Mycelium reinforced agricultural fiber bio-composites: Summary of research

USDA-ARS?s Scientific Manuscript database

Industry and the public sector have a growing interest in utilizing natural fibers, such as agricultural substrates, in the manufacture of components and products currently manufactured from fossil fuels. A patented process, developed by Ecovative Design, LLC (Ecovative), for growing fungal species ...

Long-Distance Translocation of Protein during Morphogenesis of the Fruiting Body in the Filamentous Fungus, Agaricus bisporus

PubMed Central

Woolston, Benjamin M.; Schlagnhaufer, Carl; Wilkinson, Jack; Larsen, Jeffrey; Shi, Zhixin; Mayer, Kimberly M.; Walters, Donald S.; Curtis, Wayne R.; Romaine, C. Peter

2011-01-01

Commercial cultivation of the mushroom fungus, Agaricus bisporus, utilizes a substrate consisting of a lower layer of compost and upper layer of peat. Typically, the two layers are seeded with individual mycelial inoculants representing a single genotype of A. bisporus. Studies aimed at examining the potential of this fungal species as a heterologous protein expression system have revealed unexpected contributions of the mycelial inoculants in the morphogenesis of the fruiting body. These contributions were elucidated using a dual-inoculant method whereby the two layers were differientially inoculated with transgenic β-glucuronidase (GUS) and wild-type (WT) lines. Surprisingly, use of a transgenic GUS line in the lower substrate and a WT line in the upper substrate yielded fruiting bodies expressing GUS activity while lacking the GUS transgene. Results of PCR and RT-PCR analyses for the GUS transgene and RNA transcript, respectively, suggested translocation of the GUS protein from the transgenic mycelium colonizing the lower layer into the fruiting body that developed exclusively from WT mycelium colonizing the upper layer. Effective translocation of the GUS protein depended on the use of a transgenic line in the lower layer in which the GUS gene was controlled by a vegetative mycelium-active promoter (laccase 2 and β-actin), rather than a fruiting body-active promoter (hydrophobin A). GUS-expressing fruiting bodies lacking the GUS gene had a bonafide WT genotype, confirmed by the absence of stably inherited GUS and hygromycin phosphotransferase selectable marker activities in their derived basidiospores and mycelial tissue cultures. Differientially inoculating the two substrate layers with individual lines carrying the GUS gene controlled by different tissue-preferred promoters resulted in up to a ∼3.5-fold increase in GUS activity over that obtained with a single inoculant. Our findings support the existence of a previously undescribed phenomenon of long-distance protein translocation in A. bisporus that has potential application in recombinant protein expression and biotechnological approaches for crop improvement. PMID:22163014

Actinophytocola glycyrrhizae sp. nov. isolated from the rhizosphere of Glycyrrhiza inflata.

PubMed

Cao, Chengliang; Sun, Yong; Wu, Bo; Zhao, Shuai; Yuan, Bo; Qin, Sheng; Jiang, Jihong; Huang, Ying

2018-06-25

A Gram-stain-positive, aerobic actinomycete, designated strain BMP B8152 T , was isolated from the rhizosphere of Glycyrrhiza inflata collected ashore, in Kashi, Xinjiang province, northwest PR China. A polyphasic approach was used to establish the taxonomic position of this strain. BMP B8152 T was observed to form non-fragmented substrate mycelium, and relatively scanty aerial mycelium with rod-shaped spores. Cell-wall hydrolysates contained meso-diaminopimelic acid, galactose, arabinose, glucose and rhamnose (trace). Mycolic acids were not detected. The diagnostic phospholipids were identified as diphosphatidylglycerol, phosphatidylethanolamine, hydroxyphosphatidylethanolamine, ninhydrin-positive phosphoglycolipid and phosphatidylinositol. The predominant menaquinone and fatty acid were MK-9(H4) and iso-branched hexadecanoate (iso-C16 : 0), respectively. The phylogenetic analyses based on the 16S rRNA gene sequences indicated that BMP B8152 T formed a distinct monophyletic clade clustered with Actinophytocola timorensisID05-A0653 T (98.8 % 16S rRNA gene sequence similarity), Actinophytocola oryzaeGMKU 367 T (98.6 %), Actinophytocola corallinaID06-A0464 T (98.2 %) and Actinophytocola burenkhanensisMN08-A0203 T (97.5 %). In addition, DNA-DNA hybridization values between BMP B8152 T and A. timorensisID05-A0653 T (44.2±3.6 %) and A. oryzaeGMKU 367 T (36.7±2.3 %) were well below the 70 % limit for species identification. The combined phenotypic and genotypic data indicate that the isolate represents a novel species of the genus Actinophytocola, for which the name Actinophytocola glycyrrhizae sp. nov., is proposed, with the type strain BMP B8152 T (=KCTC 49002 T =CGMCC 4.7433 T ).

Composted versus raw olive mill waste as substrates for the production of medicinal mushrooms: an assessment of selected cultivation and quality parameters.

PubMed

Zervakis, Georgios I; Koutrotsios, Georgios; Katsaris, Panagiotis

2013-01-01

Two-phase olive mill waste (TPOMW, "alperujo") is a highly biotoxic sludge-like effluent of the olive-oil milling process with a huge seasonal production. One of the treatment approaches that has so far received little attention is the use of TPOMW as substrate for the cultivation of edible mushrooms. Fifteen fungal strains belonging to five species (Basidiomycota), that is, Agrocybe cylindracea, Pleurotus cystidiosus, P. eryngii, P. ostreatus, and P. pulmonarius, were evaluated for their efficacy to colonize media composed of TPOMW, which was used either raw or composted in mixtures with wheat straw in various ratios. Qualified strains exhibited high values of biological efficiency (e.g., 120-135% for Pleurotus spp. and 125% for A. cylindracea) and productivity in subsequent cultivation experiments on substrates supplemented with 20-40% composted TPOMW or 20% raw TPOMW. Only when supplementation exceeded 60% for raw TPOMW, a negative impact was noted on mushroom yields which could be attributed to the effluent's toxicity (otherwise alleviated in the respective composted TPOMW medium). Earliness and mushroom size as well as quality parameters such as total phenolic content and antioxidant activity did not demonstrate significant differences versus the control wheat-straw substrate. The substrates hemicellulose content was negatively correlated with mycelium growth rates and yields and positively with earliness; in addition, cellulose: lignin ratio presented a positive correlation with mycelium growth and mushroom weight for A. cylindracea and with earliness for all species examined. TPOMW-based media revealed a great potential for the substitution of traditional cultivation substrates by valorizing environmentally hazardous agricultural waste.

Complete genome sequence of Intrasporangium calvum type strain (7 KIPT)

PubMed Central

Del Rio, Tijana Glavina; Chertkov, Olga; Yasawong, Montri; Lucas, Susan; Deshpande, Shweta; Cheng, Jan-Fang; Detter, Chris; Tapia, Roxanne; Han, Cliff; Goodwin, Lynne; Pitluck, Sam; Liolios, Konstantinos; Ivanova, Natalia; Mavromatis, Konstantinos; Pati, Amrita; Chen, Amy; Palaniappan, Krishna; Land, Miriam; Hauser, Loren; Chang, Yun-Juan; Jeffries, Cynthia D.; Rohde, Manfred; Pukall, Rüdiger; Sikorski, Johannes; Göker, Markus; Woyke, Tanja; Bristow, James; Eisen, Jonathan A.; Markowitz, Victor; Hugenholtz, Philip; Kyrpides, Nikos C.; Klenk, Hans-Peter; Lapidus, Alla

2010-01-01

Intrasporangium calvum Kalakoutskii et al. 1967 is the type species of the genus Intrasporangium, which belongs to the actinobacterial family Intrasporangiaceae. The species is a Gram-positive bacterium that forms a branching mycelium, which tends to break into irregular fragments. The mycelium of this strain may bear intercalary vesicles but does not contain spores. The strain described in this study is an airborne organism that was isolated from a school dining room in 1967. One particularly interesting feature of I. calvum is that the type of its menaquinone is different from all other representatives of the family Intrasporangiaceae. This is the first completed genome sequence from a member of the genus Intrasporangium and also the first sequence from the family Intrasporangiaceae. The 4,024,382 bp long genome with its 3,653 protein-coding and 57 RNA genes is a part of the Genomic Encyclopedia of Bacteria and Archaea project. PMID:21304734

[Effect of organic and inorganic toxic compounds on luminescence of luminous fungi].

PubMed

Vydriakova, G A; Gusev, A A; Medvedeva, S E

2011-01-01

The possibility of the development of the solid phase bioluminescent biotest using aerial mycelium of the luminous fungi was investigated. Effect of organic and inorganic toxic compounds (TC) at concentrations from 10(-6) to 1 mg/ml on luminescence of aerial mycelia of four species of luminous fungi-Armillaria borealis (Culture Collection of the Institute of Forest, Siberian Branch, Russian Academy of Sciences), A. mellea, A. gallica, and Lampteromyces japonicus (Fungi Collection of the Botanical Institute, Russian Academy of Sciences)--has been studied. Culture of A. mellea was shown to be most sensitive to solutions of the model TC. It was demonstrated that the sensitivity of the luminous fungi is comparable with the sensitivity of the bacteria that are used for environmental monitoring. Use of the aerial mycelium of the luminous fungi on the solid support as a test object is a promising approach in biotesting for the development of continuous biosensors for air monitoring.

Lipase of Aspergillus niger NCIM 1207: A Potential Biocatalyst for Synthesis of Isoamyl Acetate.

PubMed

Mhetras, Nutan; Patil, Sonal; Gokhale, Digambar

2010-10-01

Commercial lipase preparations and mycelium bound lipase from Aspergillus niger NCIM 1207 were used for esterification of acetic acid with isoamyl alcohol to obtain isoamyl acetate. The esterification reaction was carried out at 30°C in n-hexane with shaking at 120 rpm. Initial reaction rates, conversion efficiency and isoamyl acetate concentration obtained using Novozyme 435 were the highest. Mycelium bound lipase of A. niger NCIM 1207 produced maximal isoamyl acetate formation at an alcohol/acid ratio of 1.6. Acetic acid at higher concentrations than required for the critical alcohol/acid ratio lower than 1.3 and higher than 1.6 resulted in decreased yields of isoamyl acetate probably owing to lowering of micro-aqueous environmental pH around the enzyme leading to inhibition of enzyme activity. Mycelium bound A. niger lipase produced 80 g/l of isoamyl acetate within 96 h even though extremely less amount of enzyme activity was used for esterification. The presence of sodium sulphate during esterification reaction at higher substrate concentration resulted in increased conversion efficiency when we used mycelium bound enzyme preparations of A. niger NCIM 1207. This could be due to removal of excess water released during esterification reaction by sodium sulphate. High ester concentration (286.5 g/l) and conversion (73.5%) were obtained within 24 h using Novozyme 435 under these conditions.

Submerged Cultivation of Pleurotus sapidus with Molasses: Aroma Dilution Analyses by Means of Solid Phase Microextraction and Stir Bar Sorptive Extraction.

PubMed

Trapp, Tobias; Zajul, Martina; Ahlborn, Jenny; Stephan, Alexander; Zorn, Holger; Fraatz, Marco Alexander

2018-03-14

The basidiomycete Pleurotus sapidus (PSA) was grown in submerged cultures with molasses as substrate for the production of mycelium as a protein source for food applications. The volatilomes of the substrate, the submerged culture, and the mycelia were analyzed by gas chromatography-tandem mass spectrometry-olfactometry. For compound identification, aroma dilution analyses by means of headspace solid phase microextraction and stir bar sorptive extraction were performed via variation of the split vent flow rate. Among the most potent odorants formed by PSA were arylic compounds (e.g., p-anisaldehyde), unsaturated carbonyls (e.g., 1-octen-3-one, ( E)-2-octenal, ( E, E)-2,4-decadienal), and cyclic monoterpenoids (e.g., 3,9-epoxy- p-menth-1-ene, 3,6-dimethyl-3a,4,5,7a-tetrahydro-1-benzofuran-2(3 H)-one). Several compounds from the latter group were described for the first time in Pleurotus spp. After separation of the mycelia from the medium, the aroma compounds were mainly enriched in the culture supernatant. The sensory analysis of the mycelium correlated well with the instrumental results.

An unstructured mathematical model for growth of Pleurotus ostreatus on lignocellulosic material in solid-state fermentation systems

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sarikaya, A.; Ladisch, M.R.

1997-01-01

Inedible plant material, generated in a Controlled Ecological Life Support System (CELSS), should be recycled preferably by bioregenerative methods that utilize enzymes or micro-organisms. This material consists of hemicellulose, cellulose, and lignin with the lignin fraction representing a recalcitrant component that is not readily treated by enzymatic methods. Consequently, the white-rot fungus, Pleurotus ostreatus, is attractive since it effectively degrades lignin and produces edible mushrooms. This work describes an unstructured model for the growth of P. ostreatus in a solid-state fermentation system using lignocellulosic plant materials from Brassica napus (rapeseed) as a substrate at three different particle sizes. A logisticmore » function model based on area was found to fit the surface growth of the mycelium on the solid substrate with respect to time, whereas a model based on diameter, alone, did not fit the data as well. The difference between the two measures of growth was also evident for mycelial growth in a bioreactor designed to facilitate a slow flowrate of air through the 1.5 cm thick mat of lignocellulosic biomass particles. The result is consistent with the concept of competition of the mycelium for the substrate that surrounds it, rather than just substrate that is immediately available to single cells. This approach provides a quantitative measure of P. ostreatus growth on lignocellulosic biomass in a solid-state fermentation system. The experimental data show that the best growth is obtained for the largest particles (1 cm) of the lignocellulosic substrate. 13 refs., 6 figs., 2 tabs.« less

Composted versus Raw Olive Mill Waste as Substrates for the Production of Medicinal Mushrooms: An Assessment of Selected Cultivation and Quality Parameters

PubMed Central

Zervakis, Georgios I.; Koutrotsios, Georgios; Katsaris, Panagiotis

2013-01-01

Two-phase olive mill waste (TPOMW, “alperujo”) is a highly biotoxic sludge-like effluent of the olive-oil milling process with a huge seasonal production. One of the treatment approaches that has so far received little attention is the use of TPOMW as substrate for the cultivation of edible mushrooms. Fifteen fungal strains belonging to five species (Basidiomycota), that is, Agrocybe cylindracea, Pleurotus cystidiosus, P. eryngii, P. ostreatus, and P. pulmonarius, were evaluated for their efficacy to colonize media composed of TPOMW, which was used either raw or composted in mixtures with wheat straw in various ratios. Qualified strains exhibited high values of biological efficiency (e.g., 120–135% for Pleurotus spp. and 125% for A. cylindracea) and productivity in subsequent cultivation experiments on substrates supplemented with 20–40% composted TPOMW or 20% raw TPOMW. Only when supplementation exceeded 60% for raw TPOMW, a negative impact was noted on mushroom yields which could be attributed to the effluent's toxicity (otherwise alleviated in the respective composted TPOMW medium). Earliness and mushroom size as well as quality parameters such as total phenolic content and antioxidant activity did not demonstrate significant differences versus the control wheat-straw substrate. The substrates hemicellulose content was negatively correlated with mycelium growth rates and yields and positively with earliness; in addition, cellulose: lignin ratio presented a positive correlation with mycelium growth and mushroom weight for A. cylindracea and with earliness for all species examined. TPOMW-based media revealed a great potential for the substitution of traditional cultivation substrates by valorizing environmentally hazardous agricultural waste. PMID:24027758

Extraradical mycelium of arbuscular mycorrhizal fungi radiating from large plants depresses the growth of nearby seedlings in a nutrient deficient substrate.

PubMed

Janoušková, Martina; Rydlová, Jana; Püschel, David; Száková, Jiřina; Vosátka, Miroslav

2011-10-01

The effect of arbuscular mycorrhiza (AM) on the interaction of large plants and seedlings in an early succession situation was investigated in a greenhouse experiment using compartmented rhizoboxes. Tripleurospermum inodorum, a highly mycorrhiza-responsive early coloniser of spoil banks, was cultivated either non-mycorrhizal or inoculated with AM fungi in the central compartment of the rhizoboxes. After two months, seedlings of T. inodorum or Sisymbrium loeselii, a non-host species colonising spoil banks simultaneously with T. inodorum, were planted in lateral compartments, which were colonised by the extraradical mycelium (ERM) of the pre-cultivated T. inodorum in the inoculated treatments. The experiment comprised the comparison of two AM fungal isolates and two substrates: spoil bank soil and a mixture of this soil with sand. As expected based on the low nutrient levels in the substrates, the pre-cultivated T. inodorum plants responded positively to mycorrhiza, the response being more pronounced in phosphorus uptake than in nitrogen uptake and growth. In contrast, the growth of the seedlings, both the host and the non-host species, was inhibited in the mycorrhizal treatments. Based on the phosphorus and nitrogen concentrations in the biomass of the experimental plants, this growth inhibition was attributed to nitrogen depletion in the lateral compartments by the ERM radiating from the central compartment. The results point to an important aspect of mycorrhizal effects on the coexistence of large plants and seedlings in nutrient deficient substrates. © Springer-Verlag 2011

Potential of Pleurotus ostreatus Mycelium for Selenium Absorption

PubMed Central

Milovanović, Ivan; Brčeski, Ilija; Stajić, Mirjana; Korać, Aleksandra; Vukojević, Jelena; Knežević, Aleksandar

2014-01-01

The aim of this study was to evaluate the effect of high selenium (Se) concentrations on morphophysiological and ultrastructural properties of Pleurotus ostreatus. Mycelium growth was good in media enriched with 5.0, 10.0, and 20.0 mg L−1 of Se, concentration of 500.0 mg L−1 strongly inhibited growth, and 1000.0 mg L−1 was the minimum inhibitory concentration. Contrary to thin-walled, hyaline, branched, and anastomized hyphae with clamp-connections in the control, at Se concentrations of 100.0 and 500.0 mg L−1, they were noticeably short, frequently septed and branched, with a more intensive extracellular matrix, and without clamp-connections. At high Se concentrations, hyphae with intact membrane, without cellular contents, with a high level of vacuolization, and with numerous proteinaceous bodies were observed. Biomass yield ranged between 11.8 g L−1, in the control, and 6.8 g L−1, at an Se concentration of 100.0 mg L−1, while no production was detected at a concentration of 500.0 mg L−1. Se content in the mycelia reached a peak (938.9 μg g−1) after cultivation in the medium enriched with Se at the concentration of 20.0 mg L−1, while the highest absorption level (53.25%) was found in the medium enriched with 5.0 mg L−1 Se. PMID:25003145

The Course of Colonization of Two Different Vitis Genotypes by Plasmopara viticola Indicates Compatible and Incompatible Host-Pathogen Interactions.

PubMed

Unger, Sabine; Büche, Claudia; Boso, Susana; Kassemeyer, Hanns-Heinz

2007-07-01

ABSTRACT The course of colonization of leaf mesophyll by the causal agent of grapevine downy mildew, Plasmopara viticola, in a susceptible and a resistant grapevine genotype was examined in order to characterize the development of the pathogen in compatible and incompatible host-pathogen interactions. Within a few hours after inoculation, the pathogen was established in the susceptible Vitis vinifera cv. Müller-Thurgau and formed primary hyphae with a first haustorium. No further development occurred in the following 10 to 18 h. The next step, in which the hyphae grew and branched to colonize the intercellular space of the host tissue, was observed 1.5 days after inoculation. After 3 days, the intercostal fields were entirely filled with mycelium and sporulation was abundant under favorable environmental conditions. The first infection steps were essentially the same in the resistant V. rupestris. However, the invasive growth of P. viticola was delayed, and further development ceased before the intercostal fields were filled with mycelium.

Structural characterization of the cell wall D-glucans isolated from the mycelium of Botryosphaeria rhodina MAMB-05.

PubMed

de Lourdes Corradi da Silva, Maria; Fukuda, Eliane K; Vasconcelos, Ana Flora D; Dekker, Robert F H; Matias, Andreza C; Monteiro, Nilson K; Cardoso, Marilsa S; Barbosa, Aneli M; Silveira, Joana L M; Sassaki, Guilherme L; Carbonero, Elaine R

2008-03-17

Three D-glucans were isolated from the mycelium of the fungus Botryosphaeria rhodina MAMB-05 by sequential extraction with hot-water and hot aqueous KOH (2% w/v) followed by ethanol precipitation. Following their purification by gel permeation chromatography on Sepharose CL-4B, the structural characteristics of the D-glucans were determined by FT-IR and 13C NMR spectroscopy and, after methylation, by GC-MS. The hot-water extract produced a fraction designated Q1A that was a beta-(1-->6)-D-glucan with the following structure: [Formula: see text] The alkaline extract, when subjected to repeated freeze-thawing, yielded two fractions: K1P (insoluble) that comprised a beta-(1-->3)-D-glucan with beta-D-glucose branches at C-6 with the structure: [Formula: see text] and K1SA (soluble) consisting of a backbone chain of alpha-(1-->4)-linked D-glucopyranosyl residues substituted at O-6 with alpha-D-glucopyranosyl residues: [Formula: see text

On-target labeling of intracellular metabolites combined with chemical mapping of individual hyphae revealing cytoplasmic relocation of isotopologues.

PubMed

Hu, Jie-Bi; Chen, Yu-Chie; Urban, Pawel L

2012-06-05

A microscale analytical platform integrating microbial cell culture, isotopic labeling, along with visual and mass spectrometric imaging with single-cell resolution has been developed and applied in the monitoring of cellular metabolism in fungal mycelium. The method implements open chips with a two-dimensional surface pattern composed of hydrophobic and hydrophilic zones. Two hydrophilic islands are used as medium reservoirs, while the hydrophobic area constitutes the support for the growing aerial hyphae, which do not have direct contact with the medium. The first island, containing (12)C(6)-glucose medium, was initially inoculated with the mycelium (Neurospora crassa), and following the initial incubation period, the hyphae progressed toward the second medium island, containing an isotopically labeled substrate ((13)C(6)-glucose). The (13)C atoms were gradually incorporated into cellular metabolites, which was revealed by MALDI-MS. The fate of the chitin-biosynthesis precursor, uridine diphosphate N-acetylglucosamine (UDP-GlcNAc), was monitored by recording mass spectra with characteristic isotopic patterns, which indicated the presence of various (12)C/(13)C isotopologues. The method enabled mapping the (13)C-labeled UDP-GlcNAc in fungal mycelium and recording its redistribution in hyphae, directly on the chip.

Radiosensitivity study and radiation effects on morphology characterization of grey oyster mushroom Pleurotus sajor-caju

NASA Astrophysics Data System (ADS)

Rashid, Rosnani Abdul; Daud, Fauzi; Senafi, Sahidan; Awang, Mat Rasol; Mohamad, Azhar; Mutaat, Hassan Hamdani; Maskom, Mohd Meswan

2014-09-01

Radiosensitive dosage and morphology characterization of irradiated grey oyster mushroom Pleurotus sajor-caju by gamma rays was investigated due to effects of irradiation. In order to establish the effect, mycelium of P. sajor-caju was irradiated by gamma rays at dose 0.1 to 8.0 kGy with dose rate 0.227 Gy sec-1. The irradiation of mycelia was carried out at the radiation facility in Malaysian Nuclear Agency. The radiosensitivity study was performed by evaluating the percentage of survival irradiated mycelia. The lethal dose of the mycelium P. sajor-caju was determined at 4.0 kGy and LD50 to be equal at 2.2 kGy. The radiation effects on morphology were evaluated based on growth rate of irradiated mycelia, mycelia types, colonization period on substrate, morphology of fruit bodies and yields. The results shown growth rate of irradiated mycelium was slightly lower than the control and decreased as the dose increased. Irradiation was found can induced the primordia formation on PDA and the BE of irradiated seed is higher than to control. The irradiation is proven to be useful for generating new varieties of mushroom with commercial value to the industry.

Growth and development of Frankia spp. strain CcI3 at the single-hypha level in liquid culture.

PubMed

Huang, Ying; Benson, David R

2012-01-01

Filamentous actinobacteria from the genus Frankia grow by hyphal tip extension and branching. The growth kinetics and branching pattern of Frankia are not well studied, especially at the early stages of mycelial development. Here, we compare the growth of Frankia sp. strain CcI3 in liquid cultures with and without proteose peptone #3 (PP3) using time-lapse photomicrography and image analysis. Individual hyphae showed a pseudolinear increase in length at early stages of development, whereas at the mycelial level, the aggregate length of hyphae described an exponential rate before slowing. Growth based on optical density or microscopic observations was similar in medium with or without PP3. However, PP3 altered the pattern of mycelial development by increasing branching. Distances between the hyphal apex and first branches were on average shorter in PP3-containing media. The final interbranch distances were also shorter in PP3 medium indicating that hyphae tended to branch earlier and more often when supplemented with PP3 to give a more compact mycelium. Vesicle development in nitrogen-fixing cultures limited cell expansion as a result of vesicles truncating growth on new branches. The results provide some explanation for the growth kinetics of Frankia and some indication of how growth rates may be improved.

[Effect of the steroid molecule structure on the direction of its hydroxylation by the fungus Curvularia lunata].

PubMed

Andriushina, V A; Iaderets, V V; Stytsenko, T S; Druzhinina, A V; Voĭshvillo, N E

2013-01-01

The main and side products of hydroxylation by the C. lunata VKPM F-981 mycelium of fourteen delta(4)-3-ketosteroids of the estrane, androstane, and pregnane series and six of their delta(5)-3beta-hydroxy analogues were identified by H1 PMR spectroscopy and comparison with standard samples. The obtained experimental data are considered in terms of the triangular model of the enzyme-substrate interaction. The dependence of the direction of hydroxylation of steroid molecules and the orientation of hydroxy groups on the structure of the initial substrate was revealed.

Get a Grip: Substrate Orientation and Digital Grasping Pressures in Strepsirrhines.

PubMed

Congdon, Kimberly A; Ravosa, Matthew J

2016-01-01

Skeletal functional morphology in primates underlies many fossil interpretations. Understanding the functional correlates of arboreal grasping is central to identifying locomotor signatures in extinct primates. We tested 3 predictions linking substrate orientation and digital grasping pressures: (1) below-branch pressures are greater than above-branch and vertical-branch pressures; (2) there is no difference in pressure exerted across digits within autopods at any substrate orientation, and (3) there is no difference in pressure exerted between homologous digits across autopods at any substrate orientation. Adult males and females from 3 strepsirrhine species crossed an artificial arboreal substrate oriented for above-, below- and vertical-branch locomotion. We compared digital pressures within and across behaviors via ANOVA and Tukey's Honest Significant Difference test. Results show limited support for all predictions: below-branch pressures exceeded vertical-branch pressures and above-branch pressures for some digits and species (prediction 1), lateral digits often exerted greater pressures than medial digits (prediction 2), and pedal digits occasionally exerted greater pressures than manual digits during above-branch and vertical orientations but less often for below-branch locomotion (prediction 3). We observed functional variability across autopods, substrate and species that could underlie morphological variation within and across primates. Future work should consider the complexity of arboreality when inferring locomotor modes in fossils. © 2016 S. Karger AG, Basel.

Radiosensitivity study and radiation effects on morphology characterization of grey oyster mushroom Pleurotus sajor-caju

DOE Office of Scientific and Technical Information (OSTI.GOV)

Rashid, Rosnani Abdul; Awang, Mat Rasol; Mohamad, Azhar

2014-09-03

Radiosensitive dosage and morphology characterization of irradiated grey oyster mushroom Pleurotus sajor-caju by gamma rays was investigated due to effects of irradiation. In order to establish the effect, mycelium of P. sajor-caju was irradiated by gamma rays at dose 0.1 to 8.0 kGy with dose rate 0.227 Gy sec{sup −1}. The irradiation of mycelia was carried out at the radiation facility in Malaysian Nuclear Agency. The radiosensitivity study was performed by evaluating the percentage of survival irradiated mycelia. The lethal dose of the mycelium P. sajor-caju was determined at 4.0 kGy and LD{sub 50} to be equal at 2.2 kGy.more » The radiation effects on morphology were evaluated based on growth rate of irradiated mycelia, mycelia types, colonization period on substrate, morphology of fruit bodies and yields. The results shown growth rate of irradiated mycelium was slightly lower than the control and decreased as the dose increased. Irradiation was found can induced the primordia formation on PDA and the BE of irradiated seed is higher than to control. The irradiation is proven to be useful for generating new varieties of mushroom with commercial value to the industry.« less

Heat treatment of wheat straw by immersion in hot water decreases mushroom yield in Pleurotus ostreatus.

PubMed

Jaramillo Mejía, Santiago; Albertó, Edgardo

2013-01-01

The oyster mushroom, Pleurotus ostreatus, is cultivated worldwide. It is one of the most appreciated mushrooms due to its high nutritional value. Immersion of the substrate in hot water is one of the most popular and worldwide treatment used for mushroom farmers. It is cheap and easy to implement. To compare the yields obtained during mushroom production of P. ostreatus using different pre-treatments (immersion in hot water, sterilization by steam and the use of fungicide) to determine if they influence mushroom crop. Four different treatments of substrate (wheat straw) were carried out: (i) immersion in hot water (IHW); (ii) steam sterilization; (iii) chemical; and (iv) untreated. The residual water from the IHW treatment was used to evaluate the mycelium growth and the production of P. ostreatus. Carbendazim treatment produced highest yields (BE: 106.93%) while IHW produced the lowest BE with 75.83%. Sugars, N, P, K and Ca were found in residual water of IHW treatment. The residual water increased the mycelium growth but did not increase yields. We have proved that IHW treatment of substrate reduced yields at least 20% when compared with other straw treatments such as steam, chemical or untreated wheat straw. Nutrients like sugars, proteins and minerals were found in the residual water extract which is the resultant water where the immersion treatment is carried out. The loss of these nutrients would be the cause of yield decrease. Alternative methods to the use of IHW as treatment of the substrate should be considered to reduce economical loss. Copyright © 2012 Revista Iberoamericana de Micología. Published by Elsevier Espana. All rights reserved.

Nonomuraea glycinis sp. nov., a novel actinomycete isolated from the root of black soya bean [Glycine max (L.) Merr].

PubMed

Li, Zhilei; Song, Wei; Zhao, Junwei; Zhuang, Xiaoxin; Zhao, Yue; Wang, Xiangjing; Xiang, Wensheng

2017-12-01

A novel actinomycete, designated strain NEAU-BB2C19 T , was isolated from the root of black soya bean [Glycine max (L.) Merr] collected from Harbin, Heilongjiang Province, China, and characterized using a polyphasic approach. The strain was an aerobic, Gram-stain-positive actinomycete that formed extensively branched substrate mycelium and aerial hyphae. The predominant menaquinones were MK-9(H2) and MK-9(H0). The major cellular fatty acid profile consisted of iso-C16 : 0, 10-methyl C17 : 0 and 10-methyl C18 : 0. The polar lipid profile consisted of diphosphatidylglycerol, phosphatidylethanolamine, hydroxy-phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositol mannoside, phosphatidylglycerol and glycolipid. The DNA G+C content was 68.2±0.4 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain NEAU-BB2C19 T should be assigned to the genus Nonomuraea and formed a distinct branch with its closest neighbour Nonomuraea guangzhouensis NEAU-ZJ3 T (98.75 % 16S rRNA gene sequence similarity). The morphological and chemotaxonomic properties of the strain were also consistent with those of members of the genus Nonomuraea. A combination of DNA-DNA hybridization results and some phenotypic characteristics indicated that strain NEAU-BB2C19 T could be clearly differentiated from its closest phylogenetic relative. Thus, the strain is concluded to represent a novel species of the genus Nonomuraea, for which the name Nonomuraea glycinis sp. nov. is proposed. The type strain is NEAU-BB2C19 T (=CGMCC 4.7430 T =DSM 104838 T ).

Potential uses of spent mushroom substrate and its associated lignocellulosic enzymes.

PubMed

Phan, Chia-Wei; Sabaratnam, Vikineswary

2012-11-01

Mushroom industries generate a virtually in-exhaustible supply of a co-product called spent mushroom substrate (SMS). This is the unutilised substrate and the mushroom mycelium left after harvesting of mushrooms. As the mushroom industry is steadily growing, the volume of SMS generated annually is increasing. In recent years, the mushroom industry has faced challenges in storing and disposing the SMS. The obvious solution is to explore new applications of SMS. There has been considerable discussion recently about the potentials of using SMS for production of value-added products. One of them is production of lignocellulosic enzymes such as laccase, xylanase, lignin peroxidase, cellulase and hemicellulase. This paper reviews scientific research and practical applications of SMS as a readily available and cheap source of enzymes for bioremediation, animal feed and energy feedstock.

Properties of Acetate Kinase Isozymes and a Branched-Chain Fatty Acid Kinase from a Spirochete

PubMed Central

Harwood, Caroline S.; Canale-Parola, Ercole

1982-01-01

Spirochete MA-2, which is anaerobic, ferments glucose, forming acetate as a major product. The spirochete also ferments (but does not utilize as growth substrates) small amounts of l-leucine, l-isoleucine, and l-valine, forming the branched-chain fatty acids isovalerate, 2-methylbutyrate, and isobutyrate, respectively, as end products. Energy generated through the fermentation of these amino acids is utilized to prolong cell survival under conditions of growth substrate starvation. A branched-chain fatty acid kinase and two acetate kinase isozymes were resolved from spirochete MA-2 cell extracts. Kinase activity was followed by measuring the formation of acyl phosphate from fatty acid and ATP. The branched-chain fatty acid kinase was active with isobutyrate, 2-methylbutyrate, isovalerate, butyrate, valerate, or propionate as a substrate but not with acetate as a substrate. The acetate kinase isozymes were active with acetate and propionate as substrates but not with longer-chain fatty acids as substrates. The acetate kinase isozymes and the branched-chain fatty acid kinase differed in nucleoside triphosphate and cation specificities. Each acetate kinase isozyme had an apparent molecular weight of approximately 125,000, whereas the branched-chain fatty acid kinase had a molecular weight of approximately 76,000. These results show that spirochete MA-2 synthesizes a branched-chain fatty acid kinase specific for leucine, isoleucine, and valine fermentation. It is likely that a phosphate branched-chain amino acids is also synthesized by spirochete MA-2. Thus, in spirochete MA-2, physiological mechanisms have evolved which serve specifically to generate maintenance energy from branched-chain amino acids. PMID:6288660

Allocatelliglobosispora scoriae gen. nov., sp. nov., isolated from volcanic ash.

PubMed

Lee, Dong Wan; Lee, Soon Dong

2011-02-01

A novel actinomycete, designated strain Sco-B14(T), was isolated from volcanic ash collected near Darangshi Oreum (a parasitic or satellite volcano) in Jeju, Republic of Korea. The organism formed well-developed, branched substrate mycelium, on which short chains of non-motile spores were arranged singly or in clusters. Aerial mycelium was not produced. Globose bodies were observed. The reverse colour of colonies was light brown to brown. Diffusible pigments were produced on ISP medium 3 and oatmeal-nitrate agar. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain Sco-B14(T) formed a lineage within the family Micromonosporaceae and was distinct from established genera. The 16S rRNA gene sequence similarity of strain Sco-B14(T) to members of related genera of the family was 95.0-95.7 % to type strains of Catellatospora species, 94.7 % to Hamadaea tsunoensis IMSNU 22005(T), 94.7 % to Longispora albida K97-0003(T) and 94.0 % to Catelliglobosispora koreensis LM 042(T). 3-Hydroxydiaminopimelic acid was the diagnostic diamino acid in the cell-wall peptidoglycan. Whole-cell sugars were glucose, rhamnose, ribose, xylose, arabinose, galactose and mannose. The polar lipids included diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylinositol. The menaquinone profile contained MK-10(H(4)) (49 %), MK-9(H(4)) (24 %), MK-10(H(6)) (18 %) and MK-9(H(6)) (9 %). The predominant fatty acids were iso-C(15 : 0) and C(17 : 0). The DNA G+C content was 70.1 mol%. The combination of chemotaxonomic and phylogenetic data clearly separated the isolate from the type strains of all genera in the family Micromonosporaceae. On the basis of the phylogenetic and chemotaxonomic data presented in this paper, strain Sco-B14(T) is considered to represent a novel species of a new genus in the family Micromonosporaceae, for which the name Allocatelliglobosispora scoriae gen. nov., sp. nov. is proposed. The type strain of Allocatelliglobosispora scoriae is Sco-B14(T) (=KCTC 19661(T) =DSM 45362(T)).

A polysaccharide from cultured mycelium of Hericium erinaceus and its anti-chronic atrophic gastritis activity.

PubMed

Wang, Mingxing; Gao, Yang; Xu, Duoduo; Gao, Qipin

2015-11-01

A polysaccharide named EP-1 was found by screening cultured mycelium of Hericium erinaceus, which was extracted and subjected to precipitation with ethanol, hollow-fiber ultrafiltration and ion-exchange chromatography. The polysaccharide has a molecular weight of approximately 3100Da and is composed of glucose, mannose and galactose, thus being a heteroglycan. EP-1 has a backbone of α-d-Glc(1→3) and β-d-Glc(1→3). The β-d-Glc(1→3) and α-d-Gal-(1→3) were regarded as branches attached to the C-4 position. The α-d-Man was regarded as a terminal residue. The anti-CAG activity was evaluated in experimental systems using a cell model for identification. The polysaccharide significantly inhibited the growth of MC cells obtained from human gastric mucosa epithelium (GES-1) cells transformed by MNNG, which were used as a chronic atrophic gastritis cell model. It also interfered with the MC cells by inducing cell cycle arrest. Thus, EP-1 shows potential for the development of new functional foods and drugs. Copyright © 2015. Published by Elsevier B.V.

Nonvolatile Taste Components and Antioxidant Properties of Fruiting Body and Mycelium with High Ergothioneine Content from the Culinary-Medicinal Golden Oyster Mushroom Pleurotus citrinopileatus (Agaricomycetes).

PubMed

Lin, Shin-Yi; Chien, Shih-Chang; Wang, Sheng-Yang; Mau, Jeng-Leun

2016-01-01

Pleurotus citrinopileatus mycelium was prepared with high ergothioneine (Hi-Ergo) content and its proximate composition, nonvolatile taste components, and antioxidant properties were studied. The ergothioneine contents of fruiting bodies and Hi-Ergo and regular mycelia were 3.89, 14.57, and 0.37 mg/g dry weight, respectively. Hi-Ergo mycelium contained more dietary fiber, soluble polysaccharides, and ash but less carbohydrates, reducing sugar, fiber, and fat than regular mycelium. However, Hi-Ergo mycelium contained the smallest amounts of total sugars and polyols (47.43 mg/g dry weight). In addition, Hi-Ergo mycelium showed the most intense umami taste. On the basis of the half-maximal effective concentration values obtained, the 70% ethanolic extract from Hi-Ergo mycelium showed the most effective antioxidant activity, reducing power, and scavenging ability, whereas the fruiting body showed the most effective antioxidant activity, chelating ability, and Trolox-equivalent antioxidant capacity. Overall, Hi-Ergo mycelium could be beneficially used as a food-flavoring material or as a nutritional supplement.

Seasonal dynamics of Boletus edulis and Lactarius deliciosus extraradical mycelium in pine forests of central Spain.

PubMed

De la Varga, Herminia; Águeda, Beatriz; Ágreda, Teresa; Martínez-Peña, Fernando; Parladé, Javier; Pera, Joan

2013-07-01

The annual belowground dynamics of extraradical soil mycelium and sporocarp production of two ectomycorrhizal fungi, Boletus edulis and Lactarius deliciosus, have been studied in two different pine forests (Pinar Grande and Pinares Llanos, respectively) in Soria (central Spain). Soil samples (five per plot) were taken monthly (from September 2009 to August 2010 in Pinar Grande and from September 2010 to September 2011 in Pinares Llanos) in eight permanent plots (four for each site). B. edulis and L. deliciosus extraradical soil mycelium was quantified by real-time polymerase chain reaction, with DNA extracted from soil samples, using specific primers and TaqMan® probes. The quantities of B. edulis soil mycelium did not differ significantly between plots, but there was a significant difference over time with a maximum in February (0.1576 mg mycelium/g soil) and a minimum in October (0.0170 mg mycelium/g soil). For L. deliciosus, significant differences were detected between plots and over time. The highest amount of mycelium was found in December (1.84 mg mycelium/g soil) and the minimum in February (0.0332 mg mycelium/g soil). B. edulis mycelium quantities were positively correlated with precipitation of the current month and negatively correlated with the mean temperature of the previous month. Mycelium biomass of L. deliciosus was positively correlated with relative humidity and negatively correlated with mean temperature and radiation. No significant correlation between productivity of the plots with the soil mycelium biomass was observed for any of the two species. No correlations were found between B. edulis sporocarp production and weather parameters. Sporocarp production of L. deliciosus was positively correlated with precipitation and relative humidity and negatively correlated with maximum and minimum temperatures. Both species have similar distribution over time, presenting an annual dynamics characterized by a seasonal variability, with a clear increase on the amounts of biomass during the coldest months of the year. Soil mycelial dynamics of both species are strongly dependent on the weather.

