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Hyphal Growth (hyphal + growth)
Selected AbstractsRequirement of spermidine for developmental transitions in Aspergillus nidulansMOLECULAR MICROBIOLOGY, Issue 3 2002Yuan Jin Summary Deletion of the spermidine synthase gene in the fungus Aspergillus nidulans results in a strain, ,spdA, which requires spermidine for growth and accumulates putrescine as the sole polyamine. Vegetative growth but not sporulation or sterigmatocystin production is observed when ,spdA is grown on media supplemented with 0.05,0.10 mM exogenous spermidine. Supplementation of ,spdA with , 0.10 mM spermidine restores sterigmatocystin production and , 0.50 mM spermidine produces a phenotype with denser asexual spore production and decreased radial hyphal growth compared with the wild type. ,spdA spores germinate in unsupplemented media but germ tube growth ceases after 8 h upon which time the spores swell to approximately three times their normal diameter. Hyphal growth is resumed upon addition of 1.0 mM spermidine. Suppression of a G protein signalling pathway could not force asexual sporulation and sterigmatocystin production in ,spdA strains grown in media lacking spermidine but could force both processes in ,spdA strains supplemented with 0.05 mM spermidine. These results show that increasing levels of spermidine are required for the transitions from (i) germ tube to hyphal growth and (ii) hyphal growth to tissue differentiation and secondary metabolism. Suppression of G protein signalling can over-ride the spermidine requirement for the latter but not the former transition. [source] EVOLUTION OF REPRODUCTIVE ISOLATION WITHIN A PARASITIC FUNGAL SPECIES COMPLEXEVOLUTION, Issue 7 2007Mickael Le Gac Despite important advances in the last few years, the evolution of reproductive isolation (RI) remains an unresolved and critical gap in our understanding of speciation processes. In this study, we investigated the evolution of RI among species of the parasitic fungal species complex Microbotryum violaceum, which is responsible for anther smut disease of the Caryophyllaceae. We found no evidence for significant positive assortative mating by M. violaceum even over substantial degrees of genetic divergence, suggesting a lack of prezygotic isolation. In contrast, postzygotic isolation increased with the genetic distance between mating partners when measured as hyphal growth. Total RI, measured as the ability of the pathogen to infect and produce a diploid progeny in the host plant, was significantly and positively correlated with genetic distance, remaining below complete isolation for most of the species pairs. The results of this study, the first one on the time course of speciation in a fungus, are therefore consistent with previous works showing that RI generally evolves gradually with genetic distance, and thus presumably with time. Interestingly, prezygotic RI due to gamete recognition did not increase with genetic distance, in contrast to the pattern found in plants and animals. [source] Cloning and sequence analysis of cnaA gene encoding the catalytic subunit of calcineurin from Aspergillus oryzaeFEMS MICROBIOLOGY LETTERS, Issue 1 2001Praveen Rao Juvvadi Abstract Calcineurin has been implicated in ion-homeostasis, stress adaptation in yeast and for hyphal growth in filamentous fungi. Genomic DNA and cDNA encoding the catalytic subunit of calcineurin (cnaA) were isolated from Aspergillus oryzae. The cnaA open reading frame extended to 1727 bp and encoded a putative protein of 514 amino acids. Comparative analysis of the nucleotide sequence of cnaA genomic DNA and cDNA confirmed the presence of three introns and a highly conserved calmodulin binding domain. The deduced amino acid sequence was homologous to calcineurin A from Aspergillus nidulans (92%), Neurospora crassa (84%), human (67%), Saccharomyces cerevisiae (58%) and Schizosaccharomyces pombe (54%). Further, A. oryzae cnaA cDNA complemented S. cerevisiae calcineurin disruptant strain (,cmp1,cmp2), which was not viable in the presence of high concentrations of NaCl (1.2 M) and at alkaline pH 8.5. [source] Candida albicans ABG1 gene is involved in endocytosisFEMS YEAST RESEARCH, Issue 2 2009Verónica Veses Abstract The human fungal pathogen Candida albicans undergoes reversible morphogenetic transitions between yeast, hyphal and pseudohyphal forms. The