Search

13 páginas, 7 figuras, 1 tabla, Functional symbiosis with fungal endophytes can help plants adapt to environmental stress. Diaporthe atlantica is one of the most abundant fungal taxa associated with roots of Festuca rubra subsp. pruinosa, a grass growing in sea cliffs. This study aimed to investigate the ability of a strain of this fungus to ameliorate the impact of drought stress on tomato plants. In a greenhouse experiment, tomato plants were inoculated with Diaporthe atlantica strain EB4 and exposed to two alternative water regimes: well-watered and drought stress. Several physiological and biochemical plant parameters were evaluated. Inoculation with Diaporthe promoted plant growth in both water treatments. A significant interactive effect of Diaporthe-inoculation and water-regime showed that symbiotic plants had higher photosynthetic capacity, water-use efficiency, nutrient uptake (N, P, K, Fe and Zn), and proline content under drought stress, but not under well-watered conditions. In addition, Diaporthe improved the enzymatic antioxidant response of plants under drought, through an induced mechanism, in which catalase activity was modulated and conferred protection against reactive oxygen species generation during stress. The results support that Diaporthe atlantica plays a positive role in the modulation of tomato plant responses to drought stress by combining various processes such as improving photosynthetic capacity, nutrient uptake, enzymatic antioxidant response and osmo-protectant accumulation. Thus, drought stress in tomato can be enhanced with symbiotic fungi, This research was supported by the European Union’s H2020 research and innovation program under the Marie Sklodowska-Curie grant agreement no. 676480; by grant PID2019-109133RB-I00 funded by MCIN/AEI/10.13039/501100011033, and by project ‘CLU2019-05 — IRNASA/CSIC Unit of Excellence’ funded by the Junta de Castilla y León and co-financed by EU (ERDF ‘Europe drives our growth’)

Switched Reluctance Machines (SRM) are emerging as a possible alternative in terms of cost and supply stability to rare earth based electric vehicle traction systems. However, because of the huge amounts of energy stored and transferred back and forth between the DC source and the SRM, large DC-link capacitors must be used as buffers, which increases overall costs and size. This paper proposes a novel modulation technique which forces the exchange of energy between phases while decreasing the energy transfer between the DC bus and the SRM. This means lower DC bus currents (capacitor size and cost reduction) and lower Joule-effect conduction losses (better efficiency). The proposed modulation has been validated experimentally in a test bench and compared with the conventional torque-sharing function.

Copper is a metal ion that is required as a micronutrient for growth and proliferation. However, copper accumulation generates toxicity by multiple mechanisms, potentially leading to cell death. Due to its toxic nature at high concentrations, different chemical variants of copper have been extensively used as antifungal agents in agriculture and medicine. Most studies on copper homeostasis have been carried out in bacteria, yeast, and mammalian organisms. However, knowledge on filamentous fungi is less well documented. This review summarizes the knowledge gathered in the last few years about copper homeostasis in the filamentous fungi Aspergillus fumigatus and Aspergillus nidulans: The mechanism of action of copper, the uptake and detoxification systems, their regulation at the transcriptional level, and the role of copper homeostasis in fungal pathogenicity are presented.

Organic contaminants significantly limit the bioactivity of titanium implants, resulting in the degradation known as the ageing of titanium. To reactivate the surfaces, they can be photofunctionalized, i.e., irradiated with C-range ultraviolet (UVC) light. This descriptive in vitro study compares the effectiveness of novel light-emitting diode (LED) technology to remove contaminant hydrocarbons from three different commercially available titanium dental implants: THD, TiUnite, and SLA. The surface topography and morphology were characterized by scanning electron microscopy (SEM). The chemical compositions were analyzed by X-ray photoelectron spectroscopy (XPS), before and after the lighting treatment, by a pair of closely placed UVC (λ = 278 nm) and LED devices for 24 h. SEM analysis showed morphological differences at the macro- and micro-scopic level. XPS analysis showed a remarkable reduction in the carbon contents after the UVC treatment: from 25.6 to 19.5 C at. % (carbon atomic concentration) in the THD, from 30.2 to 20.2 C at. % in the TiUnite, from 26.1 to 19.2 C at. % in the SLA surface. Simultaneously, the concentration of oxygen and titanium increased. Therefore, LED-based UVC irradiation decontaminated titanium surfaces and improved the chemical features of them, regardless of the kind of surface.