Mismatch repair factor MSH2-MSH3 binds and alters the conformation of branched DNA structures predicted to form during genetic recombination.

PubMed

Surtees, Jennifer A; Alani, Eric

2006-07-14

Genetic studies in Saccharomyces cerevisiae predict that the mismatch repair (MMR) factor MSH2-MSH3 binds and stabilizes branched recombination intermediates that form during single strand annealing and gene conversion. To test this model, we constructed a series of DNA substrates that are predicted to form during these recombination events. We show in an electrophoretic mobility shift assay that S. cerevisiae MSH2-MSH3 specifically binds branched DNA substrates containing 3' single-stranded DNA and that ATP stimulates its release from these substrates. Chemical footprinting analyses indicate that MSH2-MSH3 specifically binds at the double-strand/single-strand junction of branched substrates, alters its conformation and opens up the junction. Therefore, MSH2-MSH3 binding to its substrates creates a unique nucleoprotein structure that may signal downstream steps in repair that include interactions with MMR and nucleotide excision repair factors.

11β-Hydroxylation of Cortexolone (Reichstein Compound S) to Hydrocortisone by Curvularia lunata Entrapped in Photo-Cross-Linked Resin Gels

PubMed Central

Sonomoto, Kenji; Hoq, M. Mozammel; Tanaka, Atsuo; Fukui, Saburo

1983-01-01

Spores of Curvularia lunata were immobilized by entrapment with photo-cross-linkable resin prepolymers and incubated to form mycelium in potato dextrose broth containing cortexolone (Reichstein compound S) as an inducer of steroid 11β-hydroxylase. In a buffer system containing 2.5% dimethyl sulfoxide, this immobilized mycelium hydroxylated cortexolone to hydrocortisone. The activity of this mycelium was comparable to the activity of free mycelium. Dimethyl sulfoxide did not inhibit hydroxylase activity at the concentration used and was effective in dissolving the product. Of the various photo-cross-linkable resin prepolymers examined, use of ENT-4000, whose main chain was polyethylene glycol 4000 (chain length, approximately 40 nm), resulted in maximum hydroxylation activity of the entrapped mycelium. The chain length of prepolymers affected markedly mycelial growth in the gels and, subsequently, the activity of the entrapped mycelium. The immobilized hydroxylation system was more stable than the system in free mycelium and could be reactivated by incubation of the entrapped mycelium in potato dextrose broth containing cortexolone. The system was tested 50 times during 100 days of operation and was found to carry out the desired transformation with overall yields of 60%. Images PMID:16346194

Do spawn storage conditions influence the colonization capacity of a wheat-straw-based substrate by Agaricus subrufescens?

PubMed

Farnet, Anne-Marie; Qasemian, Leila; Peter-Valence, Frédérique; Ruaudel, Florence; Savoie, Jean-Michel; Roussos, Sevastianos; Gaime-Perraud, Isabelle; Ziarelli, Fabio; Ferré, Élisée

2014-01-01

Storage conditions of the spawn of edible fungi are of major importance to facilitate the production of mushrooms. Here, standard storage conditions at 10°C or 15°C were used and the potential of colonization of standard European compost by the tropical species Agaricus subrufescens was assessed during the spawn running phase. Two lignocellulolytic activities, laccase and CMC-cellulase, were enhanced after storage compared to control as well as substrate transformation, as described by the aromaticity ratio and a humification ratio calculated from NMR data. This result indicates that mycelium growth probably occurred during storage at 10 or 15°C, leading to a larger amount of biomass in the inoculum. Moreover, the microbial functional diversity of the substrate was favored, showing that the electivity of the substrate was maintained. Thus, these findings indicate that recommendations for the mushroom producers can be established for A. subrufescens cultivation under European standard conditions. Copyright © 2014. Published by Elsevier SAS.

The effect of some organic substances on the mycelium of the fungus Ustilago nuda (Jens.) Rostr.

PubMed

Krátká, J

1976-01-01

Research was performed for studying the effect of some organic compounds, considered by many authors as the products ob barley seed metabolism generated after anaerobic seed treatment, on the mycelium of the fungus Ustilago nuda (Jens.) Rostr. The author examined the effectiveness of ethylacohol, acetaldehyde, acetic acid, succinic acid, lactic acid, and hydroquinone in concentrations from 1 M to 10(-6) M, and the effectiveness of extracts from disinfected seeds in doses from 10 g to 0.001 g/l. The effect of the mentioned solutions was examined as exerted on the growth of dicaryotic mycelium and on the growth of the haploid promycelium of the fungus. The dicaryotic mycelium of Ustilago nuda (Jens.) Rostr. was cultivated on potato agar with benzoic acid. The presence of the acid prevents mitosis, and the chlamydospores germinate on the nutritive medium with two fibres having binuclear cells. The haploid promycelium was cultivated on potato agar; chlamydospores germinated with one four-cell fibre, and individual cells are mononuclear and haploid. Only later, a dicarytic mycelium is created in a complex process. In all the substances used, the concentration of 1 M was found to stop further growth of mycelium. The concentration of 10(-1) M of acetic acid and hydroquinone also stopped growth, the same concentration of acetaldehyde, lactic acid, succinic acid, ethylacohol stimulated mycelium growth in comparison with the control. The concentration of 10(-6) M stimulated mycelium growth in a majority of cases. Extracts from disinfected seeds did not influence mycelium growth significantly in all cases in comparison with the control. The results were similar in the two types of mycelium.

Degradation of isoproturon and bentazone in peat- and compost-based biomixtures.

PubMed

Coppola, Laura; Pilar Castillo, Maria Del; Vischetti, Costantino

2011-01-01

The composition and properties of a biomixture used in a biobed are decisive for pesticide sorption and degradation. This study was performed to investigate the capability of compost-based substrates in mixtures with citrus peel and vine branch straw and peat-based substrates in mixtures with soil and vine branch straw at different levels in order to degrade isoproturon and bentazone. Dissipation and mineralisation rates of both pesticides were determined, and metabolic activity was followed as respiration. Compost-based substrates showed faster pesticide dissipation in the presence of lignocellulosic materials, as in garden compost and vine branch straw. The increasing content of vine branch straw in peat-based substrates does not seem to affect dissipation of the parent compounds. Low mineralisation rate was observed in all treatments. Higher pesticide degradation was observed in the lignocellulosic substrates, probably because of the development of lignin-degrading microorganisms which have shown to be robust and are able to degrade recalcitrant pesticides. Copyright © 2010 Society of Chemical Industry. Copyright © 2010 Society of Chemical Industry.

A model for growth of a single fungal hypha based on well-mixed tanks in series: simulation of nutrient and vesicle transport in aerial reproductive hyphae.

PubMed

Balmant, Wellington; Sugai-Guérios, Maura Harumi; Coradin, Juliana Hey; Krieger, Nadia; Furigo Junior, Agenor; Mitchell, David Alexander

2015-01-01

Current models that describe the extension of fungal hyphae and development of a mycelium either do not describe the role of vesicles in hyphal extension or do not correctly describe the experimentally observed profile for distribution of vesicles along the hypha. The present work uses the n-tanks-in-series approach to develop a model for hyphal extension that describes the intracellular transport of nutrient to a sub-apical zone where vesicles are formed and then transported to the tip, where tip extension occurs. The model was calibrated using experimental data from the literature for the extension of reproductive aerial hyphae of three different fungi, and was able to describe different profiles involving acceleration and deceleration of the extension rate. A sensitivity analysis showed that the supply of nutrient to the sub-apical vesicle-producing zone is a key factor influencing the rate of extension of the hypha. Although this model was used to describe the extension of a single reproductive aerial hypha, the use of the n-tanks-in-series approach to representing the hypha means that the model has the flexibility to be extended to describe the growth of other types of hyphae and the branching of hyphae to form a complete mycelium.

The transcription repressor NmrA is subject to proteolysis by three Aspergillus nidulans proteases

PubMed Central

Zhao, Xiao; Hume, Samantha L; Johnson, Christopher; Thompson, Paul; Huang, Junyong; Gray, Joe; Lamb, Heather K; Hawkins, Alastair R

2010-01-01

The role of specific cleavage of transcription repressor proteins by proteases and how this may be related to the emerging theme of dinucleotides as cellular signaling molecules is poorly characterized. The transcription repressor NmrA of Aspergillus nidulans discriminates between oxidized and reduced dinucleotides, however, dinucleotide binding has no effect on its interaction with the zinc finger in the transcription activator AreA. Protease activity in A. nidulans was assayed using NmrA as the substrate, and was absent in mycelium grown under nitrogen sufficient conditions but abundant in mycelium starved of nitrogen. One of the proteases was purified and identified as the protein Q5BAR4 encoded by the gene AN2366.2. Fluorescence confocal microscopy showed that the nuclear levels of NmrA were reduced approximately 38% when mycelium was grown on nitrate compared to ammonium and absent when starved of nitrogen. Proteolysis of NmrA occurred in an ordered manner by preferential digestion within a C-terminal surface exposed loop and subsequent digestion at other sites. NmrA digested at the C-terminal site was unable to bind to the AreA zinc finger. These data reveal a potential new layer of control of nitrogen metabolite repression by the ordered proteolytic cleavage of NmrA. NmrA digested at the C-terminal site retained the ability to bind NAD+ and showed a resistance to further digestion that was enhanced by the presence of NAD+. This is the first time that an effect of dinucleotide binding to NmrA has been demonstrated. PMID:20506376

Bioproduction of mushroom mycelium of Agaricus bisporus by commercial submerged fermentation for the production of meat analogue.

PubMed

Kim, Kyoungju; Choi, Byungsun; Lee, Inhee; Lee, Hyeyoung; Kwon, Soonhyang; Oh, Kyoungyoung; Kim, Augustine Yonghwi

2011-07-01

As worldwide interest in healthy and delicious meat analogues increases, the texture of these products has become an important indicator of quality. Mycoprotein as fungal mycelium could provide a distinctive chewing sensation; however, the unfavorable consumer perception of fungal mycelium demands the production of meat analogues with true mushroom mycelium. The industrial and economical bioprocess was developed using an inexpensive medium (30 g L(-1) sugar cane extract (SCE), 10 g L(-1) NaNO(3) and 5 g L(-1) yeast extract) and A. bisporus Suksung. The SCE was maintained at around 10 g L(-1) to minimize osmotic shock. The maximum mycelium production of 15.0 g L(-1) (dry weight) was reached within 4 days. Scanning electron microscopic analysis showed fibrous and directional structure rather than a more typical pellet structure. Meat analogues with mushroom mycelium had better textural properties, being higher in hardness, springiness, and chewiness and with preferable umami characteristics compared to meat analogues utilizing soy protein. The overall acceptance of meat analogues prepared with mycelium and soy protein, and a ground beef patty, were 5, 2 and 10, respectively. The development of an industrial bioprocess for A. bisporus mycelium allowed the production of a highly acceptable meat analogue having not only superior textural properties but also umami characteristics when compared to that of soy protein. Copyright © 2011 Society of Chemical Industry.

Formation and dispersion of mycelial pellets of Streptomyces coelicolor A3(2).

PubMed

Kim, Yul-Min; Kim, Jae-heon

2004-03-01

The pellets from a culture of Streptomyces coelicolor A3(2) that were submerged shaken were disintegrated into numerous hyphal fragments by DNase treatment. The pellets were increasingly dispersed by hyaluronidase treatment, and mycelial fragments were easily detached from the pellets. The submerged mycelium grew by forming complexes with calcium phosphate precipitates or kaolin, a soil particle. Therefore, the pellet formation of Streptomyces coelicolor A3(2) can be considered a biofilm formation, including the participation of adhesive extracellular polymers and the insoluble substrates.

Copper, zinc, and cadmium in various fractions of soil and fungi in a Swedish forest.

PubMed

Vinichuk, Mykhailo M

2013-01-01

Ectomycorrhizal fungi profoundly affect forest ecosystems through mediating nutrient uptake and maintaining forest food webs. The accumulation of metals in each transfer step from bulk soil to fungal sporocarps is not well known. The accumulation of three metals copper (Cu), zinc (Zn) and cadmium (Cd) in bulk soil, rhizosphere, soil-root interface, fungal mycelium and sporocarps of mycorrhizal fungi in a Swedish forest were compared. Concentrations of all three metals increased in the order: bulk soil < soil-root interface (or rhizosphere) < fungal mycelium < fungal sporocarps. The uptake of Cu, Zn and Cd during the entire transfer process in natural conditions between soil and sporocarps occurred against a concentration gradient. In fungal mycelium, the concentration of all three metals was about three times higher than in bulk soil, and the concentration in sporocarps was about two times higher than in mycelium. In terms of accumulation, fungi (mycelium and sporocarps) preferred Cd to Zn and Cu. Zinc concentration in sporocarps and to a lesser extent in mycelium depended on the concentration in soil, whereas, the uptake of Cu and Cd by both sporocarps and mycelium did not correlate with metal concentration in soil. Heavy metal accumulation within the fungal mycelium biomass in the top forest soil layer (0-5 cm) might account for ca. 5-9% of the total amount of Cu, 5-11% of Zn, and 16-32% of Cd. As the uptake of zinc and copper by fungi may be balanced, this implied similarities in the uptake mechanism.

Saccharopolyspora subtropica sp. nov., a thermophilic actinomycete isolated from soil of a sugar cane field.

PubMed

Wu, Hao; Liu, Bin; Pan, Shangli

2016-05-01

A novel thermophilic actinomycete, designated strain T3T, was isolated from a soil sample of a sugar cane field. The strain grew at 25-60 °C (optimum 37-50 °C), at pH 6.0-11.0 (optimum 7.0-9.0) and with 0-12.0 % (w/v) NaCl (optimum 0-7 %). The aerial mycelium was white and the vegetative mycelium was colourless to pale yellow. The substrate mycelium fragmented into rod-shaped elements after 4-5 days at 50 °C. The aerial mycelium formed flexuous chains of 5-20 spores per chain; the oval-shaped spores had spiny surfaces and were non-motile. The organism contained meso-diaminopimelic acid as the diagnostic diamino acid in the cell-wall peptidoglycan. The whole-cell sugars consisted of arabinose, galactose and ribose. The cellular fatty acid profile consisted mainly of anteiso-C17 : 0, iso-C17 : 0 and iso-C16 : 0. The quinone system was composed predominantly of MK-9(H4). The phospholipids detected were diphosphatidylglycerol, phosphatidylcholine, phosphatidylinositol, phosphatidylethanolamine, phosphatidylmethylethanolamine and ninhydrin-positive glycophospholipids. The DNA G+C content of strain T3T was 71.3 mol%. The organism showed a combination of morphological and chemotaxonomic properties typical of members of the genus Saccharopolyspora. In the 16S rRNA gene tree of Saccharopolyspora it formed a distinct phyletic line and was related most closely to Saccharopolyspora thermophila 216T. However, the phenotypic characteristics of strain T3T were significantly different from those of S. thermophila 216T and DNA-DNA hybridization revealed a low level of relatedness (28.6-32.3 %) between them. Based on the phenotypic and phylogenetic data, strain T3T represents a novel species in the genus Saccharopolyspora, for which the name Saccharopolyspora subtropica sp. nov. is proposed. The type strain is T3T ( = DSM 46801T = CGMCC 4.7206T).

Yield and size of oyster mushroom grown on rice/wheat straw basal substrate supplemented with cotton seed hull.

PubMed

Yang, Wenjie; Guo, Fengling; Wan, Zhengjie

2013-10-01

Oyster mushroom (Pleurotus ostreatus) was cultivated on rice straw basal substrate, wheat straw basal substrate, cotton seed hull basal substrate, and wheat straw or rice straw supplemented with different proportions (15%, 30%, and 45% in rice straw substrate, 20%, 30%, and 40% in wheat straw substrate) of cotton seed hull to find a cost effective substrate. The effect of autoclaved sterilized and non-sterilized substrate on growth and yield of oyster mushroom was also examined. Results indicated that for both sterilized substrate and non-sterilized substrate, oyster mushroom on rice straw and wheat basal substrate have faster mycelial growth rate, comparatively poor surface mycelial density, shorter total colonization period and days from bag opening to primordia formation, lower yield and biological efficiency, lower mushroom weight, longer stipe length and smaller cap diameter than that on cotton seed hull basal substrate. The addition of cotton seed hull to rice straw and wheat straw substrate slowed spawn running, primordial development and fruit body formation. However, increasing the amount of cotton seed hull can increase the uniformity and white of mycelium, yield and biological efficiency, and increase mushroom weight, enlarge cap diameter and shorten stipe length. Compared to the sterilized substrate, the non-sterilized substrate had comparatively higher mycelial growth rate, shorter total colonization period and days from bag opening to primordia formation. However, the non-sterilized substrate did not gave significantly higher mushroom yield and biological efficiency than the sterilized substrate, but some undesirable characteristics, i.e. smaller mushroom cap diameter and relatively long stipe length.

Comparison of Psilocybe cubensis spore and mycelium allergens.

PubMed

Helbling, A; Horner, W E; Lehrer, S B

1993-05-01

Basidiospores are an important cause of respiratory allergy in mold-sensitive atopic subjects. Collection of the large amounts of spores required for extract preparation is tedious and difficult. A desirable alternative could be mycelium grown in vitro if it is allergenically similar to spores. Therefore this study compared the allergen contents of Psilocybe cubensis spore and mycelium extracts by different techniques with the use of pooled sera from subjects who had skin test and RAST results that were positive to P. cubensis spores. Isoelectric focusing immunoprints revealed six common IgE-binding bands at isoelectric points 4.7, 5.0, 5.5, 5.6, 8.7, and 9.3. Two additional bands at isoelectric points 3.9 and 5.7 were detected only in the spore extract. Sodium dodecylsulfate-polyacrylamide gel electrophoresis immunoblots exhibited six common IgE-binding bands at 16, 35, 487, 52, 62, and 76 kd; 20 and 40 kd bands were present only in the spore extract. Although RAST and isoelectric focusing inhibition demonstrated that P. cubensis spore and mycelium extracts share many allergens, spores were allergenically more potent than mycelium. The results indicate that mycelium is a useful source of P. cubensis allergen, even though several spore allergens were not detected in mycelium.

Comparative culturing of Pleurotus spp. on coffee pulp and wheat straw: biomass production and substrate biodegradation.

PubMed

Salmones, Dulce; Mata, Gerardo; Waliszewski, Krzysztof N

2005-03-01

The results of the cultivation of six strains of Pleurotus (P. djamor (2), P. ostreatus (2) and P. pulmonarius (2)) on coffee pulp and wheat straw are presented. Metabolic activity associated with biomass of each strain was determined, as well as changes in lignin and polysaccharides (cellulose and hemicellulose), phenolic and caffeine contents in substrate samples colonized for a period of up to 36 days. Analysis were made of changes during the mycelium incubation period (16 days) and throughout different stages of fructification. Greater metabolic activity was observed in the wheat straw samples, with a significant increase between 4 and 12 days of incubation. The degradation of polysaccharide compounds was associated with the fruiting stage, while the reduction in phenolic contents was detected in both substrates samples during the first eight days of incubation. A decrease was observed in caffeine content of the coffee pulp samples during fruiting stage, which could mean that some caffeine accumulates in the fruiting bodies.

Effects of Illumination Pattern during Cultivation of Fruiting Body and Bioactive Compound Production by the Caterpillar Medicinal Mushroom, Cordyceps militaris (Ascomycetes).

PubMed

Wu, Chiu-Yeh; Liang, Zeng-Chin; Tseng, Chin-Yin; Hu, Shu-Hui

2016-01-01

We investigated the effects of light intensity in the 3 cultivation stages separately-the mycelium colonization stage, the primordial initiation stage, and the fruiting stage (in order)-on fruiting body and bioactive compound production by Cordyceps militaris. In the mycelium colonization stage, rice substrates were incubated in a spawn running room at 23°C. During the primordial initiation stage, C. militaris was grown at 18°C and illuminated 12 hours/day. In the fruiting stage the temperature was 23°C, with illumination provided 12 hours/day. The highest fruiting body yield and biological efficiency were 4.06 g dry weight/bottle and 86.83%, respectively, under 1750 ± 250 lux during the second and third stages. The cordycepin content was highest during the second and third stages under 1250 ± 250 lux. The mannitol and polysaccharide contents were highest under 1250 ± 250 and 1750 ± 250 lux during the primordial initiation stage and the fruiting stage, respectively. Thus, with controlled lighting, C. militaris can be cultivated in rice-water medium to increase fruiting body yield and bioactive compound production.

[Streptoverticillium griseoviridum n. sp., a producer of the candidin-amphotericin B group, antifungal heptaene nonaromatic antibiotic 0185].

PubMed

Konev, Iu E; Efimova, V M; Etingov, E D; Zaval'naia, N M

1978-02-01

An actinomyceteous strain LIA-0185 producing a heptaenic non-aromatic antibiotic of the candidin type was isolated from a soil sample taken in the Georgian SSR under the programme of screening antifungal antibiotics. The taxonomic study of the strain showed that it belonged to the series of viridoflavum and had the following main taxonomic features: the sporophores in the whorls, straight, remote: the aerial mycelium from yellow to dark-olive-grey; the substrate mycelium olive; the soluble pigment absent; the melanine pigment was produced on the peptone medium; the culture formed H2S; assimilated glucose, mannose, inozide and to a lesser extent fructose; did not assimilate arabinose, xylose, sucrose, lactose, ramnose and raffinose. The strain inhibited the growth of yeast and fungi, grampositive bacteria and actinomycetes and produced a complex of non-aromatic heptaenic antibiotics. The actinomycete differed from the other whorl cultures. It was classified as a new species Sv. griseoviridum sp. nov. The antibiotic complex was a mixture of 2 components, i. e. I and II present approximately in equal amounts. Component II was analogous to candidin. Component I was a new original substance.

Production of Lingzhi or Reishi medicinal mushroom, Ganoderma lucidum (W.Curt. :Fr.) P. Karst. (higher Basidiomycetes), biomass and polysaccharides by solid state cultivation.

PubMed

Berovic, Marin; Habijanic, Jozica; Boh, Bojana; Wraber, Branka; Petravic-Tominac, Vlatka

2012-01-01

Solid state cultivation of Ganoderma lucidum biomass, strain BFWS Gal 4, originally isolated from the Slovenian forest, was studied in a horizontal stirred tank reactor. Periodic mixing of N = 80 rpm, 2 min/day was used. Production of fungal polysaccharides and fungal biomass on solid substrate based on beech sawdust, olive oil, and mineral salts was studied. Optimal moisture of the solid matrix was in the range of 80% to 74%. When the moisture content dropped below 57%, the growth of the mycelium and polysaccharide production stopped, but it revived when wet air was applied in further processing. Final concentration of biomass was 0.68 mg/g of solid substrate, while proportions of extracellular and intracellular polysaccharides were 4.5 mg/g and 1.05 mg/g, respectively.

A Model for Growth of a Single Fungal Hypha Based on Well-Mixed Tanks in Series: Simulation of Nutrient and Vesicle Transport in Aerial Reproductive Hyphae

PubMed Central

Balmant, Wellington; Sugai-Guérios, Maura Harumi; Coradin, Juliana Hey; Krieger, Nadia; Furigo Junior, Agenor; Mitchell, David Alexander

2015-01-01

Current models that describe the extension of fungal hyphae and development of a mycelium either do not describe the role of vesicles in hyphal extension or do not correctly describe the experimentally observed profile for distribution of vesicles along the hypha. The present work uses the n-tanks-in-series approach to develop a model for hyphal extension that describes the intracellular transport of nutrient to a sub-apical zone where vesicles are formed and then transported to the tip, where tip extension occurs. The model was calibrated using experimental data from the literature for the extension of reproductive aerial hyphae of three different fungi, and was able to describe different profiles involving acceleration and deceleration of the extension rate. A sensitivity analysis showed that the supply of nutrient to the sub-apical vesicle-producing zone is a key factor influencing the rate of extension of the hypha. Although this model was used to describe the extension of a single reproductive aerial hypha, the use of the n-tanks-in-series approach to representing the hypha means that the model has the flexibility to be extended to describe the growth of other types of hyphae and the branching of hyphae to form a complete mycelium. PMID:25785863

Screening of micro-organisms for decolorization of melanins produced by bluestain fungi.

PubMed

Rättö, M; Chatani, M; Ritschkoff, A C; Viikari, L

2001-03-01

A total of 17 fungi and four bacteria were screened for their ability to decolorize melanin, using isolated extracellular melanin of the bluestain fungus Aureobasidium pullulans as substrate. On agar media, decolorization was observed by four fungal strains: Bjerkandera adusta VTT-D-99746, Galactomyces geotrichum VTT-D-84228, Trametes hirsuta VTT-D-95443 and Trametes versicolor VTT-D-99747. The four fungi were more efficient on nitrogen-limited medium than on complete medium. The melanin-decolorizing activity of G. geotrichum appeared to be located on the mycelium and could be liberated into the medium enzymatically.

Mycelial abundance and other factors related to truffle productivity in Tuber melanosporum-Quercus ilex orchards.

PubMed

Suz, Laura M; Martín, María P; Oliach, Daniel; Fischer, Christine R; Colinas, Carlos

2008-08-01

Relative quantification of DNA from Tuber melanosporum mycelia was performed by conventional and real-time PCR in soil from trees in three truffle orchards of different ages to determine: (1) whether burn appearance is related to the amount of T. melanosporum mycelium in soil, and (2) whether productivity onset and truffle production are related to (a) the amount of T. melanosporum mycelium in soil, (b) tree height and diameter, (c) burn extension and (d) surface rock cover. The burn seems to appear only after a certain amount of mycelium has formed. Precociously productive trees presented higher quantities of mycelium than nonproductive trees in the productivity onset study, while highly productive trees presented less quantities of mycelium than nonproductive trees in the productivity study. Trees with high but not excessive surface rock cover showed greater truffle production. Larger trees tended to display a burn earlier than smaller trees.

Growing and testing mycelium bricks as building insulation materials

NASA Astrophysics Data System (ADS)

Xing, Yangang; Brewer, Matthew; El-Gharabawy, Hoda; Griffith, Gareth; Jones, Phil

2018-02-01

In order to improve energy performance of buildings, insulation materials (such as mineral glass and rock wools, or fossil fuel-based plastic foams) are being used in increasing quantities, which may lead to potential problem with materials depletions and landfill disposal. One sustainable solution suggested is the use of bio-based, biodegradable materials. A number of attempts have been made to develop biomaterials, such as sheep wood, hemcrete or recycled papers. In this paper, a novel type of bio insulation materials - mycelium is examined. The aim is to produce mycelium materials that could be used as insulations. The bio-based material was required to have properties that matched existing alternatives, such as expanded polystyrene, in terms of physical and mechanical characteristics but with an enhanced level of biodegradability. The testing data showed mycelium bricks exhibited good thermal performance. Future work is planned to improve growing process and thermal performance of the mycelium bricks.

High-yield aqueous synthesis of multi-branched iron oxide core-gold shell nanoparticles: SERS substrate for immobilization and magnetic separation of bacteria

NASA Astrophysics Data System (ADS)

Tamer, Ugur; Onay, Aykut; Ciftci, Hakan; Bozkurt, Akif Göktuğ; Cetin, Demet; Suludere, Zekiye; Hakkı Boyacı, İsmail; Daniel, Philippe; Lagarde, Fabienne; Yaacoub, Nader; Greneche, Jean-Marc

2014-10-01

The high product yield of multi-branched core-shell Fe3- x O4@Au magnetic nanoparticles was synthesized used as magnetic separation platform and surface-enhanced Raman scattering (SERS) substrates. The multi-branched magnetic nanoparticles were prepared by a seed-mediated growth approach using magnetic gold nanospheres as the seeds and subsequent reduction of metal salt with ascorbic acid in the presence of a stabilizing agent chitosan biopolymer and silver ions. The anisotropic growth of nanoparticles was observed in the presence of chitosan polymer matrix resulting in multi-branched nanoparticles with a diameter over 100 nm, and silver ions also play a crucial role on the growth of multi-branched nanoparticles. We propose the mechanism of the formation of multi-branched nanoparticles while the properties of nanoparticles embedded in chitosan matrix are discussed. The surface morphology of nanoparticles was characterized with transmission electron microscopy, scanning electron microscopy, ultraviolet visible spectroscopy (UV-Vis), X-ray diffraction, and fourier transform infrared spectroscopy and 57Fe Mössbauer spectrometry. Additionally, the magnetic properties of the nanoparticles were also examined. We also demonstrated that the synthesized Fe3- x O4@Au multi-branched nanoparticle is capable of targeted separation of pathogens from matrix and sensing as SERS substrates.

Comparative study of metals accumulation in cultured in vitro mycelium and naturally grown fruiting bodies of Boletus badius and Cantharellus cibarius.

PubMed

Reczyński, Witold; Muszyńska, Bożena; Opoka, Włodzimierz; Smalec, Agata; Sułkowska-Ziaja, Katarzyna; Malec, Mirosław

2013-06-01

Cantharellus cibarius Fr. (chanterelle) and Boletus badius Pers. (bay bolete) harvested from natural sites in Poland were used to derive in vitro cultures. The optimal medium composition for cultures was developed. Concentrations of the chosen elements (Zn, Cu, Fe, Mg, Ni, and Cd) in mycelium samples were measured by means of atomic absorption spectrometry. Fe concentration in the analyzed mushroom materials was in the range 215.4-680.3 μg/g dry weight. Mean values of Mg were respectively (in micrograms per gram dry weight) 541.8 for mycelium of C. cibarius cultured in vitro and 1,004.1 for C. cibarius fruiting bodies and 928.9 for the mycelium of B. badius cultured in vitro and 906.4 for B. badius fruiting bodies. The mean concentrations of Zn were 442.7 μg/g dry weight in mycelium from in vitro cultures of B. badius and 172.1 in B. badius fruiting bodies and 131.9 in the case of C. cibarius in mycelium from in vitro cultures and 95.5 for the C. cibarius fruiting bodies. Cu exhibited a reversal tendency, i.e., the element concentrations in naturally grown mushrooms were significantly higher (43.57 μg/g dry weight for C. cibarius and 43.54 μg/g for B. badius) than in cultured in vitro mycelium (12.47 μg/g for C. cibarius and 4.17 μg/g for B. badius). Ni was found in lowest concentrations ranging from 0.33 to 1.88 μg/g dry weight. Toxic metal Cd was found in relatively high concentrations in naturally grown species (0.79 μg/g dry weight-1.02). The lowest was the concentration of Cd in C. cibarius mycelium from in vitro culture-0.06 μg/g dry weight-a bit higher than it was in the B. badius mycelium (0.21 μg/g).

Identification and characterization of the biotechnological potential of a wild strain of Paraconiothyrium sp.

PubMed

Arredondo-Santoyo, Marina; Vázquez-Garcidueñas, Ma Soledad; Vázquez-Marrufo, Gerardo

2018-04-30

The isolation and characterization of fungal strains from poorly described taxa allows undercover attributes of their basic biology useful for biotechnology. Here, a wild fungal strain (CMU-196) from recently described Paraconiothyrium genus was analyzed. CMU-196 was identified as Paraconiothyrium brasiliense by phylogenetic analysis of the rDNA internal transcribed spacer region (ITS). CMU-196 metabolized 57 out of 95 substrates of the Biolog FF microplates. Efficient assimilation of dextrins and glycogen indicates that CMU-196 is a good producer of amylolytic enzymes. It showed a remarkably assimilation of α-D-lactose, substrate described as inducer of cellulolytic activity but poorly assimilated by several fungi. Metabolically active mycelium of the strain decolorized broth supplemented with direct blue 71, Chicago sky blue and remazol brilliant blue R dyes. The former two dyes were also well removed from broth by mycelium inactivated by autoclaving. Both mycelia had low efficiency for removing fuchsin acid from broth and for decolorizing wastewater from the paper industry. CMU-196 strain showed extracellular laccase activity when potato dextrose broth was supplemented with Cu +2 , reaching a maximum activity of 46.8 (±0.33) U/L. Studied strain antagonized phytopathogenic Colletotrichum spp. fungi and Phytophthora spp. oomycetes in vitro, but is less effective towards Fusarium spp. fungi. CMU-196 antagonism includes overgrowing the mycelia of phytopathogens and growth inhibition, probably by hydrosoluble extracellular metabolites. The biotechnological potential of strain CMU-196 here described warrants further studies to have a more detailed knowledge of the mechanisms associated with its metabolic versatility, capacity for environmental detoxification, extracellular laccase production and antagonism against phytopathogens. This article is protected by copyright. All rights reserved. © 2018 American Institute of Chemical Engineers.

Quantitative Proteomics Analysis of Streptomyces coelicolor Development Demonstrates That Onset of Secondary Metabolism Coincides with Hypha Differentiation*

PubMed Central

Manteca, Angel; Sanchez, Jesus; Jung, Hye R.; Schwämmle, Veit; Jensen, Ole N.

2010-01-01

Streptomyces species produce many clinically important secondary metabolites, including antibiotics and antitumorals. They have a complex developmental cycle, including programmed cell death phenomena, that makes this bacterium a multicellular prokaryotic model. There are two differentiated mycelial stages: an early compartmentalized vegetative mycelium (first mycelium) and a multinucleated reproductive mycelium (second mycelium) arising after programmed cell death processes. In the present study, we made a detailed proteomics analysis of the distinct developmental stages of solid confluent Streptomyces coelicolor cultures using iTRAQ (isobaric tags for relative and absolute quantitation) labeling and LC-MS/MS. A new experimental approach was developed to obtain homogeneous samples at each developmental stage (temporal protein analysis) and also to obtain membrane and cytosolic protein fractions (spatial protein analysis). A total of 345 proteins were quantified in two biological replicates. Comparative bioinformatics analyses revealed the switch from primary to secondary metabolism between the initial compartmentalized mycelium and the multinucleated hyphae. PMID:20224110

Evaluation of the effect of process variables on the fatty acid profile of single cell oil produced by Mortierella using solid-state fermentation.

PubMed

Asadi, Seyedeh Zeinab; Khosravi-Darani, Kianoush; Nikoopour, Houshang; Bakhoda, Hossein

2015-03-01

This article reviews some of the aspects of single cell oil (SCO) production using solid-state fermentation (SSF) by fungi of the genus Mortierella. This article provides an overview of the advantages of SSF for SCO formation by the aforementioned fungus and demonstrates that the content of the polyunsaturated fatty acids (PUFA) depend on the type of fermentation media and culture conditions. Process variables that influence lipid accumulation by Mortierella spp. and the profile of the fatty acids are discussed, including incubation temperature, time, aeration, growth phase of the mycelium, particle size of the substrate, carbon to nitrogen ratio, initial moisture content and pH as well as supplementation of the substrate with nitrogen and oil. Finally, the article highlights future research trends for the scaled-up production of PUFAs in SSF.

Fungal mycelium and decomposition of needle litter in three contrasting coniferous forests

NASA Astrophysics Data System (ADS)

Virzo De Santo, Amalia; Rutigliano, Flora Angela; Berg, Björn; Fioretto, Antonietta; Puppi, Gigliola; Alfani, Anna

2002-08-01

The fungal mycelium ingrowth and the rates of mass loss and respiration of needle litter of Pinus pinea, Pinus laricio, Pinus sylvestris, and Abies alba were investigated, in three coniferous forests, over a 3-year period by means of a composite set of incubations. In the early stages, the fungal flora of the decomposing needles was dominated by dematiaceous hyphomycetes and coelomycetes. Basidiomycetes reached a peak after 6 months on pine needles, but were absent from the N-rich needles of A. alba. Soil fungi ( Penicillium, Trichoderma, Absidia, Mucor sp. pl.) became most frequent in later stages. At the end of the study period, the total mycelium amount showed the lowest values in all pine needles incubated in the P. laricio forest and the highest ones in P. pinea needles incubated in the P. pinea forest. In all data sets, as in data for boreal forests examined for comparison, the concentration of litter fungal mycelium versus litter mass loss followed a common exponential model. However, in later stages, the amount of litter fungal mycelium was very close to that of the humus at the incubation site, thus supporting the hypothesis of a logistic growth pattern. Respiration rates of decomposing litters varied with season and decreased with litter age to values close to those of the humus at the incubation site. Respiration of water-saturated litter was negatively correlated with the total mycelium concentration, and this was consistent with the observation that in far-decomposed litter only a minor fraction of the total mycelium is alive.

[What potentialities does the four-spored mushroom "Agaricus bitorquis (Quél.) Sacc." offer to the breeder?].

PubMed

Fritsche, G

1976-05-01

A. bitorquis was first taken into cultivation in 1968. It differs from A.bisporus, the only mushroom cultivated previously, in a range of properties. The claims for temperature are about 5° C higher. The fruitbodies (white smooth) are more vigorous than the sporophores of the white, scale-less strains of A.bisporus. Especially valuable characteristics which A. bitorquis brings are virus resistance, resistance to pressure, easy pickability and longer shelflife. The basidia have 4 instead of 2 spores. Consequently monosporecultures are infertile and systematic crossbreeding is a suitable breeding method.Because the hyphae do not form clamp connections, it is not possible to distinguish microscopically monocaryotic and dicaryotic mycelium. As the trials have shown, however, the compatibility of the monospore cultures can be recognized by the manner of mycelium growth on biomalt-agar. Where heterocaryotic mycelium has arisen matted, slow growing mycelium can turn into fluffy, and later on stringy fast-growing mycelium. With enough ventilation condensations of mycelium can be formed. Some combinations of monospore-cultures of different origin showed very significantly higher yields than the parental wild types, whereas other combinations of two monospore cultures were very significantly lower in yield than the parents. The combination of parental wildtypes scarcely differed in yield from the wildtype self. Regarding the course of the yield there were big differences in general.The strains also showed great variability in the shape and colour of the fruitbodies, their distribution on the bed and in other properties, such as the propensity of the mycelium to grow into the casing layer. The results are discussed.

Genotoxicity profile of erinacine A-enriched Hericium erinaceus mycelium.

PubMed

Li, I-Chen; Chen, Yen-Lien; Chen, Wan-Ping; Lee, Li-Ya; Tsai, Yueh-Ting; Chen, Chin-Chu; Chen, Chin-Shuh

2014-01-01

Hericium erinaceus ( H. erinaceus ) has a long history of usage in traditional Chinese medicine for the treatment of gastric disorders. Recently, it has become a well-established candidate in causing positive brain and nerve health-related activities by inducing nerve growth factor (NGF) from its bioactive ingredient, erinacine A. This active compound, which exists only in fermented mycelium but not in its fruiting body, increases NGF levels in astroglial cells in vitro as well as catecholamine and NGF levels in vivo . With increasing recognition of erinacine A in H. erinaceus (EAHE) mycelium improving neurodegenerative diseases, numerous products are being marketed based on these functional claims. To our knowledge, there have been no reports on the mutagenicity of EAHE prior to this paper. Hence, the present study was undertaken to determine the mutagenicity and genotoxicity effects of EAHE mycelium conducted in three standard battery of tests (reverse mutation, chromosomal aberration, and micronuclei tests) according to the latest guidelines in order to meet all international regulatory requirements and provide information on the safety of this new and promising natural remedy. Our results have indicated that EAHE mycelium did not significantly increase the number of revertant colonies in the bacterial reverse mutation test nor induce higher frequency of aberrations in the chromosome aberration test. Moreover, no statistically significant EAHE mycelium-related increase was observed in the incidence of reticulocytes per 1000 red blood cells and micronucleated reticulocytes per 1000 reticulocytes. In conclusion, the three standard battery of tests suggested that EAHE mycelium was devoid of mutagenicity and genotoxicity in the tested doses and experimental conditions.

