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Man-hong Sun - One of the best experts on this subject based on the ideXlab platform.
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identification of genes related to Chlamydospore formation in clonostachys rosea 67 1
MicrobiologyOpen, 2019Co-Authors: Zhan-bin Sun, Jun Zhang, Man-hong SunAbstract:Chlamydospores are specific structures that are of great significance to the commercialization of fungal biopesticides. To explore the genes associated with Chlamydospore formation, a biocontrol fungus Clonostachys rosea 67-1 that is capable of producing resistant spores under particular conditions was investigated by transcriptome sequencing and analysis. A total of 549,661,174 clean reads were obtained, and a series of differentially expressed genes potentially involved in fungal Chlamydospore formation were identified. At 36 hr, 67 and 117 genes were up- and downregulated in C. rosea during Chlamydospore production, compared with the control for conidiation, and 53 and 24 genes were up- and downregulated at 72 hr. GO classification suggested that the differentially expressed genes were related to cellular component, biological process, and molecular function categories. A total of 188 metabolism pathways were linked to Chlamydospore production by KEGG analysis. Sixteen differentially expressed genes were verified by reverse transcription quantitative PCR, and the expression profiles were consistent with the transcriptome data. To the best of our knowledge, it is the first report on the genes associated with Chlamydospore formation in C. rosea. The results provide insight into the molecular mechanisms underlying C. rosea sporulation, which will assist the development of fungal biocontrol agents.
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Identification of suitable reference genes during the formation of Chlamydospores in Clonostachys rosea 67-1.
MicrobiologyOpen, 2017Co-Authors: Jun Zhang, Zhan-bin Sun, Man-hong SunAbstract:Clonostachys rosea is a potential biocontrol fungus that can produce highly resistant Chlamydospores under specific conditions. To investigate the genes related to Chlamydospore formation, we identified reliable reference genes for quantification of gene expression in C. rosea 67-1 during sporulation. In this study, nine reference genes, actin (ACT), elongation factor 1 (EF1), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), histone (HIS), RNA polymerase II CTD phosphatase Fcp1 (RPP), succinate-semialdehyde dehydrogenase (SSD), TATA-binding protein (TBP), ubiquitin (UBQ), and ubiquitin-conjugating enzyme (UCE), were selected and cloned from 67-1, and their expression stability during Chlamydospore formation was determined using reverse transcription quantitative PCR and assessed using the software geNorm, NormFinder and BestKeeper. The Ct values of the candidates ranged from 19.9 to 29.7, among which HIS, ACT and SSD exhibited high expression levels. The statistical analysis showed that ACT and SSD were most stably expressed, while UBQ and GAPDH showed relatively large variations under different culture conditions. Calculation of pairwise variation value indicated that two reference genes were required for precise quantification. Finally, ACT and SSD were selected to normalize gene expression during Chlamydospore production in C. rosea 67-1. To the best of our knowledge, this is the first report of SSD as a reference gene. This study will facilitate the accurate quantification of differentially expressed genes during the generation of Chlamydospores and contribute to the investigation of the molecular mechanism underlying Chlamydospore formation in C. rosea.
Peter Staib - One of the best experts on this subject based on the ideXlab platform.
