy gene. EaRIB1 sequence is available at the GenBank Accession Number EF565374 . The putative peptide of 313-aa has a high similarity of 60–70 % with GCH II sequences from other ascomycete fungi. Gene expression and alignment studies confirmed the functional annotation of this gene. Homology model was developed with Escherichia coli (PDB 2BZ1) as template to identify the catalytic domains and to explore its functional architecture. We report here the first three-dimensional model of any fungal GCH II which due to its absence in humans assumes significance for anti-fungal drug targeting.
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spectrofluorimetric method for the estimation of total lipids in Eremothecium ashbyii fungal filaments using nile blue and avoiding interference of autofluorescent riboflavin
Journal of Microbiological Methods, 2003Co-Authors: S Vijayalakshmi, T N Karthika, Ashok Kumar Mishra, T S ChandraAbstract:A rapid, simple and sensitive spectrofluorimetric technique was developed for monitoring total lipids in hyphae of the riboflavin-overproducing fungus Eremothecium ashbyii using the fluorescent probe Nile blue in an aqueous system, avoiding the interference due to autofluorescent riboflavin. The existing methodologies for lipid estimation are tedious, requiring large biomass, solvent extraction and gravimetry. E. ashbyii is a hemiascomycete fungus which accumulates lipids in its mycelia prior to flavinogenesis. This study defines the conditions (wavelength selection and sensitivity) for the spectrofluorimetric quantification of lipids in situ in the macerated mycelia of this fungus in the presence of intracellular autofluorescent riboflavin without the need to extract the lipids from the mycelia. The fluorescent intensity was linear with the lipid concentration of the mycelia (by gravimetry) under three different growth conditions using glucose, olive oil and sunflower oil as carbon sources. This spectrofluorimetic method of lipid estimation can be applied to other fungi and microorganisms.
Cletus P Kurtzman - One of the best experts on this subject based on the ideXlab platform.
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chapter 30 Eremothecium borzi emend kurtzman 1995
The Yeasts (Fifth Edition), 2011Co-Authors: Cletus P Kurtzman, Sybren G De HoogAbstract:Publisher Summary This chapter studies the genus Eremothecium. In the determination of asexual reproduction, budding cells are absent or present, and when present, budding is multilateral on a narrow base. Cells are globose, ovoid, ellipsoidal, or cylindrical. Enteroarthric conidia are infrequently produced by one species. Pseudohyphae and true hyphae are generally present. Colonies are smooth or floccose and white, grayish, or yellow in color. In sexual reproduction it is seen that asci, which become deliquescent, form 8–32 ascospores that are fusiform or acicular. Ascospores may have a central septum, and those of some species have a tapered, terminal extension of the cell wall. Species appear to be homothallic. In physiology and biochemistry it is seen that sugars are fermented by some species. Nitrate is not assimilated. Coenzyme Q may have five, six, seven, eight, or nine isoprene units in the side chain, some of which are present in minor proportions. The chapter further examines the phylogenetic placement. The type of species mentioned is Eremothecium cymbalariae. The species accepted are Eremothecium ashbyi, Eremothecium coryli, Eremothecium cymbalariae, Eremothecium gossypii, and Eremothecium sinecaudum. The systematic discussion of the species includes synonyms, growth on 4% malt extract/0.5% yeast extract agar, growth on the surface of assimilation media, formation of ascospores, origin of the strains studied, authentic strain, systematics, ecology, biotechnology, agriculture and food, and clinical importance.
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Eremothecium borzi emend kurtzman 1995
2010Co-Authors: Cletus P Kurtzman, Sybren G De HoogAbstract:Publisher Summary This chapter studies the genus Eremothecium. In the determination of asexual reproduction, budding cells are absent or present, and when present, budding is multilateral on a narrow base. Cells are globose, ovoid, ellipsoidal, or cylindrical. Enteroarthric conidia are infrequently produced by one species. Pseudohyphae and true hyphae are generally present. Colonies are smooth or floccose and white, grayish, or yellow in color. In sexual reproduction it is seen that asci, which become deliquescent, form 8–32 ascospores that are fusiform or acicular. Ascospores may have a central septum, and those of some species have a tapered, terminal extension of the cell wall. Species appear to be homothallic. In physiology and biochemistry it is seen that sugars are fermented by some species. Nitrate is not assimilated. Coenzyme Q may have five, six, seven, eight, or nine isoprene units in the side chain, some of which are present in minor proportions. The chapter further examines the phylogenetic placement. The type of species mentioned is Eremothecium cymbalariae. The species accepted are Eremothecium ashbyi, Eremothecium coryli, Eremothecium cymbalariae, Eremothecium gossypii, and Eremothecium sinecaudum. The systematic discussion of the species includes synonyms, growth on 4% malt extract/0.5% yeast extract agar, growth on the surface of assimilation media, formation of ascospores, origin of the strains studied, authentic strain, systematics, ecology, biotechnology, agriculture and food, and clinical importance.