Out on a limb: Thermal microenvironments in the tropical forest canopy and their relevance to ants.

PubMed

Stark, Alyssa Y; Adams, Benjamin J; Fredley, Jennifer L; Yanoviak, Stephen P

2017-10-01

Small, cursorial ectotherms like ants often are immersed in the superheated air layers that develop millimeters above exposed, insolated surfaces (i.e., the thermal boundary layer). We quantified the thermal microenvironments around tree branches in the tropical rainforest canopy, and explored the effects of substrate color on the internal body temperature and species composition of arboreal ants. Branch temperatures during the day (09:00-16:00) were hottest (often > 50°C) and most variable on the upper surface, while the lowest and least variable temperatures occurred on the underside. Temperatures on black substrates declined with increasing distance above the surface in both the field and the laboratory. By contrast, a micro-scale temperature inversion occurred above white substrates. Wind events (ca. 2ms -1 ) eliminated these patterns. Internal temperatures of bodies of Cephalotes atratus workers experimentally heated in the laboratory were 6°C warmer on white vs. black substrates, and 6°C cooler than ambient in windy conditions. The composition of ant species foraging at baits differed between black-painted and unpainted tree branches, with a tendency for smaller ants to avoid the significantly hotter black surfaces. Collectively, these outcomes show that ants traversing canopy branches experience very heterogeneous thermal microenvironments that are partly influenced in predictable ways by branch surface coloration and breezy conditions. Copyright © 2017 Elsevier Ltd. All rights reserved.

Isolation and identification of gold nanoparticles synthesizing fungi from Indian Kolar Gold Field mine soil.

PubMed

Lakshmi, V Jhansi; Kannan, K P

2016-07-01

An indigenous fungal strain was isolated from Indian Kolar Gold Field mine soil. The isolate was heterothallic, branched septate, deeply floccose, fast-growing, dull green with white background conidial columnar mycelium from Aspergillus section Fumigati. Diverse metabolic patterns of the isolate exhibit high metal, thermal resistance which grews well from 28 ± 1 degrees C to 37 degrees C and pH concentration was significant on the growth of isolate. Phylogenetic analysis of 16srRNA β-Tubulin gene sequence established relationship among isolate and other taxa. Molecular identification and morphological features of fungal isolate were consistent with those of Neosartorya udagawae. Heterothallic N. udagawae FJ830683 strain was closely related to homothallic N. aureola EF661890. Fungal isolate extract synthesized narrow sized stable Gold nanoparticles (AuNPs).

Structures with three dimensional nanofences comprising single crystal segments

DOEpatents

Goyal, Amit; Wee, Sung-Hun

2013-08-27

An article includes a substrate having a surface and a nanofence supported by the surface. The nanofence includes a multiplicity of primary nanorods and branch nanorods, each of the primary nanorods being attached to said substrate, and each of the branch nanorods being attached to a primary nanorods and/or another branch nanorod. The primary and branch nanorods are arranged in a three-dimensional, interconnected, interpenetrating, grid-like network defining interstices within the nanofence. The article further includes an enveloping layer supported by the nanofence, disposed in the interstices, and forming a coating on the primary and branch nanorods. The enveloping layer has a different composition from that of the nanofence and includes a radial p-n single junction solar cell photovoltaic material and/or a radial p-n multiple junction solar cell photovoltaic material.

Biotransformation of (-)-α-pinene and geraniol to α-terpineol and p-menthane-3,8-diol by the white rot fungus, Polyporus brumalis.

PubMed

Lee, Su-Yeon; Kim, Seon-Hong; Hong, Chang-Young; Park, Se-Yeong; Choi, In-Gyu

2015-07-01

In this study, the monoterpenes, α-pinene and geraniol, were biotransformed to synthesize monoterpene alcohol compounds. Polyporus brumalis which is classified as a white rot fungus was used as a biocatalyst. Consequently α-terpineol was synthesized from α-pinene by P. brumalis mycelium, after three days. Moreover, another substrate, the acyclic monoterpenoids geraniol was transformed into the cyclic compound, p-menthane-3, 8-diol (PMD). The main metabolites, i.e., α-terpineol and PMD, are known to be bioactive monoterpene alcohol compounds. This study highlights the potential of fungal biocatalysts for monoterpene transformation.

Spatially resolving the secretome within the mycelium of the cell factory Aspergillus niger.

PubMed

Krijgsheld, Pauline; Altelaar, A F Maarten; Post, Harm; Ringrose, Jeffrey H; Müller, Wally H; Heck, Albert J R; Wösten, Han A B

2012-05-04

Aspergillus niger is an important cell factory for the industrial production of enzymes. These enzymes are released into the culture medium, from which they can be easily isolated. Here, we determined with stable isotope dimethyl labeling the secretome of five concentric zones of 7-day-old xylose-grown colonies of A. niger that had either or not been treated with cycloheximide. As expected, cycloheximide blocked secretion of proteins at the periphery of the colony. Unexpectedly, protein release was increased by cycloheximide in the intermediate and central zones of the mycelium when compared to nontreated colonies. Electron microscopy indicated that this is due to partial degradation of the cell wall. In total, 124 proteins were identified in cycloheximide-treated colonies, of which 19 secreted proteins had not been identified before. Within the pool of 124 proteins, 53 secreted proteins were absent in nontreated colonies, and additionally, 35 proteins were released ≥4-fold in the central and subperipheral zones of cycloheximide-treated colonies when compared to nontreated colonies. The composition of the secretome in each of the five concentric zones differed. This study thus describes spatial release of proteins in A. niger, which is instrumental in understanding how fungi degrade complex substrates in nature.

Production of bio-fertilizer from microwave vacuum pyrolysis of waste palm shell for cultivation of oyster mushroom (Pleurotus ostreatus)

NASA Astrophysics Data System (ADS)

Lun Nam, Wai; Huan Su, Man; Phang, Xue Yee; Chong, Min Yee; Keey Liew, Rock; Ma, Nyuk Ling; Lam, Su Shiung

2017-11-01

Microwave vacuum pyrolysis of waste palm shell (WPS) was performed to produce biochar, which was then tested as bio-fertilizer in growing Oyster mushroom (Pleurotus ostreatus). The pyrolysis approach generated a biochar containing a highly porous structure with a high BET surface area (up to 1250 m2/g) and a low moisture content (≤ 10 wt%), exhibiting desirable adsorption properties to be used as bio-fertilizer since it can act as a housing that provides many sites on which living microorganisms (mycelium or plant-growth promoting bacteria) and organic nutrients can be attached or adsorbed onto. This could in turn stimulate plant growth by increasing the availability and supply of nutrients to the targeted host plant. The results from growing Oyster mushroom using the biochar record an impressive growth rate and a monthly production of up to about 550 g of mushroom. The shorter time for mycelium growth on whole baglog (30 days) and the highest yield of Oyster mushroom (550 g) was obtained from the cultivation medium added with 20 g of biochar. Our results demonstrate that the biochar-based bio-fertilizer produce from microwave vacuum pyrolysis of WPS show exceptional promise as an alternative growing substrate for mushroom cultivation.

Optimization of the Liquid Culture Medium Composition to Obtain the Mycelium of Agaricus bisporus Rich in Essential Minerals.

PubMed

Krakowska, Agata; Reczyński, Witold; Muszyńska, Bożena

2016-09-01

Agaricus bisporus species (J.E. Lange) Imbach one of the most popular Basidiomycota species was chosen for the research because of its dietary and medicinal value. The presented herein studies included determination of essential mineral accumulation level in the mycelium of A. bisporus, cultivated on liquid cultures in the medium supplemented with addition of the chosen metals' salts. Quantitative analyses of Zn, Cu, Mg, and Fe in liquid cultures made it possible to determine the relationship between accumulation of the selected mineral in A. bisporus mycelium and the culture conditions. Monitoring of the liquid cultures and determination of the elements' concentrations in mycelium of A. bisporus were performed using the flame technique of AAS method. Concentration of Zn in the mycelium, maintained in the medium with the addition of its salt, was in a very wide range from 95.9 to 4462.0 mg/g DW. In the analyzed A. bisporus mycelium, cultured in the medium enriched with copper salt, this metal concentration changed from 89.79 to 7491.50 mg/g DW; considering Mg in liquid cultured mycelium (medium with Mg addition), its concentration has changed from 0.32 to 10.55 mg/g DW. The medium enriched with iron salts has led to bioaccumulation of Fe in mycelia of A. bisporus. Determined Fe concentration was in the range from 0.62 to 161.28 mg/g DW. The proposed method of liquid A. bisporus culturing on medium enriched with the selected macro- and microelements in proper concentrations ratio have led to obtaining maximal growth of biomass, characterized by high efficiency of the mineral accumulation. As a result, a dietary component of increased nutritive value was obtained.

Isolation, Culture and Characterization of Hirsutella sinensis Mycelium from Caterpillar Fungus Fruiting Body

PubMed Central

Ko, Yun-Fei; Liau, Jian-Ching; Lee, Chien-Sheng; Chiu, Chen-Yaw; Martel, Jan; Lin, Chuan-Sheng; Tseng, Shun-Fu; Ojcius, David M.; Lu, Chia-Chen; Lai, Hsin-Chih; Young, John D.

2017-01-01

The caterpillar fungus Ophiocordyceps sinensis (previously called Cordyceps sinensis) has been used for centuries in Asia as a tonic to improve health and longevity. Recent studies show that O. sinensis produces a wide range of biological effects on cells, laboratory animals and humans, including anti-fatigue, anti-infection, anti-inflammatory, antioxidant, and anti-tumor activities. In view of the rarity of O. sinensis fruiting bodies in nature, cultivation of its anamorph mycelium represents a useful alternative for large-scale production. However, O. sinensis fruiting bodies harvested in nature harbor several fungal contaminants, a phenomenon that led to the isolation and characterization of a large number of incorrect mycelium strains. We report here the isolation of a mycelium from a fruiting body of O. sinensis and we identify the isolate as O. sinensis’ anamorph (also called Hirsutella sinensis) based on multi-locus sequence typing of several fungal genes (ITS, nrSSU, nrLSU, RPB1, RPB2, MCM7, β-tubulin, TEF-1α, and ATP6). The main characteristics of the isolated mycelium, including its optimal growth at low temperature (16°C) and its biochemical composition, are similar to that of O. sinensis fruiting bodies, indicating that the mycelium strain characterized here may be used as a substitute for the rare and expensive O. sinensis fruiting bodies found in nature. PMID:28046129

Seasonal dynamics of extraradical mycelium and mycorrhizas in a black truffle (Tuber melanosporum) plantation.

PubMed

Queralt, Mikel; Parladé, Javier; Pera, Joan; DE Miguel, Ana María

2017-08-01

Seasonal dynamics of black truffle (Tuber melanosporum) extraradical mycelium as well as the associated mycorrhizal community have been evaluated in a 16-year-old plantation with productive and non-productive trees. Mycelium biomass was seasonally quantified by real-time PCR over two consecutive years and the correlation with environmental variables explored. Extraradical mycelium biomass varied seasonally and between the two consecutive years, being correlated with the precipitation that occurred 1 month before sampling. In addition, productive trees had more mycelium in the brûlé area than non-productive trees did. The ectomycorrhizal community composition inside the burnt areas was seasonally evaluated during a year. Ten mycorrhizal morphotypes were detected; T. melanosporum was the most abundant in productive and non-productive trees. Black truffle mycorrhizas were more abundant (mycorrhizal tips per unit of soil volume) in productive trees, and no seasonal variation was observed. The occurrence of black truffle mycorrhizas was significantly and positively correlated with the biomass of extraradical mycelium. The mycorrhizal community within the brûlé areas was significantly different between productive and non-productive trees, and no variation was detected between seasons. The assessment of the fungal vegetative structures in a mature plantation is of paramount importance to develop trufficulture methods based on the knowledge of the biological cycle of the fungus and its relationships with the associated ectomycorrhizal communities.

Effects and Mechanism of Blue Light on Monascus in Liquid Fermentation.

PubMed

Zhang, Xiaowei; Liu, Wenqing; Chen, Xiying; Cai, Junhui; Wang, Changlu; He, Weiwei

2017-03-01

The effect of light on Monascus and the underlying mechanism have received a great deal of interest for the industrial application of Monascus pigments. In this study, we have examined the effects of blue light on the culture morphology, mycelium growth, pigments, and citrinin yield of Monascus in liquid-state and oscillation fermentation, and explored the mechanism at a physiological level. It was found that blue light affected the colony morphology, the composition (chitin content), and permeability of the Monascus mycelium cell wall in static liquid culture, which indicates blue light benefits pigments secreting from aerial mycelium to culture medium. In liquid oscillation fermentation, the yields of Monascus pigments in fermentation broth (darkness 1741 U/g, blue light 2206 U/g) and mycelium (darkness 2442 U/g, blue light 1900 U/g) cultured under blue light and darkness are different. The total pigments produced per gram of Monascus mycelium under blue light was also higher (4663 U/g) than that in darkness (4352 U/g). However, the production of citrinin (88 μg/g) under blue light was evidently lower than that in darkness (150 μg/g). According to the degradation of citrinin caused by blue light and hydrogen peroxide, it can be concluded that blue light could degrade citrinin and inhibit the catalase activity of Monascus mycelium, subsequently suppressing the decomposition of hydrogen peroxide, which is the active species that degrades citrinin.

Grafting as a method for studying development in the filamentous fungus Podospora anserina.

PubMed

Silar, Philippe

2011-08-01

While grafting and transplant experiments have extensively been used to study development in animals and plants, they have seldom been employed to study fungal development. Here, grafting is used to study the interplay between mycelium and multicellular fruiting bodies during maturation in the model ascomycete Podospora anserina. Data indicate that grafts need a competent mycelium to continue their ripening. Vegetative incompatibility does not prevent transplanted fructifications to undergo development. Grafting onto mutant mycelia confirmed a previous model stating that the NADPH oxidase PaNox1 is required in the developing fruiting bodies, while the MAP kinase cascade PaMpk1 is required in the mycelium. Data also show that the IDC1 protein is required not only in the developing fruiting bodies but also in the mycelium, likely because of its role in anastomosis. Finally, entry inside the grafted fruiting bodies of a ribosomal protein tagged with GFP could be detected, suggesting that cellular components are imported from the underlying mycelium during maturation. Copyright © 2011 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

Fungal spore germination into yeast or mycelium: possible implications of dimorphism in evolution and human pathogenesis

NASA Astrophysics Data System (ADS)

Ghormade, Vandana; Deshpande, M. V.

The ability of dimorphism in fungi is conventionally regarded as a reversible change between the two vegetative forms, yeast and mycelium, in response to environmental change. A zygomycetous isolate, Benjaminiella poitrasii, exhibited yeast-mycelium transition in response to the change in temperature (37-28 °C) and decrease in glucose concentration. For the first time the presence of dimorphic response during asexual and sexual spore germination is reported under the dimorphism-triggering conditions in B. poitrasii. The zygospores germinated into budding yeast when subjected to yeast-form supporting conditions. The mycelium-form favoring conditions gave rise to true mycelium. Similarly, the asexual spores displayed a dimorphic response during germination. Our observations suggest that dimorphism is an intrinsic ability present in the vegetative, asexual, and sexual forms of the fungus. As dimorphic fungi are intermediate to the unicellular yeast and the filamentous forms, understanding of the dimorphic character could be useful to trace the evolutionary relationships among taxonomically different fungi. Moreover, the implications of spore germination during the onset of pathogenesis and in drug development for human health care are discussed.

Controlled production of Camembert-type cheeses. Part I: Microbiological and physicochemical evolutions.

PubMed

Leclercq-Perlat, Marie-Noëlle; Buono, Frédéric; Lambert, Denis; Latrille, Eric; Spinnler, Henry-Eric; Corrieu, Georges

2004-08-01

A holistic approach of a mould cheese ripening is presented. The objective was to establish relationships between the different microbiological and biochemical changes during cheese ripening. Model cheeses were prepared from pasteurized milk inoculated with Kluyveromyces lactis, Geotrichum candidum, Penicillium camemberti and Brevibacterium linens under aseptic conditions. Two cheese-making trials with efficient control of environmental parameters were carried out and showed similar ripening characteristics. K. lactis grew rapidly between days 1 and 6 (generation time around 48 h). G. candidum grew exponentially between days 4 and 10 (generation time around 4.6 d). Brevi. linens also grew exponentially but after day 6 when Pen. camemberti mycelium began developing and the pH of the rind was close to 7. Its exponential growth presented 3 phases in relation to carbon and nitrogen substrate availability. Concentrations of Pen. camemberti mycelium were not followed by viable cell count but they were evaluated visually. The viable microorganism concentrations were well correlated with the carbon substrate concentrations in the core and in the rind. The lactose concentrations were negligible after 10 d ripening, and changes in lactate quantities were correlated with fungi flora. The pH of the inner part depended on NH3. Surface pH was significantly related to NH3 concentration and to fungi growth. The acid-soluble nitrogen (ASN) and non-protein nitrogen (NPN) indexes and NH3 concentrations of the rind were low until day 6, and then increased rapidly to follow the fungi concentrations until day 45. The ASN and NPN indexes and NH3 concentrations in the core were lower than in the rind and they showed the same evolution. G. candidum and Pen. camemberti populations have a major effect on proteolysis; nevertheless, K. lactis and Brevi. linens cell lysis also had an impact on proteolysis. Viable cell counts of K. lactis, G. candidum, Pen. camemberti and Brevi. linens were correlated with the environmental conditions, with proteolytic products and with carbon substrate assimilation. NH3 diffusion from surface to the cheese core during ripening was highly suspected. Interaction phenomena between microorganisms are discussed.

Characterisation of branched gluco-oligosaccharides to study the mode-of-action of a glucoamylase from Hypocrea jecorina.

PubMed

Jonathan, M C; van Brussel, M; Scheffers, M S; Kabel, M A

2015-11-05

In the conversion of starch to fermentable glucose for bioethanol production, hydrolysis of amylopectin by α-amylases and glucoamylases is the slowest step. In this process, α-1,6-branched gluco-oligosaccharides accumulate and are slowly degraded. Glucoamylases that are able to degrade such branched oligosaccharides faster are economically beneficial. This research aimed at the isolation and characterisation of branched gluco-oligosaccharides produced from amylopectin digestion by α-amylase, to be used as substrates for comparing their degradation by glucoamylases. Branched gluco-oligosaccharides with a DP between five and twelve were purified using size exclusion chromatography. These structures were characterised after labelling with 2-aminobenzamide using UHPLC-MS(n) analysis. Further, the purified oligosaccharides were used to evaluate the mode-of-action of a glucoamylase from Hypocrea jecorina. The enzyme cleaves the α-1,4-linkage adjacent to the α-1,6-linkage at a lower rate than that of α-1,4-linkages in linear oligosaccharides. Hence, the branched gluco-oligosaccharides are a suitable substrate to evaluate glucoamylase activity on branched structures. Copyright © 2015 Elsevier Ltd. All rights reserved.

Fungus-like mycelial fossils in 2.4-billion-year-old vesicular basalt.

PubMed

Bengtson, Stefan; Rasmussen, Birger; Ivarsson, Magnus; Muhling, Janet; Broman, Curt; Marone, Federica; Stampanoni, Marco; Bekker, Andrey

2017-04-24

Fungi have recently been found to comprise a significant part of the deep biosphere in oceanic sediments and crustal rocks. Fossils occupying fractures and pores in Phanerozoic volcanics indicate that this habitat is at least 400 million years old, but its origin may be considerably older. A 2.4-billion-year-old basalt from the Palaeoproterozoic Ongeluk Formation in South Africa contains filamentous fossils in vesicles and fractures. The filaments form mycelium-like structures growing from a basal film attached to the internal rock surfaces. Filaments branch and anastomose, touch and entangle each other. They are indistinguishable from mycelial fossils found in similar deep-biosphere habitats in the Phanerozoic, where they are attributed to fungi on the basis of chemical and morphological similarities to living fungi. The Ongeluk fossils, however, are two to three times older than current age estimates of the fungal clade. Unless they represent an unknown branch of fungus-like organisms, the fossils imply that the fungal clade is considerably older than previously thought, and that fungal origin and early evolution may lie in the oceanic deep biosphere rather than on land. The Ongeluk discovery suggests that life has inhabited submarine volcanics for more than 2.4 billion years.

Carbohydrate utilization and metabolism is highly differentiated in Agaricus bisporus

PubMed Central

2013-01-01

Background Agaricus bisporus is commercially grown on compost, in which the available carbon sources consist mainly of plant-derived polysaccharides that are built out of various different constituent monosaccharides. The major constituent monosaccharides of these polysaccharides are glucose, xylose, and arabinose, while smaller amounts of galactose, glucuronic acid, rhamnose and mannose are also present. Results In this study, genes encoding putative enzymes from carbon metabolism were identified and their expression was studied in different growth stages of A. bisporus. We correlated the expression of genes encoding plant and fungal polysaccharide modifying enzymes identified in the A. bisporus genome to the soluble carbohydrates and the composition of mycelium grown compost, casing layer and fruiting bodies. Conclusions The compost grown vegetative mycelium of A. bisporus consumes a wide variety of monosaccharides. However, in fruiting bodies only hexose catabolism occurs, and no accumulation of other sugars was observed. This suggests that only hexoses or their conversion products are transported from the vegetative mycelium to the fruiting body, while the other sugars likely provide energy for growth and maintenance of the vegetative mycelium. Clear correlations were found between expression of the genes and composition of carbohydrates. Genes encoding plant cell wall polysaccharide degrading enzymes were mainly expressed in compost-grown mycelium, and largely absent in fruiting bodies. In contrast, genes encoding fungal cell wall polysaccharide modifying enzymes were expressed in both fruiting bodies and vegetative mycelium, but different gene sets were expressed in these samples. PMID:24074284

Comparative Study of Worldwide Species of Genus Lentinus (=Lentinula, Higher Basidiomycetes) Based on Linear Mycelium Growth.

PubMed

Mata, Juan Luis; Mishra, Nutan Tulapurkar

2015-01-01

Species of mushroom genus Lentinus (=Lentinula) are best known for the commercially important and extensively studied culinary-medicinal shiitake, L. edodes. A few mycelium growth studies have focused on Lentinus boryana, but information is lacking for L. raphanica and L. aciculospora, endemic to the Americas. In this study, 14 dikaryon strains representing 5 Lentinus species were grown on 5 nutritive agar media at increments of 5°C. Growth for each species was significantly slower on corn meal agar, but no differences were found among malt extract, potato dextrose, malt peptone, and yeast malt extract agars. Lentinus aciculospora and L. boryana consistently exhibited the slowest mycelium growth rates among all species and across all temperatures tested, with optima at 15°C and 20°C. The fastest mycelium growth rates for L. edodes, L. novaezelandiae, and L. raphanica occurred at 25°C. Strains of the latter continued to grow well at 30°C, whereas growth of the other 2 species declined significantly. Differences in mycelium growth rates for American strains could be partially explained by their geographic locations, indicating that understanding this physiological parameter has important ramifications for the edible mushroom industry.

Spatial distribution of sporocarps of stipitate hydnoid fungi and their belowground mycelium.

PubMed

van der Linde, Sietse; Alexander, Ian J; Anderson, Ian C

2009-09-01

Interest in stipitate hydnoid fungi of the genera Bankera, Hydnellum, Phellodon and Sarcodon has increased due to the decline in numbers of sporocarps in Europe. Conservation of these fungi is hindered by a lack of understanding of their basic ecology. In particular, a better understanding of their belowground ecology is required. Real-time PCR in conjunction with spatially explicit sampling was used to quantify the relationship between sporocarps and mycelium of Hydnellum peckii and Phellodon tomentosus. Species-specific DNA of the target species was quantified in 100 soil samples collected on a 360 x 360 cm grid at five locations where sporocarps were present. All sporocarps within the grid and up to 2 m around the grid were mapped. Sporocarp production did not occur over the whole extent of the belowground mycelium of these two species, and mycelium extended up to 330 cm away from the immediate site of sporocarp production. Spatial analyses using Kernel-smoothing and Moran's I correlograms showed that, with a single exception, there was no quantitative relationship between sporocarp distribution and the belowground abundance of mycelium. These findings have important implications for the conservation of this rare group of fungi.

Carbohydrate composition of compost during composting and mycelium growth of Agaricus bisporus.

PubMed

Jurak, Edita; Kabel, Mirjam A; Gruppen, Harry

2014-01-30

Changes of plant cell wall carbohydrate structures occurring during the process to make suitable compost for growth of Agaricus bisporus are unknown. In this paper, composition and carbohydrate structures in compost samples collected during composting and mycelium growth were analyzed. Furthermore, different extracts of compost samples were prepared with water, 1M and 4M alkali and analyzed. At the beginning of composting, 34% and after 16 days of mycelium growth 27% of dry matter was carbohydrates. Carbohydrate composition analysis showed that mainly cellulose and poorly substituted xylan chains with similar amounts and ratios of xylan building blocks were present in all phases studied. Nevertheless, xylan solubility increased 20% over the period of mycelium growth indicating partial degradation of xylan backbone. Apparently, degradation of carbohydrates occurred over the process studied by both bacteria and fungi, mainly having an effect on xylan-chain length and solubility. Copyright © 2013 Elsevier Ltd. All rights reserved.

[Biosorption of heavy metals in fluoritum decoction by fungal mycelium].

PubMed

Cui, Pei-wu; Hu, Wei; Hu, Ya-qiang; Tan, Zhao-yang

2014-09-01

To explore the biosorption technology of heavy metals in Fluoritum decoction by fungal mycelium. Four factors including fungal mycelium amount, adsorption time, pH value and temperature were employed to estimate the fungal biomass adsorption conditions for removing the heavy metals in Fluoritum decoction. Then an orthogonal experimental design was taken to optimize the biosorption process, and the removal efficiency was also evaluated. Under the optimized conditions of 1.0 g/50 mL Fluoritum decoction, 3 hours adsorption time, pH 5.0 and 40 degrees C, a result of 70.12% heavy metals removal rate was accomplished with 35.99% calcium ion loss. The study indicates that removing of heavy metals in Fluoritum decoction through fungal mycelium is feasible, and the experiment results can also provide a basis for further research on biosorption of heavy metals in traditional Chinese medicine

Isolation and Purification of Two Isoflavones from Hericium erinaceum Mycelium by High-Speed Counter-Current Chromatography.

PubMed

He, Jinzhe; Fan, Peng; Feng, Simin; Shao, Ping; Sun, Peilong

2018-03-02

High-speed counter-current chromatography (HSCCC) was used to separate and purify two isoflavones for the first time from Hericium erinaceum ( H. erinaceum ) mycelium using a two-phase solvent system composed of chloroform-dichloromethane-methanol-water (4:2:3:2, v / v / v / v ). These two isoflavones were identified as genistein (4',5,7-trihydroxyisoflavone, C 15 H 10 O₅) and daidzein (4',7-dihydroxyisoflavone, C 15 H 10 O₄), using infrared spectroscopy (IR), electro-spary ionisation mass (ESI-MS), ¹H-nuclear magnetic resonance (NMR) and 13 C-NMR spectra. About 23 mg genistein with 95.7% purity and 18 mg daidzein with 97.3% purity were isolated from 150 mg ethanolic extract of H. erinaceum mycelium. The results demonstrated that HSCCC was a feasible method to separate and purify genistein and daidzein from H. erinaceum mycelium.

Structure of FabH and factors affecting the distribution of branched fatty acids in Micrococcus luteus.

PubMed

Pereira, Jose H; Goh, Ee-Been; Keasling, Jay D; Beller, Harry R; Adams, Paul D

2012-10-01

Micrococcus luteus is a Gram-positive bacterium that produces iso- and anteiso-branched alkenes by the head-to-head condensation of fatty-acid thioesters [coenzyme A (CoA) or acyl carrier protein (ACP)]; this activity is of interest for the production of advanced biofuels. In an effort to better understand the control of the formation of branched fatty acids in M. luteus, the structure of FabH (MlFabH) was determined. FabH, or β-ketoacyl-ACP synthase III, catalyzes the initial step of fatty-acid biosynthesis: the condensation of malonyl-ACP with an acyl-CoA. Analysis of the MlFabH structure provides insights into its substrate selectivity with regard to length and branching of the acyl-CoA. The most structurally divergent region of FabH is the L9 loop region located at the dimer interface, which is involved in the formation of the acyl-binding channel and thus limits the substrate-channel size. The residue Phe336, which is positioned near the catalytic triad, appears to play a major role in branched-substrate selectivity. In addition to structural studies of MlFabH, transcriptional studies of M. luteus were also performed, focusing on the increase in the ratio of anteiso:iso-branched alkenes that was observed during the transition from early to late stationary phase. Gene-expression microarray analysis identified two genes involved in leucine and isoleucine metabolism that may explain this transition.

Heterogenic expression of genes encoding secreted proteins at the periphery of Aspergillus niger colonies.

PubMed

Vinck, Arman; de Bekker, Charissa; Ossin, Adam; Ohm, Robin A; de Vries, Ronald P; Wösten, Han A B

2011-01-01

Colonization of a substrate by fungi starts with the invasion of exploring hyphae. These hyphae secrete enzymes that degrade the organic material into small molecules that can be taken up by the fungus to serve as nutrients. We previously showed that only part of the exploring hyphae of Aspergillus niger highly express the glucoamylase gene glaA. This was an unexpected finding since all exploring hyphae are exposed to the same environmental conditions. Using GFP as a reporter, we here demonstrate that the acid amylase gene aamA, the α-glucuronidase gene aguA, and the feruloyl esterase gene faeA of A. niger are also subject to heterogenic expression within the exploring mycelium. Coexpression studies using GFP and dTomato as reporters showed that hyphae that highly express one of these genes also highly express the other genes encoding secreted proteins. Moreover, these hyphae also highly express the amylolytic regulatory gene amyR, and the glyceraldehyde-3-phosphate dehydrogenase gene gpdA. In situ hybridization demonstrated that the high expressers are characterized by a high 18S rRNA content. Taken together, it is concluded that two subpopulations of hyphae can be distinguished within the exploring mycelium of A. niger. The experimental data indicate that these subpopulations differ in their transcriptional and translational activity. © 2010 Society for Applied Microbiology and Blackwell Publishing Ltd.

Color and Streptomycetes1

PubMed Central

Pridham, Thomas G.

1965-01-01

A report summarizing the results of an international workshop on determination of color of streptomycetes is presented. The results suggest that the color systems which seem most practically appealing and effective to specialists on actinomycetes are those embracing a limited number of color names and groups. The broad groupings allow placement of isolates into reasonably well-defined categories based on color of aerial mycelium. Attempts to expand such systems (more color groups) lead to difficulties. It is common knowledge that many, if not all, of the individual groups would in these broad systems contain strains that differ in many other respects, e.g., spore-wall ornamentation, color of vegetative (substratal) mycelium, morphology of chains of spores, and numerous physiological criteria. Also, cultures of intermediate color can be found, which makes placement difficult. As it now stands, color as a criterion for characterization of streptomycetes and streptoverticillia is in questionable status. Although much useful color information can be obtained by an individual, the application of this information to that in the literature or its use in communication with other individuals leaves much to be desired. More objective methods of color determination are needed. At present, the most effective method that could be used internationally is the color-wheel system of Tresner and Backus. Furthermore, the significance of color in speciation of these organisms is an open question. Obviously, more critical work on the color problem is needed. PMID:14264847

Scanning electron microscopy observations of the interaction between Trichoderma harzianum and perithecia of Gibberella zeae.

PubMed

Inch, S; Gilbert, J

2011-01-01

Chronological events associated with the interaction between a strain of Trichoderma harzianum, T472, with known biological control activity against perithecial production of G. zeae, were studied with scanning electron microscopy to investigate the mechanisms of control. Large clusters of perithecia consisting of 5-15 perithecia formed on the autoclaved, mulched wheat straw inoculated with G. zeae alone (control) with an average of 157 perithecia per plate. Small clusters consisting of 3-6 and an average of 15 perithecia per plate perithecia formed on straw that was treated with T. harzianum. The mature perithecia from straw treated with T. harzianum produced less pigment and were lighter in color than those from the control plates. Furthermore the cells of the outer wall of these perithecia were abnormal in appearance and unevenly distributed across the surface. Immature perithecia were colonized by T. harzianum approximately 15 d after inoculation (dai) with the biocontrol agent and pathogen. Few perithecia were colonized at later stages. The affected perithecia collapsed 21 dai, compared to the perithecia in the control samples that began to collapse 28 dai. Abundant mycelium of T. harzianum was seen on the perithecia of treated samples. Perithecial structures may be resistant to penetration by the mycelium because direct penetration was not observed. Trichoderma harzianum colonized the substrate quickly and out-competed the pathogen, G. zeae.

Classification of Nonomuraea sp. ATCC 39727, an actinomycete that produces the glycopeptide antibiotic A40926, as Nonomuraea gerenzanensis sp. nov.

PubMed

Dalmastri, Claudia; Gastaldo, Luciano; Marcone, Giorgia Letizia; Binda, Elisa; Congiu, Terenzio; Marinelli, Flavia

2016-02-01

Strain ATCC 39727, which produces the antibiotic A40926 (the natural precursor of the antibiotic dalbavancin), was isolated from a soil sample collected in India, and it was originally classified as a member of the genus Actinomadura on the base of morphology and cell-wall composition. A phylogenetic analysis based on 16S rRNA gene sequences indicates that the strain forms a distinct clade within the genus Nonomuraea, and it is most closely related to Nonomuraea angiospora DSM 43173T (98.72 % similarity) and Nonomuraea jabiensis A4036T (98.69 %). The strain forms an extensively branched substrate mycelium and aerial hyphae that form spiral chains of spores with ridged surfaces. The cell wall contains meso-diaminopimelic acid and the whole-cell sugars are glucose, ribose, galactose, mannose and madurose (madurose as the diagnostic sugar). The N-acyl type of muramic acid is acetyl. The predominant menaquinone is MK-9(H4), with minor amounts of MK-9(H2), MK-9(H6) and MK-9(H0). The polar-lipid profile includes diphosphatidylglycerol, phosphatidylethanolamine, hydroxyphosphatidylethanolamine, phosphatidylmethylethanolamine, hydroxyphosphatidylmethylethanolamine, phosphatidylinositol and a series of uncharacterized phospholipids, glycolipids and phosphoglycolipids. The major cellular fatty acids are iso-C16 : 0 and 10-methyl C17 : 0. The genomic DNA G+C content is 71.2 mol%. Significant differences in the morphological, chemotaxonomic and biochemical data, together with DNA-DNA relatedness between strain ATCC 39727 and closely related type strains, clearly demonstrated that strain ATCC 39727 represents a novel species of the genus Nonomuraea, for which the name Nonomuraea gerenzanensis sp. nov. is proposed. The type strain is ATCC 39727T ( = DSM 100948T).

Quantitative detection of Lactarius deliciosus extraradical soil mycelium by real-time PCR and its application in the study of fungal persistence and interspecific competition.

PubMed

Parladé, J; Hortal, S; Pera, J; Galipienso, L

2007-01-30

Real-Time PCR has been applied to quantify extraradical soil mycelium of the edible ectomycorrhizal fungus Lactarius deliciosus in an interspecific competition experiment under greenhouse conditions. Couples of Pinus pinea seedlings inoculated with either L. deliciosus, Rhizopogon roseolus, or non-inoculated (control) were transplanted into pots filled with two types of soil in all the possible combinations. Total DNA was extracted from soil samples at 3 and 6 months after transplantation to perform real-time PCR analysis. DNA extractions from soil mixed with known amounts of mycelium of L. deliciosus were used as standards. Six months after transplantation, the percentage of mycorrhizas of L. deliciosus and seedling growth were significantly affected by the soil type. Extraradical soil mycelium of L. deliciosus was positively correlated with the final percentage of mycorrhizas and significantly affected by the sampling time and soil depth. The competition effect of R. roseolus was not significant for any of the measured parameters, probably due to the sharp decrease of the mycorrhizal colonization by this fungus. We conclude that real-time PCR is a powerful technique for extraradical mycelium quantification in studies aimed at evaluating the persistence of introduced strains of L. deliciosus in field plantations.

A Broadband High-Gain Bi-Layer Log-Periodic Dipole Array (LPDA) for Ultra High Frequency (UHF) Conformal Load Bearing Antenna Structures (CLAS) Applications

DTIC Science & Technology

2014-08-01

AFRL-RQ-WP-TR-2014-0212 University of South Carolina Department of Electrical Engineering Columbia, SC 29208 Structures Technology Branch...S2603-04-C01. Cleared for Public Release - Case Number: . Nicholas Bishop and M. Ali are with the Department of Electrical Engineering, University of...Lower substrate Upper substrate Foam core Coax Feed tube LPDA traces Coax inner conductor Feed tube Copper plate Input 88ABW-2014-3668, 8

Protective Effects of Hericium erinaceus Mycelium and Its Isolated Erinacine A against Ischemia-Injury-Induced Neuronal Cell Death via the Inhibition of iNOS/p38 MAPK and Nitrotyrosine

PubMed Central

Lee, Kam-Fai; Chen, Jiann-Hwa; Teng, Chih-Chuan; Shen, Chien-Heng; Hsieh, Meng-Chiao; Lu, Chien-Chang; Lee, Ko-Chao; Lee, Li-Ya; Chen, Wan-Ping; Chen, Chin-Chu; Huang, Wen-Shih; Kuo, Hsing-Chun

2014-01-01

Hericium erinaceus, an edible mushroom, has been demonstrated to potentiate the effects of numerous biological activities. The aim of this study was to investigate whether H. erinaceus mycelium could act as an anti-inflammatory agent to bring about neuroprotection using a model of global ischemic stroke and the mechanisms involved. Rats were treated with H. erinaceus mycelium and its isolated diterpenoid derivative, erinacine A, after ischemia reperfusion brain injuries caused by the occlusion of the two common carotid arteries. The production of inflammatory cytokines in serum and the infracted volume of the brain were measured. The proteins from the stroke animal model (SAM) were evaluated to determine the effect of H. erinaceus mycelium. H. erinaceus mycelium reduced the total infarcted volumes by 22% and 44% at a concentration of 50 and 300 mg/kg, respectively, compared to the SAM group. The levels of acute inflammatory cytokines, including interleukin-1β, interleukin-6 and tumor necrosis factor á, were all reduced by erinacine A. Levels of nitrotyrosine-containing proteins, phosphorylation of p38 MAPK and CCAAT enhancer-binding protein (C/EBP) and homologous protein (CHOP) expression were attenuated by erinacine A. Moreover, the modulation of ischemia injury factors present in the SAM model by erinacine A seemed to result in the suppression of reactive nitrogen species and the downregulation of inducible NO synthase (iNOS), p38 MAPK and CHOP. These findings confirm the nerve-growth properties of Hericium erinaceus mycelium, which include the prevention of ischemic injury to neurons; this protective effect seems to be involved in the in vivo activity of iNOS, p38 MAPK and CHOP. PMID:25167134

Protective effects of Hericium erinaceus mycelium and its isolated erinacine A against ischemia-injury-induced neuronal cell death via the inhibition of iNOS/p38 MAPK and nitrotyrosine.