16 p.-7 fig., Tolerance of microorganisms to abiotic stress is enabled by regulatory mechanisms that coordinate the expression and activity of resistance genes. Alkalinity and high salt concentrations are major environmental physicochemical stresses. Here, we analyzed the roles of sodium-extrusion family (ENA) transporters EnaA, EnaB and EnaC in the response to these stress conditions in the filamentous fungus Aspergillus nidulans. While EnaC has a minor role, EnaB is a key element for tolerance to Na+ and Li+ toxicity. Adaptation to alkaline pH requires the concerted action of EnaB with EnaA. Accordingly, expression of enaA and enaB was induced by Na+, Li+ and pH 8. These expression patterns are altered in a sltADelta background and completely inhibited in a mutant expressing non-functional PacC protein (palH72). However, a constitutively active PacC form was not sufficient to restore maximum enaA expression. In agreement with their predicted role as membrane ATPases, EnaA localized to the plasma membrane while EnaB accumulated at structures resembling the endoplasmic reticulum. Overall, results suggest different PacC- and SltA-dependent roles for EnaB in pH and salt homeostasis, acting in coordination with EnaA at pH 8 but independently under salt stress., Work at CIB-CSIC was funded by MINECO grants BFU2012-33142 and BFU2015-66806-R, as well as MICIU/AEI grant RTI2018-094263-B-100 to EAE (partially supported by FEDER, EU). Work at UPV/EHU was funded by Basque Government grants IT599-13 to UU and Elkartek19/72 to Prof. María Teresa Dueñas, as well as grants PPGA19/08 and GIU19/014 to O.E. AM-I held a predoctoral FPI fellowship from the Ministerio de Educación y Ciencia associated with Grant BIO2006-00556 to Miguel A. Peñalva, and a research contract associated with grant IT599-13 to UU.

FluG is a long recognized early regulator of asexual development in

The demand for more reliable and efficient electric machines and drives is constantly growing in the renewable energy and transport electrification sectors. Such drive systems are usually fed by semiconductor switch-based inverters, which, unlike balanced pure sine-wave AC sources, produce large-amplitude, high-frequency common-mode voltage (CMV) waveforms, as a result of the application of pulse-width modulation (PWM). This can lead to a number of issues, such as high electromagnetic interference, deterioration of stator winding insulation, and leakage current flow through motor bearings, which dramatically reduce the life-cycle of machines and drives. Thus, this topic has been extensively investigated in the scientific literature, where either corrective or preventive mitigation approaches have been proposed. The former attempt to relieve the damage produced, whereas the latter tackle the problem at its root, by minimizing or eliminating the CMV produced by the inverter. This work provides a comprehensive review of the major CMV mitigation/elimination solutions, with emphasis on preventive actions, in the form of inverter topology variants and/or advanced modulation techniques. A wide picture of this subject is provided to researchers and field engineers, with valuable information and practical hints for the design and development of high-performance electric drive systems. Indeed, an in-depth analysis of the most recent literature clearly shows that best results are obtained by conveniently combining alternative topologies and modulation techniques, which, in some cases, make it possible to completely suppress the CMV component.

In Aspergillus nidulans, asexual differentiation requires the presence of the transcription factor FlbB at the cell tip and apical nuclei. Understanding the relationship between these two pools is crucial for elucidating the biochemical processes mediating conidia production. Tip-to-nucleus communication was demonstrated by photo-convertible FlbB::Dendra2 visualization. Tip localization of FlbB depends on Cys382 in the C-terminus and the bZIP DNA-binding domain in the N-terminus. FlbE, a critical FlbB interactor, binds the bZIP domain. Furthermore, the absence of FlbE results in loss of tip localization but not nuclear accumulation. flbE deletion also abrogates transcriptional activity indicating that FlbB gains transcriptional competence from interactions with FlbE at the tip. Finally, a bipartite nuclear localization signal is required for nuclear localization of FlbB. Those motifs of FlbB may play various roles in the sequence of events necessary for the distribution and activation of this transcriptionally active developmental factor. The tip accumulation, FlbE-dependent activation, transport and nuclear import sketch out a process of relaying an environmentally triggered signal from the tip to the nuclei. As the first known instance of transcription factor-mediated tip-to-nucleus communication in filamentous fungi, this provides a general framework for analyses focused on elucidating the set of molecular mechanisms coupling apical signals to transcriptional events.