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Global Transcriptome Sequencing Identifies Chlamydospore Specific Markers in Candida albicans and
2016Co-Authors: Candida Dubliniensis, Katja Palige, Ronny Martin, Francesco Citiulo, Derek J. Sullivan, Johann Weber, Claudia Staib, Bernhard Hube, Peter StaibAbstract:Candida albicans and Candida dubliniensis are pathogenic fungi that are highly related but differ in virulence and in some phenotypic traits. During in vitro growth on certain nutrient-poor media, C. albicans and C. dubliniensis are the only yeast species which are able to produce Chlamydospores, large thick-walled cells of unknown function. Interestingly, only C. dubliniensis forms pseudohyphae with abundant Chlamydospores when grown on Staib medium, while C. albicans grows exclusively as a budding yeast. In order to further our understanding of Chlamydospore development and assembly, we compared the global transcriptional profile of both species during growth in liquid Staib medium by RNA sequencing. W
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Chlamydospore formation in Candida albicans and Candida dubliniensis– an enigmatic developmental programme
Mycoses, 2007Co-Authors: Peter Staib, Joachim MorschhauserAbstract:Chlamydospore formation has served for a long time for identification of the human fungal pathogen Candida albicans, but the biological function of these structures still remains a secret. They have been proposed to allow survival in harsh environmental conditions, but this assumption remains to be proven. Chlamydospores are produced only by the two closely related species C. albicans and Candida dubliniensis, whose natural habitats are humans and warm-blooded animals, but not by other Candida species that are also found outside animal hosts. However, no role in the pathogenesis of Candida infections has been assigned to these unusual cells and only a limited number of studies have been conducted in the past to unravel their function. The development of new molecular tools and the recent discovery of mating in C. albicans have also restimulated investigations to understand the morphogenesis and function of Chlamydospores. The finding that Chlamydospore formation is differentially controlled by certain environmental signals in C. albicans and C. dubliniensis has opened new approaches to study the regulation of this morphogenetic programme. These studies have already identified genes and signalling pathways that are required for Chlamydospore production and should lead to a detailed understanding of this fascinating developmental process.
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Liquid growth conditions for abundant Chlamydospore formation in Candida dubliniensis
Mycoses, 2005Co-Authors: Peter Staib, Joachim MorschhauserAbstract:Staib liquid medium (syn. Guizotia abyssinica creatinine medium) was evaluated for the induction of Chlamydospores in the yeast species Candida dubliniensis. During growth in this liquid medium C. dubliniensis produced abundant Chlamydospores at the tips of pseudohyphal elements, whereas C. albicans produced only round to oval blastospores. Chlamydospore formation could be induced with the same efficiency by use of a liquid growth medium in which Helianthus annuus (sunflower) seeds, another plant of the family Compositae, were substituted for Guizotia abyssinica plant seeds. Liquid growth culture conditions for rapid and abundant formation of Chlamydospores in C. dubliniensis should facilitate their enrichment and their biochemical and genetic analysis.
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differential expression of the nrg1 repressor controls species specific regulation of Chlamydospore development in candida albicans and candida dubliniensis
Molecular Microbiology, 2004Co-Authors: Peter Staib, Joachim MorschhauserAbstract:Summary Candida albicans and Candida dubliniensis are opportunistic fungal pathogens that are closely related but differ in their epidemiology and in some phenotypic characteristics, including certain virulence-related traits. A comparison of these two species at the molecular level could therefore provide new insights into the biology and pathogenicity of Candida. Both species share the ability to produce Chlamydospores, but only C. dubliniensis forms pseudohyphae with abundant Chlamydospores on Staib agar (syn. Guizotia abyssinica creatinine agar), on which C. albicans grows as a budding yeast. To understand the basis of this species-specific, differential regulation of morphogenetic development, we set out to identify C. albicans genes that repress Chlamydospore formation under these conditions. A C. albicans genomic library was integrated into the C. dubliniensis genome and transformants were screened for clones in which filamentation and/or Chlamydospore production on Staib agar was suppressed. This screen identified two genes, CaNRG1 and CaPDE2, encoding a general transcriptional repressor and a high affinity cAMP phosphodiesterase, respectively. Expression of CaNRG1 in C. dubliniensis repressed pseudohyphae and Chlamydospore formation, whereas expression of CaPDE2 only reduced the extent of filamentous growth but did not affect Chlamydospore formation. We found that C. dubliniensis, but not C. albicans, specifically downregulates NRG1 expression on Staib medium to allow Chlamydospore development. Artificial overexpression of CdNRG1 suppressed pseudohyphal growth and production of Chlamydospores in C. dubliniensis. Conversely, deletion of CaNRG1 in C. albicans resulted in Chlamydospore formation on Staib agar, confirming its central role in the regulation of this morphogenetic process. Our results demonstrate that differential regulation of a single gene, NRG1, in C. albicans and C. dubliniensis is responsible for their species-specific response to environmental signals that induce Chlamydospore development.