PubMed

Lee, Kam-Fai; Chen, Jiann-Hwa; Teng, Chih-Chuan; Shen, Chien-Heng; Hsieh, Meng-Chiao; Lu, Chien-Chang; Lee, Ko-Chao; Lee, Li-Ya; Chen, Wan-Ping; Chen, Chin-Chu; Huang, Wen-Shih; Kuo, Hsing-Chun

2014-08-27

Hericium erinaceus, an edible mushroom, has been demonstrated to potentiate the effects of numerous biological activities. The aim of this study was to investigate whether H. erinaceus mycelium could act as an anti-inflammatory agent to bring about neuroprotection using a model of global ischemic stroke and the mechanisms involved. Rats were treated with H. erinaceus mycelium and its isolated diterpenoid derivative, erinacine A, after ischemia reperfusion brain injuries caused by the occlusion of the two common carotid arteries. The production of inflammatory cytokines in serum and the infracted volume of the brain were measured. The proteins from the stroke animal model (SAM) were evaluated to determine the effect of H. erinaceus mycelium. H. erinaceus mycelium reduced the total infarcted volumes by 22% and 44% at a concentration of 50 and 300 mg/kg, respectively, compared to the SAM group. The levels of acute inflammatory cytokines, including interleukin-1β, interleukin-6 and tumor necrosis factor á, were all reduced by erinacine A. Levels of nitrotyrosine-containing proteins, phosphorylation of p38 MAPK and CCAAT enhancer-binding protein (C/EBP) and homologous protein (CHOP) expression were attenuated by erinacine A. Moreover, the modulation of ischemia injury factors present in the SAM model by erinacine A seemed to result in the suppression of reactive nitrogen species and the downregulation of inducible NO synthase (iNOS), p38 MAPK and CHOP. These findings confirm the nerve-growth properties of Hericium erinaceus mycelium, which include the prevention of ischemic injury to neurons; this protective effect seems to be involved in the in vivo activity of iNOS, p38 MAPK and CHOP.

Quality of Bread Supplemented with Antrodia
salmonea-Fermented Grains

PubMed Central

Chien, Rao-Chi; Ulziijargal, Enkhjargal

2016-01-01

Summary Fermented grains of buckwheat, oat, embryo rice and wheat, which were prepared by solid-state fermentation with Antrodia salmonea, and the mycelium was used to substitute 7% of wheat flour to make bread. No difference in proximate composition, texture profile and contents of non-volatile taste components was observed among bread samples. White bread and bread supplemented with mycelium and fermented grains looked different. Bread supplemented with fermented grains had similar thermal properties, which differed from those of white bread and bread supplemented with mycelium. Bread supplemented with fermented grains contained substantial mass fractions (on dry mass basis) of adenosine (0.92–1.96 µg/g), ergosterol (24.53–30.12 µg/g), ergothioneine (2.16–3.18 µg/g) and γ-aminobutyric acid (2.20–2.45 µg/g). In addition, bread supplemented with mycelium contained lovastatin (0.43 µg/g). White bread and bread supplemented with fermented grains had similar sensory results. Overall, fermented grains could be incorporated into bread to provide beneficial effects. PMID:27904408

Exposure of Metarhizium acridum mycelium to light induces tolerance to UV-B radiation.

PubMed

Brancini, Guilherme T P; Rangel, Drauzio E N; Braga, Gilberto Ú L

2016-03-01

Metarhizium acridum is an entomopathogenic fungus commonly used as a bioinsecticide. The conidium is the fungal stage normally employed as field inoculum in biological control programs and must survive under field conditions such as high ultraviolet-B (UV-B) exposure. Light, which is an important stimulus for many fungi, has been shown to induce the production of M. robertsii conidia with increased stress tolerance. Here we show that a two-hour exposure to white or blue/UV-A light of fast-growing mycelium induces tolerance to subsequent UV-B irradiation. Red light, however, does not have the same effect. In addition, we established that this induction can take place with as little as 1 min of white-light exposure. This brief illumination scheme could be relevant in future studies of M. acridum photobiology and for the production of UV-B resistant mycelium used in mycelium-based formulations for biological control. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Glutamic acid promotes monacolin K production and monacolin K biosynthetic gene cluster expression in Monascus.

PubMed

Zhang, Chan; Liang, Jian; Yang, Le; Chai, Shiyuan; Zhang, Chenxi; Sun, Baoguo; Wang, Chengtao

2017-12-01

This study investigated the effects of glutamic acid on production of monacolin K and expression of the monacolin K biosynthetic gene cluster. When Monascus M1 was grown in glutamic medium instead of in the original medium, monacolin K production increased from 48.4 to 215.4 mg l -1 , monacolin K production increased by 3.5 times. Glutamic acid enhanced monacolin K production by upregulating the expression of mokB-mokI; on day 8, the expression level of mokA tended to decrease by Reverse Transcription-polymerase Chain Reaction. Our findings demonstrated that mokA was not a key gene responsible for the quantity of monacolin K production in the presence of glutamic acid. Observation of Monascus mycelium morphology using Scanning Electron Microscope showed glutamic acid significantly increased the content of Monascus mycelium, altered the permeability of Monascus mycelium, enhanced secretion of monacolin K from the cell, and reduced the monacolin K content in Monascus mycelium, thereby enhancing monacolin K production.

Influence of culture medium growth variables on Ganoderma lucidum exopolysaccharides structural features.

PubMed

Fraga, Irene; Coutinho, João; Bezerra, Rui M; Dias, Albino A; Marques, Guilhermina; Nunes, Fernando M

2014-10-13

In this work the effect of carbon and nitrogen levels and initial pH of the wheat extract culture medium of submerged culture of Ganoderma lucidum on the amount, purity and structural features of exopolysaccharides (EPS) were studied. A low peptone level (1.65 g L(-1)) favored mycelium biomass, EPS purity, but a higher supply of peptone (4.80 g L(-1)) is needed for maximum EPS production. The carbohydrate composition of the EPS and structural features also changed significantly according to the different growing conditions, being observed significant differences in the (1 → 3)/(1 → 4)-Glcp ratio and also on the branching degree of EPS. As the biological activities of EPS are highly dependent on the polysaccharide structural features, this variability can have implications on the EPS biological activities, but can also be used advantageously to produce tailor made polysaccharides with specific applications. Copyright © 2014 Elsevier Ltd. All rights reserved.

Three New Soil-inhabiting Species of Trichoderma in the Stromaticum Clade with Test of Their Antagonism to Pathogens.

PubMed

Chen, Kai; Zhuang, Wen-Ying

2017-09-01

Trichoderma is a dominant component of the soil mycoflora. During the field investigations of northern, central, and southwestern China, three new species in the Stromaticum clade were encountered from soil, and named as T. hebeiense, T. sichuanense, and T. verticillatum. Their phylogenetic positions were determined by analyses of the combined two genes: partial sequences of translation elongation factor 1-alpha and the second largest RNA polymerase subunit-encoding genes. Distinctions between the new species and their close relatives were discussed. Trichoderma hebeiense appeared as a separate terminal branch. The species is distinctive by its oblong conidia and aggregated pustules in culture. Trichoderma sichuanense features in concentric colony and produces numerous clean exudates on aerial mycelium in culture. Trichoderma verticillatum is characterized by its verticillium-like synanamorph and production of abundant chlamydospores. In vitro antagonism towards the new species was tested by dual culture technique.

Mycoparasitism studies of Trichoderma harzianum against Sclerotinia sclerotiorum: evaluation of antagonism and expression of cell wall-degrading enzymes genes.

PubMed

Troian, Rogério Fraga; Steindorff, Andrei Stecca; Ramada, Marcelo Henrique Soller; Arruda, Walquiria; Ulhoa, Cirano José

2014-10-01

Trichoderma spp. are known for their biocontrol activity against several plant pathogens. A specific isolate of Trichoderma harzianum, 303/02, has the potential to inhibit the growth of Sclerotinia sclerotiorum, an important agent involved in several crop diseases. In this study, the interaction between T. harzianum 303/02 and mycelia, sclerotia and apothecia of S. sclerotiorum was studied by scanning electron microscopy. RT-qPCR was used to examine the expression of 11 genes potentially involved in biocontrol. T. harzianum 303/02 parasitizes S. sclerotiorum by forming branches that coil around the hyphae. The fungus multiplied abundantly at the sclerotia and apothecia surface, forming a dense mycelium that penetrated the inner surface of these structures. The levels of gene expression varied according to the type of structure with which T. harzianum was interacting. The data also showed the presence of synergistic action between the cell-wall degrading enzymes.

Alteration in the ultrastructural morphology of mycelial hyphae and the dynamics of transcriptional activity of lytic enzyme genes during basidiomycete morphogenesis.

PubMed

Vetchinkina, Elena; Kupryashina, Maria; Gorshkov, Vladimir; Ageeva, Marina; Gogolev, Yuri; Nikitina, Valentina

2017-04-01

The morphogenesis of macromycetes is a complex multilevel process resulting in a set of molecular-genetic, physiological-biochemical, and morphological-ultrastructural changes in the cells. When the xylotrophic basidiomycetes Lentinus edodes, Grifola frondosa, and Ganoderma lucidum were grown on wood waste as the substrate, the ultrastructural morphology of the mycelial hyphal cell walls differed considerably between mycelium and morphostructures. As the macromycetes passed from vegetative to generative development, the expression of the tyr1, tyr2, chi1, chi2, exg1, exg2, and exg3 genes was activated. These genes encode enzymes such as tyrosinase, chitinase, and glucanase, which play essential roles in cell wall growth and morphogenesis.

Patterns of diffusibility of lignin and carbohydrate degrading systems in wood-rotting fungi

DOE Office of Scientific and Technical Information (OSTI.GOV)

Rosenberg, S.L.

1979-09-01

In an attempt to identify organisms that produce diffusible lignin-degrading systems, a culturing apparatus was constructed which contained two compartments separated by a bacteriological membrane filter. Lignin-degrading fungi were grown with lignocellulose in one compartment, and diffusion channels were maintained through the membrane to sterile lignocellulose in the adjoining compartment. For the fungi tested, both lignin and carbohydrate were degraded when the mycelium and the substrate were in physical contact, but only carbohydrate was degraded significantly in the adjoining compartment containing sterile lignocellulose. Two organisms, Coriolus versicolor and Trichoderma reesii QM 9414 displayed slight diffusible lignin-degrading activity. Some fungi producedmore » more diffusible carbohydrate-degrading activity than others.« less

Intraspecific ploidy variation: A hidden, minor player in plant-soil-mycorrhizal fungi interactions.

PubMed

Sudová, Radka; Pánková, Hana; Rydlová, Jana; Münzbergová, Zuzana; Suda, Jan

2014-01-01

Genome duplication and arbuscular mycorrhizal (AM) symbiosis are ubiquitous in angiosperms. While the significance of each of these phenomena separately has been intensively studied, their interaction remains to be understood. Three diploid and three hexaploid populations of Aster amellus (Asteraceae) were characterized in terms of the soil conditions in situ and mycorrhizal root colonization. In a greenhouse experiment, the effects of ploidy level, substrate conditions, and AM fungi on plant performance were then separated by growing noninoculated plants or plants inoculated with AM fungi in substrates native to either the diploids or hexaploids. The diploids inhabited nutritionally richer sites but did not differ from hexaploid plants in the level of mycorrhizal root colonization in situ. In the experiment, hexaploids generally performed better than the diploids. This intercytotype growth difference was enhanced by soil fertility, with hexaploids benefiting more from nutritionally richer substrate than the diploids. AM inoculation was crucial for plant growth and phosphorus uptake. The interaction between ploidy level and AM inoculation significantly influenced only dry mass of roots, phosphorus concentrations in shoot biomass, and the length of the extraradical mycelium in the nonsterile substrates. Our results support the idea that polyploidy can affect the mycorrhizal growth response of host plants. Nevertheless, the effects of the interaction between ploidy and inoculation were weaker than the main effects of these factors.

Improved deoxyribozymes for synthesis of covalently branched DNA and RNA.

PubMed

Lee, Christine S; Mui, Timothy P; Silverman, Scott K

2011-01-01

A covalently branched nucleic acid can be synthesized by joining the 2'-hydroxyl of the branch-site ribonucleotide of a DNA or RNA strand to the activated 5'-phosphorus of a separate DNA or RNA strand. We have previously used deoxyribozymes to synthesize several types of branched nucleic acids for experiments in biotechnology and biochemistry. Here, we report in vitro selection experiments to identify improved deoxyribozymes for synthesis of branched DNA and RNA. Each of the new deoxyribozymes requires Mn²(+) as a cofactor, rather than Mg²(+) as used by our previous branch-forming deoxyribozymes, and each has an initially random region of 40 rather than 22 or fewer combined nucleotides. The deoxyribozymes all function by forming a three-helix-junction (3HJ) complex with their two oligonucleotide substrates. For synthesis of branched DNA, the best new deoxyribozyme, 8LV13, has k(obs) on the order of 0.1 min⁻¹, which is about two orders of magnitude faster than our previously identified 15HA9 deoxyribozyme. 8LV13 also functions at closer-to-neutral pH than does 15HA9 (pH 7.5 versus 9.0) and has useful tolerance for many DNA substrate sequences. For synthesis of branched RNA, two new deoxyribozymes, 8LX1 and 8LX6, were identified with broad sequence tolerances and substantial activity at pH 7.5, versus pH 9.0 for many of our previous deoxyribozymes that form branched RNA. These experiments provide new, and in key aspects improved, practical catalysts for preparation of synthetic branched DNA and RNA.

ANALYSIS AND EVALUATION OF MYCELIUM REINFORCED NATURAL FIBER BIO-COMPOSITES

USDA-ARS?s Scientific Manuscript database

There is a need for biodegradable alternatives to the inert plastics and expanded foams that are common in both the manufacturing process and device componentry. The material in this study is a bio-composite patented by Ecovative Design LLC. The manufacturer's bio-composite utilizes fungal mycelium ...

Substrate-induced inactivation of the OXA2 beta-lactamase.

PubMed Central

Ledent, P; Frère, J M

1993-01-01

The hydrolysis time courses of 22 beta-lactam antibiotics by the class D OXA2 beta-lactamase were studied. Among these, only three appeared to correspond to the integrated Henri-Michaelis equation. 'Burst' kinetics, implying branched pathways, were observed with most penicillins, cephalosporins and with flomoxef and imipenem. Kinetic parameters characteristic of the different phases of the hydrolysis were determined for some substrates. Mechanisms generally accepted to explain such reversible partial inactivations involving branches at either the free enzyme or the acyl-enzyme were inadequate to explain the enzyme behaviour. The hydrolysis of imipenem was characterized by the occurrence of two 'bursts', and that of nitrocefin by a partial substrate-induced inactivation complicated by a competitive inhibition by the hydrolysis product. PMID:8240304

Evaluation of the infection process by Lecanicillium fungicola in Agaricus bisporus by scanning electron microscopy.

PubMed

Santana Nunes, Janaira; Rocha de Brito, Manuela; Cunha Zied, Diego; Aparecida das Graças Leite, Eloisa; Souza Dias, Eustáquio; Alves, Eduardo

Lecanicillium fungicola causes dry bubble disease in Agaricus bisporus mushrooms leading to significant economic losses in commercial production. To monitor the infection process of L. fungicola in Brazilian strains of A. bisporus. The interaction between the mycelium of L. fungicola (LF.1) and three strains of A. bisporus (ABI 7, ABI 11/14 and ABI 11/21) was studied. Electron microscopy and X-ray microanalyses of vegetative growth and basidiocarp infection were evaluated. Micrographs show that the vegetative mycelium of the Brazilian strains of A. bisporus is not infected by the parasite. The images show that the pathogen can interlace the hyphae of A. bisporus without causing damage, which contributes to the presence of L. fungicola during the substrate colonization, allowing their presence during primordial formation of A. bisporus. In the basidiocarp, germ tubes form within 16h of infection with L. fungicola and the beginning of penetration takes place within 18h, both without the formation of specialized structures. Scanning electron microscopy enabled the process of colonization and reproduction to be observed within the formation of phialides, conidiophores and verticils of L. fungicola. The formation of calcium oxalate crystals by the pathogen was also visible using the X-ray microanalysis, both at the hyphae in the Petri plate and at basidiocarp infection site. Copyright © 2016 Asociación Española de Micología. Publicado por Elsevier España, S.L.U. All rights reserved.

Electrical stimulation in white oyster mushroom (Pleurotus florida) production

NASA Astrophysics Data System (ADS)

Roshita, I.; Nurfazira, K. M. P.; Fern, C. Shi; Ain, M. S. Nur

2017-09-01

White oyster mushroom (Pleurotus florida) is an edible mushroom that gained popularity due to its nutritional values, low production cost and ease of cultivation. There are several research reported on the mushroom fruiting bodies which were actively developed when applying electrical shock treatment. This study was aimed to investigate the effects of different electrical voltages on the growth and yield of white oyster mushroom (Pleurotus florida). Five different electrical voltages had been applied during spawning period which were 6V, 9V, 12V, 15V and mushroom bags without any treatment served as control. Treatment at 6V showed the highest rate for mycelium growth while 15V took the shortest time for fruiting body formation. However, no significant different (P>0.05) among all the treatments was observed for the time taken for the mycelium to fill-up the bag and pinhead emergence. The total fresh weight and percentage of biological efficiency for treatment at 9V showed higher values compared to control. Treatment at 9V also showed the largest pileus diameter and the most firm in the pileus texture. Meanwhile, treatment at 6V showed the highest a* value (redness). In addition, different electrical voltage treatments applied did not show any significant effect on substrate utilization efficiency, colour L* and b* values. In conclusion, among all the electrical treatments applied, 9V could be considered as the best treatment to enhance the yield of white oyster mushroom.

Simulated nitrogen deposition affects wood decomposition by cord-forming fungi.

PubMed

Bebber, Daniel P; Watkinson, Sarah C; Boddy, Lynne; Darrah, Peter R

2011-12-01

Anthropogenic nitrogen (N) deposition affects many natural processes, including forest litter decomposition. Saprotrophic fungi are the only organisms capable of completely decomposing lignocellulosic (woody) litter in temperate ecosystems, and therefore the responses of fungi to N deposition are critical in understanding the effects of global change on the forest carbon cycle. Plant litter decomposition under elevated N has been intensively studied, with varying results. The complexity of forest floor biota and variability in litter quality have obscured N-elevation effects on decomposers. Field experiments often utilize standardized substrates and N-levels, but few studies have controlled the decay organisms. Decomposition of beech (Fagus sylvatica) blocks inoculated with two cord-forming basidiomycete fungi, Hypholoma fasciculare and Phanerochaete velutina, was compared experimentally under realistic levels of simulated N deposition at Wytham Wood, Oxfordshire, UK. Mass loss was greater with P. velutina than with H. fasciculare, and with N treatment than in the control. Decomposition was accompanied by growth of the fungal mycelium and increasing N concentration in the remaining wood. We attribute the N effect on wood decay to the response of cord-forming wood decay fungi to N availability. Previous studies demonstrated the capacity of these fungi to scavenge and import N to decaying wood via a translocating network of mycelium. This study shows that small increases in N availability can increase wood decomposition by these organisms. Dead wood is an important carbon store and habitat. The responses of wood decomposers to anthropogenic N deposition should be considered in models of forest carbon dynamics.

Application of solid-phase extraction to agar-supported fermentation.

PubMed

Le Goff, Géraldine; Adelin, Emilie; Cortial, Sylvie; Servy, Claudine; Ouazzani, Jamal

2013-09-01

Agar-supported fermentation (Ag-SF), a variant of solid-state fermentation, has recently been improved by the development of a dedicated 2 m(2) scale pilot facility, Platotex. We investigated the application of solid-phase extraction (SPE) to Ag-SF in order to increase yields and minimize the contamination of the extracts with agar constituents. The selection of the appropriate resin was conducted on liquid-state fermentation and Diaion HP-20 exhibited the highest recovery yield and selectivity for the metabolites of the model fungal strains Phomopsis sp. and Fusarium sp. SPE applied to Ag-SF resulted in a particular compartmentalization of the culture. The mycelium that requires oxygen to grow migrates to the top layer and formed a thick biofilm. The resin beads intercalate between the agar surface and the mycelium layer, and trap directly the compounds secreted by the mycelium through a "solid-solid extraction" (SSE) process. The resin/mycelium layer is easily recovered by scraping the surface and the target metabolites extracted by methanol. Ag-SF associated to SSE represents an ideal compromise for the production of bioactive secondary metabolites with limited economic and environmental impact.

Enhancement of sludge dewaterability with filamentous fungi Talaromyces flavus S1 by depletion of extracellular polymeric substances or mycelium entrapment.

PubMed

Liu, He; Shi, Jiasheng; Xu, Xiaoyu; Zhan, Xinmin; Fu, Bo; Li, Yifei

2017-12-01

This study was conducted to explore the mechanism of dewaterability improvement of waste activated sludge by the filamentous fungus Talaromyces flavus S1. When the fungal spores were inoculated to the sterilized sludge, the sludge dewaterability was significantly improved by 48.1% and the reasons can be attributed to sludge pellet formation and degradation of extracellular polymeric substances, in particular the slime-EPS and loosely-bound EPS (LB-EPS). With the addition of fungal mycelium into the either sterilized sludge or non-sterilized sludge, the values of CST decreased by 74.0% and 43.7%, respectively, suggesting the fungal mycelium can improve the sludge dewaterability. After conditioned by the mycelium, the sludge cake by the diaphragm filter press was thicker and showed less water content than the control sludge. The results in this study demonstrated that the Talaromyces flavus S1 can serve as an environmentally friendly biological dewatering agent and has a promising application potential in the future. Copyright © 2017 Elsevier Ltd. All rights reserved.

Optimization of laccase production by Trametes versicolor cultivated on industrial waste.

PubMed

Tišma, Marina; Znidaršič-Plazl, Polona; Vasić-Rački, Durđa; Zelić, Bruno

2012-01-01

Laccases are very interesting biocatalysts for several industrial applications. Its production by different white-rot fungi can be stimulated by a variety of inducing substrates, and the use of lignocellulosic wastes or industrial by-products is one of the possible approaches to reduce production costs. In this work, various industrial wastes were tested for laccase production by Trametes versicolor MZKI G-99. Solid waste from chemomechanical treatment facility of a paper manufacturing plant showed the highest potential for laccase production. Enzyme production during submerged cultivation of T. versicolor on the chosen industrial waste has been further improved by medium optimization using genetic algorithm. Concentrations of five components in the medium were optimized within 60 shake-flasks experiments, where the highest laccase activity of 2,378 U dm(-3) was achieved. Waste from the paper industry containing microparticles of CaCO(3) was found to stimulate the formation of freely dispersed mycelium and laccase production during submerged cultivation of T. versicolor. It was proven to be a safe and inexpensive substrate for commercial production of laccase and might be more widely applicable for metabolite production by filamentous fungi.

Activity of Extracts from Submerged Cultured Mycelium of Winter Mushroom, Flammulina velutipes (Agaricomycetes), on the Immune System In Vitro.

PubMed

Kashina, Svetlana; Villavicencio, Lerida Liss Flores; Zaina, Silvio; Ordaz, Marco Balleza; Sabanero, Gloria Barbosa; Fujiyoshi, Victor Tsutsumi; Lopez, Myrna Sabanero

2016-01-01

Extracts from submerged cultured mycelium of two strains of Flammulina velutipes, a popular culinary mushroom, were obtained by ultrasound and tested in vitro to determine their activity in innate immunity (monocytes/ macrophages). In addition, polyclonal antibodies against the extracts were produced. Both extracts have similar glycoproteins that contain mannose and glucose but have different glycoproteins with galactoseamine units. Two novel immunogenic glycoproteins with molecular weights of 32 and 25 kDa have been revealed. It is thought that these proteins are produced only by submerged cultured mycelium. Both extracts show immune-enhancing activity based on the significant modification of various parameters such as cytokine production, phagocytosis, and reactive oxygen species production.

Production of laccase by Coriolus versicolor and its application in decolorization of dyestuffs: (II). Decolorization of dyes by laccase containing fermentation broth with or without self-immobilized mycelia.

PubMed

Lin, Jian-Ping; Lian, Wei; Xia, Li-Ming; Cen, Pei-Lin

2003-01-01

The capability of decolorization for commercial dyes by Coriolus versicolor fermentation broth containing laccase with or without immobilized mycelium was evaluated. With cell-free fermentation broth containing laccase, high decolorization ratio was achieved foracid orange 7, but not for the other dyes concerned. The immobilized mycelium was proved to be more efficient than the cell-free system. All the four dyestuffs studied were found being decolourized with certain extent by immobilized mycelium. The repeated-batch decolorization was carried out with satisfactory results. The experimental data showed that the continuous decolorization of wastewater from a printing and dyeing industry was possible by using the self-immobilized C. versicolor.

Mushroom as a product and their role in mycoremediation

PubMed Central

2014-01-01

Mushroom has been used for consumption as product for a long time due to their flavor and richness in protein. Mushrooms are also known as mycoremediation tool because of their use in remediation of different types of pollutants. Mycoremediation relies on the efficient enzymes, produced by mushroom, for the degradation of various types of substrate and pollutants. Besides waste degradation, mushroom produced a vendible product for consumption. However, sometimes they absorb the pollutant in their mycelium (biosorption process) and cannot be consumed due to absorbed toxicants. This article reviews the achievement and current status of mycoremediation technology based on mushroom cultivation for the remediation of waste and also emphasizes on the importance of mushroom as product. This critical review is also focused on the safety aspects of mushroom cultivation on waste. PMID:24949264

Dynamics of Penicillium camemberti growth quantified by real-time PCR on Camembert-type cheeses under different conditions of temperature and relative humidity.

PubMed

Leclercq-Perlat, Marie-Noëlle; Picque, Daniel; Martin Del Campo Barba, Sandra Teresita; Monnet, Christophe

2013-06-01

Penicillium camemberti plays a major role in the flavor and appearance of Camembert-type cheeses. However, little is known about its mycelium growth kinetics during ripening. We monitored the growth of P. camemberti mycelium in Camembert-type cheeses using real-time PCR in 4 ripening runs, performed at 2 temperatures (8 and 16°C) and 2 relative humidities (88 and 98%). These findings were compared with P. camemberti quantification by spore concentration. During the first phase, the mycelium grew but no spores were produced, regardless of the ripening conditions. During the second phase, which began when lactose was depleted, the concentration of spores increased, especially in the cheeses ripened at 16°C. Sporulation was associated with a large decrease in the mycelial concentration in the cheeses ripened at 16°C and 98% relative humidity. It was hypothesized that lactose is the main energy source for the growth of P. camemberti mycelium at the beginning of ripening and that its depletion would trigger stress, resulting in sporulation. Copyright © 2013 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Enhancing growth and yield of grey oyster mushroom (Pleurotus sajor-caju) using sound treatment at different intervals

NASA Astrophysics Data System (ADS)

Roshita, I.; Mukhlis, A. Amir; Ain, M. S. Nur; Fern, C. Shi; Zarina, Z.

2017-09-01

Mushrooms are famous for their use as source of nutrient and medicinal purposes. Wild mushrooms grew in a large number in the nature after a heavy down pour. Some believes that the thunderstorm and lightning can have effects on the growth of mushrooms. Hence, this study was conducted to investigate the impact of different acoustic sound treatment intervals towards the growth of grey oyster mushroom (Pleurotus sajor-caju). Five different sound treatment intervals involved which were; no treatment (control), 5-day, 10-day, 15-day and 20 day. The variables investigated were mycelium growth rate, growth stage performance (durations for mycelium filling up the bags, pinhead emergence and fruiting bodies formation), yield (number of fruiting bodies, total weight of fruiting bodies and percentage of biological efficiency) and physical analyses (pileus size, colour and texture). There were significant differences (P<0.05) observed in the mycelium growth rate, mycelium filling up the bags and number of fruiting bodies formation among different treatment intervals. As conclusion, the sound treated at different intervals have significant impact on the growth and yield of mushroom production where treatment at 5-day intervals was found to be the best treatment interval.

Mycelium growth stimulation of the desert truffle Terfezia claveryi chatin by β-cyclodextrin.

PubMed

López-Nicolás, José Manuel; Pérez-Gilabert, Manuela; García-Carmona, Francisco; Lozano-Carrillo, María Cecilia; Morte, Asunción

2013-01-01

The commercial value of Terfezia claveryi, an edible desert truffle with important gastronomic, nutritional, and antioxidant properties, has led to growing interest in its cultivation. The erratic and slow growth of T. claveryi mycelium in vitro represents an impairment to obtain mycorrhizal plants, and it makes necessary to find a new culture medium able to overcome these drawbacks. In this work, we analyze the effect of cyclodextrins (CDs) on the growth of T. claveryi mycelium. Different parameters, including colony diameter, growth rate, and colony fresh weight, were evaluated, both in the presence and absence of these encapsulant agents. The results obtained confirm the ability of CDs to stimulate the growth of T. claveryi mycelium when present in the culture medium. A similar effect was observed when CDs were added to the culture medium of Tuber melanosporum. Three natural (α-, β-, and γ) and two modified (hydroxypropil-β and methyl-β) CDs were assayed. The best results were obtained with β-cyclodextrin, but no improvement was observed with its chemically modified derivatives. CDs complex the different compounds present in the culture medium which impair mycelial growth. © 2013 American Institute of Chemical Engineers.

Ultrastructural changes in the mycelium of Hericium erinaceum (Bull.; Fr.) Pers. under selenium-induced oxidative stress.

PubMed

Ślusarczyk, Joanna; Kuraś, Mieczysław; Malinowska, Eliza; Skalicka-Woźniak, Krystyna; Głowniak, Kazimierz

2014-10-01

In this study we examined the influence of various forms of selenium (organic and inorganic) on the vivacity of Hericium erinaceum mycelium and structural changes and ultrastructure occurring during its development in submerged culture. The mycelium was grown on sodium selenite (Na₂SeO₃), Selol (with 20 and 50 g kg⁻¹ Se, respectively) and a mixture of Na₂SeO₃ and Selol. Samples of the mycelium were collected on day 3 and day 24 of the incubation and viewed under an electron microscope. Selol at concentration 20 g kg⁻¹ did not cause any damage to the cell ultrastructure, but it contributed to the thickening of the cell wall, which implied an influence on polysaccharide production. In the other cases, degradation changes appeared in the protoplasm and the thickness of the cell wall did not increase. The nature of the effect exerted by various sources of selenium in the culture medium on the formation of polysaccharides probably results from the differences in their chemical composition and differences in the toxicity of these compounds towards the cells, but is also connected with the decomposition of the wall surrounding degraded fungal cells. © 2014 Society of Chemical Industry.

Crystal structure of full-length Mycobacterium tuberculosis H37Rv glycogen branching enzyme: insights of N-terminal beta-sandwich in substrate specificity and enzymatic activity.

PubMed

Pal, Kuntal; Kumar, Shiva; Sharma, Shikha; Garg, Saurabh Kumar; Alam, Mohammad Suhail; Xu, H Eric; Agrawal, Pushpa; Swaminathan, Kunchithapadam

2010-07-02

The open reading frame Rv1326c of Mycobacterium tuberculosis (Mtb) H37Rv encodes for an alpha-1,4-glucan branching enzyme (MtbGlgB, EC 2.4.1.18, Uniprot entry Q10625). This enzyme belongs to glycoside hydrolase (GH) family 13 and catalyzes the branching of a linear glucose chain during glycogenesis by cleaving a 1-->4 bond and making a new 1-->6 bond. Here, we show the crystal structure of full-length MtbGlgB (MtbGlgBWT) at 2.33-A resolution. MtbGlgBWT contains four domains: N1 beta-sandwich, N2 beta-sandwich, a central (beta/alpha)(8) domain that houses the catalytic site, and a C-terminal beta-sandwich. We have assayed the amylase activity with amylose and starch as substrates and the glycogen branching activity using amylose as a substrate for MtbGlgBWT and the N1 domain-deleted (the first 108 residues deleted) MtbDelta108GlgB protein. The N1 beta-sandwich, which is formed by the first 105 amino acids and superimposes well with the N2 beta-sandwich, is shown to have an influence in substrate binding in the amylase assay. Also, we have checked and shown that several GH13 family inhibitors are ineffective against MtbGlgBWT and MtbDelta108GlgB. We propose a two-step reaction mechanism, for the amylase activity (1-->4 bond breakage) and isomerization (1-->6 bond formation), which occurs in the same catalytic pocket. The structural and functional properties of MtbGlgB and MtbDelta108GlgB are compared with those of the N-terminal 112-amino acid-deleted Escherichia coli GlgB (ECDelta112GlgB).

Aligned Carbon Nanotube Carpets on Carbon Substrates for High Power Electronic Applications

DTIC Science & Technology

2016-06-01

SiOx by a vapor-solid-solid mechanism ,” J. Am. Chem. Soc., vol. 133, pp. 197–199, 2011. [146] B. Liu, W. Ren, C. Liu, C.-H. Sun , L. Gao, S. Li, C... Mechanical and Thermal Systems Branch Power and Control Division JUNE 2016 Interim Report DISTRIBUTION STATEMENT A: Approved for public release...Advisor Program Engineer Mechanical and Thermal Systems Branch Mechanical and Thermal Systems Branch Power and Control Division Power and Control

Branching out in locomotion: the mechanics of perch use in birds and primates.

PubMed

Bonser, R H

1999-06-01

Many animals use thin perches, such as the branches of trees, as locomotory substrates. In this paper, I have reviewed the literature concerned with measurements of locomotory forces made by birds and primates on thin and flexible substrates. Through a knowledge of the locomotory forces exerted by animals when using different substrates, the mechanical cost of their use can be established. We are just beginning to learn about the magnitude and patterns of force production in various branch-using vertebrates, primarily as a result of the development of instrumented perches. Instrumented perches have been designed to measure the forces produced by birds and primates when leaping from rigid and flexible horizontal and flexible vertical perches, and also from instrumented handgrips during brachiation. The development of these techniques for birds and primates allows us to compare the way in which they use perches as locomotory substrates. In both birds and primates, the magnitudes of landing forces are smaller than those during take-off. Two explanations have been proposed; the difference is either a consequence of perch compliance or it is a strategic decision to be cautious of 'new' perches. Leaps from flexible perches may be somewhat inefficient because considerable energy is dissipated in bending the perch, and this energy may remain unrecovered when the animal leaves contact with the perch.

RADIOLYSIS OF ORGANIC COMPOUNDS IN THE ADSORBED STATE

DOEpatents

Sutherland, J.W.; Allen, A.O.

1961-10-01

>A method of forming branch chained hydrocarbons by means of energetic penetrating radiation is described. A solid zeolite substrate is admixed with a cobalt ion and is irradiated with a hydrocarbon adsorbed therein. Upon irradiation with gamma rays, there is an increased yield of branched and lower molecular straight chain compounds. (AEC)

A Novel Branching Enzyme of the GH-57 Family in the Hyperthermophilic Archaeon Thermococcus kodakaraensis KOD1

PubMed Central

Murakami, Taira; Kanai, Tamotsu; Takata, Hiroki; Kuriki, Takashi; Imanaka, Tadayuki

2006-01-01

Branching enzyme (BE) catalyzes formation of the branch points in glycogen and amylopectin by cleavage of the α-1,4 linkage and its subsequent transfer to the α-1,6 position. We have identified a novel BE encoded by an uncharacterized open reading frame (TK1436) of the hyperthermophilic archaeon Thermococcus kodakaraensis KOD1. TK1436 encodes a conserved protein showing similarity to members of glycoside hydrolase family 57 (GH-57 family). At the C terminus of the TK1436 protein, two copies of a helix-hairpin-helix (HhH) motif were found. TK1436 orthologs are distributed in archaea of the order Thermococcales, cyanobacteria, some actinobacteria, and a few other bacterial species. When recombinant TK1436 protein was incubated with amylose used as the substrate, a product peak was detected by high-performance anion-exchange chromatography, eluting more slowly than the substrate. Isoamylase treatment of the reaction mixture significantly increased the level of short-chain α-glucans, indicating that the reaction product contained many α-1,6 branching points. The TK1436 protein showed an optimal pH of 7.0, an optimal temperature of 70°C, and thermostability up to 90°C, as determined by the iodine-staining assay. These properties were the same when a protein devoid of HhH motifs (the TK1436ΔH protein) was used. The average molecular weight of branched glucan after reaction with the TK1436ΔH protein was over 100 times larger than that of the starting substrate. These results clearly indicate that TK1436 encodes a structurally novel BE belonging to the GH-57 family. Identification of an overlooked BE species provides new insights into glycogen biosynthesis in microorganisms. PMID:16885460

Re-engineering specificity in 1,3-1, 4-β-glucanase to accept branched xyloglucan substrates.

PubMed

Addington, Trevor; Calisto, Barbara; Alfonso-Prieto, Mercedes; Rovira, Carme; Fita, Ignasi; Planas, Antoni

2011-02-01

Family 16 carbohydrate active enzyme members Bacillus licheniformis 1,3-1,4-β-glucanase and Populus tremula x tremuloides xyloglucan endotransglycosylase (XET16-34) are highly structurally related but display different substrate specificities. Although the first binds linear gluco-oligosaccharides, the second binds branched xylogluco-oligosaccharides. Prior engineered nucleophile mutants of both enzymes are glycosynthases that catalyze the condensation between a glycosyl fluoride donor and a glycoside acceptor. With the aim of expanding the glycosynthase technology to produce designer oligosaccharides consisting of hybrids between branched xylogluco- and linear gluco-oligosaccharides, enzyme engineering on the negative subsites of 1,3-1,4-β-glucanase to accept branched substrates has been undertaken. Removal of the 1,3-1,4-β-glucanase major loop and replacement with that of XET16-34 to open the binding cleft resulted in a folded protein, which still maintained some β-glucan hydrolase activity, but the corresponding nucleophile mutant did not display glycosynthase activity with either linear or branched glycosyl donors. Next, point mutations of the 1,3-1,4-β-glucanase β-sheets forming the binding site cleft were mutated to resemble XET16-34 residues. The final chimeric protein acquired binding affinity for xyloglucan and did not bind β-glucan. Therefore, binding specificity has been re-engineered, but affinity was low and the nucleophile mutant of the chimeric enzyme did not show glycosynthase activity to produce the target hybrid oligosaccharides. Structural analysis by X-ray crystallography explains these results in terms of changes in the protein structure and highlights further engineering approaches toward introducing the desired activity. © 2010 Wiley-Liss, Inc.

The physiological effects of multi-walled carbon nanotubes (MWCNTs) on conidia and the development of the entomopathogenic fungus, Metarhizium anisopliae (Metsch.) Sorok.

PubMed

Gorczyca, Anna; Kasprowicz, Marek J; Lemek, Tadeusz

2014-01-01

The aim of the study was an in vitro evaluation of the effect of MWCNTs on the conidia of two strains of entomopathogenic fungus, Metarhizium anisopliae. The study made use of water suspensions of MWCNTs (concentration ∼ 3 mg·mL(-1)) made from commercial nanotubes and centrifuged. The conidia were placed in contact with nanotubes for 240 h. An assessment of MWCNT influence on conidia was performed after 1, 24, 72 and 240 h and focused on the linear growth of vegetative mycelium derived from these conidia, mycelium sporulation in subcultures and pathogenicity. Using TEM imaging, it was demonstrated that carbon nanotubes are able to damage cell membranes of the examined fungi conidia. However, the absence was noted of a significantly fungistatic effect of both MWCNT suspensions on the examined strains with respect to the physiological features in question. The increase in vegetative mycelium effected by spores after contact with MWCNTs was characterized by a slight modification in relation to the control. There was no strong trend (inhibition - stimulation), in relation to the effect of the tested suspension of carbon nanotubes, on the development of the vegetative mycelium in in vitro culture. Sporulation of the mycelium after completion of the culture only occurred in one case (strain Ma73F and culture of spores after 24-h contact with MWCNTs) significantly more intensely than in the controls. With respect to pathogenicity for test insects compared to the control strain, Ma73F spores grown from the longest contact with nanotubes suspensions performed significantly better. On the basis of the calculated of mycelium index growth rates and the time of death of the test insects (LT50), it was found that the adverse effects of water suspension MWCNTs on the spores of M. anisopliae were applied after a short contact with biological material. This indicates unfavorable physical rather than chemical effects on the tested cell. Over time, nanotube aggregation in water suspensions led to changes in their influence on the cells under examination.

Fungicides affect the production of extracellular mucilaginous material (ECMM) and the peripheral growth unit (PGU) in two wood-rotting basidiomycetes.

PubMed

Vesentini, Damiano; Dickinson, David J; Murphy, Richard J

2006-10-01

This study shows the effect of two fungicides on the production of extracellular mucilaginous material (ECMM) in two wood-rotting basidiomycetes and identifies a mechanism that might be responsible for the variation observed. Increasing concentrations of the fungicides copper sulphate (CuSO4) and cyproconazole in the growth medium increased the proportion of ECMM in the biomass of Trametes versicolor and Gloeophyllum trabeum. These fungicides also caused a reduction in the length of the peripheral growth unit (PGU) of the mycelia leading to a more highly branched morphology and a larger number of hyphal tips, the sites for active secretion of ECMM, per unit length of mycelium. It is postulated that both in T. versicolor and G. trabeum this change in growth leads to the increases observed in the proportion of ECMM in the total biomass. The implications of these results are discussed with a view to a potential protective role of ECMM against stress and toxic environments.