Microbial cells interact with the environment by adapting to external changes. Signal transduction pathways participate in both sensing and responding in the form of modification of gene expression patterns, enabling cell survival. The filamentous fungal-specific SltA pathway regulates tolerance to alkalinity, elevated cation concentrations and, as shown in this work, also stress conditions induced by borates. Growth of sltA− mutants is inhibited by increasing millimolar concentrations of boric acid or borax (sodium tetraborate). In an attempt to identify genes required for boron-stress response, we determined the boric acid or borax-dependent expression of sbtA and sbtB, orthologs of Saccharomyces cerevisiae bor1, and a reduction in their transcript levels in a ΔsltA mutant. Deletion of sbtA, but mainly that of sbtB, decreased the tolerance to boric acid or borax. In contrast, null mutants of genes coding for additional transporters of the Solute Carrier (SLC) family, sB, sbtD or sbtE, showed an unaltered growth pattern under the same stress conditions. Taken together, our results suggest that the SltA pathway induces, through SbtA and SbtB, the export of toxic concentrations of borates, which have largely recognized antimicrobial properties., Work at CIB-CSIC was funded by MINECO project BFU2012-33142 and MINECO/FEDER/EU project BFU2015-66806-R to E.A.E. Work at the UPV/EHU lab was funded by the Basque Government (grant IT599-13 to U.U.) and UPV/EHU (grant EHUA15/08 to O.E.). M.V. held a contract under project BFU2012-33142., We acknowledge support by the CSIC Open Access Publication Initiative through its Unit of Information Resources for Research (URICI).

The mycelium is an organised cellular network that develops according to a functionally coherent plan. As it expands, the mycelium is capable of modulating the relative abundance of different cell types to suit the prevailing environmental conditions. This versatile pattern of multicellular development involves sophisticated environmental sensing and intercellular communication systems that have barely been recognised. This review describes an insight into our current understanding of the signalling molecules and mechanisms that take part in the ordered and timely emergence of various cell types and their biological significance. The prospects that this emerging knowledge may offer for the sustainable control of fungal colonisation or dispersal will also be considered.

In this study, the authors show that it is particularly advantageous to use the fractional-order hold (FROH) instead of the zero-order hold (ZOH) in the digital control of lightly damped resonant systems, as far as transient energy consumption is concerned. First, they show that for a typical digital controller choice FROH reconstruction may well produce a continuous-time signal with less associated energy than ZOH reconstruction. Second, they focus on lightly damped resonant systems, where high energy savings are likely to occur because of the initially swinging nature of the discrete-time control signals driving these systems. Then, they propose an easy procedure to tune the FROH, which is not bound to z-domain controller design, and is therefore radically different from most, if not all, procedures published so far. Finally, they provide some simulation-based application examples confirming the significant control energy reduction attained, in excess of 50%, while not hindering the output performance.

This paper presents an exhaustive experimental study on the performance of a new design of generalized sampled-data hold function (GSHF) introduced in Ugalde, Barcena, and Basterretxea (2012) . First a simple tuning procedure is developed for the GSHF to improve the intersample response while not damaging the plant output response. Next the reconstruction algorithm is enhanced so that the GSHF can remove the steady-state errors that dry friction causes in positioning systems. The experiments conducted on a small-scale lightly damped resonant system show that significant energy savings can be achieved in regulation with no extra hardware, and that no additional computational load results, except having to call the D/A converter more than once per sampling period.

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 C(2) H(2) 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.