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Eremothecium borzi emend kurtzman
The Yeasts (Fourth Edition)#R##N#A Taxonomic Study, 1998Co-Authors: G S De Hoog, Cletus P Kurtzman, H J Phaff, M W MillerAbstract:Publisher Summary This chapter focuses on Eremothecium genus and its constituent species. The budding cells of this genus are absent or present, and when present, budding is multilateral on a narrow base. The cells are globose, ovoidal, ellipsoidal, or cylindroidal. Its asci, which become deliquescent, form 8-32 ascospores that are fusiform or acicular. Ascospores may have a central septum, and those of some species have a tapered, terminal extension of the cell wall. The sugars are fermented by some species and nitrate is not assimilated. The constituent species of this genus include Eremothecium ashbyi, Eremothecium coryli, Eremothecium cymbalari, and Eremothecium gossypii. The asci of Eremothecium ashbyi are ellipsoidal to fusiform, and occur mostly intercalary and in long chains. The ascospores are evenly distributed in the ascus, and are hyaline, narrow, sickleshaped, rounded at the tip, but with a basal spine. The Asci in Eremothecium coryli are long, curved cylindrical cells with rounded ends, although they are sometimes of irregular shape. Examination under the light microscope reveals the presence of vacuoles and a difference of refringence between the two halves of the spores. When stained with the Ziehl–Neelsen or the Schaeffer–Fulton malachite green technique, the half supporting the whip-like appendage is not acid fast and does not retain the malachite green.
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relationships among the genera ashbya Eremothecium holleya and nematospora determined from rdna sequence divergence
Journal of Industrial Microbiology & Biotechnology, 1995Co-Authors: Cletus P KurtzmanAbstract:Species of the generaAshbya, Eremothecium, Holleya, andNematospora were compared from extent of divergence in a 580-nucleotide region near the 5′ end of the large subunit (26S) ribosomal DNA gene. The four genera are closely related and comprise a subclade of the hemiascomycetes. Because the taxa show little divergence, it is proposed that all be placed in the genusEremothecium. The family Eremotheciaceae, fam. nov., is proposed.
Jurgen Wendland - One of the best experts on this subject based on the ideXlab platform.
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chromosome number reduction in Eremothecium coryli by two telomere to telomere fusions
Genome Biology and Evolution, 2014Co-Authors: Jurgen Wendland, Andrea WaltherAbstract:The genus Eremothecium belongs to the Saccharomyces complex of pre-whole-genome duplication (WGD) yeasts and contains both dimorphic and filamentous species. We established the 9.1-Mb draft genome of Eremothecium coryli, which encodes 4,682 genes, 186 tRNA genes, and harbors several Ty3 transposons as well as more than 60 remnants of transposition events (LTRs). The initial de novo assembly resulted in 19 scaffolds, which were assembled based on synteny to other Eremothecium genomes into six chromosomes. Interestingly, we identified eight E. coryli loci that bear centromeres in the closely related species E. cymbalariae. Two of these E. coryli loci, CEN1 and CEN8, however, lack conserved DNA elements and did not convey centromere function in a plasmid stability assay. Correspondingly, using a comparative genomics approach we identified two telomere-to-telomere fusion events in E. coryli as the cause of chromosome number reduction from eight to six chromosomes. Finally, with the genome sequences of E. coryli, E. cymbalariae, and Ashbya gossypii a reconstruction of three complete chromosomes of an Eremothecium ancestor revealed that E. coryli is more syntenic to this ancestor than the other Eremothecium species.