Metal release and sequestration from black slate mediated by a laccase of Schizophyllum commune.

PubMed

Kirtzel, Julia; Scherwietes, Eric Leon; Merten, Dirk; Krause, Katrin; Kothe, Erika

2018-06-25

Schizophyllum commune is a filamentous basidiomycete which can degrade complex organic macromolecules like lignin by the secretion of a large repertoire of enzymes. One of these white rot enzymes, laccase, exhibits a broad substrate specificity and is able to oxidize a variety of substances including carbonaceous rocks. To investigate the role of laccase in bioweathering, laccase gene lcc2 was overexpressed, and the influence on weathering of black slate, originating from a former alum mine in Schmiedefeld, Germany, was examined. The metal release from the rock material was enhanced, associated with a partial metal accumulation into the mycelium. A sequestration of metals could be shown with fluorescent staining methods, and an accumulation of Zn, Cd, and Pb was visualized in different cell organelles. Additionally, we could show an increased metal resistance of the laccase overexpressing strain.

The application of biotic elicitor on Artemisia annua L. to increase artemisinin content

NASA Astrophysics Data System (ADS)

Darwati, I.; Manohara, D.; Rohimatun; Nurhayati, H.

2018-01-01

Artemisinin-based Combination Therapy (ACT) has been recommended by WHO as an alternative to treat malaria overcoming drug resistance. The secondary metabolic products in plants, including artemisinin, can be increased by utilizing biotic elicitor from fungi. The research was conducted in Gunung Putri Research Installation, Cipanas, West Java from 2010 to 2011. Phytophthora sp. from eggplant and Colletotrichum sp. from Artemisia annua were applied as biotic elicitor. The types of biotic elicitor applied to the plants were 1) the medium of potato dextrose broth were inoculated with fungi and harvested after 10 days (filtrate), 2) powdery mycelium of both fungi. There were 16 treatments: control negative, control positive (uninoculated medium) 1%, 2%, 3% (v/v)], Phytophthora sp. filtrate [1, 2% and 3% (v/v)], Colletotrichum sp. filtrate [1, 2% and 3% (v/v)], Phytophthora sp. mycelium [1%, 2% and 3% (w/v)], Colletotrichum sp mycelium [1%, 2% and 3% (w/v)]. The elicitor application increased plant production by 26.21% and artemisinin yield by 72% compared to control. Furthermore, the artemisinin production of the plants treated with medium inoculated with 2% filtrate of Phytophthora sp (FP2) (25.19 kg/ha) and 1% powdery mycelium of Colletotrichum sp (MC1) (26.42 kg/ha) were higher than control (K) (11.17 kg/ha).

Trichoderma harzianum might impact phosphorus transport by arbuscular mycorrhizal fungi.

PubMed

De Jaeger, Nathalie; de la Providencia, Ivan E; de Boulois, Hervé Dupré; Declerck, Stéphane

2011-09-01

Trichoderma sp. is a biocontrol agent active against plant pathogens via mechanisms such as mycoparasitism. Recently, it was demonstrated that Trichoderma harzianum was able to parasitize the mycelium of an arbuscular mycorrhizal (AM) fungus, thus affecting its viability. Here, we question whether this mycoparasitism may reduce the capacity of Glomus sp. to transport phosphorus ((33)P) to its host plant in an in vitro culture system. (33)P was measured in the plant and in the fungal mycelium in the presence/absence of T. harzianum. The viability and metabolic activity of the extraradical mycelium was measured via succinate dehydrogenase and alkaline phosphatase staining. Our study demonstrated an increased uptake of (33)P by the AM fungus in the presence of T. harzianum, possibly related to a stress reaction caused by mycoparasitism. In addition, the disruption of AM extraradical hyphae in the presence of T. harzianum affected the (33)P translocation within the AM fungal mycelium and consequently the transfer of (33)P to the host plant. The effects of T. harzianum on Glomus sp. may thus impact the growth and function of AM fungi and also indirectly plant performance by influencing the source-sink relationship between the two partners of the symbiosis. © 2011 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

Synthesis and excellent field emission properties of three-dimensional branched GaN nanowire homostructures

NASA Astrophysics Data System (ADS)

Li, Enling; Sun, Lihe; Cui, Zhen; Ma, Deming; Shi, Wei; Wang, Xiaolin

2016-10-01

Three-dimensional branched GaN nanowire homostructures have been synthesized on the Si substrate via a two-step approach by chemical vapor deposition. Structural characterization reveals that the single crystal GaN nanowire trunks have hexagonal wurtzite characteristics and grow along the [0001] direction, while the homoepitaxial single crystal branches grow in a radial direction from the six-sided surfaces of the trunks. The field emission measurements demonstrate that the branched GaN nanowire homostructures have excellent field emission properties, with low turn-on field at 2.35 V/μm, a high field enhancement factor of 2938, and long emission current stability. This indicates that the present branched GaN nanowire homostructures will become valuable for practical field emission applications.

Anatomy of the β-branching enzyme of polyketide biosynthesis and its interaction with an acyl-ACP substrate.

PubMed

Maloney, Finn P; Gerwick, Lena; Gerwick, William H; Sherman, David H; Smith, Janet L

2016-09-13

Alkyl branching at the β position of a polyketide intermediate is an important variation on canonical polyketide natural product biosynthesis. The branching enzyme, 3-hydroxy-3-methylglutaryl synthase (HMGS), catalyzes the aldol addition of an acyl donor to a β-keto-polyketide intermediate acceptor. HMGS is highly selective for two specialized acyl carrier proteins (ACPs) that deliver the donor and acceptor substrates. The HMGS from the curacin A biosynthetic pathway (CurD) was examined to establish the basis for ACP selectivity. The donor ACP (CurB) had high affinity for the enzyme (Kd = 0.5 μM) and could not be substituted by the acceptor ACP. High-resolution crystal structures of HMGS alone and in complex with its donor ACP reveal a tight interaction that depends on exquisite surface shape and charge complementarity between the proteins. Selectivity is explained by HMGS binding to an unusual surface cleft on the donor ACP, in a manner that would exclude the acceptor ACP. Within the active site, HMGS discriminates between pre- and postreaction states of the donor ACP. The free phosphopantetheine (Ppant) cofactor of ACP occupies a conserved pocket that excludes the acetyl-Ppant substrate. In comparison with HMG-CoA (CoA) synthase, the homologous enzyme from primary metabolism, HMGS has several differences at the active site entrance, including a flexible-loop insertion, which may account for the specificity of one enzyme for substrates delivered by ACP and the other by CoA.

Growth of Pleurotus ostreatus on wheat straw and wheat-grain-based media: Biochemical aspects and preparation of mushroom inoculum.

PubMed

Sainos, E; Díaz-Godínez, G; Loera, O; Montiel-González, A M; Sánchez, C

2006-10-01

Mycelial growth, intracellular activity of proteases, laccases and beta-1,3-glucanases, and cytoplasmic protein were evaluated in the vegetative phase of Pleurotus ostreatus grown on wheat straw and in wheat-grain-based media in Petri dishes and in bottles. The productivity of the wheat straw and wheat-grain-based spawn in cylindrical polyethylene bags containing 5 kg of chopped straw was also determined. We observed high activity of proteases and high content of intracellular protein in cultures grown on wheat straw. This suggests that the proteases are not secreted into the medium and that the protein is an important cellular reserve. On the contrary, cultures grown on wheat straw secreted laccases into the medium, which could be induced by this substrate. P. ostreatus grown on media prepared with a combination of wheat straw and wheat grain showed a high radial growth rate in Petri dishes and a high level of mycelial growth in bottles. The productivities of wheat straw and wheat-grain-based spawn were similar. Our results show that cheaper and more productive mushroom spawn can be prepared by developing the mycelium on wheat straw and wheat-grain-based substrates.

Sudden substrate dilution induces a higher rate of citric acid production by Aspergillus niger.

PubMed Central

Legisa, M; Gradisnik-Grapulin, M

1995-01-01

On the basis of the present knowledge of Aspergillus niger metabolism during citric acid fermentation, an idea on how to improve the process was formed. Initially, a higher sucrose concentration was used for the germination of spores, which caused a higher intracellular level of the osmoregulator, glycerol, to be present. When citric acid started to be excreted into the medium, the substrate was suddenly diluted. Optimization of this procedure resulted in a nearly tripled volumetric rate (grams per liter per hour) of acid production, while the overall fermentation time was halved compared with the usual batch process. Yet, a characteristic delay was observed at the start of the acid excretion after the dilution. Hypo-osmotic shock caused a prominent elevation of intracellular cyclic AMP levels. Simultaneously, the specific activity of 6-phosphofructo-1-kinase increased significantly, probably due to phosphorylation of the protein molecule by cyclic AMP-dependent protein kinase. Specific 6-phosphofructo-1-kinase activity was much higher in the treated than in the normally growing mycelium. The metabolic flow through glycolysis was expected to be higher, which should contribute to a higher volumetric rate of acid production. PMID:7618885

Sudden substrate dilution induces a higher rate of citric acid production by Aspergillus niger.

PubMed

Legisa, M; Gradisnik-Grapulin, M

1995-07-01

On the basis of the present knowledge of Aspergillus niger metabolism during citric acid fermentation, an idea on how to improve the process was formed. Initially, a higher sucrose concentration was used for the germination of spores, which caused a higher intracellular level of the osmoregulator, glycerol, to be present. When citric acid started to be excreted into the medium, the substrate was suddenly diluted. Optimization of this procedure resulted in a nearly tripled volumetric rate (grams per liter per hour) of acid production, while the overall fermentation time was halved compared with the usual batch process. Yet, a characteristic delay was observed at the start of the acid excretion after the dilution. Hypo-osmotic shock caused a prominent elevation of intracellular cyclic AMP levels. Simultaneously, the specific activity of 6-phosphofructo-1-kinase increased significantly, probably due to phosphorylation of the protein molecule by cyclic AMP-dependent protein kinase. Specific 6-phosphofructo-1-kinase activity was much higher in the treated than in the normally growing mycelium. The metabolic flow through glycolysis was expected to be higher, which should contribute to a higher volumetric rate of acid production.

Defense Response and Suppression of Phytophthora Blight Disease of Pepper by Water Extract from Spent Mushroom Substrate of Lentinula edodes

PubMed Central

Kang, Dae-Sun; Min, Kyong-Jin; Kwak, A-Min; Lee, Sang-Yeop; Kang, Hee-Wan

2017-01-01

The spent mushroom substrate (SMS) of Lentinula edodes that was derived from sawdust bag cultivation was used as materials for controlling Phytophthora blight disease of pepper. Water extract from SMS (WESMS) of L. edodes inhibited mycelial growth of Phytophthora capsici, suppressed Phytophthora blight disease of pepper seedlings by 65% and promoted growth of the plant over 30%. In high performance liquid chromatography (HPLC) analysis, oxalic acid was detected as the main organic acid compound in WESMS and inhibited the fungal mycelium at a minimum concentration of 200 mg/l. In quantitative real-time PCR, the transcriptional expression of CaBPR1 (PR protein 1), CaBGLU (β-1,3-glucanase), CaPR-4 (PR protein 4), and CaPR-10 (PR protein 10) were significantly enhanced on WESMS and DL-β-aminobutyric acid (BABA) treated pepper leaves. In addition, the salicylic acid content was also increased 4 to 6 folds in the WESMS and BABA treated pepper leaves compared to water treated leaf sample. These findings suggest that WESMS of L. edodes suppress Phytophthora blight disease of pepper through multiple effects including antifungal activity, plant growth promotion, and defense gene induction. PMID:28592945

Defense Response and Suppression of Phytophthora Blight Disease of Pepper by Water Extract from Spent Mushroom Substrate of Lentinula edodes.

PubMed

Kang, Dae-Sun; Min, Kyong-Jin; Kwak, A-Min; Lee, Sang-Yeop; Kang, Hee-Wan

2017-06-01

The spent mushroom substrate (SMS) of Lentinula edodes that was derived from sawdust bag cultivation was used as materials for controlling Phytophthora blight disease of pepper. Water extract from SMS (WESMS) of L. edodes inhibited mycelial growth of Phytophthora capsici , suppressed Phytophthora blight disease of pepper seedlings by 65% and promoted growth of the plant over 30%. In high performance liquid chromatography (HPLC) analysis, oxalic acid was detected as the main organic acid compound in WESMS and inhibited the fungal mycelium at a minimum concentration of 200 mg/l. In quantitative real-time PCR, the transcriptional expression of CaBPR1 (PR protein 1), CaBGLU (β-1,3-glucanase), CaPR-4 (PR protein 4), and CaPR-10 (PR protein 10) were significantly enhanced on WESMS and DL-β-aminobutyric acid (BABA) treated pepper leaves. In addition, the salicylic acid content was also increased 4 to 6 folds in the WESMS and BABA treated pepper leaves compared to water treated leaf sample. These findings suggest that WESMS of L. edodes suppress Phytophthora blight disease of pepper through multiple effects including antifungal activity, plant growth promotion, and defense gene induction.

Proteinase pattern in Trametes versicolor in response to carbon and nitrogen starvation.

PubMed

Staszczak, M; Nowak, G

1984-01-01

In stationary cultures of Trametes versicolor seven proteinase bands were revealed by electrophoresis in mycelium and five in the medium. Under conditions of nitrogen starvation the number of bands in mycelium was unchanged whereas one extracellular proteinase was missing. In the case of carbon starvation one new intracellular proteinase activity appeared and one extracellular activity disappeared. Moreover, in all starved cultures distinct differences in the intensity of particular bands were observed.

Axenic culture and encapsulation of the intraradical forms of Glomus spp.

PubMed

Strullu, D G; Romand, C; Plenchette, C

1991-05-01

In recent years there have been many attempts to cultivate in vitro vesicular-arbuscular mycorrhizal (VAM) fungi which are obligate symbionts. Resting spores extracted from soils are often used as inoculum. Mycorrhizal root pieces are also used for inoculation but the role of intra-radical structures has not been clearly established. On agar medium vegetative mycelium was regenerated from individual intra-radical vesicles and from hyphae extracted by enzymatic maceration. After cell penetration, the mycelium probably accumulates substances which allow growth of VAM fungi in pure culture. When associated with tomato roots, this mycelium forms typical mycorrhizae. Encapsulation stabilized the biological properties of mycorrhizal roots and isolated vesicles. The immobilization also preserved the infectivity of the intra-radical hyphae and vesicles. After 25 years of exclusive utilization of resting spores as starting material for axenic and dual cultures of VAM fungi, it appears that intra-radical vesicles may be preferable propagules.

Fate of Carbohydrates and Lignin during Composting and Mycelium Growth of Agaricus bisporus on Wheat Straw Based Compost

PubMed Central

Jurak, Edita; Punt, Arjen M.; Arts, Wim; Kabel, Mirjam A.; Gruppen, Harry

2015-01-01

In wheat straw based composting, enabling growth of Agaricus bisporus mushrooms, it is unknown to which extent the carbohydrate-lignin matrix changes and how much is metabolized. In this paper we report yields and remaining structures of the major components. During the Phase II of composting 50% of both xylan and cellulose were metabolized by microbial activity, while lignin structures were unaltered. During A. bisporus’ mycelium growth (Phase III) carbohydrates were only slightly consumed and xylan was found to be partially degraded. At the same time, lignin was metabolized for 45% based on pyrolysis GC/MS. Remaining lignin was found to be modified by an increase in the ratio of syringyl (S) to guaiacyl (G) units from 0.5 to 0.7 during mycelium growth, while fewer decorations on the phenolic skeleton of both S and G units remained. PMID:26436656

Genome-wide Identification and characterization of circular RNAs in the rice blast fungus Magnaporthe oryzae.

PubMed

Yuan, Jialan; Wang, Zhao; Xing, Junjie; Yang, Qingyong; Chen, Xiao-Lin

2018-04-30

Numerous circRNAs have been identified in different organisms, but little attention has been addressed on fungal circRNAs. Here, we identified a total of 8,848 circRNAs from the model plant pathogenic fungus M. oryzae. 5,840 circRNAs were identified from mycelium, 2,721 circRNAs from conidium, while only 287 circRNAs from both tissues. This indicated that most of the M. oryzae circRNAs were specifically expressed in mycelium or in conidium. Parental genes of circRNAs in mycelium were enriched in basic metabolisms required for normal growth, while in conidium, they were enriched in biogenesis of storages potentially used for infection. M. oryzae circRNAs could also bind to miRNAs, suggesting they may also function as sponges in fungi. This study suggested M. oryzae circRNAs could play important roles in regulation of growth and development.

SACE_0012, a TetR-family transcriptional regulator, affects the morphogenesis of Saccharopolyspora erythraea.

PubMed

Yin, Xiaojuan; Xu, Xinqiang; Wu, Hang; Yuan, Li; Huang, Xunduan; Zhang, Buchang

2013-12-01

Saccharopolyspora erythraea, a mycelium-forming actinomycete, produces a clinically important antibiotic erythromycin. Extensive investigations have provided insights into erythromycin biosynthesis in S. erythraea, but knowledge of its morphogenesis remains limited. By gene inactivation and complementation strategies, the TetR-family transcriptional regulator SACE_0012 was identified to be a negative regulator of mycelium formation of S. erythraea A226. Detected by quantitative real-time PCR, the relative transcription of SACE_7115, the amfC homolog for an aerial mycelium formation protein, was dramatically increased in SACE_0012 mutant, whereas erythromycin biosynthetic gene eryA, a pleiotropic regulatory gene bldD, and the genes SACE_2141, SACE_6464, SACE_6040, that are the homologs to the sporulation regulators WhiA, WhiB, WhiG, were not differentially expressed. SACE_0012 disruption could not restore its defect of aerial development in bldD mutant, and also did not further accelerate the mycelium formation in the mutant of SACE_7040 gene, that was previously identified to be a morphogenesis repressor. Furthermore, the transcriptional level of SACE_0012 had not markedly changed in bldD and SACE_7040 mutant over A226. Taken together, these results suggest that SACE_0012 is a negative regulator of S. erythraea morphogenesis by mainly increasing the transcription of amfC gene, independently of the BldD regulatory system.

Biochemical Characterization of Extracellular Cellulase from Tuber maculatum Mycelium Produced Under Submerged Fermentation.

PubMed

Bedade, Dattatray K; Singhal, Rekha S; Turunen, Ossi; Deska, Jan; Shamekh, Salem

2017-02-01

Interaction of truffle mycelium with the host plant involves the excretion of extracellular enzymes. The ability of Tuber maculatum mycelium to produce an extracellular cellulase during submerged fermentation was demonstrated for the first time. T. maculatum mycelia were isolated and tested for extracellular cellulase production at variable pH on solid agar medium, and the highest activity was observed at pH 7.0. Furthermore, T. maculatum was subjected to submerged fermentation in basal salt medium for cellulase production. Under optimized conditions using sodium carboxymethyl cellulose (0.5 % w/v) as carbon source and an initial pH of 7.0, the enzyme production yielded 1.70 U/mL of cellulase in the cell-free supernatant after 7 days of incubation time. The optimum of the obtained cellulase's activity was at pH 5.0 and a temperature of 50 °C. The enzyme showed good thermostability at 50 °C by retaining 99 % of its maximal activity over an incubation time of 100 min. The cellulase activity was inhibited by Fe 2+ and slightly activated by Mn 2+ and Cu 2+ at 1 mM concentration. The results indicated that truffle mycelium is utilizing cellulosic energy source in the root system, and the optimal conditions are those existing in the acidic Finnish soil.

Adapting High-Resolution Respirometry to Glucose-Limited Steady State Mycelium of the Filamentous Fungus Penicillium ochrochloron: Method Development and Standardisation

PubMed Central

Schinagl, Christoph W.; Vrabl, Pamela; Burgstaller, Wolfgang

2016-01-01

Fungal electron transport systems (ETS) are branched, involving alternative NADH dehydrogenases and an alternative terminal oxidase. These alternative respiratory enzymes were reported to play a role in pathogenesis, production of antibiotics and excretion of organic acids. The activity of these alternative respiratory enzymes strongly depends on environmental conditions. Functional analysis of fungal ETS under highly standardised conditions for cultivation, sample processing and respirometric assay are still lacking. We developed a highly standardised protocol to explore in vivo the ETS—and in particular the alternative oxidase—in Penicillium ochrochloron. This included cultivation in glucose-limited chemostat (to achieve a defined and reproducible physiological state), direct transfer without any manipulation of a broth sample to the respirometer (to maintain the physiological state in the respirometer as close as possible to that in the chemostat), and high-resolution respirometry (small sample volume and high measuring accuracy). This protocol was aimed at avoiding any changes in the physiological phenotype due to the high phenotypic plasticity of filamentous fungi. A stable oxygen consumption (< 5% change in 20 minutes) was only possible with glucose limited chemostat mycelium and a direct transfer of a broth sample into the respirometer. Steady state respiration was 29% below its maximum respiratory capacity. Additionally to a rotenone-sensitive complex I and most probably a functioning complex III, the ETS of P. ochrochloron also contained a cyanide-sensitive terminal oxidase (complex IV). Activity of alternative oxidase was present constitutively. The degree of inhibition strongly depended on the sequence of inhibitor addition. This suggested, as postulated for plants, that the alternative terminal oxidase was in dynamic equilibrium with complex IV—independent of the rate of electron flux. This means that the onset of activity does not depend on a complete saturation or inhibition of the cytochrome pathway. PMID:26771937

Dielectric Waveguides Splitter and Hybrid/Isolator for Bidirectional Link

NASA Technical Reports Server (NTRS)

Tang, Adrian Joseph (Inventor); Chattopadhyay, Goutam (Inventor); Chahat, Nacer E. (Inventor); Decrossas, Emmanuel (Inventor)

2016-01-01

A system, method, device, and apparatus provide a dielectric waveguide splitter/bi-directional link. A dielectric substrate fabricated into a first Y-junction waveguide with a first port splitting into a first branch leading to a second port and a second branch leading to a third port. An angle between the first branch and the second branch is below ninety degrees (90.degree.). The dielectric waveguide splitter enables millimeter-wave (mmWave) transmission between the first port and the second port while reducing feedback of the mmWave between the second and third port. Two Y-junction waveguides may be fabricated back-to-back to provide simultaneous bidirectional mmWave transmission at a single frequency.

Chemically Functionalized Carbon Nanotubes as Substrates for Neuronal Growth

PubMed Central

Hu, Hui; Ni, Yingchun; Montana, Vedrana; Haddon, Robert C.; Parpura, Vladimir

2009-01-01

We report the use of chemically modified carbon nanotubes as a substrate for cultured neurons. The morphological features of neurons that directly reflect their potential capability in synaptic transmission are characterized. The chemical properties of carbon nanotubes are systematically varied by attaching different functional groups that confer known characteristics to the substrate. By manipulating the charge carried by functionalized carbon nanotubes we are able to control the outgrowth and branching pattern of neuronal processes. PMID:21394241

Geothermomicrobium terrae gen. nov., sp. nov., a novel member of the family Thermoactinomycetaceae.

PubMed

Zhou, En-Min; Yu, Tian-Tian; Liu, Lan; Ming, Hong; Yin, Yi-Rui; Dong, Lei; Tseng, Min; Nie, Guo-Xing; Li, Wen-Jun

2014-09-01

Strains YIM 77562(T) and YIM 77580, two novel Gram-staining-positive, filamentous bacterial isolates, were recovered from the Rehai geothermal field, Tengchong, Yunnan province, south-west China. Good growth was observed at 50-55 °C and pH 7.0. Aerial mycelium was absent on all media tested. Substrate mycelium was well-developed, long and moderately flexuous, and formed abundant, single, warty, ornamented endospores. Phylogenetic analysis of the 16S rRNA gene sequences of the two strains indicated that they belong to the family Thermoactinomycetaceae. Similarity levels between the 16S rRNA gene sequences of the two strains and those of type strains of members of the Thermoactinomycetaceae were 88.33-93.24 %; the highest sequence similarity was with Hazenella coriacea DSM 45707(T). In both strains, the predominant menaquinone was MK-7, the diagnostic diamino acid was meso-diaminopimelic acid and the major cellular fatty acids were iso-C14 : 0, iso-C15 : 0 and iso-C16 : 0. The major polar lipids were diphosphatidylglycerol, phosphatidylmethylethanolamine, unidentified polar lipids and unidentified phospholipids. The genomic DNA G+C contents of strains YIM 77562(T) and YIM 77580 were 45.5 and 44.2 mol%, respectively. DNA-DNA relatedness data suggest that the two isolates represent a single species. Based on phylogenetic analyses and physiological and biochemical characteristics, it is proposed that the two strains represent a single novel species in a new genus, Geothermomicrobium terrae gen. nov., sp. nov. The type strain of Geothermomicrobium terrae is YIM 77562(T) ( = CCTCC AA 2011022(T) = JCM 18057(T)). © 2014 IUMS.

Streptomyces humi sp. nov., an actinobacterium isolated from soil of a mangrove forest.

PubMed

Zainal, Nurullhudda; Ser, Hooi-Leng; Yin, Wai-Fong; Tee, Kok-Keng; Lee, Learn-Han; Chan, Kok-Gan

2016-03-01

A novel Streptomyces strain, MUSC 119(T), was isolated from a soil collected from a mangrove forest. Cells of MUSC 119(T) stained Gram-positive and formed light brownish grey aerial mycelium and grayish yellowish brown substrate mycelium on ISP 2 medium. A polyphasic approach was used to determine the taxonomic status of strain MUSC 119(T), which shows a range of phylogenetic and chemotaxonomic properties consistent with those of the genus Streptomyces. The cell wall peptidoglycan consisted of LL-diaminopimelic acid. The predominant menaquinones were identified as MK-9(H8), MK-9(H6) and MK-9(H4). The polar lipid profile consisted of phosphatidylinositol, phosphatidylethanolamine, glycolipids, diphosphatidylglycerol and four phospholipids. The predominant cellular fatty acids were anteiso-C15:0, iso-C16:0, and anteiso-C17:0. The cell wall sugars were glucose, mannose, ribose and rhamnose. The phylogenetic analysis based on 16S rRNA gene sequence similarity showed that strain MUSC119(T) to be closely related to Streptomyces rhizophilus JR-41(T) (99.0 % sequence similarity), S. panaciradicis 1MR-8(T) (98.9 %), S. gramineus JR-43(T) (98.8 %) and S. graminisoli JR-19(T) (98.7 %). These results suggest that MUSC 119(T) should be placed within the genus Streptomyces. DNA-DNA relatedness values between MUSC 119(T) to closely related strains ranged from 14.5 ± 1.3 to 27.5 ± 0.7 %. The G+C content was determined to be 72.6 mol %. The polyphasic study of MUSC 119(T) showed that this strain represents a novel species, for which the name Streptomyces humi sp. nov. is proposed. The type strain of S. humi is MUSC 119(T) (=DSM 42174(T) = MCCC 1K00505(T)).

Quantitative Analysis of Filament Branch Orientation in Listeria Actin Comet Tails.

PubMed

Jasnin, Marion; Crevenna, Alvaro H

2016-02-23

Several bacterial and viral pathogens hijack the host actin cytoskeleton machinery to facilitate spread and infection. In particular, Listeria uses Arp2/3-mediated actin filament nucleation at the bacterial surface to generate a branched network that will help propel the bacteria. However, the mechanism of force generation remains elusive due to the lack of high-resolution three-dimensional structural data on the spatial organization of the actin mother and daughter (i.e., branch) filaments within this network. Here, we have explored the three-dimensional structure of Listeria actin tails in Xenopus laevis egg extracts using cryo-electron tomography. We found that the architecture of Listeria actin tails is shared between those formed in cells and in cell extracts. Both contained nanoscopic bundles along the plane of the substrate, where the bacterium lies, and upright filaments (also called Z filaments), both oriented tangentially to the bacterial cell wall. Here, we were able to identify actin filament intersections, which likely correspond to branches, within the tails. A quantitative analysis of putative Arp2/3-mediated branches in the actin network showed that mother filaments lie on the plane of the substrate, whereas daughter filaments have random deviations out of this plane. Moreover, the analysis revealed that branches are randomly oriented with respect to the bacterial surface. Therefore, the actin filament network does not push directly toward the surface but rather accumulates, building up stress around the Listeria surface. Our results favor a mechanism of force generation for Listeria movement where the stress is released into propulsive motion. Copyright © 2016 Biophysical Society. Published by Elsevier Inc. All rights reserved.

The effect of Mirabilis jalapa leaves biopesticide treatment on the mycelium growth of entomopathogenic fungi Beauveria bassiana inside the larvae body Crocidolomia binotalis

NASA Astrophysics Data System (ADS)

Pramita, Mia; Anggraeni, Tjandra

2015-09-01

Pest control with biological method (biopesticide and entomopathogenic fungi) is an alternative program to reduce application of chemical insecticide. Biopesticide of Mirabilis jalapa leaves has been discovered rich in secondary metabolites which has antifeedant activity that can provide physiological interference in insect larvae and the generation numbers[1]. Entomopathogenic fungi Beauveria bassiana has potential to control pest populations[2]. The growth of mycelium B. bassiana may interfere metabolism process inside the host body. Otherwise, B. bassiana produce toxins such as beauvericin that can increase mortality of pest. Combination of M. jalapa and B. bassiana reduce LT50 on C. binotalis larvae[3]. Thus, this study aims to determine influence of provision of biopesticide M. jalapa leaves on growth of mycelium entomopathogenic fungi B. bassiana inside larvae body C. binotalis and to detect the presence of beauvericin in vivo. Third instar larvae of C. binotalis were divided into a control, fungal and combination group. The combination group was given biopesticide and fungi. The concentration of biopesticide was 0.8% (w/v) and concentration of fungi spores was 107 spores/ml. Spores (vol. 5µl) done topically to larvae in interval 6 hours after treatment of biopesticide on non-pesticide cabbage leaves. Afterwards, histological observations performed at 24, 48, 72, 96 hours after treatment. The result show of emergence hyphae and mycelium growth inside lumen of larvae midgut on combination group faster than fungal group. This is thought to be caused by the influence of secondary metabolites of biopesticide M. jalapa leaves. In addition, beauviricin is detectable both of fungal and combination group. Thus, it can be concluded that treatment of biopesticide from M. jalapa leaves can accelerate on growth of mycelium entomopathogenic fungi B. bassiana inside the larvae body C. binotalis and toxic of B. bassiana such as beauvericin was detected on fungal and combination group.

Multi-branched gold nanostars with fractal structure for SERS detection of the pesticide thiram

NASA Astrophysics Data System (ADS)

Zhu, Jian; Liu, Mei-Jin; Li, Jian-Jun; Li, Xin; Zhao, Jun-Wu

2018-01-01

The surface-enhanced Raman scattering (SERS) activity of multi-branched gold nanostars with fractal structure has been investigated for trace detection of pesticide thiram. Raman spectrum results show that the gold nanostars substrate can produce about 102 fold stronger signal than the thiram alone with the thiram concentration increase of 103 times and 1.4 fold stronger signal than the gold nanostars without fractal feature. In the detection procedure, the most prominent SERS peak at 1376 cm- 1 has been chosen to characterize and quantify the concentration of thiram. Experimental results indicate this Raman substrate based on fractal gold nanostars exhibits excellent selective probing performance for thiram with a detection limit as low as 10- 10 M in solution and 0.24 ng/cm2 in apple peels. Interference experiment results show that the effects from the interfering pesticides could be neglected in the detection procedure. Therefore, the gold nanostars as a SERS substrate have excellent sensitivity and selectivity.

Characterization of a novel debranching enzyme from Nostoc punctiforme possessing a high specificity for long branched chains

DOE Office of Scientific and Technical Information (OSTI.GOV)

Choi, Ji-Hye; Lee, Heeseob; Kim, Young-Wan

2009-01-09

A novel debranching enzyme from Nostoc punctiforme PCC73102 (NPDE) exhibits hydrolysis activity toward both {alpha}-(1,6)- and {alpha}-(1,4)-glucosidic linkages. The action patterns of NPDE revealed that branched chains are released first, and the resulting maltooligosaccharides are then hydrolyzed. Analysis of the reaction with maltooligosaccharide substrates labeled with {sup 14}C-glucose at the reducing end shows that NPDE specifically liberates glucose from the reducing end. Kinetic analyses showed that the hydrolytic activity of NPDE is greatly affected by the length of the substrate. The catalytic efficiency of NPDE increased considerably upon using substrates that can occupy at least eight glycone subsites such asmore » maltononaose and maltooctaosyl-{alpha}-(1,6)-{beta}-cyclodextrin. These results imply that NPDE has a unique subsite structure consisting of -8 to +1 subsites. Given its unique subsite structure, side chains shorter than maltooctaose in amylopectin were resistant to hydrolysis by NPDE, and the population of longer side chains was reduced.« less

An Extracellular Cell-Attached Pullulanase Confers Branched α-Glucan Utilization in Human Gut Lactobacillus acidophilus.

PubMed

Møller, Marie S; Goh, Yong Jun; Rasmussen, Kasper Bøwig; Cypryk, Wojciech; Celebioglu, Hasan Ufuk; Klaenhammer, Todd R; Svensson, Birte; Abou Hachem, Maher

2017-06-15

Of the few predicted extracellular glycan-active enzymes, glycoside hydrolase family 13 subfamily 14 (GH13_14) pullulanases are the most common in human gut lactobacilli. These enzymes share a unique modular organization, not observed in other bacteria, featuring a catalytic module, two starch binding modules, a domain of unknown function, and a C-terminal surface layer association protein (SLAP) domain. Here, we explore the specificity of a representative of this group of pullulanases, Lactobacillus acidophilus Pul13_14 ( La Pul13_14), and its role in branched α-glucan metabolism in the well-characterized Lactobacillus acidophilus NCFM, which is widely used as a probiotic. Growth experiments with L. acidophilus NCFM on starch-derived branched substrates revealed a preference for α-glucans with short branches of about two to three glucosyl moieties over amylopectin with longer branches. Cell-attached debranching activity was measurable in the presence of α-glucans but was repressed by glucose. The debranching activity is conferred exclusively by La Pul13_14 and is abolished in a mutant strain lacking a functional La Pul13_14 gene. Hydrolysis kinetics of recombinant La Pul13_14 confirmed the preference for short-branched α-glucan oligomers consistent with the growth data. Curiously, this enzyme displayed the highest catalytic efficiency and the lowest K m reported for a pullulanase. Inhibition kinetics revealed mixed inhibition by β-cyclodextrin, suggesting the presence of additional glucan binding sites besides the active site of the enzyme, which may contribute to the unprecedented substrate affinity. The enzyme also displays high thermostability and higher activity in the acidic pH range, reflecting adaptation to the physiologically challenging conditions in the human gut. IMPORTANCE Starch is one of the most abundant glycans in the human diet. Branched α-1,6-glucans in dietary starch and glycogen are nondegradable by human enzymes and constitute a metabolic resource for the gut microbiota. The role of health-beneficial lactobacilli prevalent in the human small intestine in starch metabolism remains unexplored in contrast to colonic bacterial residents. This study highlights the pivotal role of debranching enzymes in the breakdown of starchy branched α-glucan oligomers (α-limit dextrins) by human gut lactobacilli exemplified by Lactobacillus acidophilus NCFM, which is one of the best-characterized strains used as probiotics. Our data bring novel insight into the metabolic preference of L. acidophilus for α-glucans with short α-1,6-branches. The unprecedented affinity of the debranching enzyme that confers growth on these substrates reflects its adaptation to the nutrient-competitive gut ecological niche and constitutes a potential advantage in cross-feeding from human and bacterial dietary starch metabolism. Copyright © 2017 American Society for Microbiology.

An Extracellular Cell-Attached Pullulanase Confers Branched α-Glucan Utilization in Human Gut Lactobacillus acidophilus

PubMed Central

Møller, Marie S.; Rasmussen, Kasper Bøwig; Cypryk, Wojciech; Celebioglu, Hasan Ufuk; Klaenhammer, Todd R.; Svensson, Birte

2017-01-01

ABSTRACT Of the few predicted extracellular glycan-active enzymes, glycoside hydrolase family 13 subfamily 14 (GH13_14) pullulanases are the most common in human gut lactobacilli. These enzymes share a unique modular organization, not observed in other bacteria, featuring a catalytic module, two starch binding modules, a domain of unknown function, and a C-terminal surface layer association protein (SLAP) domain. Here, we explore the specificity of a representative of this group of pullulanases, Lactobacillus acidophilus Pul13_14 (LaPul13_14), and its role in branched α-glucan metabolism in the well-characterized Lactobacillus acidophilus NCFM, which is widely used as a probiotic. Growth experiments with L. acidophilus NCFM on starch-derived branched substrates revealed a preference for α-glucans with short branches of about two to three glucosyl moieties over amylopectin with longer branches. Cell-attached debranching activity was measurable in the presence of α-glucans but was repressed by glucose. The debranching activity is conferred exclusively by LaPul13_14 and is abolished in a mutant strain lacking a functional LaPul13_14 gene. Hydrolysis kinetics of recombinant LaPul13_14 confirmed the preference for short-branched α-glucan oligomers consistent with the growth data. Curiously, this enzyme displayed the highest catalytic efficiency and the lowest Km reported for a pullulanase. Inhibition kinetics revealed mixed inhibition by β-cyclodextrin, suggesting the presence of additional glucan binding sites besides the active site of the enzyme, which may contribute to the unprecedented substrate affinity. The enzyme also displays high thermostability and higher activity in the acidic pH range, reflecting adaptation to the physiologically challenging conditions in the human gut. IMPORTANCE Starch is one of the most abundant glycans in the human diet. Branched α-1,6-glucans in dietary starch and glycogen are nondegradable by human enzymes and constitute a metabolic resource for the gut microbiota. The role of health-beneficial lactobacilli prevalent in the human small intestine in starch metabolism remains unexplored in contrast to colonic bacterial residents. This study highlights the pivotal role of debranching enzymes in the breakdown of starchy branched α-glucan oligomers (α-limit dextrins) by human gut lactobacilli exemplified by Lactobacillus acidophilus NCFM, which is one of the best-characterized strains used as probiotics. Our data bring novel insight into the metabolic preference of L. acidophilus for α-glucans with short α-1,6-branches. The unprecedented affinity of the debranching enzyme that confers growth on these substrates reflects its adaptation to the nutrient-competitive gut ecological niche and constitutes a potential advantage in cross-feeding from human and bacterial dietary starch metabolism. PMID:28411221

The influences of sugars and plant growth regulators on β-glucan synthesis of G. lucidum mycelium in submerged culture

NASA Astrophysics Data System (ADS)

Thao, Cao Phuong; Tien, Le Thi Thuy

2017-09-01

β - glucan is intracellular polysaccharide (IPS), extracted from Ganoderma lucidum mycelium that can enhance human immune respond. This study aimed to stimulate the production of β - glucan in G. lucidum mycelium through optimating the carbonhydrates and plant rowth regulators in submerged culture. The results showed that the stimulation or inhibition of IPS production as well as β - glucan biosynthesis could be adjusted depend on the type and concentration of carbonhydrates and plant growth regulators. The supplement of lactose 80 g/L and BA 1 mg/L in medium could cause the highest IPS production (644.478 mg/g DW) and β - glucan increased up to 0.15/DW, that raised twice as much as without plant growth regulators. Futhermore, the optimation of other environmental elements were figured out were completely dark and 150 rpm on rotary shaker. This result could be used as premise for production of β - glucan in pilot.

Detection of White Root Disease (Rigidoporus Microporus) in Various Soil Types in the Rubber Plantations Based on The Serological Reaction

NASA Astrophysics Data System (ADS)

Indriani Dalimunthe, Cici; Tistama, Radite; Wahyuni, Sri

2017-12-01

The Conventional detection of White Root Disease (Rigidoporus microporus, WRD) still uses the visual method based on an abnormal color of leaf or mycelium growth on the tap root neck. The method was less effective and less efficient. The serological technique uses yolk chicken antibodies induced by immunization with mycelium extract. The purpose of this research was to examine the consistency of selected antibodies in detecting root fungi at various soil types in the rubber plantations. This research used a Completely Randomized Design non-factorial with twelve (12) treatments and two (2) replications. The results showed that the antibodies could detect WRD in various soils types. The serological detection was higher precisely than visual observation. The development of WRD mycelium varies depending on the soil types and it was different in the each estate area. In addition, this research is expected to get a serology kit to detect early symptoms of WRD in the rubber plants.