Fungal colonies expand through apically extending peripheral hyphae. These long and relatively unbranched leaders leave behind cells that show a higher degree of branching and lower growth rates. Towards the centre of the colony, quiescent, but viable cells sustain processes such as transport of water, solutes and organelles, and support the concerted genesis of specialized reproductive structures. A survey of chemical signals controlling changes in hyphal growth and branching of Aspergillus nidulans in solid minimal medium revealed that progressive accumulation of bicarbonate from the periphery to the centre of the colony effected a dual modulation of both parameters. At areas immediately behind the periphery, bicarbonate induced progressive branching without a significant change in the specific growth rate. At distal and central regions, increased branching was accompanied by progressive inhibition of the specific growth rate. The results provide new evidence that chemical cues participate in the control of colony morphology and function, which are likely to vary with changing environmental conditions.

Aspergillus nidulans switches from vegetative growth to conidiation when aerial hyphae make contact with the atmosphere, or are subjected to specific environmental stress. The activation of the central conidiation pathway led by the transcription factor brlA is a critical milestone in this morphogenetic transition. A number of upstream developmental activators (UDAs), expressed in vegetative cells, are required for this process to occur in conjunction with cessation of vegetative growth. Mutants affected in these factors remain aconidial (fluffy) with low brlA expression levels (flb). In this report, we describe FlbE as a UDA containing two conserved but hitherto uncharacterized domains, which functions in close association with putative transcription factor FlbB. Both UDAs are functionally interdependent, and colocalize at the hypha tip in an actin cytoskeleton-dependent manner. Moreover, bimolecular fluorescence studies show that they physically interact in vivo. These findings add evidence in favour of the existence of a signalling complex at or near the Spitzenkörper as an important part of the machinery controlling the morphogenetic transition between vegetative growth and conidiation.

In Aspergillus nidulans, asexual differentiation requires the presence of the transcription factor FlbB at the cell tip and apical nuclei. Understanding the relationship between these two pools is crucial for elucidating the biochemical processes mediating conidia production. Tip-to-nucleus communication was demonstrated by photo-convertible FlbB::Dendra2 visualization. Tip localization of FlbB depends on Cys382 in the C-terminus and the bZIP DNA-binding domain in the N-terminus. FlbE, a critical FlbB interactor, binds the bZIP domain. Furthermore, the absence of FlbE results in loss of tip localization but not nuclear accumulation. flbE deletion also abrogates transcriptional activity indicating that FlbB gains transcriptional competence from interactions with FlbE at the tip. Finally, a bipartite nuclear localization signal is required for nuclear localization of FlbB. Those motifs of FlbB may play various roles in the sequence of events necessary for the distribution and activation of this transcriptionally active developmental factor. The tip accumulation, FlbE-dependent activation, transport and nuclear import sketch out a process of relaying an environmentally triggered signal from the tip to the nuclei. As the first known instance of transcription factor-mediated tip-to-nucleus communication in filamentous fungi, this provides a general framework for analyses focused on elucidating the set of molecular mechanisms coupling apical signals to transcriptional events.

In the model fungus Aspergillus nidulans, asexual development is induced from vegetative hyphae by a set of early regulators including the bZIP-type transcription factor FlbB. To determine the range of genes under the influence of the transcriptional activity of FlbB and to characterize their role in fungal development, we sequenced and compared the transcriptomes of a ΔflbB mutant and its isogenic wild-type strain at different developmental stages. Results confirmed the activating role of FlbB on downstream regulators of conidiation such as flbD and brlA. However, FlbB has additional functions beyond the induction of asexual development. Among the changes observed, absence of a functional FlbB caused induction of the dba cluster and synthesis of a secondary metabolite with bactericidal properties. In addition, a new transcriptional target of FlbB was unveiled, urdA, that codes for a putative transcription factor that represses premature sexual development. Taken together, our results indicate that the activators of asexual development simultaneously exert a role on other cellular functions, including an inhibitory effect on the sexual cycle, and reinforce the hypothesis that mutually exclusive metabolic and cellular patterns are associated with different morphogenetic programs.