fungal vacuole actively participates in differentiation processes and plays a key role supporting hyphal growth. The ABG1 gene of C. albicans encodes an essential protein located in the vacuolar membranes of both yeast and hyphae. Using fluorescence microscopy of a green fluorescent protein-tagged version of Abg1p, a fraction of the protein was detected in hyphal tips, not associated with vacuolar membranes. Live cell imaging of emerging germ tubes showed that Abg1p migrated to the polarized growth site and colocalized with endocytic vesicles. Phenotypic analysis of a methionine-regulated conditional mutant confirmed that Abg1p is involved in endocytosis. [source] Transcriptional profiling of the early stages of germination in Candida albicans by real-time RT-PCRFEMS YEAST RESEARCH, Issue 3 2004Mika Toyoda Abstract By using real-time RT-PCR, we profiled the expression of CGR1, CaMSI3, EFG1, NRG1, and TUP1 in Candida albicans strains JCM9061 and CAI4 under several conditions, including induction of morphological transition, heat shock, and treatment with calcium inhibitors. Expression of CaMSI3 changed under these growth conditions except during heat shock. CGR1 expression increased during the early stages of hyphal growth in JCM9061, while expression was strain-dependent during heat shock. Both EFG1 and NRG1 were similarly expressed under hypha-inducing conditions and heat shock. Expression of TUP1 was slightly different from the expression of EFG1 or NRG1. [source] Gluconeogenesis in Candida albicansFEMS YEAST RESEARCH, Issue 3 2002D. Eschrich Abstract According to different metabolic situations in various stages of Candida albicans pathogenesis the regulation of carbohydrate metabolism was investigated. We report the genetic characterization of all major C. albicans gluconeogenic and glyoxylate cycle genes (fructose-1,6-bisphosphatase, PEP carboxykinase, malate synthase and isocitrate lyase) which were isolated after functional complementation of the corresponding Saccharomyces cerevisiae deletion mutants. Remarkably, the regulation of the heterologously expressed C. albicans gluconeogenic and glyoxylate cycle genes was similar to that of the homologous S. cerevisiae genes. A C. albicans,Cafbp1 deletion strain failed to utilize non-fermentable carbon sources but hyphal growth was not affected. Our results show that regulation of gluconeogenesis in C. albicans is similar to that of S. cerevisiae and that the current knowledge on how gluconeogenesis is regulated will facilitate the physiological understanding of C. albicans. [source] Light and Electron Microscopy of the Compatible Interaction Between Arabidopsis and the Downy Mildew Pathogen Peronospora parasiticaJOURNAL OF PHYTOPATHOLOGY, Issue 6 2003E. M. Soylu Abstract In this study, we focused on compatible interactions between Peronospora parasitica isolate Emoy-2 and wild-type (Oy-0) and mutant (Ws- eds1) Arabidopsis thaliana accessions by using light and transmission electron microscopy (TEM). Light microscopy of compatible interactions revealed that conidia germinated and penetrated through the anticlinal cell walls of two epidermal cells. Rapid spreading of the hyphal growth with formation of numerous haustoria within the mesophyll cells was subsequently followed by profuse sporulation in the absence of host cell necrosis on both wild-type and mutant accessions. TEM observations revealed that coenocytic intercellular hyphae ramified and spread intercellularly throughout the host tissue forming several haustoria in host mesophyll cells. Intracellular haustoria were lobed with the diameter of 6,7 ,m. Each haustorium was connected to intercellular hyphae in the absence of apparent haustorial neck. The cytoplasm of the haustorium included the organelles characteristic of the pathogen. Callose-like deposits were frequently observed at sites of penetration around the proximal region of the haustorial neck. Apart from a few callose ensheatments, no obvious response was observed in host cells following formation of haustoria. Most of mesophyll cells contained normal haustoria and the host cytoplasm displayed a high degree of structural integrity. Absence of host cell wall alteration and cell death in penetrated host cell of both accessions suggest that the pathogen exerts considerable control over basic cellular processes and in this respect, response to this biotroph