Potential Physiologies of Deep Branches on the Tree of Life with Deep Subsurface Samples from IODP Leg 347: Baltic Sea Paleoenvironment

NASA Astrophysics Data System (ADS)

Lloyd, K. G.; Bird, J. T.; Shumaker, A.

2014-12-01

Very little is known about how evolutionary branches that are distantly related to cultured microorganisms make a living in the deep subsurface marine environment. Here, sediments are cut-off from surface inputs of organic substrates for tens of thousands of years; yet somehow support a diverse population of microorganisms. We examined the potential metabolic and ecological roles of uncultured archaea and bacteria in IODP Leg 347: Baltic Sea Paleoenvironment samples, using quantitative PCR holes 60B, 63E, 65C, and 59C and single cell genomic analysis for hole 60B. We quantified changes in total archaea and bacteria, as well as deeply-branching archaeal taxa with depth. These sediment cores alternate between high and low salinities, following a glacial cycle. This allows changes in the quantities of these groups to be placed in the context of potentially vastly different organic matter sources. In addition, single cells were isolated, and their genomes were amplified and sequenced to allow a deeper look into potential physiologies of uncultured deeply-branching organisms found up to 86 meters deep in marine sediments. Together, these data provide deeper insight into the relationship between microorganisms and their organic matter substrates in this extreme environments.

Branch pattern of starch internal structure influences the glucogenesis by mucosal Nt-maltase-glucoamylase.

PubMed

Lin, Amy Hui-Mei; Ao, Zihua; Quezada-Calvillo, Roberto; Nichols, Buford L; Lin, Chi-Tien; Hamaker, Bruce R

2014-10-13

To produce sufficient amounts of glucose from food starch, both α-amylase and mucosal α-glucosidases are required. We found previously that the digestion rate of starch is influenced by its susceptibility to mucosal α-glucosidases. In the present study, six starches and one glycogen were pre-hydrolyzed by α-amylase for various time periods, and then further hydrolyzed with the mucosal α-glucosidase, the N-terminal subunit of maltase-glucoamylase (Nt-MGAM), to generate free glucose. Results showed that α-amylase amplified the Nt-MGAM glucogenesis, and that the amplifications differed in various substrates. The amount of branches within α-amylase hydrolysate substrates was highly related to the rate of Nt-MGAM glucogenesis. After de-branching, the hydrolysates showed three fractions, Fraction 1, 2, and 3, in size exclusion chromatographs. We found that the α-amylase hydrolysates with higher quantity of the Fraction 3 (molecules with relatively short chain-length) and shorter average chain-length of this fraction had lower rates of Nt-MGAM glucogenesis. This study revealed that the branch pattern of α-amylase hydrolysates modulates glucose release by Nt-MGAM. It further supported the hypothesis that the internal structure of starch affects its digestibility at the mucosal α-glucosidase level. Published by Elsevier Ltd.

Diagonal gaits in the feathertail glider Acrobates pygmaeus (Acrobatidae, Diprotodontia): Insights for the evolution of primate quadrupedalism.

PubMed

Karantanis, Nikolaos-Evangelos; Youlatos, Dionisios; Rychlik, Leszek

2015-09-01

Research on primate origins has revolved around arboreality and, more specifically, the adaptations that are linked to safe navigation in the fine-branch niche. To this end, extant non-primate mammals have been used as models to assess the significance of these adaptations. However, the size of these models is larger than that estimated for early primates. In contrast, the feathertail marsupial glider Acrobates pygmaeus, with a body mass of 12 g, a clawless opposable hallux, and terminal branch feeding habits appears more suited to modeling behavioral adaptations to the small branch milieu. Analysis of video recordings of 18 feathertail gliders walking on poles of variable diameter and inclination revealed that they preferentially used diagonal sequence gaits, fast velocities and low duty factors. Diagonal gaits did not correlate to duty factor, but increased as substrate size decreased, and from descending to ascending locomotion. Furthermore, the duty factor index increased in more diagonal gaits and ascending locomotion. Finally, velocities were lower on smaller substrates, and were mainly regulated by stride frequency and, to a lesser degree, stride length. Feathertail glider gaits displayed noteworthy behavioral convergences with primate quadrupedalism, but some of these results need additional investigation. Despite any discrepancies, these features appear to be favorable for quadrupedal progression on small branches, providing a selective advantage for navigating within a fine branch niche and highlighting the importance of small body size in early primate evolution. Copyright © 2015 Elsevier Ltd. All rights reserved.

Tempeh: a mold-modified indigenous fermented food made from soybeans and/or cereal grains.

PubMed

Hachmeister, K A; Fung, D Y

1993-01-01

A variety of indigenous fermented foods exist today; however, tempeh has been one of the most widely accepted and researched mold-modified fermented products. Tempeh is a traditional fermented food made from soaked and cooked soybeans inoculated with a mold, usually of the genus Rhizopus. After fermentation has occurred, the soybeans are bound together into a compact cake by dense cottony mycelium. An important function of the mold in the fermentation process is the synthesis of enzymes, which hydrolyze soybean constituents and contribute to the development of a desirable texture, flavor, and aroma of the product. Enzymatic hydrolysis also may decrease or eliminate antinutritional constituents; consequently, the nutritional quality of the fermented product may be improved. Current technology and new scientific advancements have enabled researchers to examine specific strains of Rhizopus and new substrates such as cereal grains. Because Kansas produces numerous cereal grains, production of a fermented tempeh-like product using wheat, sorghum (milo), oats, rye, barley, corn, and triticale is a definite possibility for generating a Kansas Value-Added Product. In this study, several different tempeh-like products were produced using various cereal grains inoculated with Rhizopus oligosporus NRRL 2549 or R. oligosporus NRRL 2710. Grains used included hard red winter wheat, triticale, yellow sorghum (milo), and red sorghum (milo). The grain source as well as the strain of R. oligosporus used influenced the product's appearance, flavor, and patty integrity. Results showed that R. oligosporus NRRL 2549 produced more mycelium at a more rapid rate than did the R. oligosporus NRRL 2710 strain. The combination of red sorghum and R. oligosporus NRRL 2549 yielded a product with good patty texture, aroma, and appearance. Furthermore, the red sorghum fermented product was well suited for slicing. On the other hand, yellow sorghum inoculated with either R. oligosporus NRRL 2549 or R. oligosporus NRRL 2710 failed to produce an organoleptically suitable product. Triticale also was found to be an unacceptable substrate for the production of a tempeh-like product. Although the fermented wheat product had a desirable aroma and flavor, it lacked patty integrity and crumbled when sliced. Further research is needed to evaluate the economic significance and industrial applications of these tempeh-like products.

Influence of inorganic and organic selenium on number of living mycelial cells and their ultrastructure in culture of Hericium erinaceum (Bull.: Fr. Pers.).

PubMed

Slusarczyk, Joanna; Malinowska, Eliza; Krzyczkowski, W; Kuraś, M

2013-03-01

Mycelium of the white-rot fungus (Hericium erinaceum (Bull.: Fr. Pers.) produces polysaccharides showing anticancer and immunostimulating activity. In our previous works, we have shown that organic selenitetriglycerides (Selol) contribute to the increase of biosynthesis of exopolysaccharides (EPS) having antioxidative properties and containing large amounts of selenium. The present work is a study of influence of inorganic and organic form of selenium on viability of H. erinaceum mycelium and on ultrastructural changes taking place during its development in submerged culture. The mycelium was grown on media containing sodium selenite (Na2SeO3), a mixture of Na2SeO3 + Selol2% and on control medium (no selenium added). It was shown that mycelium cultured for 3 days in control conditions on standard media contained almost 100% of living cells, with over 80% after 24 days. Treatment with 100 ppm of Na2SeO3 lowered the number of viable cells to 11.8% and 9.1% after 3 and 24 days, respectively. The addition of 2% Selol caused the amounts of living cells to remain at ca 90%. Apparently, Selol helped the cells to cope with the toxic activity of inorganic selenium ions. The addition of sodium selenite induced degradative changes in cell organelles. Such changes were not observed in the case of Na2SeO3 + Selol mixture, in which case cells contained numerous ribosomes and small lipid bodies.

In situ and in vitro colonization of Cathaya argyrophylla (Pinaceae) by ectomycorrhizal fungi.

PubMed

Vaario, Lu-Min; Xing, Shu-Tang; Xie, Zong-Qiang; Lun, Zhi-Ming; Sun, Xue; Li, Yu Hua

2006-03-01

Cathaya argyrophylla, a critically endangered conifer, is found to grow at four isolated areas located in subtropical mountains of China. To examine the involvement and usefulness of mycorrhizas for sustaining the population of this tree, we compared the root system, morphology, and structure of mycorrhizal roots of C. argyrophylla, which were collected from a natural stand and an artificial stand, each grown at a different location. More mycorrhizal roots were found for trees from an artificial stand. The presence of extramatrical mycelium, mantle, and Hartig net revealed that C. argyrophylla formed an ectomycorrhizal association in both sampling sites. Starch granules were found in mycorrhizal roots collected only from a natural stand. The aseptic synthesis of C. argyrophylla and Cenococcum geophilum was established for the first time in vitro. Typical ectomycorrhizas formed on seedlings on RM medium containing 0.1 g/l glucose, 5 weeks after inoculation. By light microscopy, the synthesized mycorrhizas showed a thin mantle from which emanated extramatrical hyphae and highly branched Hartig net. A simple, rapid, and convenient mycorrhiza synthesis system was developed, which facilitates further studies on ectomycorrhizal development of C. argyrophylla.

Fungi associated with black mould on baobab trees in southern Africa.

PubMed

Cruywagen, Elsie M; Crous, Pedro W; Roux, Jolanda; Slippers, Bernard; Wingfield, Michael J

2015-07-01

There have been numerous reports in the scientific and popular literature suggesting that African baobab (Adansonia digitata) trees are dying, with symptoms including a black mould on their bark. The aim of this study was to determine the identity of the fungi causing this black mould and to consider whether they might be affecting the health of trees. The fungi were identified by sequencing directly from mycelium on the infected tissue as well as from cultures on agar. Sequence data for the ITS region of the rDNA resulted in the identification of four fungi including Aureobasidium pullulans, Toxicocladosporium irritans and a new species of Rachicladosporium described here as Rachicladosporium africanum. A single isolate of an unknown Cladosporium sp. was also found. These fungi, referred to here as black mould, are not true sooty mould fungi and they were shown to penetrate below the bark of infected tissue, causing a distinct host reaction. Although infections can lead to dieback of small twigs on severely infected branches, the mould was not found to kill trees.

An Exploration into the Bacterial Community under Different Pasteurization Conditions during Substrate Preparation (Composting-Phase II) for Agaricus bisporus Cultivation.

PubMed

Vieira, Fabricio Rocha; Pecchia, John Andrew

2018-02-01

Substrate preparation (i.e., composting) for Agaricus bisporus cultivation is the most critical point of mushroom production. Among many factors involved in the composting process, the microbial ecology of the system is the underlying drive of composting and can be influenced by composting management techniques. Pasteurization temperature at the beginning of phase II, in theory, may influence the bacterial community and subsequently the "selectivity" and nutrition of the final substrate. Therefore, this hypothesis was tested by simulation in bioreactors under different pasteurization conditions (57 °C/6 h, 60 °C/2 h, and 68 °C/2 h), simulating conditions adopted by many producers. Bacterial diversity, based on 16S ribosomal RNA obtained by high-throughput sequencing and classified in operational taxonomic units (OTUs), was greater than previously reported using culture-dependent methods. Alpha diversity estimators show a lower diversity of OTUs under a high-temperature pasteurization condition. Bacillales order shows a relatively higher OTU abundance under a high-pasteurization temperature, which also was related to high ammonia emission measurements. On the other hand, beta diversity analysis showed no significantly changes in the bacterial community structure under different conditions. Agaricus bisporus mycelium growth during a standard spawn run period was significantly slower in the compost pasteurized at high temperature. Since the bacterial community structure was not greatly affected by different pasteurization conditions but by-products left (e.g., ammonia) at the end of compost conditioning varied, further studies need to be conducted to determine the functional role of the microbial communities found during substrate preparation for Agaricus bisporus cultivation.

Mycelium and polysaccharide production of Agaricus blazei Murrill by submerged fermentation.

PubMed

Lin, Jr-Hui; Yang, Shang-Shyng

2006-04-01

Over the last decade, Agaricus blazei Murrill has been studied and developed as a novel functional food in Japan, Korea, China, and Taiwan. Due to the low yields, the fruiting bodies of A. blazei Murrill are relatively expensive, and a cheap and stable source of A. blazei Murrill mycelium for commercial purposes is highly desirable. Culture media and conditions were investigated with a view to reducing the cost and improving the mycelium and polysaccharide production of A. blazei Murrill by submerged fermentation. Thirty six isolates of A. blazei Murrill were isolated from 22 fruiting bodies produced in Taiwan, and 16 of them could be successfully cultivated on mannitol-egg yolk-polymyxin medium. The isolates were identified by species-specific polymerase chain reaction (PCR) and optimized for the culture media and conditions by submerged fermentation for mycelium and polysaccharide production. Some properties of polysaccharide extract were also investigated. All of the PCR products with species-specific primers showed high identity and matched the internal transcribed spacer 1 sequences of A. blazei Murrill. The phylogenic tree of A. blazei Murrill isolates generated from random amplified polymorphic DNAs arranged all samples into 3 groups and 2 independent cases. The optimal culture media of mycelium production in submerged fermentation were 5% malt extract, 0.1% yeast extract, and 0.5% peptone at pH 6.0, while the optimal culture conditions were 200 mL medium in 500 mL Hinton flask, shaking at 90 rpm for 3 days and then shifting to 105 rpm for 5 days at 27 degrees C. Each liter of A. blazei Murrill M72 yielded 10.83 +/- 0.24 g dried mycelia weight and each liter of A. blazei Murrill M152 produced 0.251 +/- 0.004 g crude polysaccharide (3.03 +/- 0.05% of dried mycelia weight). Crude polysaccharide of A. blazei Murrill M162 contained 82.27-99.14% of total sugar and less than 1.63% of protein; it had 4 major molecular weight components (274.1, 32.7, 7.5, and 2.1 kDa, respectively), with the 2.1 kDa portion possibly a beta-(1,3)-glucan. These results show that selection of media and conditions can be employed in order to improve the mycelium and polysaccharide production of A. blazei Murrill M72 or M152 by submerged fermentation. Mycelia and polysaccharide production of A. blazei Murrill with submerged fermentation is potentially feasible.

In Vitro Fermentation of Linear and α-1,2-Branched Dextrans by the Human Fecal Microbiota▿

PubMed Central

Sarbini, Shahrul R.; Kolida, Sofia; Naeye, Thierry; Einerhand, Alexandra; Brison, Yoann; Remaud-Simeon, Magali; Monsan, Pierre; Gibson, Glenn R.; Rastall, Robert A.

2011-01-01

The role of structure and molecular weight in fermentation selectivity in linear α-1,6 dextrans and dextrans with α-1,2 branching was investigated. Fermentation by gut bacteria was determined in anaerobic, pH-controlled fecal batch cultures after 36 h. Inulin (1%, wt/vol), which is a known prebiotic, was used as a control. Samples were obtained at 0, 10, 24, and 36 h of fermentation for bacterial enumeration by fluorescent in situ hybridization and short-chain fatty acid analyses. The gas production of the substrate fermentation was investigated in non-pH-controlled, fecal batch culture tubes after 36 h. Linear and branched 1-kDa dextrans produced significant increases in Bifidobacterium populations. The degree of α-1,2 branching did not influence the Bifidobacterium populations; however, α-1,2 branching increased the dietary fiber content, implying a decrease in digestibility. Other measured bacteria were unaffected by the test substrates except for the Bacteroides-Prevotella group, the growth levels of which were increased on inulin and 6- and 70-kDa dextrans, and the Faecalibacterium prausnitzii group, the growth levels of which were decreased on inulin and 1-kDa dextrans. A considerable increase in short-chain fatty acid concentration was measured following the fermentation of all dextrans and inulin. Gas production rates were similar among all dextrans tested but were significantly slower than that for inulin. The linear 1-kDa dextran produced lower total gas and shorter time to attain maximal gas production compared to those of the 70-kDa dextran (branched) and inulin. These findings indicate that dextrans induce a selective effect on the gut flora, short-chain fatty acids, and gas production depending on their length. PMID:21666027

alpha-1,4-Glucan lyase, a new class of starch/glycogen degrading enzyme. III. Substrate specificity, mode of action, and cleavage mechanism.

PubMed

Yu, S; Ahmad, T; Kenne, L; Pedersén, M

1995-05-11

The alpha-1,4-glucan lyase (EC 4.2.2.-), purified from the red alga Gracilariopsis lemaneiformis, is a single polypeptide with a molecular mass of 116,654 Da as determined by matrix-assisted laser-desorption mass spectrometry. It degraded maltose, maltosaccharides, amylose, amylopectin and glycogen, forming 1,5-anhydro-D-fructose from the non-reducing end groups. The substrate specificity, mode of action, and cleavage mechanism of the enzyme were studied by using various naturally occurring and synthesized substrates. This enzyme was highly specific for the alpha-1,4-D-glucosidic bond. When a linear alpha-1,4-glucan was used as substrate, the enzyme split the substrate from the non-reducing end and released 1,5-anhydro-D-fructose successively until only one glucose unit was left. When a branched pentasaccharide of 6(2)-alpha-maltosylmaltotriose, obtained from glycogen by alpha-amylase limitation, was used as substrate, the glucose group in the 4-position of the 4,6-branched residue was not cleaved off. Using maltoheptaose as substrate and following the reaction with HPLC and 1H-NMR spectroscopy, it was found that the action mode of the lyase followed a multichain attack mechanism. 1H- and 13C-NMR spectroscopic studies on unlabelled and labelled amylose (1-2H, 2-2H, 1-13C) as substrates indicated that the lyase cleaved the C-(1')-O(4) bond forming a double bond between C-1' and C-2', thus forming the enol form of 1,5-anhydro-D-fructose. It also indicated that the catalytic process of the lyase involved proton exchanges among C-1, C-2, C-3 and the solvent.

Ergot alkaloids produced by submerged cultures of Claviceps zizaniae.

PubMed

Kantorová, Michaela; Kolínská, Renata; Pazoutová, Sylvie; Honzátko, Ales; Havlícek, Vladimír; Flieger, Miroslav

2002-07-01

Two ergopeptine alkaloids, alpha-ergocryptine (1) and its C(8) epimer alpha-ergocryptinine, have been isolated from the mycelium and fermentation broth of submerged cultures of Claviceps zizaniae CCM 8240. The structure of 1 was determined by HPLC/positive ion APCI MS and NMR analysis. Alkaloid concentrations of 10 microg/mL in 14-day-old fermentation broth and 1 mg/g of dry mycelium mass were found. These results are of considerable biotechnological interest since these were the only detectable alkaloids produced. Toxicity of naturally occurring sclerotia of C. zizaniae cannot be excluded.

Biodegradation of Oil Slicks on the Marine Environment.

DTIC Science & Technology

1976-11-01

Ransenula, Candida, Rho4o~torula and Penicillium . The substrate range of these organisms wasevaluated on 11 aliphatic , alicyclic and aromatic...and aromatic compounds (TR 1, ],97C). On the basis of the above substrate range tests and growth, characteristic., a Flavobacteri um sp . and a...incubation. ~ -Paraff Ins served as the bestsubstrates for both organisms, but Plavobacteriua sp . exhibited higher rates of mineralization. Branching

Progenitor outgrowth from the niche in Drosophila trachea is guided by FGF from decaying branches.

PubMed

Chen, Feng; Krasnow, Mark A

2014-01-10

Although there has been progress identifying adult stem and progenitor cells and the signals that control their proliferation and differentiation, little is known about the substrates and signals that guide them out of their niche. By examining Drosophila tracheal outgrowth during metamorphosis, we show that progenitors follow a stereotyped path out of the niche, tracking along a subset of tracheal branches destined for destruction. The embryonic tracheal inducer branchless FGF (fibroblast growth factor) is expressed dynamically just ahead of progenitor outgrowth in decaying branches. Knockdown of branchless abrogates progenitor outgrowth, whereas misexpression redirects it. Thus, reactivation of an embryonic tracheal inducer in decaying branches directs outgrowth of progenitors that replace them. This explains how the structure of a newly generated tissue is coordinated with that of the old.

Effect of single-strand break on branch migration and folding dynamics of Holliday junctions.

PubMed

Palets, Dmytro; Lushnikov, Alexander Y; Karymov, Mikhail A; Lyubchenko, Yuri L

2010-09-22

The Holliday junction (HJ), or four-way junction, is a central intermediate state of DNA for homologous genetic recombination and other genetic processes such as replication and repair. Branch migration is the process by which the exchange of homologous DNA regions occurs, and it can be spontaneous or driven by proteins. Unfolding of the HJ is required for branch migration. Our previous single-molecule fluorescence studies led to a model according to which branch migration is a stepwise process consisting of consecutive migration and folding steps. Folding of the HJ in one of the folded conformations terminates the branch migration phase. At the same time, in the unfolded state HJ rapidly migrates over entire homology region of the HJ in one hop. This process can be affected by irregularities in the DNA double helical structure, so mismatches almost terminate a spontaneous branch migration. Single-stranded breaks or nicks are the most ubiquitous defects in the DNA helix; however, to date, their effect on the HJ branch migration has not been studied. In addition, although nicked HJs are specific substrates for a number of enzymes involved in DNA recombination and repair, the role of this substrate specificity remains unclear. Our main goal in this work was to study the effect of nicks on the efficiency of HJ branch migration and the dynamics of the HJ. To accomplish this goal, we applied two single-molecule methods: atomic force microscopy and fluorescence resonance energy transfer. The atomic force microscopy data show that the nick does not prevent branch migration, but it does decrease the probability that the HJ will pass the DNA lesion. The single-molecule fluorescence resonance energy transfer approaches were instrumental in detailing the effects of nicks. These studies reveal a dramatic change of the HJ dynamics. The nick changes the structure and conformational dynamics of the junctions, leading to conformations with geometries that are different from those for the intact HJ. On the basis of these data, we propose a model of branch migration in which the propensity of the junction to unfold decreases the lifetimes of folded states, thereby increasing the frequency of junction fluctuations between the folded states. Copyright © 2010 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Quantification of extraradical soil mycelium and ectomycorrhizas of Boletus edulis in a Scots pine forest with variable sporocarp productivity.

PubMed

De la Varga, Herminia; Agueda, Beatriz; Martínez-Peña, Fernando; Parladé, Javier; Pera, Joan

2012-01-01

The availability of most edible ectomycorrhizal mushrooms depends on their natural fructification. Sporocarp formation of these fungi is linked to habitat characteristics and climate conditions, but these data alone do not explain all the trends of fungal fruiting and dynamics. It could be hypothesized that the amount of soil mycelia could also be related to the production of carpophores. Soil samples (five cylinders of 250 cm(3) per plot) were taken monthly, from September to November, in five fenced permanent plots (5 × 5 m) in Pinar Grande (Soria, Spain), a Pinus sylvestris stand situated in the north of the Sistema Ibérico mountain range. Plots were chosen to establish a gradient of Boletus edulis productivity from 0 to 38.5 kg/ha year, according to the mean fresh weight of sporocarps collected during the last 10 years. B. edulis ectomycorrhizal root tips were identified in each soil sample according to its morphology and counted. DNA extractions were performed with the PowerSoil(TM) DNA Isolation Kit and quantification of extraradical soil mycelium by real-time polymerase chain reaction using specific primers and a TaqMan® probe. The concentration of soil mycelium of B. edulis (mg mycelium/g soil) did not differ significantly between plots (p = 0.1397), and sampling time (p = 0.7643) within the fructification period. The number of mycorrhizal short roots per soil volume showed significant differences between the plots (p = 0.0050) and the three sampling times (p < 0.0001). No significant correlation between the number of mycorrhizas and the productivity of the plot (kg of B. edulis/ha year) was detected (p = 0.615). A statistically significant positive correlation (p = 0.0481) was detected between the concentration of mycelia of B. edulis in the soil samples and the presence of short roots mycorrhizal with B. edulis in these samples. The productivity of the plots, in terms of sporocarps produced during the last 10 years, was not correlated either with the concentration of soil mycelium or with the presence or abundance of ectomycorrhizas.

ςBldN, an Extracytoplasmic Function RNA Polymerase Sigma Factor Required for Aerial Mycelium Formation in Streptomyces coelicolor A3(2)

PubMed Central

Bibb, Maureen J.; Molle, Virginie; Buttner, Mark J.

2000-01-01

Sporulation mutants of Streptomyces coelicolor appear white because they are defective in the synthesis of the gray polyketide spore pigment, and such white (whi) mutants have been used to define 13 sporulation loci. whiN, one of five new whi loci identified in a recent screen of NTG (N-methyl-N′-nitro-N-nitrosoguanidine)-induced whi strains (N. J. Ryding et al., J. Bacteriol. 181:5419–5425, 1999), was defined by two mutants, R112 and R650. R650 produced frequent spores that were longer than those of the wild type. In contrast, R112 produced long, straight, undifferentiated hyphae, although rare spore chains were observed, sometimes showing highly irregular septum placement. Subcloning and sequencing showed that whiN encodes a member of the extracytoplasmic function subfamily of RNA polymerase sigma factors and that the sigma factor has an unusual N-terminal extension of approximately 86 residues that is not present in other sigma factors. A constructed whiN null mutant failed to form aerial mycelium (the “bald” phenotype) and, as a consequence, whiN was renamed bldN. This observation was not totally unexpected because, on some media, the R112 point mutant produced substantially less aerial mycelium than its parent, M145. The bldN null mutant did not fit simply into the extracellular signaling cascade proposed for S. coelicolor bld mutants. Expression of bldN was analyzed during colony development in wild-type and aerial mycelium-deficient bld strains. bldN was transcribed from a single promoter, bldNp. bldN transcription was developmentally regulated, commencing approximately at the time of aerial mycelium formation, and depended on bldG and bldH, but not on bldA, bldB, bldC, bldF, bldK, or bldJ or on bldN itself. Transcription from the p1 promoter of the response-regulator gene bldM depended on bldN in vivo, and the bldMp1 promoter was shown to be a direct biochemical target for ςBldN holoenzyme in vitro. PMID:10913095

Fluctuations in the population density of Gram-negative bacteria in a chernozem in the course of a succession initiated by moistening and chitin and cellulose introduction

NASA Astrophysics Data System (ADS)

Polyanskaya, L. M.; Ivanov, K. E.; Zvyagintsev, D. G.

2012-10-01

The role has been studied of Gram-negative bacteria in the destruction of polymers widely spread in soils: chitin and cellulose. The introduction of chitin had no positive effect on the population density of Gram-negative bacteria, but it advanced the date of their appearance: the maximum population density of Gram-negative bacteria was recorded not on the 7th-15th day as in the control but much earlier, on the 3rd-7th day of the experiment. Consequently, the introduction of chitin as an additional source of nutrition promoted revealing of the Gram-negative bacteria already at the early stages of the succession. In the course of the succession, when the fungal mycelium begins to die off, the actinomycetic mycelium increases in length, i.e., Gram-negative bacteria are replaced at this stage with Gram-positive ones, the leading role among which belongs to actinomycetes. The growth rate of Gram-negative bacteria is higher than that of actinomycetes, so they start chitin utilization at the early stages of the succession, whereas actinomycetes dominate at the late stages. The population density of Gram-negative bacteria was lower under the anaerobic conditions as compared with that in the aerobic ones. The population density of Gram-negative bacteria in the lower layer of the A horizon and in the B horizon was slightly higher only in the case of the chitin introduction. When cellulose was introduced into the soil under aerobic conditions, the population density of Gram-negative bacteria in all the layers of the A horizon was maximal from the 14th to the 22nd day of the experiment. Cellulose was utilized in the soil mostly by fungi, and this was suggested by the increase of the length of the fungal mycelium. Simultaneously, an increase in the length of the actinomycetal mycelium was observed, as these organisms also perform cellulose hydrolysis in soils. The Gram-negative bacteria began to develop at the stage of the fungal mycelium destruction, which indirectly confirmed the chitinolytic activity of these bacteria.

Propagules of arbuscular mycorrhizal fungi in a secondary dry forest of Oaxaca, Mexico.

PubMed

Guadarrama, Patricia; Castillo-Argüero, Silvia; Ramos-Zapata, José A; Camargo-Ricalde, Sara L; Alvarez-Sánchez, Javier

2008-03-01

Plant cover loss due to changes in land use promotes a decrease in spore diversity of arbuscular mycorrhizal fungi (AMF), viable mycelium and, therefore, in AMF colonization, this has an influence in community diversity and, as a consequence, in its recovery. To evaluate different AMF propagules, nine plots in a tropical dry forest with secondary vegetation were selected: 0, 1, 7, 10, 14, 18, 22, 25, and 27 years after abandonment in Nizanda, Oaxaca, Mexico. The secondary vegetation with different stages of development is a consequence of slash and burn agriculture, and posterior abandonment. Soil samples (six per plot) were collected and percentage of AMF field colonization, extrarradical mycelium, viable spore density, infectivity and most probable number (MPN) ofAMF propagules were quantified through a bioassay. Means for field colonization ranged between 40% and 70%, mean of total mycelium length was 15.7 +/- 1.88 mg(-1) dry soil, with significant differences between plots; however, more than 40% of extracted mycelium was not viable, between 60 and 456 spores in 100 g of dry soil were recorded, but more than 64% showed some kind of damage. Infectivity values fluctuated between 20% and 50%, while MPN showed a mean value of 85.42 +/- 44.17 propagules (100 g dry soil). We conclude that secondary communities generated by elimination of vegetation with agricultural purposes in a dry forest in Nizanda do not show elimination of propagules, probably as a consequence of the low input agriculture practices in this area, which may encourage natural regeneration.

Comparison of methodologies for conidia production by Alternaria alternata from citrus

PubMed Central

Carvalho, Daniel D. C.; Alves, Eduardo; Batista, Tereza R. S.; Camargos, Renato B.; Lopes, Eloísa A. G. L.

2008-01-01

Conidia production is a problem in the study of Alternaria alternata from citrus. Thus, this study aimed to compare existing methodologies for conidial production of A. alternata isolated from Ponkan tangerine (2 isolates), Cravo lemon (1 isolate), Pêra orange (2 isolates) and Murcott tangor (1 isolate). The methodologies used were conidia production with 12 and 24 hours under white fluorescent light, evaluation with 24 and 48 hours after applying fungal mycelium stress technique, cold stress followed by injury of mycelium and evaluation with 24 hours, using healthy vegetable tissue and the use of black fluorescent near ultraviolet (NUV) lamp. Satisfactory result was obtained with A. alternata isolate from Murcott tangor, with the production of 2.8 × 105 conidia mL-1, when fungal mycelium was stressed (Petri dish with 66.66% of fungi growth) and subsequently 24 h of growth. The use of white light (24 h) and black fluorescent NUV lamp also induced expressive conidia production by one isolate of Ponkan tangerine, which produced 17.2 × 105 and 10.1 × 105conidia mL-1 and another of Murcott tangor, which produced 13.9 × 105 and 10.1 × 105 conidia mL-1, respectively. The remaining methodologies analyzed in this study were not able to induce conidia production in satisfactory quantity. The use of both mycelium stress technique and white light (24 h) and black fluorescent NUV lamp allowed the production of enough quantities of conidia to be used in vitro (detection of fungitoxic substances) and in vivo (pathogenicity test) assays, respectively. PMID:24031309

Erinacine A-Enriched Hericium erinaceus Mycelium Produces Antidepressant-Like Effects through Modulating BDNF/PI3K/Akt/GSK-3β Signaling in Mice.

PubMed

Chiu, Chun-Hung; Chyau, Charng-Cherng; Chen, Chin-Chu; Lee, Li-Ya; Chen, Wan-Ping; Liu, Jia-Ling; Lin, Wen-Hsin; Mong, Mei-Chin

2018-01-24

Antidepressant-like effects of ethanolic extract of Hericium erinaceus (HE) mycelium enriched in erinacine A on depressive mice challenged by repeated restraint stress (RS) were examined. HE at 100, 200 or 400 mg/kg body weight/day was orally given to mice for four weeks. After two weeks of HE administration, all mice except the control group went through with 14 days of RS protocol. Stressed mice exhibited various behavioral alterations, such as extending immobility time in the tail suspension test (TST) and forced swimming test (FST), and increasing the number of entries in open arm (POAE) and the time spent in the open arm (PTOA). Moreover, the levels of norepinephrine (NE), dopamine (DA) and serotonin (5-HT) were decreased in the stressed mice, while the levels of interleukin (IL)-6 and tumor necrosis factor (TNF)-α were increased. These changes were significantly inverted by the administration of HE, especially at the dose of 200 or 400 mg/kg body weight/day. Additionally, HE was shown to activate the BDNF/TrkB/PI3K/Akt/GSK-3β pathways and block the NF-κB signals in mice. Taken together, erinacine A-enriched HE mycelium could reverse the depressive-like behavior caused by RS and was accompanied by the modulation of monoamine neurotransmitters as well as pro-inflammatory cytokines, and regulation of BDNF pathways. Therefore, erinacine A-enriched HE mycelium could be an attractive agent for the treatment of depressive disorders.

Erinacine A-Enriched Hericium erinaceus Mycelium Produces Antidepressant-Like Effects through Modulating BDNF/PI3K/Akt/GSK-3β Signaling in Mice

PubMed Central

Chiu, Chun-Hung; Chyau, Charng-Cherng; Chen, Chin-Chu; Lee, Li-Ya; Chen, Wan-Ping; Liu, Jia-Ling; Lin, Wen-Hsin; Mong, Mei-Chin

2018-01-01

Antidepressant-like effects of ethanolic extract of Hericium erinaceus (HE) mycelium enriched in erinacine A on depressive mice challenged by repeated restraint stress (RS) were examined. HE at 100, 200 or 400 mg/kg body weight/day was orally given to mice for four weeks. After two weeks of HE administration, all mice except the control group went through with 14 days of RS protocol. Stressed mice exhibited various behavioral alterations, such as extending immobility time in the tail suspension test (TST) and forced swimming test (FST), and increasing the number of entries in open arm (POAE) and the time spent in the open arm (PTOA). Moreover, the levels of norepinephrine (NE), dopamine (DA) and serotonin (5-HT) were decreased in the stressed mice, while the levels of interleukin (IL)-6 and tumor necrosis factor (TNF)-α were increased. These changes were significantly inverted by the administration of HE, especially at the dose of 200 or 400 mg/kg body weight/day. Additionally, HE was shown to activate the BDNF/TrkB/PI3K/Akt/GSK-3β pathways and block the NF-κB signals in mice. Taken together, erinacine A-enriched HE mycelium could reverse the depressive-like behavior caused by RS and was accompanied by the modulation of monoamine neurotransmitters as well as pro-inflammatory cytokines, and regulation of BDNF pathways. Therefore, erinacine A-enriched HE mycelium could be an attractive agent for the treatment of depressive disorders. PMID:29364170

Ufd2p synthesizes branched ubiquitin chains to promote the degradation of substrates modified with atypical chains

PubMed Central

Liu, Chao; Liu, Weixiao; Ye, Yihong; Li, Wei

2017-01-01

Ubiquitination of a subset of proteins by ubiquitin chain elongation factors (E4), represented by Ufd2p in Saccharomyces cerevisiae, is a pivotal regulator for many biological processes. However, the mechanism of Ufd2p-mediated ubiquitination is largely unclear. Here, we show that Ufd2p catalyses K48-linked multi-monoubiquitination on K29-linked ubiquitin chains assembled by the ubiquitin ligase (Ufd4p), resulting in branched ubiquitin chains. This reaction depends on the interaction of K29-linked ubiquitin chains with two N-terminal loops of Ufd2p. Only following the addition of K48-linked ubiquitin to substrates modified with K29-linked ubiquitin chains, can the substrates be escorted to the proteasome for degradation. We demonstrate that this ubiquitin chain linkage switching reaction is essential for ERAD, oleic acid and acid pH resistance in yeast. Thus, our results suggest that Ufd2p functions by switching ubiquitin chain linkages to allow the degradation of proteins modified with a ubiquitin linkage, which is normally not targeted to the proteasome. PMID:28165462

Transformation of Mycelial and Yeast Forms of Paracoccidioides brasiliensis in Cultures and in Experimental Inoculations

PubMed Central

Carbonell, Luis M.; Rodríguez, Joaquín

1965-01-01

Carbonell, Luis M. (Instituto Venezolano de Investigaciones Científicas, Caracas, Venezuela), and Joaquín Rodríguez. Transformation of mycelial and yeast forms of Paracoccidioides brasiliensis in cultures and in experimental inoculations. J. Bacteriol. 90:504–510. 1965.—Experimental transformations of mycelial to yeast and yeast to mycelial forms in culture, and mycelial to yeast forms in tissue, were studied. All the transitional forms that appeared in culture were also seen in tissue, but in fewer number. Most of the hyphae in culture were transformed into yeast, but only a few in tissue. Yeast appeared in testicle around the 3rd day after inoculation, but on the 10th day in subcutaneous tissue. Pathogenicity of mycelium was high, since yeast was found in almost all of the organs inoculated with mycelium. Histologically, an acute inflammation occurred first, owing to the inoculation of mycelium, followed by a giant-cell granuloma with abundant hyphae detritus. These giant cells almost disappeared about 10 days after inoculation, giving place to a second giant-cell granuloma with yeast forms. Images PMID:14329466

Development of actinomycetes in brown semidesert soil under low water pressure

NASA Astrophysics Data System (ADS)

Zvyagintsev, D. G.; Zenova, G. M.; Sudnitsyn, I. I.; Gracheva, T. A.; Lapygina, E. E.; Napol'skaya, K. R.; Sydnitsyna, A. E.

2012-07-01

Under laboratory conditions, the spores of a xerotolerant Streptomyces odorifera strain germinated in brown semidesert soil even at extremely low soil water pressure ( P = -96.4 MPa, -964 atm, a w 0.50); the plantlets increased in length and formed mycelium, on which a new generation of spores was produced (a complete development cycle of the actinomycetes—from a spore to the formation of new spores—passed). The duration of the first cycles of the actinomycetes' development varied from 13 days at P = -27 atm to 57 days at P = -964 atm and was directly proportional to the absolute value of the soil water pressure ( P). In the first cycles of the actinomycetes' development, the rate of increase of the concentration of the germinated spores and mycelium, as well as the logarithms of the mycelium-to-germinated spore concentration ratios, was inversely proportional to the logarithm of P. These relationships indicated that the energy state of the water determined its availability to soil biota and, hence, the activity of its physiological and biochemical processes.

Seeds as natural matrices for immobilization of Aspergillus niger mycelium producing pectinases.

PubMed

Fiedurek, J; Szczodrak, J; Rogalski, J

1995-04-01

A simple method for the immobilization of Aspergillus niger mycelium producing polygalacturonase (PG) and pectinesterase (PE) is described. Fungal conidia were immobilized on wheat, rye, barley, peas, buckwheat and mustards seeds. Spongy mycelia overgrowing the seed surfaces on mineral medium with pectin produced extracellular PG and PE; the highest production was reached on the wheat carrier. Some of the variables influencing the enzymatic activity have been optimized. After every 24 h, a culture liquid with 6.8-7.8 U of PG ml-1 and 7.0-10.1 U of PE ml-1 was obtained. This procedure also made possible repeated batch enzyme production and, as many as eight subsequent 24-h batches could be fermented by using the same carrier without any loss of PG activity. The addition of sodium orthovanadate (1 mmol) into the medium with pectin caused a significant increase in PG and PE activity produced by free cells of A. niger (by 1.59-fold and 1.67-fold respectively), and only 0.47-fold of PG activity in case of the immobilized mycelium.