The filamentous fungus Penicillium cyclopium conidiates in the presence of calcium ions in submerged culture without nutrient limitation according to a precisely timed program. Conidiation could be prematurely induced in a nutritionally sufficient medium which had previously supported growth, suggesting that a metabolite which influenced the process was produced. A diterpenoid with conidiation-inducing activity, conidiogenone, was purified from the culture medium, along with conidiogenol, a putative derivative with delayed activity. Contrary to previous thought, the presence of calcium was demonstrated to only decrease the threshold concentration of conidiogenone required for the induction to proceed. In light of these results, a mechanism of conidiation induction is presented according to which the mycelium produces a conidiation inducer (conidiogenone) that accumulates extracellularly. When a threshold concentration is reached, induction likely takes place by interaction with a specific cellular receptor. The results indicate that conidiogenone is both sufficient and necessary to induce conidiation.

The amount of acid or base consumed in yeast cultures has been recently assigned to the pathway of nitrogen assimilation under respiratory conditions with no contribution by carbon metabolism (Castrillo et al., 1995). In this investigation, experiments under respiro- fermentative conditions have shown that production or consumption of ethanol does not contribute significantly to the specific rate of proton production (qH + ), thus ex- tending the previously obtained relationships for all aerobic conditions in which other major acid/base con- tributions are not involved. Tests in batch and chemostat culture confirm the validity of qH + as a formal control parameter in aerobic fermentations. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 58: 445-450, 1998.

32 p.-6 fig.-2 tab., Morphogenesis encompasses programmed changes in gene expression that lead to the development of specialized cell types. In the model fungus Aspergillus nidulans, asexual development involves the formation of characteristic cell types, collectively known as the conidiophore. With the aim of determining the transcriptional changes that occur upon induction of asexual development, we have applied massive mRNA sequencing to compare the expression pattern of 19-h-old submerged vegetative cells (hyphae) with that of similar hyphae after exposure to the air for 5 h. We found that the expression of 2,222 (20.3%) of the predicted 10,943 A. nidulans transcripts was significantly modified after air exposure, 2,035 being downregulated and 187 upregulated. The activation during this transition of genes that belong specifically to the asexual developmental pathway was confirmed. Another remarkable quantitative change occurred in the expression of genes involved in carbon or nitrogen primary metabolism. Genes participating in polar growth or sexual development were transcriptionally repressed, as were those belonging to the HogA/SakA stress response mitogen-activated protein (MAP) kinase pathway. We also identified significant expression changes in several genes purportedly involved in redox balance, transmembrane transport, secondary metabolite production, or transcriptional regulation, mainly binuclear-zinc cluster transcription factors. Genes coding for these four activities were usually grouped in metabolic clusters, which may bring regulatory implications for the induction of asexual development. These results provide a blueprint for further stage-specific gene expression studies during conidiophore development. © 2013, American Society for Microbiology. All Rights Reserved., This work has been supported by the Basque Government through grant (IT393-10) and the Ministerio de Economía y Competitividad (formerly Ministerio de Ciencia e Innovación) through grant (BFU2010-17528) to U.U., grant (BFU2009- 08701) to E.A.E. and grants from the German Science Foundation (DFG Fi 459), the Fonds der Chemischen Industrie, the Baden-Württemberg Stiftung, and the Centre for Functional Nanostructures to R.F. A. G. is now a contract researcher from The University of The Basque Country. J.R. was a postdoctoral fellow of the Ministerio de Ciencia e Innovación. O.E is a contract researcher associated to grant BFU2010- 17528.

Whey proteins were hydrolyzed in a membrane recycle reactor (MRR) in the presence of Alcalase 0.6L under varying enzyme to substrate ratios and residence times. Substrate conversion was directly dependent on these two parameters, and at a fixed enzyme-substrate ratio of 10%, conversion levels could be controlled through residence time, resulting in peptide permeates with homogeneous molecular and functional properties. In addition, recycling of the enzyme and elimination of peptide products resulted in much improved enzyme yields and process productivity levels. However, the process was only operational for 7 h, given membrane fouling and enzyme inactivation phenomena whose contribution occurred in a time-dependent manner. Changes in process design leading to increased reactor stability were examined, and the system is considered to provide a suitable basis for the production of peptides with specific functional and nutritional properties at the industrial level in the future.