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genome evolution in the Eremothecium clade of the saccharomyces complex revealed by comparative genomics
G3: Genes Genomes Genetics, 2011Co-Authors: Jurgen Wendland, Andrea WaltherAbstract:We used comparative genomics to elucidate the genome evolution within the pre–whole-genome duplication genus Eremothecium. To this end, we sequenced and assembled the complete genome of Eremothecium cymbalariae, a filamentous ascomycete representing the Eremothecium type strain. Genome annotation indicated 4712 gene models and 143 tRNAs. We compared the E. cymbalariae genome with that of its relative, the riboflavin overproducer Ashbya (Eremothecium) gossypii, and the reconstructed yeast ancestor. Decisive changes in the Eremothecium lineage leading to the evolution of the A. gossypii genome include the reduction from eight to seven chromosomes, the downsizing of the genome by removal of 10% or 900 kb of DNA, mostly in intergenic regions, the loss of a TY3-Gypsy–type transposable element, the re-arrangement of mating-type loci, and a massive increase of its GC content. Key species-specific events are the loss of MNN1-family of mannosyltransferases required to add the terminal fourth and fifth α-1,3-linked mannose residue to O-linked glycans and genes of the Ehrlich pathway in E. cymbalariae and the loss of ZMM-family of meiosis-specific proteins and acquisition of riboflavin overproduction in A. gossypii. This reveals that within the Saccharomyces complex genome, evolution is not only based on genome duplication with subsequent gene deletions and chromosomal rearrangements but also on fungi associated with specific environments (e.g. involving fungal-insect interactions as in Eremothecium), which have encountered challenges that may be reflected both in genome streamlining and their biosynthetic potential.
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a molecular toolbox for manipulating Eremothecium coryli
Microbiological Research, 2007Co-Authors: Selina Gastmann, Alexander Dunkler, Andrea Walther, Keith Klein, Jurgen WendlandAbstract:Summary The genus Eremothecium contains dimorphic and filamentous fungal species, most notably Eremothecium sinecaudum (Holleya sinecauda), a dimorphic plant pathogen, which was isolated from mustard seeds, and Eremothecium gossypii (Ashbya gossypii), a filamentous fungus, which is well known for its ability to produce riboflavin. In this study, we present the initial molecular characterization of another Eremothecium species classified as Eremothecium coryli. E.coryli is a dimorphic fungus. We have developed, based on previously described reagents, a transformation system for E. coryli using kanMX and NATMX3 as dominant selectable marker genes on freely replicating plasmids conferring resistance to the antibiotics G418 and nourseothricin, respectively. As reporter genes we could introduce lacZ and GFP, which were controlled either by the AgTEF1 promoter or by regulatable MET promoters derived from the A. gossypii and Saccharomyces cerevisiae MET3 genes. These newly established tools will allow a detailed comparison of different growth modes in filamentous or dimorphic species within the genus Eremothecium. & 2007 Elsevier GmbH. All rights reserved.
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Initial molecular characterization of a novel Rho-type GTPase RhoH in the filamentous ascomycete Ashbya gossypii
Current Genetics, 2005Co-Authors: Andrea Walther, Jurgen WendlandAbstract:The organization of the actin cytoskeleton is of central importance in determining cell shape and morphogenesis. Rho-type GTP-binding proteins are known for their role to activate downstream effector proteins that act as scaffolds and direct the assembly of actin filaments. We have identified a novel Rho-type GTPase, RHOH , in the filamentous ascomycete Ashbya gossypii and also in one of its closest relatives, Holleya sinecauda both species belonging to the genus Eremothecium . RHOH is a paralog of AgRHO1 and was generated by tandem duplication in an evolutionary ancestor but is not found in Saccharomyces cerevisae . Deletion of RHOH was carried out in both A. gossypii and H. sinecauda and showed that this gene is non-essential. However, a cell wall defect was observed that led to increased cell lysis at the hyphal tip and an increased sensitivity against the cell wall agents calcofluor and SDS in the AgrhoH strain. These results suggested at least partially overlapping functions between RhoH and Rho1 without excluding any spilt functions of these proteins. Deletion of both the RHOH and RHO1 genes in A. gossypii exacerbated the mutant rho1 phenotype and resulted in germination deficient spores. This suggests that Rho1/RhoH -in contrast to Cdc42- are required at the early steps of spore germination.
A E Kalingan - One of the best experts on this subject based on the ideXlab platform.