oomycete differs considerably from responses to other pathogens such as necrotrophs. [source] Antifungal Activity of a Bowman,Birk-type Trypsin Inhibitor from Wheat KernelJOURNAL OF PHYTOPATHOLOGY, Issue 7-8 2000G. Chilosi A trypsin inhibitor from wheat kernel (WTI) was found to have a strong antifungal activity against a number of pathogenic fungi and to inhibit fungal trypsin-like activity. WTI inhibited in vitro spore germination and hyphal growth of pathogens, with protein concentration required for 50% growth inhibition (IC50) values ranging from 111.7 to above 500 ,g/ml. As observed by electron microscopy, WTI determined morphological alterations represented by hyphal growth inhibition and branching. One of the fungal species tested, Botrytis cinerea produced a trypsin-like protease, which was inhibited by the trypsin inhibitor. WTI, as well as other seed defence proteins, appear to be an important resistance factor in wheat kernels during rest and early germination when plants are particularly exposed to attack by potential soil-borne pathogens. Zusammenfassung Ein Trypsinhemmer aus Weizenkörnern (WTI) zeigte eine starke antifungale Aktivität gegenüber verschiedenen pathogenen Pilzen und hemmte deren trypsinähnliche Aktivität. WTI hemmte in vitro die Sporenkeimung und das Hyphenwachstum der Pathogene, wobei die IC50 -Werte zwischen 111,7 und mehr als 500 ,g/ml lagen. Elektronenmikroskopische Untersuchungen zeigten, dai WTI morphologische Veränderungen bewirkte, die aus einer Hemmung des Hyphenwachstums und einer veränderten Verzweigung bestanden. Eine der untersuchten Pilzarten, Botrytis cinerea, bildete eine trypsinähnliche Protease, die durch den Trypsininhibitor gehemmt wurde. Ebenso wie andere sameneigene Abwehrproteine scheint WTI während der Keimruhe und in den frühen Stadien der Keimung, wenn die Pflanzen gegenüber möglichen bodenbürtigen Pathogenen besonders exponiert sind, ein wichtiger Resistenzfaktor in Weizenkörnern zu sein. [source] FlbC is a putative nuclear C2H2 transcription factor regulating development in Aspergillus nidulansMOLECULAR MICROBIOLOGY, Issue 5 2010Nak-Jung Kwon Summary Asexual development (conidiation) in Aspergillus is governed by multiple regulators. Here, we characterize the upstream developmental activator FlbC in Aspergillus nidulans. flbC mRNA is detectable throughout the life cycle, at relatively high levels during vegetative growth, early asexual and late sexual developmental phases. The deletion of flbC causes a delay/reduction in conidiation, brlA and vosA expression, and conidial germination. While overexpression of flbC (OEflbC) does not elaborate conidiophores, it inhibits hyphal growth and activates expression of brlA, abaA and vosA, but not wetA. FlbC is conserved in filamentous Ascomycetes containing two C2H2 zinc fingers at the C-terminus and a putative activation domain at the N-terminus. FlbC localizes in the nuclei of both hyphae and developmental cells. Localization and expression of FlbC are not affected by the absence of FlbB or FlbE, and vice versa. Importantly, overexpression of flbC causes growth inhibition and activation of abaA and vosA in the absence of brlA and abaA respectively. In vitro DNA-binding assay reveals that FlbC binds to the brlA, abaA and vosA, but not the wetA, promoters. In summary, FlbC is a putative nuclear transcription factor necessary for proper activation of conidiation, and its balanced activity is crucial for governing growth and development in A. nidulans. [source] CPCR1, but not its interacting transcription factor AcFKH1, controls fungal arthrospore formation in Acremonium chrysogenumMOLECULAR MICROBIOLOGY, Issue 5 2005Birgit Hoff Summary Fungal morphogenesis and secondary metabolism are frequently associated; however, the molecular determinants connecting both processes remain largely undefined. Here we demonstrate that CPCR1 (cephalosporin C regulator 1 from Acremonium chrysogenum), a member of the winged helix/regulator factor X (RFX) transcription factor family that regulates cephalosporin C biosynthesis, also controls morphological development in the ,-lactam producer A. chrysogenum. The use of a disruption strain, multicopy strains as well as several recombinant control strains revealed that CPCR1 is required for hyphal fragmentation, and thus the formation of arthrospores. In a ,cpcR1 disruption strain that exhibits only hyphal