Substrate specificities and intracellular distributions of three N-glycan processing enzymes functioning at a key branch point in the insect N-glycosylation pathway.

PubMed

Geisler, Christoph; Jarvis, Donald L

2012-03-02

Man(α1-6)[GlcNAc(β1-2)Man(α1-3)]ManGlcNAc(2) is a key branch point intermediate in the insect N-glycosylation pathway because it can be either trimmed by a processing β-N-acetylglucosaminidase (FDL) to produce paucimannosidic N-glycans or elongated by N-acetylglucosaminyltransferase II (GNT-II) to produce complex N-glycans. N-acetylglucosaminyltransferase I (GNT-I) contributes to branch point intermediate production and can potentially reverse the FDL trimming reaction. However, there has been no concerted effort to evaluate the relationships among these three enzymes in any single insect system. Hence, we extended our previous studies on Spodoptera frugiperda (Sf) FDL to include GNT-I and -II. Sf-GNT-I and -II cDNAs were isolated, the predicted protein sequences were analyzed, and both gene products were expressed and their acceptor substrate specificities and intracellular localizations were determined. Sf-GNT-I transferred N-acetylglucosamine to Man(5)GlcNAc(2), Man(3)GlcNAc(2), and GlcNAc(β1-2)Man(α1-6)[Man(α1-3)]ManGlcNAc(2), demonstrating its role in branch point intermediate production and its ability to reverse FDL trimming. Sf-GNT-II only transferred N-acetylglucosamine to Man(α1-6)[GlcNAc(β1-2)Man(α1-3)]ManGlcNAc(2), demonstrating that it initiates complex N-glycan production, but cannot use Man(3)GlcNAc(2) to produce hybrid or complex structures. Fluorescently tagged Sf-GNT-I and -II co-localized with an endogenous Sf Golgi marker and Sf-FDL co-localized with Sf-GNT-I and -II, indicating that all three enzymes are Golgi resident proteins. Unexpectedly, fluorescently tagged Drosophila melanogaster FDL also co-localized with Sf-GNT-I and an endogenous Drosophila Golgi marker, indicating that it is a Golgi resident enzyme in insect cells. Thus, the substrate specificities and physical juxtapositioning of GNT-I, GNT-II, and FDL support the idea that these enzymes function at the N-glycan processing branch point and are major factors determining the net outcome of the insect cell N-glycosylation pathway.

Early development of Moniliophthora perniciosa basidiomata and developmentally regulated genes

PubMed Central

2009-01-01

Background The hemibiotrophic fungus Moniliophthora perniciosa is the causal agent of Witches' broom, a disease of Theobroma cacao. The pathogen life cycle ends with the production of basidiocarps in dead tissues of the infected host. This structure generates millions of basidiospores that reinfect young tissues of the same or other plants. A deeper understanding of the mechanisms underlying the sexual phase of this fungus may help develop chemical, biological or genetic strategies to control the disease. Results Mycelium was morphologically analyzed prior to emergence of basidiomata by stereomicroscopy, light microscopy and scanning electron microscopy. The morphological changes in the mycelium before fructification show a pattern similar to other members of the order Agaricales. Changes and appearance of hyphae forming a surface layer by fusion were correlated with primordia emergence. The stages of hyphal nodules, aggregation, initial primordium and differentiated primordium were detected. The morphological analysis also allowed conclusions on morphogenetic aspects. To analyze the genes involved in basidiomata development, the expression of some selected EST genes from a non-normalized cDNA library, representative of the fruiting stage of M. perniciosa, was evaluated. A macroarray analysis was performed with 192 selected clones and hybridized with two distinct RNA pools extracted from mycelium in different phases of basidiomata formation. This analysis showed two groups of up and down-regulated genes in primordial phases of mycelia. Hydrophobin coding, glucose transporter, Rho-GEF, Rheb, extensin precursor and cytochrome p450 monooxygenase genes were grouped among the up-regulated. In the down-regulated group relevant genes clustered coding calmodulin, lanosterol 14 alpha demethylase and PIM1. In addition, 12 genes with more detailed expression profiles were analyzed by RT-qPCR. One aegerolysin gene had a peak of expression in mycelium with primordia and a second in basidiomata, confirming their distinctiveness. The number of transcripts of the gene for plerototolysin B increased in reddish-pink mycelium and indicated an activation of the initial basidiomata production even at this culturing stage. Expression of the glucose transporter gene increased in mycelium after the stress, coinciding with a decrease of adenylate cyclase gene transcription. This indicated that nutrient uptake can be an important signal to trigger fruiting in this fungus. Conclusion The identification of genes with increased expression in this phase of the life cycle of M. perniciosa opens up new possibilities of controlling fungus spread as well as of genetic studies of biological processes that lead to basidiomycete fruiting. This is the first comparative morphologic study of the early development both in vivo and in vitro of M. perniciosa basidiomata and the first description of genes expressed at this stage of the fungal life cycle. PMID:19653910

Relation between germination and mycelium growth of individual fungal spores.

PubMed

Gougouli, Maria; Koutsoumanis, Konstantinos P

2013-02-15

The relation between germination time and lag time of mycelium growth of individual spores was studied by combining microscopic and macroscopic techniques. The radial growth of a large number (100-200) of Penicillium expansum and Aspergillus niger mycelia originating from single spores was monitored macroscopically at isothermal conditions ranging from 0 to 30°C and 10 to 41.5°C, respectively. The radial growth curve for each mycelium was fitted to a linear model for the estimation of mycelium lag time. The results showed that the lag time varied significantly among single spores. The cumulative frequency distributions of the lag times were fitted to the modified Gompertz model and compared with the respective distributions for the germination time, which were obtained microscopically. The distributions of the measured mycelium lag time were found to be similar to the germination time distributions under the same conditions but shifted in time with the lag times showing a significant delay compared to germination times. A numerical comparison was also performed based on the distribution parameters λ(m) and λ(g), which indicate the time required from the spores to start the germination process and the completion of the lag phase, respectively. The relative differences %(λ(m)-λ(g))/λ(m) were not found to be significantly affected by temperatures tested with mean values of 72.5±5.1 and 60.7±2.1 for P. expansum for A. niger, respectively. In order to investigate the source of the above difference, a time-lapse microscopy method was developed providing videos with the behavior of single fungal spore from germination until mycelium formation. The distances of the apexes of the first germ tubes that emerged from the swollen spore were measured in each frame of the videos and these data were expressed as a function of time. The results showed that in the early hyphal development, the measured radii appear to increase exponentially, until a certain time, where growth becomes linear. The two phases of hyphal development can explain the difference between germination and lag time. Since the lag time is estimated from the extrapolation of the regression line of the linear part of the graph only, its value is significantly higher than the germination time, t(G). The relation of germination and lag time was further investigated by comparing their temperature dependence using the Cardinal Model with Inflection. The estimated values of the cardinal parameters (T(min), T(opt), and T(max)) for 1/λ(g) were found to be very close to the respective values for 1/λ(m), indicating similar temperature dependence between them. Copyright © 2012 Elsevier B.V. All rights reserved.

Synthesis of 1,2-cis-2-C-branched aryl-C-glucosides via desulfurization of carbohydrate based hemithioacetals

PubMed Central

Mebrahtu, Fanuel M; Manana, Mandlenkosi M; Madumo, Kagiso; Sokamisa, Mokela S

2015-01-01

Summary 1-C and 2-C-branched carbohydrates are present as substructures in a number of biologically important compounds. Although the synthesis of such carbohydrate derivatives is extensively studied, the synthesis of 1,2-cis-2-C-branched C-, S-, and N-glycosides is less explored. In this article a synthetic strategy for the synthesis of 1,2-cis-2-C-branched-aryl-C-glucosides is reported via a hydrogenolytic desulfurization of suitably orientated carbohydrate based hemithioacetals. 1,2-cis-2-Hydroxymethyl and 2-carbaldehyde of aryl-C-glucosides have been synthesized using the current strategy in very good yields. The 2-carbaldehyde-aryl-C-glucosides have been identified as suitable substrates for the stereospecific preparation of 2,3-unsaturated-aryl-C-glycosides (Ferrier products). PMID:26124859

Progenitor Outgrowth from the Niche in Drosophila Trachea Is Guided by FGF from Decaying Branches

PubMed Central

Chen, Feng; Krasnow, Mark A.

2014-01-01

Although there has been progress identifying adult stem and progenitor cells and the signals that control their proliferation and differentiation, little is known about the substrates and signals that guide them out of their niche. By examining Drosophila tracheal outgrowth during metamorphosis, we show that progenitors follow a stereotyped path out of the niche, tracking along a subset of tracheal branches destined for destruction. The embryonic tracheal inducer branchless FGF (fibroblast growth factor) is expressed dynamically just ahead of progenitor outgrowth in decaying branches. Knockdown of branchless abrogates progenitor outgrowth, whereas misexpression redirects it. Thus, reactivation of an embryonic tracheal inducer in decaying branches directs outgrowth of progenitors that replace them. This explains how the structure of a newly generated tissue is coordinated with that of the old. PMID:24408434

Substrate-Directed Hydroacylation: Rh-Catalyzed Coupling of Vinyl Phenols and Non-Chelating Aldehydes

PubMed Central

Murphy, Stephen K.; Bruch, Achim

2014-01-01

We report a protocol for branched-selective hydroacylation of vinylphenols with aryl, alkenyl and alkyl aldehydes. This cross-coupling yields α-aryl ketones that can be cyclized to benzofurans, and it enables access to eupomatenoid natural products in four steps or less from eugenol. Excellent reactivity and high levels of branched regioselectivity are obtained. We propose that aldehyde decarbonylation is overcome by using an anionic directing group on the olefin and a small bite-angle diphosphine ligand. PMID:24478146

Synthesis and photoelectrochemical properties of a novel CuO/ZnO nanorod photocathode for solar hydrogen generation

NASA Astrophysics Data System (ADS)

Shaislamov, Ulugbek; Lee, Heon-Ju

2016-10-01

Here, we present a facile synthesis method and photoelectrochemical characterizations of a p-type CuO-nanorod array photoelectrode with ZnO nanorod branches. Vertically-aligned CuO nanorods were synthesized by using direct oxidation of metallic Cu nanorods grown on a Cu substrate by using a facile template-assisted electrodeposition method. The formed CuONR/ZnONB hierarchically-structured photoelectrode exhibited remarkable photoelectrodechemical performance and outstanding stability compared to the CuO NR photoelectrode without ZnO NR branches. Morphological, optical and electrochemical characterizations were carried out in order to examine the effects of ZnO nanorod branches on the stability and the overall electrochemical performance of the electrode.

Enzymes involved in branched-chain amino acid metabolism in humans.

PubMed

Adeva-Andany, María M; López-Maside, Laura; Donapetry-García, Cristóbal; Fernández-Fernández, Carlos; Sixto-Leal, Cristina

2017-06-01

Branched-chain amino acids (leucine, isoleucine and valine) are structurally related to branched-chain fatty acids. Leucine is 2-amino-4-methyl-pentanoic acid, isoleucine is 2-amino-3-methyl-pentanoic acid, and valine is 2-amino-3-methyl-butanoic acid. Similar to fatty acid oxidation, leucine and isoleucine produce acetyl-coA. Additionally, leucine generates acetoacetate and isoleucine yields propionyl-coA. Valine oxidation produces propionyl-coA, which is converted into methylmalonyl-coA and succinyl-coA. Branched-chain aminotransferase catalyzes the first reaction in the catabolic pathway of branched-chain amino acids, a reversible transamination that converts branched-chain amino acids into branched-chain ketoacids. Simultaneously, glutamate is converted in 2-ketoglutarate. The branched-chain ketoacid dehydrogenase complex catalyzes the irreversible oxidative decarboxylation of branched-chain ketoacids to produce branched-chain acyl-coA intermediates, which then follow separate catabolic pathways. Human tissue distribution and function of most of the enzymes involved in branched-chain amino acid catabolism is unknown. Congenital deficiencies of the enzymes involved in branched-chain amino acid metabolism are generally rare disorders. Some of them are associated with reduced pyruvate dehydrogenase complex activity and respiratory chain dysfunction that may contribute to their clinical phenotype. The biochemical phenotype is characterized by accumulation of the substrate to the deficient enzyme and its carnitine and/or glycine derivatives. It was established at the beginning of the twentieth century that the plasma level of the branched-chain amino acids is increased in conditions associated with insulin resistance such as obesity and diabetes mellitus. However, the potential clinical relevance of this elevation is uncertain.

[Degradation of fluorene and fluoranthene by the basidiomycete Pleurotus ostreatus].

PubMed

Pozdnyakova, N N; Chernyshova, M P; Grinev, V S; Landesman, E O; Koroleva, O V; Turkovskaya, O V

2016-01-01

The dependence of the degree of fluorene and fluoranthene degradation by the fungus Pleurotus ostreatus D1 on the culture medium composition has been studied. Polycyclic aromatic hydrocarbons (PAHs) have been transformed in Kirk’s medium (under conditions of laccase production) with the formation of a quinone metabolite and 9-fluorenone upon the use of fluoranthene and fluorene as substrates, respectively. More complete degradation with the formation of an intermediate metabolite, phthalic acid that has undergone subsequent utilization, has occurred in basidiomycete-rich medium (under the production of both laccase and versatile peroxidase). The formation of phthalic acid as a metabolite of fluoranthene degradation by lignolytic fungi has been revealed for the first time. The data allow the supposition that both extracellular laccase and laccase on the mycelium surface can participate in the initial stages of PAH metabolism, while versatile peroxidase is necessary for the oxidation of the formed metabolites. A scheme of fluorene metabolism by Pleurotus ostreatus D1 is suggested.

The medicinal Agaricus mushroom cultivated in Brazil: biology, cultivation and non-medicinal valorisation.

PubMed

Largeteau, Michèle L; Llarena-Hernández, Régulo Carlos; Regnault-Roger, Catherine; Savoie, Jean-Michel

2011-12-01

Sun mushroom is a cultivated mushroom extensively studied for its medicinal properties for several years and literature abounds on the topic. Besides, agronomical aspects were investigated in Brazil, the country the mushroom comes from, and some studies focus on the biology of the fungus. This review aimed to present an overview of the non-medicinal knowledge on the mushroom. Areas of commercial production and marketing trends are presented. Its specific fragrance, taste, nutritional value and potential use of extracts as food additives are compared to those of the most cultivated fungi and laboratory models. The interest of the mushroom for lignocellulosic enzyme production and source of biomolecules for the control of plant pathogens are shown. Investigation of genetic variability among cultivars is reported. Growing and storage of mycelium, as well as cultivation conditions (substrate and casing generally based on local products; indoor and outdoor cultivation; diseases and disorders) are described and compared to knowledge on Agaricus bisporus.

Compost Grown Agaricus bisporus Lacks the Ability to Degrade and Consume Highly Substituted Xylan Fragments

PubMed Central

de Vries, Ronald P.; Gruppen, Harry; Kabel, Mirjam A.

2015-01-01

The fungus Agaricus bisporus is commercially grown for the production of edible mushrooms. This cultivation occurs on compost, but not all of this substrate is consumed by the fungus. To determine why certain fractions remain unused, carbohydrate degrading enzymes, water-extracted from mushroom-grown compost at different stages of mycelium growth and fruiting body formation, were analyzed for their ability to degrade a range of polysaccharides. Mainly endo-xylanase, endo-glucanase, β-xylosidase and β-glucanase activities were determined in the compost extracts obtained during mushroom growth. Interestingly, arabinofuranosidase activity able to remove arabinosyl residues from doubly substituted xylose residues and α-glucuronidase activity were not detected in the compost enzyme extracts. This correlates with the observed accumulation of arabinosyl and glucuronic acid substituents on the xylan backbone in the compost towards the end of the cultivation. Hence, it was concluded that compost grown A. bisporus lacks the ability to degrade and consume highly substituted xylan fragments. PMID:26237450

Complete genome sequence of Actinosynnema mirum type strain (101T)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Land, Miriam; Lapidus, Alla; Mayilraj, Shanmugam

2009-05-20

Actinosynnema mirum Hasegawa et al. 1978 is the type species of the genus, and is of phylogenetic interest because of its central phylogenetic location in the Actino-synnemataceae, a rapidly growing family within the actinobacterial suborder Pseudo-nocardineae. A. mirum is characterized by its motile spores borne on synnemata and as a producer of nocardicin antibiotics. It is capable of growing aerobically and under a moderate CO2 atmosphere. The strain is a Gram-positive, aerial and substrate mycelium producing bacterium, originally isolated from a grass blade collected from the Raritan River, New Jersey. Here we describe the features of this organism, together withmore » the complete genome sequence and annotation. This is the first complete genome sequence of a member of the family Actinosynnemataceae, and only the second sequence from the actinobacterial suborder Pseudonocardineae. The 8,248,144 bp long single replicon genome with its 7100 protein-coding and 77 RNA genes is part of the Genomic Encyclopedia of Bacteria and Archaea project.« less

Effects of Cultivating Conditions on the Water Soluble Polysaccharides Content of Ganoderma lucidum Mycelium in Submerged Flask Culture

NASA Astrophysics Data System (ADS)

Rosyida, V. T.; Hayati, S. N.; Apriyana, W.; Darsih, C.; Hernawan; Poeloengasih, C. D.

2017-12-01

The carcinostatic substance in Ganoderma lucidum (Fr.) Karst (Polyporaceae) is a water soluble polysaccharides (WSP) which might be useful in immunotherapy. Attempt to produce effective substances from cultured mycelia is important to carry out since solid cultivation is a time consuming and quality fluctuating. The effects of cultivating conditions on the water soluble polysaccharides content of G. Lucidum mycelium were investigated in submerged flask cultures. Culture from fruiting bodies was maintained on potato dextrose-agar slope. Slopes were inoculated and incubated at 30°C for 7 days, and stored at 4°C. The flask experiments were performed in 100 ml erlenmeyer flasks containing 20 ml of the sterilized media. Actively growing mycelia (1 piece, 5 mm X 5 mm) from a newly prepared slant culture (about 7 days incubation at 30°C) were inoculated into the flask. The pH was measured and adjusted to the desired value by addition of either 4 M HCl or 2.5 M NaOH. Incubation temperature were 20, 25, and 30°C. At the end of inoculation period (14 days) mycelium consisting of individual pellets was harvested and wash for the analysis. WSP content was analysed using phenol-sulfuric acid method. The optimal initial pH for metabolite production would depend on the culture medium. Generally, high values of pH, such as 9, negatively affect both cell growth and WSP production. The optimum temperature range for the high G. lucidum mycelium and WSP production were found to be 25 - 30 °C at pH values 5 - 7 in both of media.

Effect of fungal mycelia on the HPLC-UV and UV-vis spectrophotometric assessment of mycelium-bound epoxide hydrolase using glycidyl phenyl ether.

PubMed

Dolcet, Marta M; Torres, Mercè; Canela, Ramon

2016-06-25

The use of mycelia as biocatalysts has technical and economic advantages. However, there are several difficulties in obtaining accurate results in mycelium-catalysed reactions. Firstly, sample extraction, indispensable because of the presence of mycelia, can bring into the extract components with a similar structure to that of the analyte of interest; secondly, mycelia can influence the recovery of the analyte. We prepared calibration standards of 3-phenoxy-1,2-propanediol (PPD) in the pure solvent and in the presence of mycelia (spiked before or after extraction) from five fungi (Aspergillus niger, Aspergillus tubingensis, Penicillium aurantiogriseum, Penicillium sp. and Aspergillus terreus). The quantification of PPD was carried out by HPLC-UV and UV-vis spectrophotometry. The manuscript shows that the last method is as accurate as the HPLC method. However, the colorimetric method led to a higher data throughput, which allowed the study of more samples in a shorter time. Matrix effects were evaluated visually from the plotted calibration data and statistically by simultaneously comparing the intercept and slope of calibration curves performed with solvent, post-extraction spiked standards and pre-extraction spiked standards. Significant differences were found between the post- and pre-extraction spiked matrix-matched functions. Pre-extraction spiked matrix-matched functions based on A. tubingensis mycelia, selected as the reference, were validated and used to compensate for low recoveries. These validated functions were successfully applied to the quantification of PPD achieved during the hydrolysis of glycidyl phenyl ether by mycelium-bound epoxide hydrolases and equivalent hydrolysis yields were determined by HPLC-UV and UV-vis spectrophotometry. This study may serve as starting point to implement matrix effects evaluation when mycelium-bound epoxide hydrolases are studied. Copyright © 2016 Elsevier B.V. All rights reserved.

Growth inhibition of fungus Phycomyces blakesleeanus by anion channel inhibitors anthracene-9-carboxylic and niflumic acid attained through decrease in cellular respiration and energy metabolites.

PubMed

Stanić, Marina; Križak, Strahinja; Jovanović, Mirna; Pajić, Tanja; Ćirić, Ana; Žižić, Milan; Zakrzewska, Joanna; Antić, Tijana Cvetić; Todorović, Nataša; Živić, Miroslav

2017-03-01

Increasing resistance of fungal strains to known fungicides has prompted identification of new candidates for fungicides among substances previously used for other purposes. We have tested the effects of known anion channel inhibitors anthracene-9-carboxylic acid (A9C) and niflumic acid (NFA) on growth, energy metabolism and anionic current of mycelium of fungus Phycomyces blakesleeanus. Both inhibitors significantly decreased growth and respiration of mycelium, but complete inhibition was only achieved by 100 and 500 µM NFA for growth and respiration, respectively. A9C had no effect on respiration of human NCI-H460 cell line and very little effect on cucumber root sprout clippings, which nominates this inhibitor for further investigation as a potential new fungicide. Effects of A9C and NFA on respiration of isolated mitochondria of P. blakesleeanus were significantly smaller, which indicates that their inhibitory effect on respiration of mycelium is indirect. NMR spectroscopy showed that both A9C and NFA decrease the levels of ATP and polyphosphates in the mycelium of P. blakesleeanus, but only A9C caused intracellular acidification. Outwardly rectifying, fast inactivating instantaneous anionic current (ORIC) was also reduced to 33±5 and 21±3 % of its pre-treatment size by A9C and NFA, respectively, but only in the absence of ATP. It can be assumed from our results that the regulation of ORIC is tightly linked to cellular energy metabolism in P. blakesleeanus, and the decrease in ATP and polyphosphate levels could be a direct cause of growth inhibition.

Crystal Structure of Full-length Mycobacterium tuberculosis H37Rv Glycogen Branching Enzyme; Insights of N-Terminal [beta]-Sandwich in Sustrate Specifity and Enzymatic Activity

DOE Office of Scientific and Technical Information (OSTI.GOV)

Pal, Kuntal; Kumar, Shiva; Sharma, Shikha

2010-07-13

The open reading frame Rv1326c of Mycobacterium tuberculosis (Mtb) H37Rv encodes for an {alpha}-1,4-glucan branching enzyme (MtbGlgB, EC 2.4.1.18, Uniprot entry Q10625). This enzyme belongs to glycoside hydrolase (GH) family 13 and catalyzes the branching of a linear glucose chain during glycogenesis by cleaving a 1 {yields} 4 bond and making a new 1 {yields} 6 bond. Here, we show the crystal structure of full-length MtbGlgB (MtbGlgBWT) at 2.33-{angstrom} resolution. MtbGlgBWT contains four domains: N1 {beta}-sandwich, N2 {beta}-sandwich, a central ({beta}/{alpha}){sub 8} domain that houses the catalytic site, and a C-terminal {beta}-sandwich. We have assayed the amylase activity with amylosemore » and starch as substrates and the glycogen branching activity using amylose as a substrate for MtbGlgBWT and the N1 domain-deleted (the first 108 residues deleted) Mtb{Delta}108GlgB protein. The N1 {beta}-sandwich, which is formed by the first 105 amino acids and superimposes well with the N2 {beta}-sandwich, is shown to have an influence in substrate binding in the amylase assay. Also, we have checked and shown that several GH13 family inhibitors are ineffective against MtbGlgBWT and Mtb{Delta}108GlgB. We propose a two-step reaction mechanism, for the amylase activity (1 {yields} 4 bond breakage) and isomerization (1 {yields} 6 bond formation), which occurs in the same catalytic pocket. The structural and functional properties of MtbGlgB and Mtb{Delta}108GlgB are compared with those of the N-terminal 112-amino acid-deleted Escherichia coli GlgB (EC{Delta}112GlgB).« less

Hierarchical Si/ZnO trunk-branch nanostructure for photocurrent enhancement

PubMed Central

2014-01-01

Hierarchical Si/ZnO trunk-branch nanostructures (NSs) have been synthesized by hot wire assisted chemical vapor deposition method for trunk Si nanowires (NWs) on indium tin oxide (ITO) substrate and followed by the vapor transport condensation (VTC) method for zinc oxide (ZnO) nanorods (NRs) which was laterally grown from each Si nanowires (NWs). A spin coating method has been used for zinc oxide (ZnO) seeding. This method is better compared with other group where they used sputtering method for the same process. The sputtering method only results in the growth of ZnO NRs on top of the Si trunk. Our method shows improvement by having the growth evenly distributed on the lateral sides and caps of the Si trunks, resulting in pine-leave-like NSs. Field emission scanning electron microscope image shows the hierarchical nanostructures resembling the shape of the leaves of pine trees. Single crystalline structure for the ZnO branch grown laterally from the crystalline Si trunk has been identified by using a lattice-resolved transmission electron microscope. A preliminary photoelectrochemical (PEC) cell testing has been setup to characterize the photocurrent of sole array of ZnO NR growth by both hydrothermal-grown (HTG) method and VTC method on ITO substrates. VTC-grown ZnO NRs showed greater photocurrent effect due to its better structural properties. The measured photocurrent was also compared with the array of hierarchical Si/ZnO trunk-branch NSs. The cell with the array of Si/ZnO trunk-branch NSs revealed four-fold magnitude enhancement in photocurrent density compared with the sole array of ZnO NRs obtain from VTC processes. PMID:25246872

Histone acetyltransferase TGF-1 regulates Trichoderma atroviride secondary metabolism and mycoparasitism.

PubMed

Gómez-Rodríguez, Elida Yazmín; Uresti-Rivera, Edith Elena; Patrón-Soberano, Olga Araceli; Islas-Osuna, María Auxiliadora; Flores-Martínez, Alberto; Riego-Ruiz, Lina; Rosales-Saavedra, María Teresa; Casas-Flores, Sergio

2018-01-01

Some filamentous fungi of the Trichoderma genus are used as biocontrol agents against airborne and soilborne phytopathogens. The proposed mechanism by which Trichoderma spp. antagonizes phytopathogens is through the release of lytic enzymes, antimicrobial compounds, mycoparasitism, and the induction of systemic disease-resistance in plants. Here we analyzed the role of TGF-1 (Trichoderma Gcn Five-1), a histone acetyltransferase of Trichoderma atroviride, in mycoparasitism and antibiosis against the phytopathogen Rhizoctonia solani. Trichostatin A (TSA), a histone deacetylase inhibitor that promotes histone acetylation, slightly affected T. atroviride and R. solani growth, but not the growth of the mycoparasite over R. solani. Application of TSA to the liquid medium induced synthesis of antimicrobial compounds. Expression analysis of the mycoparasitism-related genes ech-42 and prb-1, which encode an endochitinase and a proteinase, as well as the secondary metabolism-related genes pbs-1 and tps-1, which encode a peptaibol synthetase and a terpene synthase, respectively, showed that they were regulated by TSA. A T. atroviride strain harboring a deletion of tgf-1 gene showed slow growth, thinner and less branched hyphae than the wild-type strain, whereas its ability to coil around the R. solani hyphae was not affected. Δtgf-1 presented a diminished capacity to grow over R. solani, but the ability of its mycelium -free culture filtrates (MFCF) to inhibit the phytopathogen growth was enhanced. Intriguingly, addition of TSA to the culture medium reverted the enhanced inhibition growth of Δtgf-1 MFCF on R. solani at levels compared to the wild-type MFCF grown in medium amended with TSA. The presence of R. solani mycelium in the culture medium induced similar proteinase activity in a Δtgf-1 compared to the wild-type, whereas the chitinolytic activity was higher in a Δtgf-1 mutant in the absence of R. solani, compared to the parental strain. Expression of mycoparasitism- and secondary metabolism-related genes in Δtgf-1 was differentially regulated in the presence or absence of R. solani. These results indicate that histone acetylation may play important roles in the biocontrol mechanisms of T. atroviride.

Histone acetyltransferase TGF-1 regulates Trichoderma atroviride secondary metabolism and mycoparasitism

PubMed Central

Patrón-Soberano, Olga Araceli; Islas-Osuna, María Auxiliadora; Flores-Martínez, Alberto; Riego-Ruiz, Lina; Rosales-Saavedra, María Teresa

2018-01-01

Some filamentous fungi of the Trichoderma genus are used as biocontrol agents against airborne and soilborne phytopathogens. The proposed mechanism by which Trichoderma spp. antagonizes phytopathogens is through the release of lytic enzymes, antimicrobial compounds, mycoparasitism, and the induction of systemic disease-resistance in plants. Here we analyzed the role of TGF-1 (Trichoderma Gcn Five-1), a histone acetyltransferase of Trichoderma atroviride, in mycoparasitism and antibiosis against the phytopathogen Rhizoctonia solani. Trichostatin A (TSA), a histone deacetylase inhibitor that promotes histone acetylation, slightly affected T. atroviride and R. solani growth, but not the growth of the mycoparasite over R. solani. Application of TSA to the liquid medium induced synthesis of antimicrobial compounds. Expression analysis of the mycoparasitism-related genes ech-42 and prb-1, which encode an endochitinase and a proteinase, as well as the secondary metabolism-related genes pbs-1 and tps-1, which encode a peptaibol synthetase and a terpene synthase, respectively, showed that they were regulated by TSA. A T. atroviride strain harboring a deletion of tgf-1 gene showed slow growth, thinner and less branched hyphae than the wild-type strain, whereas its ability to coil around the R. solani hyphae was not affected. Δtgf-1 presented a diminished capacity to grow over R. solani, but the ability of its mycelium -free culture filtrates (MFCF) to inhibit the phytopathogen growth was enhanced. Intriguingly, addition of TSA to the culture medium reverted the enhanced inhibition growth of Δtgf-1 MFCF on R. solani at levels compared to the wild-type MFCF grown in medium amended with TSA. The presence of R. solani mycelium in the culture medium induced similar proteinase activity in a Δtgf-1 compared to the wild-type, whereas the chitinolytic activity was higher in a Δtgf-1 mutant in the absence of R. solani, compared to the parental strain. Expression of mycoparasitism- and secondary metabolism-related genes in Δtgf-1 was differentially regulated in the presence or absence of R. solani. These results indicate that histone acetylation may play important roles in the biocontrol mechanisms of T. atroviride. PMID:29708970

Streptomyces pini sp. nov., an actinomycete isolated from phylloplane of pine (Pinus sylvestris L.) needle-like leaves.

PubMed

Madhaiyan, Munusamy; Poonguzhali, Selvaraj; Saravanan, Venkatakrishnan Sivaraj; Duraipandiyan, Veeramuthu; Al-Dhabi, Naif Abdullah; Pragatheswari, Dhandapani; Santhanakrishnan, Palani; Kim, Soo-Jin; Weon, Hang-Yeon; Kwon, Soon-Wo

2016-10-01

A novel siderophore-producing actinomycete, designated PL19T, was isolated from the Scots-pine needle-like leaves collected from TNAU campus, Coimbatore, India. The isolate was chemoorganotrophic in nutrition and able to grow at 30 °C, and the optimum pH and NaCl facilitated the growth pH 6-11 and 0-8 % (w/v), respectively. The cells are filamentous and the mycelia formed are basically of wide and intricately branched substrate mycelium from which aerial mycelia arises, later gets differentiated into spores that are warty and arranged spirally. The 16S rRNA gene of strain PL19T was sequenced and was highly similar to the type strains of species of the genus Streptomyces, including Streptomyces barkulensis RC1831T (98.8 % pairwise similarity), Streptomyces fenghuangensis GIMN4.003T (98.2 %), Streptomyces nanhaiensis SCSIO 01248T (98.0 %), Streptomyces radiopugnans R97T (97.9 %), Streptomyces atacamensis C60T (97.8 %) and Streptomyces macrosporus NBRC 14749T (97.2 %), all of which were subjected to taxonomical characterization using a polyphasic approach. The strains showed unique carbon utilization patterns, and it possesses iso-C16 : 0 anteiso-C15 : 0 and anteiso-C17 : 0 as a major cellular fatty acids. The cell-wall was dominated with ll-type diaminopimelic acid, and the menaquinone type was MK-9(H6, H8). These chemotaxonomic evidences placed strain PL19T within the genus Streptomyces. The determination of G+C ratio (69.5 mol%) and DNA-DNA hybridization values (13.4-31.8 % with the phylogenetically related species) helped in further hierarchical classification of strain PL19T. Based on morphological, physiological and chemotaxonomic data as well as DNA-DNA hybridization values, strain PL19T could be distinguished from the evolutionarily closest species currently available. All these collective data show that strain PL19T represents a novel species of the genus Streptomyces, for which the name Streptomyces pini sp. nov. is proposed. The type strain is PL19T (=NRRL B-24728T=ICMP 17783T).

Identification of Multiple Phosphorylation Sites on Maize Endosperm Starch Branching Enzyme IIb, a Key Enzyme in Amylopectin Biosynthesis

PubMed Central

Makhmoudova, Amina; Williams, Declan; Brewer, Dyanne; Massey, Sarah; Patterson, Jenelle; Silva, Anjali; Vassall, Kenrick A.; Liu, Fushan; Subedi, Sanjeena; Harauz, George; Siu, K. W. Michael; Tetlow, Ian J.; Emes, Michael J.

2014-01-01

Starch branching enzyme IIb (SBEIIb) plays a crucial role in amylopectin biosynthesis in maize endosperm by defining the structural and functional properties of storage starch and is regulated by protein phosphorylation. Native and recombinant maize SBEIIb were used as substrates for amyloplast protein kinases to identify phosphorylation sites on the protein. A multidisciplinary approach involving bioinformatics, site-directed mutagenesis, and mass spectrometry identified three phosphorylation sites at Ser residues: Ser649, Ser286, and Ser297. Two Ca2+-dependent protein kinase activities were partially purified from amyloplasts, termed K1, responsible for Ser649 and Ser286 phosphorylation, and K2, responsible for Ser649 and Ser297 phosphorylation. The Ser286 and Ser297 phosphorylation sites are conserved in all plant branching enzymes and are located at opposite openings of the 8-stranded parallel β-barrel of the active site, which is involved with substrate binding and catalysis. Molecular dynamics simulation analysis indicates that phospho-Ser297 forms a stable salt bridge with Arg665, part of a conserved Cys-containing domain in plant branching enzymes. Ser649 conservation appears confined to the enzyme in cereals and is not universal, and is presumably associated with functions specific to seed storage. The implications of SBEIIb phosphorylation are considered in terms of the role of the enzyme and the importance of starch biosynthesis for yield and biotechnological application. PMID:24550386

Mathematical modeling of the "plant community -soil-like substrate -gas exchange with the human" closed ecosystem

NASA Astrophysics Data System (ADS)

Barkhatov, Yuri; Gubanov, Vladimir; Tikhomirov, Alexander A.; Degermendzhy, Andrey G.

A mathematical model of the "plant community -soil-like substrate -gas exchange with the human" experimental biological life support system (BLSS) has been constructed to predict its functioning and estimate feasibility of controlling it. The mathematical model consists of three compartments -two `phytotron' models (with wheat and radish) and the `mycotron' model (for mushrooms). The following components are included in the model: edible mushrooms (mushroom fruit bodies and mycelium); wheat; radish; straw (processed by mycelium); dead organic matter in the phytotron (separately for the wheat unit and for the radish unit); worms; worms' coprolites; vermicompost used as a soil-like substrate (SLS); bacterial microflora; min-eral nitrogen, phosphorus and iron; products of the system intended for humans (wheat grains, radish roots and mushroom fruit bodies); oxygen and carbon dioxide. Under continuous gas exchange, the mass exchange between the compartments occurs at the harvesting time. The conveyor character of the closed ecosystem functioning has been taken into account -the num-ber of culture age groups can be regulated (in experiments -4 and 8 age groups). The conveyor cycle duration can be regulated as well. The module is designed for the food and gas exchange requirements of 1/30 of a virtually present human. Aim of model analysis is determination of investigation direction in real experimental BLSS. The model allows doing dynamic calcu-lations of closure coefficient based on the main elements taken into account in the model and evaluating all dynamic components of the system under different conditions and modes of its operation, especially under the conditions that can hardly be created experimentally. One of the sustainability conditions can be long-duration functioning of the system under the light-ing that is far from the optimum. The mathematical model of the system can demonstrate variants of its sustainable functioning or ruin under various critical conditions probable for the LSS. An example is loss of part of green plant biomass. Model calculations have been done for different variants of loss of wheat biomass. We estimated the ability of the model to predict the optimal number of age groups in the LSS plant conveyor. This is an essential parameter, because if the number is too low, the total mass of the system components will vary and if it is too high, the system will be too complicated and costly. A high value of this parameter can also be interpreted as approximation to biosphere models. Dynamics of closure coefficient for the nitrogen and carbon loops was investigated for different variants of the BLSS. The system with biological utilization of the wheat straw has the highest closure coefficient, reaching 0.96, and can be used as a prototype of the BLSS of a new generation, with an essentially closed material cycling.

Anti-Gastric Ulcer Activity of Polysaccharide Fraction Isolated from Mycelium Culture of Lion's Mane Medicinal Mushroom, Hericium erinaceus (Higher Basidiomycetes).

PubMed

Wang, Mingxing; Konishi, Tetsuya; Gao, Yang; Xu, Duoduo; Gao, Qipin

2015-01-01

Hericium erinaceus is a culinary-medicinal mushroom that is used in traditional medicine, in folk medicine, and as medicinal cuisine in Asian countries such as China, Japan, and Korea. H. erinaceus exhibits various pharmacological properties, such as anti-cancer, immunomodulation, anti-dementia, and anti-gastric ulcer effects. The extracts of the fruiting body of H. erinaceus demonstrate anti-gastritis activity. However, the active principle in the extract, as well as the mechanism to treat gastric ulcers, remains uncertain. The current study aims to identify the active component, with anti-gastric ulcer function, from the extracts of the H. erinaceus mycelium culture. In the experiment, anti-gastric ulcer activity was evaluated using an ethanol-induced ulcer model in mice and with an 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay using MC cell lines. The results suggest that the polysaccharide fraction can significantly decrease the ulcerated area compared with the control group and the effect is fairly dose dependent, irrespective of animal or cell experiments. These results indicate that the polysaccharide fraction is the active component of the H. erinaceus mycelium culture, which protects against gastric ulcers.

Mycelial antineoplastic activity of Agaricus blazei.

PubMed

Bertéli, Míria Benetati Delgado; Umeo, Suzana Harue; Bertéli, André; do Valle, Juliana Silveira; Linde, Giani Andrea; Colauto, Nelson Barros

2014-08-01

Basidiocarp of Agaricus blazei (=Agaricus brasiliensis; =Agaricus subrufescens) is used as teas or capsules due to its antineoplastic effect but there are few reports of using mycelium for this purpose. The objective of this study was to evaluate the antineoplastic activity on sarcoma 180 cells implanted in mice of two forms of preparation of the mycelium from two A. blazei strains grown in culture medium with different concentrations of isolated soy protein. Mycelia were grown in Pontecorvo medium with different concentrations of isolated soybean protein (ISP). Mycelial hot water extract, moistened mycelial powder, hot water extract of green tea, Ifosfamida(®) (ifosfamide drug), and saline solution were administered daily by gavage in mice with sarcoma 180 cells to evaluate antineoplastic activity. It was concluded that antineoplastic activity was the same for both strains, except when used as moistened mycelial powder, which rules out the use of mycelial powder in capsules. Mycelial hot water extract had high antineoplastic activity with lower metabolic demand on the spleen and maintenance of normal blood parameters. Mycelial growth in different ISP concentrations had the same antineoplastic activity. Also the vegetative mycelium was as effective as the basidiocarp for sarcoma 180 tumor inhibition. Green tea was as effective as mycelial hot water extract.

Production of a COX-2 inhibitor, 2,4,5-trimethoxybenzaldehyde, with submerged cultured Antrodia camphorata.