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influence of type and concentration of flavinogenic factors on production of riboflavin by Eremothecium ashbyii nrrl 1363
Bioresource Technology, 2002Co-Authors: A E Kalingan, Chungmin LiaoAbstract:Abstract A study was conducted to determine the effect of various low cost organic wastes as flavinogenic factors and the various concentrations at which they induced flavinogenecity resulting in higher yields of riboflavin. A high-yielding riboflavin strain; Eremothecium ashbyii NRRL 1363 was chosen to determine the flavinogenicity. Carbon source at 50 g l −1 (dextrose equivalents) of molasses and nitrogen source at 50 g l −1 (weight/volume) of peanut seed cake were found to be optimal levels to yield higher riboflavin. Among the organic wastes, (beef extract, hog casings, blood meal, fish meal) hog casings in association with fish meal supported the highest yield of riboflavin. Among the different recovery processes studied, a vacuum drying process was the most efficient allowing maximum yield, followed by drying at 90 °C and freeze-drying. It is apparent from this study that inexpensive or waste organic materials could induce E. ashbyii to synthesize and secrete riboflavin at higher levels in the medium and this could be purified using a vacuum drying process. This bioconversion process allows us to recycle the biomaterials and produce a value-added product of economic importance.
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the kinetics of riboflavin secretion by Eremothecium ashbyii nrrl 1363
Bioprocess Engineering, 1998Co-Authors: A E KalinganAbstract:For riboflavin production a highly flavinogenic strain Eremothecium ashbyii is used. Vitamins are classified as chemical substances that control and effect the physiological processes; Riboflavin is one among them. The deficiency of riboflavin in human beings results in the cracking of lips and corners of mouth (cheilosis); nerve tissues are affected. For riboflavin production a highly flavinogenic strain Eremothecium ashbyii NRRL 1363 was used. Investigations were conducted in shake flask using inexpensive and abundantly available raw materials. Among the stimulants, a combination of hog casings and beef extract stimulated the highest and promoted the maximum riboflavin yield followed by the combination of fish meal and beef extract. The fermented broth (an enriched, riboflavin concentrate) can be directly used as a feed grade riboflavin. To upgrade it to pharmaceutical grade further investigations are required.
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application of agro industrial by products for riboflavin production by Eremothecium ashbyii nrrl 1363
Applied Microbiology and Biotechnology, 1997Co-Authors: A E Kalingan, M R V KrishnanAbstract:Riboflavin production is significantly determined by the type and initial concentration of the carbon and nitrogen sources and also by other flavinogenic stimulants. Using an optimum carbon and nitrogen concentration, an industrial fermentation medium has been designed with molasses as the carbon source and peanut seed cake as the nitrogen source. In addition the stimulatory effect of some of the low-cost agro-industrial by-products on riboflavin yield was investigated.
Andrea Walther - One of the best experts on this subject based on the ideXlab platform.
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chromosome number reduction in Eremothecium coryli by two telomere to telomere fusions
Genome Biology and Evolution, 2014Co-Authors: Jurgen Wendland, Andrea WaltherAbstract:The genus Eremothecium belongs to the Saccharomyces complex of pre-whole-genome duplication (WGD) yeasts and contains both dimorphic and filamentous species. We established the 9.1-Mb draft genome of Eremothecium coryli, which encodes 4,682 genes, 186 tRNA genes, and harbors several Ty3 transposons as well as more than 60 remnants of transposition events (LTRs). The initial de novo assembly resulted in 19 scaffolds, which were assembled based on synteny to other Eremothecium genomes into six chromosomes. Interestingly, we identified eight E. coryli loci that bear centromeres in the closely related species E. cymbalariae. Two of these E. coryli loci, CEN1 and CEN8, however, lack conserved DNA elements and did not convey centromere function in a plasmid stability assay. Correspondingly, using a comparative genomics approach we identified two telomere-to-telomere fusion events in E. coryli as the cause of chromosome number reduction from eight to six chromosomes. Finally, with the genome sequences of E. coryli, E. cymbalariae, and Ashbya gossypii a reconstruction of three complete chromosomes of an Eremothecium ancestor revealed that E. coryli is more syntenic to this ancestor than the other Eremothecium species.