growth, the wild-type cpcR1 gene was able to restore arthrospore formation; a phenomenon not observed for ,cpcR1 derivatives or non-related genes. The intracellular expression of cpcR1, and control genes (pcbC, egfp) was determined by in vivo monitoring of fluorescent protein fusions. Further, the role of the forkhead transcription factor AcFKH1, which directly interacts with CPCR1, was studied by generating an Acfkh1 knockout strain. In contrast to CPCR1, AcFKH1 is not directly involved in the fragmentation of hyphae. Instead, the presence of AcFKH1 seems to be necessary for CPCR1 function in A. chrysogenum morphogenesis, as overexpression of a functional cpcR1 gene in a ,Acfkh1 background has no effect on arthrospore formation. Moreover, strains lacking Acfkh1 exhibit defects in cell separation, indicating an involvement of the forkhead transcription factor in mycelial growth of A. chrysogenum. Our data offer the potential to control fungal growth in biotechnical processes that require defined morphological stages for optimal production yields. [source] Cell cycle regulator Cdc14 is expressed during sporulation but not hyphal growth in the fungus-like oomycete Phytophthora infestansMOLECULAR MICROBIOLOGY, Issue 2 2003Audrey M. V. Ah Fong Summary Cdc14 proteins are important regulators of mitosis and the cell cycle. These phosphatases have been studied previously only in yeasts and metazoans, which grow by fission or budding. Here we describe a homologue (piCdc14) from the oomycete Phytophthora infestans, a primitive eukaryote lacking a classical cell cycle. PiCdc14 complements a cdc14ts mutant of Saccharomyces cerevisiae and may function like other Cdc14 proteins, but displays a strikingly different pattern of expression. Whereas previously studied Cdc14 genes are constitutively transcribed, piCdc14 is not expressed during normal growth but instead only during asexual sporulation. In transformants of P. infestans expressing a fusion between the piCdc14 promoter and the ,-glucuronidase reporter, expression was first detected in sporangiophore initials, persisted in sporangiophores bearing immature sporangia, and later became restricted to mature sporangia. After germination, expression ended a few hours before the resumption of mitosis in hyphae emerged from the spores. Homology-dependent silencing experiments supported an essential role of piCdc14 in sporulation. It is proposed that the function of piCdc14 may be to synchronise nuclear behaviour during sporulation and maintain dormancy in spores until germination. These results help illuminate the process of sporulation in oomycetes and the evolution of the cell cycle in eukaryotes. [source] Requirement of spermidine for developmental transitions in Aspergillus nidulansMOLECULAR MICROBIOLOGY, Issue 3 2002Yuan Jin Summary Deletion of the spermidine synthase gene in the fungus Aspergillus nidulans results in a strain, ,spdA, which requires spermidine for growth and accumulates putrescine as the sole polyamine. Vegetative growth but not sporulation or sterigmatocystin production is observed when ,spdA is grown on media supplemented with 0.05,0.10 mM exogenous spermidine. Supplementation of ,spdA with , 0.10 mM spermidine restores sterigmatocystin production and , 0.50 mM spermidine produces a phenotype with denser asexual spore production and decreased radial hyphal growth compared with the wild type. ,spdA spores germinate in unsupplemented media but germ tube growth ceases after 8 h upon which time the spores swell to approximately three times their normal diameter. Hyphal growth is resumed upon addition of 1.0 mM spermidine. Suppression of a G protein signalling pathway could not force asexual sporulation and sterigmatocystin production in ,spdA strains grown in media lacking spermidine but could force both processes in ,spdA strains supplemented with 0.05 mM spermidine. These results show that increasing levels of spermidine are required for the transitions from (i) germ tube to hyphal growth and (ii) hyphal growth to tissue differentiation and secondary metabolism. Suppression of G protein signalling can over-ride the spermidine requirement for the latter but not the former transition. [source] Dynamic distribution of BIMGPP1 in living hyphae of Aspergillus indicates a novel role in septum formationMOLECULAR MICROBIOLOGY, Issue 5 2002H. Fox Summary Mutation of bimG, the