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Chlamydospore formation on staib agar as a species specific characteristic of candida dubliniensis
Mycoses, 1999Co-Authors: Peter Staib, Joachim MorschhauserAbstract:Staib agar (Syn. Guizotia abyssinica creatinine agar) was evaluated for differentiation between the highly related yeast species Candida albicans and Candida dubliniensis. On these agar plates C. dubliniensis formed rough colonies due to mycelial growth and produced abundant Chlamydospores whereas C. albicans grew only in smooth colonies and without Chlamydospore formation. The rough colonies of C. dubliniensis could be readily distinguished from the smooth C. albicans colonies. These results demonstrate that, under certain growth conditions, mycelial growth with Chlamydospore formation is a species-specific marker that can be used for the identification of C. dubliniensis.
Joachim Morschhauser - One of the best experts on this subject based on the ideXlab platform.
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Chlamydospore formation in Candida albicans and Candida dubliniensis– an enigmatic developmental programme
Mycoses, 2007Co-Authors: Peter Staib, Joachim MorschhauserAbstract:Chlamydospore formation has served for a long time for identification of the human fungal pathogen Candida albicans, but the biological function of these structures still remains a secret. They have been proposed to allow survival in harsh environmental conditions, but this assumption remains to be proven. Chlamydospores are produced only by the two closely related species C. albicans and Candida dubliniensis, whose natural habitats are humans and warm-blooded animals, but not by other Candida species that are also found outside animal hosts. However, no role in the pathogenesis of Candida infections has been assigned to these unusual cells and only a limited number of studies have been conducted in the past to unravel their function. The development of new molecular tools and the recent discovery of mating in C. albicans have also restimulated investigations to understand the morphogenesis and function of Chlamydospores. The finding that Chlamydospore formation is differentially controlled by certain environmental signals in C. albicans and C. dubliniensis has opened new approaches to study the regulation of this morphogenetic programme. These studies have already identified genes and signalling pathways that are required for Chlamydospore production and should lead to a detailed understanding of this fascinating developmental process.
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Liquid growth conditions for abundant Chlamydospore formation in Candida dubliniensis
Mycoses, 2005Co-Authors: Peter Staib, Joachim MorschhauserAbstract:Staib liquid medium (syn. Guizotia abyssinica creatinine medium) was evaluated for the induction of Chlamydospores in the yeast species Candida dubliniensis. During growth in this liquid medium C. dubliniensis produced abundant Chlamydospores at the tips of pseudohyphal elements, whereas C. albicans produced only round to oval blastospores. Chlamydospore formation could be induced with the same efficiency by use of a liquid growth medium in which Helianthus annuus (sunflower) seeds, another plant of the family Compositae, were substituted for Guizotia abyssinica plant seeds. Liquid growth culture conditions for rapid and abundant formation of Chlamydospores in C. dubliniensis should facilitate their enrichment and their biochemical and genetic analysis.