When growing Aspergillus nidulans hyphae encounter the atmosphere, they initiate a morphogenetic program leading to the production of spores. Mutants that are defective in the fluG gene fail to undergo sporulation because they lack an endogenous diffusible factor that purportedly accumulates on aerial hyphae, thus signaling the initiation of development. In this study, the defect could be reversed by adding culture extracts from a wild-type strain onto a mutant colony. Moreover, a bioassay-guided purification of the active culture extract resulted in the identification of the active agent as dehydroaustinol. However, this meroterpenoid was active only when administered in conjunction with the orsellinic acid derivative diorcinol. These two compounds formed an adduct that was detected by HRMS in an LC-MS experiment. The diorcinol-dehydroaustinol adduct prevented crystal formation of the signal on the surface of aerial hyphae and on an artificially prepared aqueous film and also increased the signal lipophilicity.

A whey protein hydrolysate was prepared by incubation of reconstituted whey or a whey protein concentrate with Alcalase 0.6L. The proteolytic degradation of α-lactalbumin and β-lactoglobulin initially resulted in 6-kDa and, later, 2.5-kDa degradation products, quickly followed by the appearance of multiple peptides of 1 kDa or smaller. The hydrolysate showed a steady increase in solubility and a biphasic change in foaming characteristics with decreasing peptide size. At the highest degree of hydrolysis achieved (22%), the majority of the peptides were smaller than 1 kDa and could be efficiently assimilated by the yeast Kluyveromyces marxianus growing in a defined medium.

15 páginas, 2 figuras -- PAGS nros. 704-718, Development in the ascomycete A. nidulans is principally determined by environmental signals. Adaptability to oxidative stimuli can derive in changes of growth patterns and/or the activation of sexual or asexual reproductive cycles but this model fungus might also respond to high osmotic or salt concentrations, the redox state, the availability and quantity of carbon or nitrogen sources and the degree or quality of illumination. Since each cell within the colony follows a single morphogenetic program at a time, all these environmental cues might be sensed and integrated into a limited number of intracellular signals which, finally, would activate the required morphogenetic program and repress the others. This signaling mainly occurs through stress response pathways. The present review aims to summarize the available knowledge on how these pathways transduce environmental stimuli to mediate morphological changes in Aspergillus nidulans., This work was supported by the Spanish Ministerio de Educación y Ciencia through grant BFU2009-08701 to E.A.E, and by the Basque Government through grant IT393- 10 to U.U. O.E. is a contract researcher of The University of The Basque Country

Germination of Aspergillus nidulans conidia in liquid cultures was progressively inhibited at inoculum loads above 1 × 105 conidia mL−1. High conidial densities also inhibited growth of neighbouring mycelia. The eight-carbon oxylipin 1-octen-3-ol was identified as the main inhibitor in a fraction also containing 3-octanone and 3-octanol. These three oxylipins also increased the conidiation rate of dark-grown surface cultures, but had no effect on liquid cultures. 3-octanone was the most conidiogenic compound. The action of 3-octanone required functional forms of developmental activators fluG, flbB-D and brlA, and was not additive to the conidiogenic effect of stress stimuli such as osmotic stress or carbon starvation. Oxylipins were produced shortly after hyphae made contact with the atmosphere and were most effective on aerial mycelia, indicating that they perform their signalling function in the gas phase.

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 C(2) H(2) 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.

The isolation and structure determination of conidiogenol and conidiogenone, tetracyclic diterpenes with a novel carbon skeleton, from extracts of the fermentation broth of Penicillium cyclopium is reported. Conidiogenol and conidiogenone are potent and selective inducers of conidiogenesis in P. cyclopium in liquid culture, and relay information about the environmental conditions to the producing organism. (C) 2002 Elsevier Science Ltd. All rights reserved.