major protein phosphatase 1 gene in Aspergillus nidulans, causes multiple cell cycle and hyphal growth defects that are associated with overphosphorylation of subcellular components. We have used functional translational fusions with the green fluorescent protein (GFP) to show that BIMG has at least four discrete locations within growing hyphae. Three of these locations, the hyphal tip, the spindle pole body and the nucleus, correlate with previously known requirements for bimGPP1 in mitosis and hyphal growth and are highly dynamic. BIMG-GFP in the hyphal tip seemed to be associated with the plasma membrane and formed a collar of fluorescence within the apical dome. The distribution of nuclear BIMG-GFP varied depending on nutritional conditions; on poor medium, it concentrated more in the nucleolus than in the nucleoplasm, whereas on rich medium, it was more evenly distributed between the two nuclear regions. The association of BIMG-GFP with developing septa was transient, and we present evidence that BIMG phosphatase plays a direct role in septum formation, distinct from its role in mitosis. We conclude that, by being physically present at several sites, the BIMG phosphatase has roles in multiple cellular processes. [source] Effects of repeated low-dose UVB irradiation on the hyphal growth of Candida albicansMYCOSES, Issue 1 2006J. Brasch Summary Ultraviolet B light (UVB) can have negative phototropic effects on fungi. Candida albicans is often found on human skin exposed to UVB. Therefore, it is of medical interest to know whether a negative phototropic response to UVB irradiation can support an invasive growth of this potentially dangerous agent. In our study we investigated how repeated irradiation with low doses of UVB can influence the hyphal growth of C. albicans. Six randomly chosen strains of C. albicans were tested. Formation of hyphae was induced and maintained within transparent agar plates. The fungi were exposed to UVB three times daily for 7 days from either the obverse or the reverse side during incubation. The wavelength spectrum was in the range of 310,315 nm, single doses were between 0.0018 and 0.432 J cm,2. After 7 days the morphology and growth direction of C. albicans cells were determined microscopically. All six strains showed a common and dose-dependent response to UVB irradiation: the progression of hyphal growth was inhibited, no phototropic effects were seen and as a new finding an increased formation of blastospores was observed. We conclude that an irradiation of human skin colonized by C. albicans with doses of UVB that can occur under natural or artificial conditions is unlikely to trigger skin invasion by C. albicans. [source] Arbuscular mycorrhizal fungi can transfer substantial amounts of nitrogen to their host plant from organic materialNEW PHYTOLOGIST, Issue 1 2009Joanne Leigh Summary ,,Nitrogen (N) capture by arbuscular mycorrhizal (AM) fungi from organic material is a recently discovered phenomenon. This study investigated the ability of two Glomus species to transfer N from organic material to host plants and examined whether the ability to capture N is related to fungal hyphal growth. ,,Experimental microcosms had two compartments; these contained either a single plant of Plantago lanceolata inoculated with Glomus hoi or Glomus intraradices, or a patch of dried shoot material labelled with 15N and 13carbon (C). In one treatment, hyphae, but not roots, were allowed access to the patch; in the other treatment, access by both hyphae and roots was prevented. ,,When allowed, fungi proliferated in the patch and captured N but not C, although G. intraradices transferred more N than G. hoi to the plant. Plants colonized with G. intraradices had a higher concentration of N than controls. ,,Up to one-third of the patch N was captured by the AM fungi and transferred to the plant, while c. 20% of plant N may have been patch derived. These findings indicate that uptake from organic N could be important in AM symbiosis for both plant and fungal partners and that some AM fungi may acquire inorganic N from organic sources. [source] Orchid mycorrhiza: implications of a mycophagous life styleOIKOS, Issue 3 2009Hanne N. Rasmussen Orchid mycorrhiza probably affects about 25,000 plant species and thus roughly one tenth of all higher plants. Histologically, this symbiosis resembles other kinds of