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genome evolution in the Eremothecium clade of the saccharomyces complex revealed by comparative genomics
G3: Genes Genomes Genetics, 2011Co-Authors: Jurgen Wendland, Andrea WaltherAbstract:We used comparative genomics to elucidate the genome evolution within the pre–whole-genome duplication genus Eremothecium. To this end, we sequenced and assembled the complete genome of Eremothecium cymbalariae, a filamentous ascomycete representing the Eremothecium type strain. Genome annotation indicated 4712 gene models and 143 tRNAs. We compared the E. cymbalariae genome with that of its relative, the riboflavin overproducer Ashbya (Eremothecium) gossypii, and the reconstructed yeast ancestor. Decisive changes in the Eremothecium lineage leading to the evolution of the A. gossypii genome include the reduction from eight to seven chromosomes, the downsizing of the genome by removal of 10% or 900 kb of DNA, mostly in intergenic regions, the loss of a TY3-Gypsy–type transposable element, the re-arrangement of mating-type loci, and a massive increase of its GC content. Key species-specific events are the loss of MNN1-family of mannosyltransferases required to add the terminal fourth and fifth α-1,3-linked mannose residue to O-linked glycans and genes of the Ehrlich pathway in E. cymbalariae and the loss of ZMM-family of meiosis-specific proteins and acquisition of riboflavin overproduction in A. gossypii. This reveals that within the Saccharomyces complex genome, evolution is not only based on genome duplication with subsequent gene deletions and chromosomal rearrangements but also on fungi associated with specific environments (e.g. involving fungal-insect interactions as in Eremothecium), which have encountered challenges that may be reflected both in genome streamlining and their biosynthetic potential.
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a molecular toolbox for manipulating Eremothecium coryli
Microbiological Research, 2007Co-Authors: Selina Gastmann, Alexander Dunkler, Andrea Walther, Keith Klein, Jurgen WendlandAbstract:Summary The genus Eremothecium contains dimorphic and filamentous fungal species, most notably Eremothecium sinecaudum (Holleya sinecauda), a dimorphic plant pathogen, which was isolated from mustard seeds, and Eremothecium gossypii (Ashbya gossypii), a filamentous fungus, which is well known for its ability to produce riboflavin. In this study, we present the initial molecular characterization of another Eremothecium species classified as Eremothecium coryli. E.coryli is a dimorphic fungus. We have developed, based on previously described reagents, a transformation system for E. coryli using kanMX and NATMX3 as dominant selectable marker genes on freely replicating plasmids conferring resistance to the antibiotics G418 and nourseothricin, respectively. As reporter genes we could introduce lacZ and GFP, which were controlled either by the AgTEF1 promoter or by regulatable MET promoters derived from the A. gossypii and Saccharomyces cerevisiae MET3 genes. These newly established tools will allow a detailed comparison of different growth modes in filamentous or dimorphic species within the genus Eremothecium. & 2007 Elsevier GmbH. All rights reserved.
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Initial molecular characterization of a novel Rho-type GTPase RhoH in the filamentous ascomycete Ashbya gossypii
Current Genetics, 2005Co-Authors: Andrea Walther, Jurgen WendlandAbstract:The organization of the actin cytoskeleton is of central importance in determining cell shape and morphogenesis. Rho-type GTP-binding proteins are known for their role to activate downstream effector proteins that act as scaffolds and direct the assembly of actin filaments. We have identified a novel Rho-type GTPase, RHOH , in the filamentous ascomycete Ashbya gossypii and also in one of its closest relatives, Holleya sinecauda both species belonging to the genus Eremothecium . RHOH is a paralog of AgRHO1 and was generated by tandem duplication in an evolutionary ancestor but is not found in Saccharomyces cerevisae . Deletion of RHOH was carried out in both A. gossypii and H. sinecauda and showed that this gene is non-essential. However, a cell wall defect was observed that led to increased cell lysis at the hyphal tip and an increased sensitivity against the cell wall agents calcofluor and SDS in the AgrhoH strain. These results suggested at least partially overlapping functions between RhoH and Rho1 without excluding any spilt functions of these proteins. Deletion of both the RHOH and RHO1 genes in A. gossypii exacerbated the mutant rho1 phenotype and resulted in germination deficient spores. This suggests that Rho1/RhoH -in contrast to Cdc42- are required at the early steps of spore germination.