12 páginas, 7 figuras, 2 tablas -- PAGS nros. 3934-3945, Asexual development in the filamentous fungus Aspergillus nidulans is governed by the timely expression and cellular localization of multiple transcription factors. Hence, factors mediating import and export across the nuclear pore complexes (karyopherins) are expected to play a key role in coordinating the developmental programme. Here we characterize KapI, a putative homologue of the Saccharomyces cerevisiae Kap121/Pse1p karyopherin. KapI is a non-essential importin-β-like protein located in the nucleus during vegetative growth and conidiophore development. The ΔkapI phenotype is aconidial with many aerial hyphae. This phenotype can be suppressed under abiotic stress. In this regard, it resembles that of the null allele of the bZIP transcription factor FlbB. However a ΔflbB; ΔkapI double mutant exhibited an additive phenotype with totally impaired conidiation, unresponsive to abiotic stress. In contrast to ΔflbB, the null kapI mutant is not a fluffy-low-bristle expression mutant. Taken together the findings indicate that KapI is required during asexual development, mediating the nuclear transport of factors acting in a different pathway(s) from those involving the upstream developmental activators, This work was supported by the Spanish Ministerio de Educación y Ciencia through grants BFU2006-04185 and BFU2004-03499/BMC to E. A. E. and U. O. U., respectively, and by the University of The Basque Country (UPV/EHU) for grant GIU08/32 to U. U. A. G. and E. H.-G. held predoctoral fellowships from the Basque Government. A. M.-I. held a predoctoral FPI fellowship from the Ministerio de Educación y Ciencia. O. E. was a contract researcher of the CIC Nanogune, and currently holds a research contract associated with grant BFU2006-04185 at the CIB

15 páginas, 6 figuras, 3 tablas -- PAGS nros. 775-789, Conidiophore formation in Aspergillus nidulans involves a developmental programme in which vegetative hyphae give rise to an ordered succession of differentiated cells: foot cell, stalk, vesicle, metulae, phialides and conidia. The developmental transition requires factors that are expressed in vegetative hyphae that activate the expression of the main regulator of conidiation, BrlA. One such element is the bZIP-type transcription factor FlbB. We found that flbB- mutants show defective branching patterns and are susceptible to autolysis under high sorbitol or sucrose concentrations, revealing a role in vegetative growth. In addition, FlbB plays a role in conidiophore initiation, as its upregulation reduces conidiophore vesicle swelling and generates a reduced number of metulae. FlbB was located at the tip of growing metulae, following a similar pattern as described in vegetative hyphae. In wild-type strains, the transition from metulae to phialides could be reversed to generate vegetative hyphae, indicating the existence of a specific control point at this stage of conidiophore formation. The combined evidence points to FlbB as a key factor in the transition to asexual development, playing a role at various control points in which the process could be reversed, Top of pageDiscussionEduardo A. Espeso wishes to thank Ministerio de Educación y Ciencia for support through Grant BFU2006-04185. Unai Ugalde wishes to thank Ministerio de Educación y Ciencia for Grant BFU2004-03499/BMC, and University of The Basque Country (UPV/EHU) for Grant GIU08/32. Oier Etxebeste is a contract researcher of the CIC Nanogune. Erika Herrero-García was a contract researcher under the research programme of Gipuzkoako Foru Aldundia/Diputación Foral de Gipuzkoa. Aitor Garzia and Ana B. Rodríguez-Urra held predoctoral fellowships from Basque Government and UPV/EHU respectively. Lidia Araújo-Bazán held a predoctoral fellowship from the Ministerio de Educación y Ciencia

A procedure for the production of plasma membrane vesicles, consisting of enzyme-assisted physical homogenization, differential centrifugation and partitioning in an aqueous polymer two phase system resulted in a fraction containing highly purified plasma membranes, as determined by vanadate-inhibited H + ATPase and near absence of intracellular membrane marker activities. H + ATPase latency tests indicated that at least 90% of the vesicles were oriented with the apoplastic surface facing outward (right-side-out orientation) in the upper phase, while inside out vesicles and other microsomal membrane material collected in the lower phase.

Summary: A new simple procedure for the production of protoplasts of Penicillium cyclopium with high regeneration rates and efficient transport activity is described, involving the use of a preparation of Novozym 234 with very low protease activity. The combination of a heat pretreatment at 55 °C for 15 min with the use of the protease inhibitor aprotinin resulted in a 97% reduction of Novozym 234 protease activity with respect to untreated controls. Polysaccharide-hydrolysing activity was inhibited much less, to 60% of the untreated Novozym 234 level. Protoplasts could be successfully produced with the new low-protease Novozym 234 preparation, showing a threefold increase in regeneration capacity compared to control protoplasts obtained with the original preparation. The rates of 3-O-methylglucose uptake and the capacity to accumulate this sugar analogue were also higher in protoplasts obtained by the new method.