endomycorrhiza, the fungal hyphae growing within living plant cells. Considerable evidence, however, suggests that it is not a two-way exchange relationship and thus not potentially mutualistic, such as the wide-spread endomycorrhiza between plants and Glomalean fungi, known as arbuscular mycorrhiza. During the achlorophyllous seedling stage orchids are obligately dependent on the fungi; some species remain so through life, while others establish photosynthesis but to varying degrees remain facultatively dependent of /responsive to fungal infection as adults. None of the fungi involved are so far known to depend on the symbiosis with orchids. Transfer of organic carbon compounds from hyphae to the orchid has been demonstrated repeatedly, but it is not clear to what extent this takes place during a biotrophic phase while the intracellular hyphae remain intact, or during the subsequent extensive degradation of the hyphal coils. The advantage of viewing orchid mycorrhiza basically as a unilateral mycophagous relationship, in spite of hypothetical beneficial spin-offs to the mycobiont, is that it provides a conceptual framework similar to that of other parasitic or fungivore relationships; mechanisms known in such relationships could be searched for in future studies of the orchid,fungus symbiosis. These could include mechanisms for recognition, attraction and selection of fungi, physiological regulation of internal hyphal growth, breakdown, and material transfer, nutritional consequences of the plant's preference(s) and trophic changes, fungal avoidance mechanisms, and consequences at population and ecosystem levels. A whole range of possible life strategies becomes apparent that could support divergent evolution and lead to the proliferation of species that has indeed occurred in the orchid family. We outline some of the possible physiological mechanisms and ecological implications of this approach. [source] Effects of temperature on germination and hyphal growth from conidia of Ramularia rhei and Ascochyta rhei, causing spot diseases of rhubarb (Rheum rhaponticum)PLANT PATHOLOGY, Issue 5 2006Y. Zhao Rhubarb leaf and petiole spot disease, caused by Ramularia rhei and Ascochyta rhei, has gradually become more noticeable in the UK field crop. Conidial germination and subsequent colony growth of R. rhei and A. rhei were investigated under in vitro conditions on potato dextrose agar and in vivo on leaf discs. Results indicated that the two fungi responded differently to temperature. Ramularia rhei was better adapted to temperatures , 25°C, with an optimum around 20°C, whereas A. rhei was more adapted to temperatures , 15°C, with an optimum > 25°C. Overall, conidia of R. rhei germinated and subsequent colonies grew at greater rates than those of A. rhei on leaf discs at temperatures , 25°C. These results indicated that it is important to identify the causal agent of leaf and petiole spot diseases in rhubarb field crops in order to estimate disease risks accurately. [source] Induction of Cell Wall Thickening by the Antifungal Compound Dihydromaltophilin Disrupts Fungal Growth and is Mediated by Sphingolipid BiosynthesisTHE JOURNAL OF EUKARYOTIC MICROBIOLOGY, Issue 2 2009SHAOJIE LI ABSTRACT. Dihydromaltophilin (heat-stable antifungal factor [HSAF]) is an antifungal metabolite produced in Lysobacter enzymogenes biocontrol strain C3. This compound induces cell wall thickening in Aspergillus nidulans. Here we show that the cell wall thickening is a general response to HSAF in diverse fungal species. In the A. nidulans model, the thickened cell wall negatively affects hyphal growth. Growth of HSAF-pre-treated hyphae failed to resume at hyphal tips with thick cell wall and the actin cable could not re-polarize at the thickened region of the cell wall, even after the treated hyphae were transferred to drug-free medium. Moreover, HSAF-induced cell wall thickening is mediated by sphingolipid synthesis: HSAF failed to induce cell wall thickening in the absence of ceramide synthase BarA and the sphingolipid synthesis inhibitor myriocin was able to suppress HSAF-induced cell wall thickening. The thickened cell wall could be digested by chitinase suggesting that chitin contributes to the HSAF-induced thickening. Furthermore, HSAF treatment activated the transcription of two chitin synthase encoding genes chsB and chsC. [source] | ||||||||||||||||||||||