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differential expression of the nrg1 repressor controls species specific regulation of Chlamydospore development in candida albicans and candida dubliniensis
Molecular Microbiology, 2004Co-Authors: Peter Staib, Joachim MorschhauserAbstract:Summary Candida albicans and Candida dubliniensis are opportunistic fungal pathogens that are closely related but differ in their epidemiology and in some phenotypic characteristics, including certain virulence-related traits. A comparison of these two species at the molecular level could therefore provide new insights into the biology and pathogenicity of Candida. Both species share the ability to produce Chlamydospores, but only C. dubliniensis forms pseudohyphae with abundant Chlamydospores on Staib agar (syn. Guizotia abyssinica creatinine agar), on which C. albicans grows as a budding yeast. To understand the basis of this species-specific, differential regulation of morphogenetic development, we set out to identify C. albicans genes that repress Chlamydospore formation under these conditions. A C. albicans genomic library was integrated into the C. dubliniensis genome and transformants were screened for clones in which filamentation and/or Chlamydospore production on Staib agar was suppressed. This screen identified two genes, CaNRG1 and CaPDE2, encoding a general transcriptional repressor and a high affinity cAMP phosphodiesterase, respectively. Expression of CaNRG1 in C. dubliniensis repressed pseudohyphae and Chlamydospore formation, whereas expression of CaPDE2 only reduced the extent of filamentous growth but did not affect Chlamydospore formation. We found that C. dubliniensis, but not C. albicans, specifically downregulates NRG1 expression on Staib medium to allow Chlamydospore development. Artificial overexpression of CdNRG1 suppressed pseudohyphal growth and production of Chlamydospores in C. dubliniensis. Conversely, deletion of CaNRG1 in C. albicans resulted in Chlamydospore formation on Staib agar, confirming its central role in the regulation of this morphogenetic process. Our results demonstrate that differential regulation of a single gene, NRG1, in C. albicans and C. dubliniensis is responsible for their species-specific response to environmental signals that induce Chlamydospore development.
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Chlamydospore formation on staib agar as a species specific characteristic of candida dubliniensis
Mycoses, 1999Co-Authors: Peter Staib, Joachim MorschhauserAbstract:Staib agar (Syn. Guizotia abyssinica creatinine agar) was evaluated for differentiation between the highly related yeast species Candida albicans and Candida dubliniensis. On these agar plates C. dubliniensis formed rough colonies due to mycelial growth and produced abundant Chlamydospores whereas C. albicans grew only in smooth colonies and without Chlamydospore formation. The rough colonies of C. dubliniensis could be readily distinguished from the smooth C. albicans colonies. These results demonstrate that, under certain growth conditions, mycelial growth with Chlamydospore formation is a species-specific marker that can be used for the identification of C. dubliniensis.
Zhan-bin Sun - One of the best experts on this subject based on the ideXlab platform.
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identification of genes related to Chlamydospore formation in clonostachys rosea 67 1
MicrobiologyOpen, 2019Co-Authors: Zhan-bin Sun, Jun Zhang, Man-hong SunAbstract:Chlamydospores are specific structures that are of great significance to the commercialization of fungal biopesticides. To explore the genes associated with Chlamydospore formation, a biocontrol fungus Clonostachys rosea 67-1 that is capable of producing resistant spores under particular conditions was investigated by transcriptome sequencing and analysis. A total of 549,661,174 clean reads were obtained, and a series of differentially expressed genes potentially involved in fungal Chlamydospore formation were identified. At 36 hr, 67 and 117 genes were up- and downregulated in C. rosea during Chlamydospore production, compared with the control for conidiation, and 53 and 24 genes were up- and downregulated at 72 hr. GO classification suggested that the differentially expressed genes were related to cellular component, biological process, and molecular function categories. A total of 188 metabolism pathways were linked to Chlamydospore production by KEGG analysis. Sixteen differentially expressed genes were verified by reverse transcription quantitative PCR, and the expression profiles were consistent with the transcriptome data. To the best of our knowledge, it is the first report on the genes associated with Chlamydospore formation in C. rosea. The results provide insight into the molecular mechanisms underlying C. rosea sporulation, which will assist the development of fungal biocontrol agents.
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Identification of suitable reference genes during the formation of Chlamydospores in Clonostachys rosea 67-1.