11 páginas, 8 figuras, 3 tablas -- PAGS nros. 38-48, The fungal colony is a complex multicellular unit consisting of various cell types and functions. Asexual spore formation (conidiation) is integrated through sensory and regulatory elements into the general morphogenetic plan, in which the activation of the transcription factor BrlA is the first determining step. A number of early regulatory elements acting upstream of BrlA (fluG and flbA-E) have been identified, but their functional relations remain to be further investigated. In this report we describe FlbB as a putative basic-zipper-type transcription factor restricted to filamentous fungi. FlbB accumulates at the hyphal apex during early vegetative growth but is later found in apical nuclei, suggesting that an activating modification triggers nuclear import. Moreover, proper temporal and quantitative expression of FlbB is a prerequisite for brlA transcription, and misscheduled overexpression inhibits conidiation. We also present evidence that FlbB activation results in the production of a second diffusible signal, acting downstream from the FluG factor, to induce conidiation, The work carried out at UW-Madison was supported by Hatch (WIS04667) and National Science Foundation (MCB-0421863 and IOS-0640067) grants to J.H.Y. E.A.E thanks Ministerio de Educación y Ciencia for support through grant BFU2006-04185. U.U. thanks Ministerio de Educación y Ciencia for grant BFU2004-03499/BMC and UPV/EHU for grant GIU05/36. O.E received a doctoral grant from UPV/EHU, and A.G. was a contract researcher under the research program of Gipuzkoako Foru Aldundia/Diputación Foral de Gipuzkoa. The work was also supported by the Max-Planck-Institute for Terrestrial Microbiology, Marburg, Germany

Asexual spores or conidia are dispersive propagules produced as an alternative to vegetative growth by a diverse group of filamentous fungi. The cellular development programmes which govern conidiation have been intensely studied in the last few decades, although important gaps stand in the way of our understanding of this phenomenon, namely in the areas of the environmental sensing mechanisms and signal transduction pathways. The aim of this review is to summarize the current advances in conidiation induction in the genus Penicillium, and to put them into context with the state of our knowledge stemming from work in widely studied fungal model systems.

Publisher Summary This chapter provides an overview of the key aspects of fungal single cell protein (SCP) production, consumption and marketing, presents the most up to date developments of this interdisciplinary field, and discusses the future prospects of fungal SCP. Substitution of animal products by foods of plant origin is not totally exempt from these dangers, as the use of pesticides or the appearance of carcinogens, such as nitrosamines have already been reported in the past. SCP products for animal and human nutrition are safer in this respect, since the components from which, they are produced are easily controlled, and their genetic background is well known. They represent a line of non-conventional substitutes, which will continue to have a market for these reasons. The continued research on the production of microorganisms for animal and human consumption will undoubtedly find application in the future. This research should also incorporate the development of recombinant strains from nonconventional GRAS (Generally Regarded as a Safe) yeasts and fungi.

A plasma membrane fraction was obtained by the combined use of differential centrifugation and aqueous polymer two-phase partitioning techniques. Vanadate-inhibited ATPase and glucan synthase activities were highly enriched in this fraction, although the presence of ATPase activity which was not inhibited by vanadate, nitrate, molybdate, anyimycin A or azide was also detected. Other intracellular membrane marker activities were present at very low or undetectable levels. A further separation step using Percoll density gradient centrifugation resulted in the separation of a fraction which exclusively contained vanadate-inhibited ATPase activity, and was enriched with silicotungstic-acid-staining membrane material. Latency tests performed on the plasma membrane markers showed that the membrane vesicles were in the right-side-out orientation.

The conversion of whey to single-cell protein by yeasts was investigated. The most suitable organism tested wasKluyveromycesmarxianus NCYC 1424. The efficiency of whey conversion to biomass was directly related to higher oxygen availability in the medium.