MicrobiologyOpen, 2017Co-Authors: Jun Zhang, Zhan-bin Sun, Man-hong SunAbstract:Clonostachys rosea is a potential biocontrol fungus that can produce highly resistant Chlamydospores under specific conditions. To investigate the genes related to Chlamydospore formation, we identified reliable reference genes for quantification of gene expression in C. rosea 67-1 during sporulation. In this study, nine reference genes, actin (ACT), elongation factor 1 (EF1), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), histone (HIS), RNA polymerase II CTD phosphatase Fcp1 (RPP), succinate-semialdehyde dehydrogenase (SSD), TATA-binding protein (TBP), ubiquitin (UBQ), and ubiquitin-conjugating enzyme (UCE), were selected and cloned from 67-1, and their expression stability during Chlamydospore formation was determined using reverse transcription quantitative PCR and assessed using the software geNorm, NormFinder and BestKeeper. The Ct values of the candidates ranged from 19.9 to 29.7, among which HIS, ACT and SSD exhibited high expression levels. The statistical analysis showed that ACT and SSD were most stably expressed, while UBQ and GAPDH showed relatively large variations under different culture conditions. Calculation of pairwise variation value indicated that two reference genes were required for precise quantification. Finally, ACT and SSD were selected to normalize gene expression during Chlamydospore production in C. rosea 67-1. To the best of our knowledge, this is the first report of SSD as a reference gene. This study will facilitate the accurate quantification of differentially expressed genes during the generation of Chlamydospores and contribute to the investigation of the molecular mechanism underlying Chlamydospore formation in C. rosea.
Jun Zhang - One of the best experts on this subject based on the ideXlab platform.
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identification of genes related to Chlamydospore formation in clonostachys rosea 67 1
MicrobiologyOpen, 2019Co-Authors: Zhan-bin Sun, Jun Zhang, Man-hong SunAbstract:Chlamydospores are specific structures that are of great significance to the commercialization of fungal biopesticides. To explore the genes associated with Chlamydospore formation, a biocontrol fungus Clonostachys rosea 67-1 that is capable of producing resistant spores under particular conditions was investigated by transcriptome sequencing and analysis. A total of 549,661,174 clean reads were obtained, and a series of differentially expressed genes potentially involved in fungal Chlamydospore formation were identified. At 36 hr, 67 and 117 genes were up- and downregulated in C. rosea during Chlamydospore production, compared with the control for conidiation, and 53 and 24 genes were up- and downregulated at 72 hr. GO classification suggested that the differentially expressed genes were related to cellular component, biological process, and molecular function categories. A total of 188 metabolism pathways were linked to Chlamydospore production by KEGG analysis. Sixteen differentially expressed genes were verified by reverse transcription quantitative PCR, and the expression profiles were consistent with the transcriptome data. To the best of our knowledge, it is the first report on the genes associated with Chlamydospore formation in C. rosea. The results provide insight into the molecular mechanisms underlying C. rosea sporulation, which will assist the development of fungal biocontrol agents.
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Identification of suitable reference genes during the formation of Chlamydospores in Clonostachys rosea 67-1.
MicrobiologyOpen, 2017Co-Authors: Jun Zhang, Zhan-bin Sun, Man-hong SunAbstract:Clonostachys rosea is a potential biocontrol fungus that can produce highly resistant Chlamydospores under specific conditions. To investigate the genes related to Chlamydospore formation, we identified reliable reference genes for quantification of gene expression in C. rosea 67-1 during sporulation. In this study, nine reference genes, actin (ACT), elongation factor 1 (EF1), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), histone (HIS), RNA polymerase II CTD phosphatase Fcp1 (RPP), succinate-semialdehyde dehydrogenase (SSD), TATA-binding protein (TBP), ubiquitin (UBQ), and ubiquitin-conjugating enzyme (UCE), were selected and cloned from 67-1, and their expression stability during Chlamydospore formation was determined using reverse transcription quantitative PCR and assessed using the software geNorm, NormFinder and BestKeeper. The Ct values of the candidates ranged from 19.9 to 29.7, among which HIS, ACT and SSD exhibited high expression levels. The statistical analysis showed that ACT and SSD were most stably expressed, while UBQ and GAPDH showed relatively large variations under different culture conditions. Calculation of pairwise variation value indicated that two reference genes were required for precise quantification. Finally, ACT and SSD were selected to normalize gene expression during Chlamydospore production in C. rosea 67-1. To the best of our knowledge, this is the first report of SSD as a reference gene. This study will facilitate the accurate quantification of differentially expressed genes during the generation of Chlamydospores and contribute to the investigation of the molecular mechanism underlying Chlamydospore formation in C. rosea.