PubMed

Chen, C-C; Chyau, C-C; Hseu, T-H

2007-04-01

To investigate the active ingredient in fruiting bodies and to produce it with cultured mycelium in Antrodia camphorata (BCRC 35398). The volatile components from the fruiting bodies, the liquid cultured broth of A. camphorata and Cinnamomum kanehirae wood were separately isolated by steam distillation-solvent extraction and identified by gas chromatography-mass spectrometry. In the fruiting bodies, a COX-2 inhibitor 2,4,5-trimethoxybenzaldehyde (TMBA) was found to be the most abundant constituent, but was totally absent in its cultured broth and its natural host, C. kanehirae wood. On feeding with the acid-digested sawdust of C. kanehirae wood or vanillin to the broth for culture, TMBA was produced in both cultured broths. The TMBA identified in fruiting bodies was an active ingredient whose functions consisted with the reported experiences of this mushroom. Feeding vanillin to culture broth could produce TMBA containing mycelium product like its fruiting bodies did. This study found an active ingredient in fruiting bodies of A. camphorata and elucidated this compound derived from digested sawdust of C. kanehirae wood. A feasible method was also developed to produce TMBA containing mycelium by feeding vanillin.

Botanical pesticides effect from shells of bean’s cashew nut on biological agents of trichoderma sp. and gliocladium sp.

NASA Astrophysics Data System (ADS)

Bande, L. O. S.; Mariadi; Gusnawaty, HS; Nuriadi; Trisulpa, L.; Rahmania

2018-02-01

A shell of cashew nut (Anacardium occidentanle) has contained Cashew Nut Shell Liquid (CNSL) that is used as botanical pesticides. CNSL oil consists of active substance such as anacardat acid, cardol and cardanol. Utilization of the pesticides from shells of cashew nut to control pests and diseases of plants would be affected on biological agents. The objective of this research was to investigate pesticides inhibition on the increase of mycelium Trichoderma sp. and Gliocladium sp. by in vitro method. The tested concentration sample consisted of 0.0% (control), 2.5%, 7.5% and 10.0% in PDA media. The results of this research showed that 2.5% botanical pesticides concentration could minimize mycelium of Trichoderma sp. and Gliocladium sp. 22.73% and 21.04% respectively and also the increase shells of cashew extract could be affected the increase of mycelium inhibition. The extract with 2.5% concentration was the recommended concentration to control of fruit rot diseases and if concentration was 10.0% then its inhibition become 54.98% and 49.35%, respectively. The results proved that uncontrolled utilization of the pesticides could be affected on decrease of Trichoderma sp. and Gliocladium sp. growth.

Using FT-NIR spectroscopy technique to determine arginine content in fermented Cordyceps sinensis mycelium.

PubMed

Xie, Chuanqi; Xu, Ning; Shao, Yongni; He, Yong

2015-01-01

This research investigated the feasibility of using Fourier transform near-infrared (FT-NIR) spectral technique for determining arginine content in fermented Cordyceps sinensis (C. sinensis) mycelium. Three different models were carried out to predict the arginine content. Wavenumber selection methods such as competitive adaptive reweighted sampling (CARS) and successive projections algorithm (SPA) were used to identify the most important wavenumbers and reduce the high dimensionality of the raw spectral data. Only a few wavenumbers were selected by CARS and CARS-SPA as the optimal wavenumbers, respectively. Among the prediction models, CARS-least squares-support vector machine (CARS-LS-SVM) model performed best with the highest values of the coefficient of determination of prediction (Rp(2)=0.8370) and residual predictive deviation (RPD=2.4741), the lowest value of root mean square error of prediction (RMSEP=0.0841). Moreover, the number of the input variables was forty-five, which only accounts for 2.04% of that of the full wavenumbers. The results showed that FT-NIR spectral technique has the potential to be an objective and non-destructive method to detect arginine content in fermented C. sinensis mycelium. Copyright © 2015 Elsevier B.V. All rights reserved.

Production and Multiplication of Native Compost Fungal Activator by Using Different Substrates and Its Influence on Growth and Development of Capsicum chinensis Jacq. “Bhut Jolokia”

PubMed Central

Parkash, Vipin; Saikia, Ankur Jyoti

2015-01-01

In vitro experiment was carried out to see the effect of saw dusts of Pinus kesiya, Shorea robusta, and Callicarpa arborea on Trichoderma harzianum, isolate TH-13 mass production, along with its biotization effect on Capsicum chinensis Jacq. “Bhut Jolokia.” Early mycelium initiation (2 days) occurred in S. robusta followed by P. kesiya and C. arborea (3.5 days). The sporulation was observed earlier in S. robusta (100% after 6 days) than P. kesiya (33.4% after 8 days) and C. arborea (16.7% after 9 days) but no sporulation was observed in control. The complete sporulation was also earlier in S. robusta (100% after 10 days) than P. kesiya (33.4% after 15 days) and C. arborea (16.4% after 18 days). Higher conidial yield (86 × 106) was also in S. robusta than P. kesiya (70 × 106) and C. arborea (45 × 106), respectively. The increase in height (60–70 cm), number of leaves (600–650), and yield of chili (120–150 fruits) were also more in inoculated C. chinensis seedlings than control. It is concluded that S. robusta saw dust is the best substrate for mass production of compost fungal activator and can be used in nursery practices for quality stock production of various crops/plantations. PMID:25632354

Valorization of spent oyster mushroom substrate and laccase recovery through successive solid state cultivation of Pleurotus, Ganoderma, and Lentinula strains.

PubMed

Economou, Christina N; Diamantopoulou, Panagiota A; Philippoussis, Antonios N

2017-06-01

Spent mushroom substrate (SMS) of Pleurotus ostreatus was supplemented with wheat bran and soybean flour in various proportions to obtain C/N ratios of 10, 20, and 30, and their effect was evaluated in successive cultivation of Pleurotus ostreatus, Pleurotus pulmonarius, Ganoderma adspersum, Ganoderma resinaceum, and Lentinula edodes strains with respect to mycelium growth rate, biomass concentration, recovery of the enzyme laccase and crude exopolysaccharides, and also with additional fruiting body production. All fungi showed the highest growth rate on unamended SMS (C/N 30), with G. resinaceum being the fastest colonizer (Kr = 9.84 mm day -1 ), while biomass concentration maximized at C/N 10. Moreover, supplementation affected positively laccase activity, with P. pulmonarius furnishing the highest value (44,363.22 U g -1 ) at C/N 20. On the contrary, L. edodes growth, fruiting, and laccase secretion were not favored by SMS supplementation. Fruiting body formation was promoted at C/N 30 for Ganoderma and at C/N 20 for Pleurotus species. Exopolysaccharide production of further studied Pleurotus strains was favored at a C/N 20 ratio, at the initial stage of SMS colonization. The obtained results support the potential effective utilization of supplemented SMS for laccase production from Ganoderma spp. and for new fruiting body production of Pleurotus spp.

Phosphatase-mediated bioprecipitation of lead by soil fungi.

PubMed

Liang, Xinjin; Kierans, Martin; Ceci, Andrea; Hillier, Stephen; Gadd, Geoffrey Michael

2016-01-01

Geoactive soil fungi were examined for their ability to release inorganic phosphate (Pi ) and mediate lead bioprecipitation during growth on organic phosphate substrates. Aspergillus niger and Paecilomyces javanicus grew in 5 mM Pb(NO3)2-containing media amended with glycerol 2-phosphate (G2P) or phytic acid (PyA) as sole P sources, and liberated Pi into the medium. This resulted in almost complete removal of Pb from solution and extensive precipitation of lead-containing minerals around the biomass, confirming the importance of the mycelium as a reactive network for biomineralization. The minerals were identified as pyromorphite (Pb5(PO4)3Cl), only produced by P. javanicus, and lead oxalate (PbC2O4), produced by A. niger and P. javanicus. Geochemical modelling of lead and lead mineral speciation as a function of pH and oxalate closely correlated with experimental conditions and data. Two main lead biomineralization mechanisms were therefore distinguished: pyromorphite formation depending on organic phosphate hydrolysis and lead oxalate formation depending on oxalate excretion. This also indicated species specificity in biomineralization depending on nutrition and physiology. Our findings provide further understanding of lead geomycology and organic phosphates as a biomineralization substrate, and are also relevant to metal immobilization biotechnologies for bioremediation, metal and P biorecovery, and utilization of waste organic phosphates. © 2015 Society for Applied Microbiology and John Wiley & Sons Ltd.

Biodegradation kinetics of thin-stillage treatment by Aspergillus awamori and characterization of recovered chitosan.

PubMed

Ray, S Ghosh; Ghangrekar, M M

2016-02-01

An attempt has been made to provide solution for distillery wastewater using fungal pretreatment followed by an anaerobic process to achieve higher organic matter removal, which is a challenge at present with currently adopted technologies. Submerged growth kinetics of distillery wastewater supernatant by Aspergillus awamori was also evaluated. The proposed kinetic models using a logistic equation for fungal growth and the Leudeking-Piret equation for product formation were validated experimentally, and substrate consumption equation was derived using estimated kinetic coefficients. Up to 59.6 % chemical oxygen demand (COD) and 70 % total organic carbon (TOC) removals were observed in 96 h of fungal incubation. Maximum specific growth rate of fungi, coefficient of biomass yield on substrate and growth-associated product formation coefficient were estimated to be 0.07 ± 0.01 h(-1), 0.614 kg biomass/kg utilized COD and 0.215 kg CO2/kg utilized TOC, respectively. The chitosan recovery of 0.072-0.078 kg/kg of dry mycelium was obtained using dilute sulphuric acid extraction, showing high purity and characteristic chitosan properties according to FTIR and XRD analyses. After anaerobic treatment of the fungal pretreated effluent with COD concentration of 7.920 ± 0.120 kg COD/m(3) (organic loading rate of 3.28 kg COD/m(3) day), overall COD reduction of 91.07 % was achieved from distillery wastewater.

Novel polyelectrolytes

NASA Technical Reports Server (NTRS)

Rembaum, Alan (Inventor); Yen, Shiao-Ping Siao (Inventor)

1978-01-01

Cationic polyelectrolytes are formed by the polymerization in absence of oxygen of a monomer of the general formula: ##STR1## where x is 3 or more than 6 and Z is I, Br or Cl to form high charge density linear polymers. Segments of the linear polymer may be attached to or formed in the presence of polyfunctional reactive tertiary amines or halogen polymeric substrates or polyfunctional lower molecular reactive polyfunctional substrates to form branched or star polyelectrolytes by a quaternization polymerization reaction.

Interactions between the arbuscular mycorrhizal (AM) fungus Glomus intraradices and nontransformed tomato roots of either wild-type or AM-defective phenotypes in monoxenic cultures.

PubMed

Bago, Alberto; Cano, Custodia; Toussaint, Jean-Patrick; Smith, Sally; Dickson, Sandy

2006-09-01

Monoxenic symbioses between the arbuscular mycorrhizal (AM) fungus Glomus intraradices and two nontransformed tomato root organ cultures (ROCs) were established. Wild-type tomato ROC from cultivar "RioGrande 76R" was employed as a control for mycorrhizal colonization and compared with its mutant line (rmc), which exhibits a highly reduced mycorrhizal colonization (rmc) phenotype. Structural features of the two root lines were similar when grown either in soil or under in vitro conditions, indicating that neither monoxenic culturing nor the rmc mutation affected root development or behavior. Colonization by G. intraradices in monoxenic culture of the wild-type line was low (<10%) but supported extensive development of extraradical mycelium, branched absorbing structures, and spores. The reduced colonization of rmc under monoxenic conditions (0.6%) was similar to that observed previously in soil. Extraradical development of runner hyphae was low and proportional to internal colonization. Few spores were produced. These results might suggest that carbon transfer may be modified in the rmc mutant. Our results support the usefulness of monoxenically obtained mycorrhizas for investigation of AM colonization and intraradical symbiotic functioning.

Electrochemical method of controlling thiolate coverage on a conductive substrate such as gold

NASA Technical Reports Server (NTRS)

Porter, Marc D. (Inventor); Weisshaar, Duane E. (Inventor)

1998-01-01

An electrochemical method for forming a partial monomolecular layer of a predetermined extent of coverage of a thiolate of the formula, XRS--, therein R can be a linear or branched chain hydrocarbon or an aromatic or the like and X can be any compatible end group, e.g., OH, COOH, CH.sub.3 or the like, upon a substrate such as gold, which involves applying in an electrochemical system a constant voltage preselected to yield the desired predetermined extent of coverage.

Electrochemical method of controlling thiolate coverage on a conductive substrate such as gold

DOEpatents

Porter, Marc D.; Weisshaar, Duane E.

1998-10-27

An electrochemical method for forming a partial monomolecular layer of a predetermined extent of coverage of a thiolate of the formula, XRS--, therein R can be a linear or branched chain hydrocarbon or an aromatic or the like and X can be any compatible end group, e.g., OH, COOH, CH.sub.3 or the like, upon a substrate such as gold, which involves applying in an electrochemical system a constant voltage preselected to yield the desired predetermined extent of coverage.

Electrochemical method of controlling thiolate coverage on a conductive substrate such as gold

DOEpatents

Porter, Marc D.; Weisshaar, Duane E.

1997-06-03

An electrochemical method for forming a partial monomolecular layer of a predetermined extent of coverage of a thiolate of the formula, XRS.sup.-, wherein R can be a linear or branched chain hydrocarbon or an aromatic or the like and X can be any compatible end group, e.g., OH, COOH, CH.sub.3 or the like, upon a substrate such as gold, which involves applying in an electrochemical system a constant voltage preselected to yield the desired predetermined extent of coverage.

BCAA Metabolism and NH3 Homeostasis.

PubMed

Conway, M E; Hutson, S M

2016-01-01

The branched chain amino acids (BCAA) are essential amino acids required not only for growth and development, but also as nutrient signals and as nitrogen donors to neurotransmitter synthesis and glutamate/glutamine cycling. Transamination and oxidative decarboxylation of the BCAAs are catalysed by the branched-chain aminotransferase proteins (BCATm, mitochondrial and BCATc, cytosolic) and the branched-chain α-keto acid dehydrogenase enzyme complex (BCKDC), respectively. These proteins show tissue, cell compartmentation, and protein-protein interactions, which call for substrate shuttling or channelling and nitrogen transfer for oxidation to occur. Efficient regulation of these pathways is mediated through the redox environment and phosphorylation in response to dietary and hormonal stimuli. The wide distribution of these proteins allows for effective BCAA utilisation. We discuss how BCAT, BCKDC, and glutamate dehydrogenase operate in supramolecular complexes, allowing for efficient channelling of substrates. The role of BCAAs in brain metabolism is highlighted in rodent and human brain, where differential expression of BCATm indicates differences in nitrogen metabolism between species. Finally, we introduce a new role for BCAT, where a change in function is triggered by oxidation of its redox-active switch. Our understanding of how BCAA metabolism and nitrogen transfer is regulated is important as many studies now point to BCAA metabolic dysregulation in metabolic and neurodegenerative conditions.

Morphology-controlled cactus-like branched anatase TiO2 arrays with high light-harvesting efficiency for dye-sensitized solar cells

NASA Astrophysics Data System (ADS)

Wu, Wu-Qiang; Rao, Hua-Shang; Feng, Hao-Lin; Guo, Xin-Dong; Su, Cheng-Yong; Kuang, Dai-Bin

2014-08-01

The present work establishes a facile process for one-step hydrothermal growth of vertically aligned anatase cactus-like branched TiO2 (CBT) arrays on a transparent conducting oxide (TCO) substrate. Various CBT morphologies are obtained by adjusting the potassium titanium oxide oxalate (PTO) reactant concentration (from 0.05 M to 0.15 M) and this yields a morphologically-controllable branched TiO2 arrays geometry. The CBT arrays consist of a vertically oriented nanowire (NW) or nanosheet (NS) stem and a host of short nanorod (NR) branches. The hierarchical CBT arrays demonstrate their excellent candidatures as photoanodes, which are capable of exhibiting high light-harvesting efficiency in dye-sensitized solar cells (DSSCs). Consequently, DSSCs based on 7 μm long optimized CBT arrays (0.05 M PTO), which are assembled with high density and high aspect-ratio NR branches, exhibit an impressive power conversion efficiency of 6.43% under AM 1.5G one sun illumination. The high performance can be attributed to the prominent light-harvesting efficiency, resulting from larger surface area and superior light-scattering capability.

Antitubercular Activity of Mycelium-Associated Ganoderma Lanostanoids.

PubMed

Isaka, Masahiko; Chinthanom, Panida; Sappan, Malipan; Supothina, Sumalee; Vichai, Vanicha; Danwisetkanjana, Kannawat; Boonpratuang, Thitiya; Hyde, Kevin D; Choeyklin, Rattaket

2017-05-26

In a continuation of our research into antitubercular lanostane triterpenoids from submerged cultures of Ganoderma species, three strains, Ganoderma orbiforme BCC 22325, Ganoderma sp. BCC 60695, and Ganoderma australe BCC 22314, have been investigated. Fourteen new lanostane triterpenoids, together with 35 known compounds, were isolated. Antitubercular activities of these mycelium-associated Ganoderma lanostanoids against Mycobacterium tuberculosis H37Ra were evaluated. Taken together with the assay data of previously isolated compounds, structure-activity relationships of the antitubercular activity are proposed. Most importantly, 3β- and 15α-acetoxy groups were shown to be critical for antimycobacterial activity. The most potent compound was (24E)-3β,15α-diacetoxylanosta-7,9(11),24-trien-26-oic acid (35).

Quantitative proteomics links metabolic pathways to specific developmental stages of the plant-pathogenic oomycete Phytophthora capsici.

PubMed

Pang, Zhili; Srivastava, Vaibhav; Liu, Xili; Bulone, Vincent

2017-04-01

The oomycete Phytophthora capsici is a plant pathogen responsible for important losses to vegetable production worldwide. Its asexual reproduction plays an important role in the rapid propagation and spread of the disease in the field. A global proteomics study was conducted to compare two key asexual life stages of P. capsici, i.e. the mycelium and cysts, to identify stage-specific biochemical processes. A total of 1200 proteins was identified using qualitative and quantitative proteomics. The transcript abundance of some of the enriched proteins was also analysed by quantitative real-time polymerase chain reaction. Seventy-three proteins exhibited different levels of abundance between the mycelium and cysts. The proteins enriched in the mycelium are mainly associated with glycolysis, the tricarboxylic acid (or citric acid) cycle and the pentose phosphate pathway, providing the energy required for the biosynthesis of cellular building blocks and hyphal growth. In contrast, the proteins that are predominant in cysts are essentially involved in fatty acid degradation, suggesting that the early infection stage of the pathogen relies primarily on fatty acid degradation for energy production. The data provide a better understanding of P. capsici biology and suggest potential metabolic targets at the two different developmental stages for disease control. © 2016 BSPP AND JOHN WILEY & SONS LTD.

Effect of dry mycelium of Penicillium chrysogenum fertilizer on soil microbial community composition, enzyme activities and snap bean growth.

PubMed

Wang, Bing; Liu, Huiling; Cai, Chen; Thabit, Mohamed; Wang, Pu; Li, Guomin; Duan, Ziheng

2016-10-01

The dry mycelium fertilizer (DMF) was produced from penicillin fermentation fungi mycelium (PFFM) following an acid-heating pretreatment to degrade the residual penicillin. In this study, it was applied into soil as fertilizer to investigate its effects on soil properties, phytotoxicity, microbial community composition, enzyme activities, and growth of snap bean in greenhouse. As the results show, pH, total nitrogen, total phosphorus, total potassium, and organic matter of soil with DMF treatments were generally higher than CON treatment. In addition, the applied DMF did not cause heavy metal and residual drug pollution of the modified soil. The lowest GI values (<0.3) were recorded at DMF8 (36 kg DMF/plat) on the first days after applying the fertilizer, indicating that severe phytotoxicity appeared in the DMF8-modified soil. Results of microbial population and enzyme activities illustrated that DMF was rapidly decomposed and the decomposition process significantly affected microbial growth and enzyme activities. The DMF-modified soil phytotoxicity decreased at the late fertilization time. DMF1 was considered as the optimum amount of DMF dose based on principal component analysis scores. Plant height and plant yield of snap bean were remarkably enhanced with the optimum DMF dose.

Antigenotoxic Effect of Trametes spp. Extracts against DNA Damage on Human Peripheral White Blood Cells

PubMed Central

Živković, Lada; Stajić, Mirjana; Vukojević, Jelena; Milovanović, Ivan; Spremo-Potparević, Biljana

2015-01-01

Trametes species have been used for thousands of years in traditional and conventional medicine for the treatment of various types of diseases. The goal was to evaluate possible antigenotoxic effects of mycelium and basidiocarp extracts of selected Trametes species and to assess dependence on their antioxidant potential. Trametes versicolor, T. hirsuta, and T. gibbosa were the species studied. Antigenotoxic potentials of extracts were assessed on human peripheral white blood cells with basidiocarp and mycelium extracts of the species. The alkaline comet test was used for detection of DNA strand breaks and alkali-labile sites, as well as the extent of DNA migration. DPPH assay was used to estimate antioxidative properties of extracts. Fruiting body extracts of T. versicolor and T. gibbosa as well as T. hirsuta extracts, except that at 20.0 mg/mL, were not genotoxic agents. T. versicolor extract had at 5.0 mg/mL the greatest antigenotoxic effect in both pre- and posttreatment of leukocytes. The mycelium extracts of the three species had no genotoxic activity and significant antigenotoxic effect against H2O2-induced DNA damage, both in pre- and posttreatment. The results suggest that extracts of these three species could be considered as strong antigenotoxic agents able to stimulate genoprotective response of cells. PMID:26258163

Proteome analysis provides insight into the regulation of bioactive metabolites in Hericium erinaceus.

PubMed

Zeng, Xu; Ling, Hong; Yang, Jianwen; Chen, Juan; Guo, Shunxing

2018-05-05

Hericium erinaceus, a famous edible mushroom, is also a well-known traditional medicinal fungus. To date, a large number of bioactive metabolites with antitumor, antibacterial, and immune-boosting effects were isolated from the free-living mycelium and fruiting body of H. erinaceus. Here we used the proteomic approach to explore proteins involved in the regulation of bioactive metabolites, including terpenoid, polyketide, sterol and etc. RESULTS: Using mass spectrometry, a total of 2543 unique proteins were identified using H. erinaceus genome, of which 2449, 1855, 1533 and 690 proteins were successfully annotated in Nr, KOG, KEGG and GO databases. Among them, 722 proteins were differentially expressed (528 up- and 194 down-regulated) in fruiting body compared with mycelium. Most of differentially expressed proteins were putatively involved in energy metabolism, molecular signaling, and secondary metabolism. Additionally, numerous proteins involved in terpenoid, polyketide, and sterol biosynthesis were identified. Our data revealed that proteins involved in polyketide biosynthesis were up-regulated in the fruiting body, while some proteins in mevalonate (MEP) pathway from terpenoid biosynthesis were generally up-regulated in mycelium. The present study suggested that the differential regulation of biosynthesis genes could produce various bioactive metabolites with pharmacological effects in H. erinaceus. Copyright © 2017. Published by Elsevier B.V.

Enhancement of the neuroprotective activity of Hericium erinaceus mycelium co-cultivated with Allium sativum extract.

PubMed

Kim, Myong Ki; Choi, Woon Yong; Lee, Hyeon Yong

2015-02-01

This study investigated the neuroprotective effects of Hericium erinaceus mycelium enriched with garlic extract (HGE) on rat pheochromocytoma nerve cells (PC12). The survival rates of the PC12 nerve cells and the neurite-bearing cells after the addition of HGE were estimated as 3.5 × 10(3) viable cells/ml and 2.3 × 10(3) viable cells/ml, respectively, which were 50% and 30% higher, respectively, compared with the untreated group. For the in vivo ischemia experiments, after treatment with the HGE extract, the hippocampal CA1 region was more strongly stained (>20%) than the control group, and the HGE extract also promoted higher staining levels than HFB, HM and HGEF, and even the garlic extract. This result indicates that HGE must have neuroprotective effects. Furthermore, HGE greatly decreased p21 gene expression to approximately 70% of the control and decreased p21 gene expression to even lower levels compared with HM, HGEF and the garlic extract. This work suggests that a synergistic effect of the H. erinaceus mycelium and the garlic extract (mainly allicin) exist because the amount of allicin in HGE (5.81 µg/ml) was lower than the garlic extract itself (6.89 µg/ml).

Branchfall as a Demographic Filter for Epiphyte Communities: Lessons from Forest Floor-Based Sampling

PubMed Central

Sarmento Cabral, Juliano; Petter, Gunnar; Mendieta-Leiva, Glenda; Wagner, Katrin; Zotz, Gerhard; Kreft, Holger

2015-01-01

Local variation in the abundance and richness of vascular epiphytes is often attributed to environmental characteristics such as substrate and microclimate. Less is known, however, about the impacts of tree and branch turnover on epiphyte communities. To address this issue, we surveyed branches and epiphytes found on the forest floor in 96 transects in two forests (Atlantic rainforest in Brazil and Caribbean rainforest in Panama). In the Brazilian forest, we additionally distinguished between edge and core study sites. We quantified branch abundance, epiphyte abundance, richness and proportion of adults to investigate the trends of these variables over branch diameter. Branches <2 cm in diameter comprised >90% of all branches on the forest floor. Abundance and richness of fallen epiphytes per transect were highest in the Brazilian core transects and lowest in the Panamanian transects. The majority of epiphytes on the floor (c. 65%) were found attached to branches. At all three study sites, branch abundance and branch diameter were negatively correlated, whereas epiphyte abundance and richness per branch, as well as the proportion of adults were positively correlated with branch diameter. The relationship between branch diameter and absolute epiphyte abundance or richness differed between study sites, which might be explained by differences in forest structure and dynamics. In the Panamanian forest, epiphytes had been previously inventoried, allowing an evaluation of our surveying method by comparing canopy and forest floor samplings. Individuals found on the forest floor corresponded to 13% of all individuals on branches <10 cm in diameter (including crowns), with abundance, richness and composition trends on forest floor reflecting canopy trends. We argue that forest floor surveys provide useful floristic and, most notably, demographic information particularly on epiphytes occurring on the thinnest branches, which are least accessible. Here, branchfall acts as an important demographic filter structuring epiphyte communities. PMID:26083417

Design principles of autocatalytic cycles constrain enzyme kinetics and force low substrate saturation at flux branch points

PubMed Central

Barenholz, Uri; Davidi, Dan; Reznik, Ed; Bar-On, Yinon; Antonovsky, Niv; Noor, Elad; Milo, Ron

2017-01-01

A set of chemical reactions that require a metabolite to synthesize more of that metabolite is an autocatalytic cycle. Here, we show that most of the reactions in the core of central carbon metabolism are part of compact autocatalytic cycles. Such metabolic designs must meet specific conditions to support stable fluxes, hence avoiding depletion of intermediate metabolites. As such, they are subjected to constraints that may seem counter-intuitive: the enzymes of branch reactions out of the cycle must be overexpressed and the affinity of these enzymes to their substrates must be relatively weak. We use recent quantitative proteomics and fluxomics measurements to show that the above conditions hold for functioning cycles in central carbon metabolism of E. coli. This work demonstrates that the topology of a metabolic network can shape kinetic parameters of enzymes and lead to seemingly wasteful enzyme usage. DOI: http://dx.doi.org/10.7554/eLife.20667.001 PMID:28169831

Application of HPLC to study the kinetics of a branched bi-enzyme system consisting of hypoxanthine-guanine phosphoribosyltransferase and xanthine oxidase--an important biochemical system to evaluate the efficiency of the anticancer drug 6-mercaptopurine in ALL cell line.

PubMed

Kalra, Sukirti; Paul, Manash K; Balaram, Hemalatha; Mukhopadhyay, Anup Kumar

2007-05-01

The thiopurine antimetabolite 6-mercaptopurine (6MP) is an important chemotherapeutic drug in the conventional treatment of childhood acute lymphoblastic leukemia (ALL). 6MP is mainly catabolized by both hypoxanthine-guanine phosphoribosyltransferase (HGPRT) and xanthine oxidase (XOD) to form thioinosinic monophosphate (TIMP) (therapeutically active metabolite) and 6-thiouric acid (6TUA) (inactive metabolite), respectively. The activity of both the enzymes varies among ALL patients governing the active and the inactive metabolite profile within the immature lymphocytes. Therefore, an attempt was made to study the kinetic nature of the branched bi-enzyme system acting on 6MP and to quantitate TIMP and 6TUA formed when the two enzymes are present in equal and variable ratios. The quantification of the branched kinetics using spectrophotometric method presents problem due to the closely apposed lambda(max) of the substrates and products. Hence, employing an HPLC method, the quantification of the products was done with the progress of time. The limit of quantification (LOQ) of substrate was found to be 10nM and for products as 50 nM. The limit of detection (LOD) was found to be 1 nM for the substrate and the products. The method exhibited linearity in the range of 0.01-100 microM for 6MP and 0.05-100 microM for both 6TUA and TIMP. The amount of TIMP formed was higher than that of 6TUA in the bi-enzyme system when both the enzymes were present in equivalent enzymatic ratio. It was further found that enzymatic ratios play an important role in determining the amounts of TIMP and 6TUA. This method was further validated using actively growing T-ALL cell line (Jurkat) to study the branched kinetics, wherein it was observed that treatment of 50 microM 6MP led to the generation of 12 microM TIMP and 0.8 microM 6TUA in 6 h at 37 degrees C.

Steam explosion treatment for ethanol production from branches pruned from pear trees by simultaneous saccharification and fermentation.

PubMed

Sasaki, Chizuru; Okumura, Ryosuke; Asada, Chikako; Nakamura, Yoshitoshi

2014-01-01

This study investigated the production of ethanol from unutilized branches pruned from pear trees by steam explosion pretreatment. Steam pressures of 25, 35, and 45 atm were applied for 5 min, followed by enzymatic saccharification of the extracted residues with cellulase (Cellic CTec2). High glucose recoveries, of 93.3, 99.7, and 87.1%, of the total sugar derived from the cellulose were obtained from water- and methanol-extracted residues after steam explosion at 25, 35, and 45 tm, respectively. These values corresponded to 34.9, 34.3, and 27.1 g of glucose per 100 g of dry steam-exploded branches. Simultaneous saccharification and fermentation experiments were done on water-extracted residues and water- and methanol-extracted residues by Kluyveromyces marxianus NBRC 1777. An overall highest theoretical ethanol yield of 76% of the total sugar derived from cellulose was achieved when 100 g/L of water- and methanol-washed residues from 35 atm-exploded pear branches was used as substrate.

Short-term effects of hurricane disturbance on food availability for migrant songbirds during autumn stopover

USGS Publications Warehouse

Dobbs, R.C.; Barrow, W.C.; Jeske, C.W.; Dimiceli, J.; Michot, T.C.; Beck, J.W.

2009-01-01

Understanding the consequences of hurricanes on the food resources available to neotropical-nearctic migrant songbirds may provide important insight into the effects of hurricanes on migratory populations. During autumn migration 2006 we investigated the foraging ecology of two species of insectivorous migrants, Blue-gray Gnatcatcher (Polioptila caerulea) and Yellow Warbler (Dendroica petechia), and the availability of their foraging substrates and arthropod food resources in two coastal forests in western Louisiana, which were impacted to different degrees by Hurricane Rita in autumn 2005. Both migrant species attacked prey on bark substrates significantly more frequently, and on live foliage less frequently, in severely damaged forest than in lightly damaged forest (??2 tests, P < 0.05). However, both species attacked prey on bark less than expected given its availability (i.e., migrants avoided bark), and attacked prey on live foliage more than expected given its availability (i.e., migrants selected live foliage), in severely damaged forest (??2 tests, P < 0.03). Branch-clipping revealed that arthropod biomass on live hackberry (Celtis laevigata) and sweet acacia (Acacia farnesiana) branches was significantly higher in severely damaged forest than in lightly damaged forest (Mann-Whitney test, P < 0.01). However, because live foliage was significantly less available in severely damaged forest, overall food availability for migrants was lower in severely damaged forest than in lightly damaged forest. Migrant use of, and arthropod biomass on, bark and live-foliage substrates were thus dependent on the availability of those substrates, which differed between sites as a result of hurricane-related habitat disturbance. These results demonstrate that severe hurricane disturbance reduces food availability for insectivorous songbirds during migratory stopover by reducing the availability of preferred foraging substrates. ?? 2009 The Society of Wetland Scientists.

Basidiospore and Protoplast Regeneration from Raised Fruiting Bodies of Pathogenic Ganoderma boninense.

PubMed

Govender, Nisha T; Mahmood, Maziah; Seman, Idris A; Mui-Yun, Wong

2016-08-26

Ganoderma boninense, a phytopathogenic white rot fungus had sought minimal genetic characterizations despite huge biotechnological potentials. Thus, efficient collection of fruiting body, basidiospore and protoplast of G. boninense is described. Matured basidiocarp raised under the glasshouse conditions yielded a total of 8.3 × 104 basidiospores/ml using the low speed centrifugation technique. Mycelium aged 3-day-old treated under an incubation period of 3 h in lysing enzyme from Trichoderma harzianum (10 mg/ml) suspended in osmotic stabilizer (0.6 M potassium chloride and 20 mM dipotassium phosphate buffer) yielded the highest number of viable protoplasts (8.9 × 106 single colonies) among all possible combinations tested (regeneration media, age of mycelium, osmotic stabilizer, digestive enzyme and incubation period).

[Correlations between the characters of the mycelium vegetative growth and the formation of the fruiting body of Ganoderma luciderm].

PubMed

Wang, Shu-fang; He, Xiu-feng; He, Hui-xia; Zhu, Ping

2006-01-01

To select a proper Ganoderma luciderm strain for the fruiting body production. The strains were cultivated on the agar media and in the liquid media, respectively. Then the strains were inoculated onto the solid medium made from agricultural products (such as wheat bran, corn powder, wood meal, etc.) and cultured for a certain period. Strains, which were easier to produce polyporic tissues at the vegetative growth stage, would be more quickly to form fruiting body with high quality and yield of the spores. Appearance of the polyporic tissues at the mycelium vegetative growth stage could be used as a marker for the strain selection for the G. luciderm substituted cultivation.

Characterization of the functional interactions of plastidial starch phosphorylase and starch branching enzymes from rice endosperm during reserve starch biosynthesis.

PubMed

Nakamura, Yasunori; Ono, Masami; Sawada, Takayuki; Crofts, Naoko; Fujita, Naoko; Steup, Martin

2017-11-01

Functional interactions of plastidial phosphorylase (Pho1) and starch branching enzymes (BEs) from the developing rice endosperm are the focus of this study. In the presence of both Pho1 and BE, the same branched primer molecule is elongated and further branched almost simultaneously even at very low glucan concentrations present in the purified enzyme preparations. By contrast, in the absence of any BE, glucans are not, to any significant extent, elongated by Pho1. Based on our in vitro data, in the developing rice endosperm, Pho1 appears to be weakly associated with any of the BE isozymes. By using fluorophore-labeled malto-oligosaccharides, we identified maltose as the smallest possible primer for elongation by Pho1. Linear dextrins act as carbohydrate substrates for BEs. By functionally interacting with a BE, Pho1 performs two essential functions during the initiation of starch biosynthesis in the rice endosperm: First, it elongates maltodextrins up to a degree of polymerization of at least 60. Second, by closely interacting with BEs, Pho1 is able to elongate branched glucans efficiently and thereby synthesizes branched carbohydrates essential for the initiation of amylopectin biosynthesis. Copyright © 2017 Elsevier B.V. All rights reserved.

Fragrance release from the surface of branched poly (amide)s.

PubMed

Aulenta, Francesca; Drew, Michael G B; Foster, Alison; Hayes, Wayne; Rannard, Steven; Thornthwaite, David W; Youngs, Tristan G A

2005-01-31

Enzymes are powerful tools in organic synthesis that are able to catalyse a wide variety of selective chemical transformations under mild and environmentally friendly conditions. Enzymes such as the lipases have also found applications in the synthesis and degradation of polymeric materials. However, the use of these natural catalysts in the synthesis and the post-synthetic modification of dendrimers and hyperbranched molecules is an application of chemistry yet to be explored extensively. In this study the use of two hydrolytic enzymes, a lipase from Candida cylindracea and a cutinase from Fusarium solani pisii, were investigated in the selective cleavage of ester groups situated on the peripheral layer of two families of branched polyamides. These branched polyamides were conjugated to simple fragrances citronellol and L-menthol via ester linkages. Hydrolysis of the ester linkage between the fragrances and the branched polyamide support was carried out in aqueous buffered systems at slightly basic pH values under the optimum operative conditions for the enzymes used. These preliminary qualitative investigations revealed that partial cleavage of the ester functionalities from the branched polyamide support had occurred. However, the ability of the enzymes to interact with the substrates decreased considerably as the branching density, the rigidity of the structure and the bulkiness of the polyamide-fragrance conjugates increased.

Electrochemical method of controlling thiolate coverage on a conductive substrate such as gold

DOEpatents

Porter, M.D.; Weisshaar, D.E.

1998-10-27

An electrochemical method is described for forming a partial monomolecular layer of a predetermined extent of coverage of a thiolate of the formula, XRS-, therein R can be a linear or branched chain hydrocarbon or an aromatic or the like and X can be any compatible end group, e.g., OH, COOH, CH{sub 3} or the like, upon a substrate such as gold, which involves applying in an electrochemical system a constant voltage preselected to yield the desired predetermined extent of coverage. 13 figs.

Antagonistic evaluation of Trichodermaviride and Aspergillusflavus against wood-decay fungus Pleurotusostreatus

NASA Astrophysics Data System (ADS)

Erwin; Iskandar, E.; Husien, N.

2018-04-01

Pleurotusostreatus cultivation is often limited by the presence of antagonistic microorganisms that can also grow together on a variety of substrates that are used as growth media of P. ostreatus. As a preliminary study, this research aims to evaluate the antagonism of Trichodermaviride and Aspergillusflavus in interacting with the P.ostreatus fungus by in-vitro. An antagonism test using a dual culture method of these fungi with 1 month incubation time on Poteto Dextrose Agar (PDA) media was performed. Evaluation of the antagonistic properties of these fungi is conducted through macroscopic and microscopic observations. Macroscopically, the growth of T. viride and A. flavus mycelium showed that both colonies were more dominant than P. ostreatus colonies. Microscopically, hyphae of T. viride and A. flavus indicate the presence of very strong micoparasitic properties that characterized by attachment and convolution of both hyphae to P. ostreatus hyphae. It was concluded thatthe growth of T. viride and A. flavus fungi was more dominant and had potential as an inhibiting agent of P. ostreatus fungal growth, respectively, in dual culture on PDA media.

Ectomycorrhizal fungi as an alternative to the use of chemical fertilisers in nursery production of Pinus pinaster.

PubMed

Sousa, Nadine R; Franco, Albina R; Oliveira, Rui S; Castro, Paula M L

2012-03-01

Addition of fertilisers is a common practice in nursery production of conifer seedlings. The aim of this study was to evaluate whether ectomycorrhizal (ECM) fungi can be an alternative to the use of chemical fertilisers in the nursery production of Pinus pinaster. A greenhouse nursery experiment was conducted by inoculating seedlings obtained from seeds of P. pinaster plus trees with a range of compatible ECM fungi: (1) Thelephora terrestris, (2) Rhizopogon vulgaris, (3) a mixture of Pisolithus tinctorius and Scleroderma citrinum, and (4) a mixture of Suillus bovinus, Laccaria laccata and Lactarius deterrimus, using forest soil as substrate. Plant development was assessed at two levels of N-P-K fertiliser (0 or 600 mg/seedling). Inoculation with a mixture of mycelium from S. bovinus, L. laccata and L. deterrimus and with a mixture of spores of P. tinctorius and S. citrinum improved plant growth and nutrition, without the need of fertiliser. Results indicate that selected ECM fungi can be a beneficial biotechnological tool in nursery production of P. pinaster. Copyright © 2010 Elsevier Ltd. All rights reserved.

Spatial differentiation of gene expression in Aspergillus niger colony grown for sugar beet pulp utilization

PubMed Central

Benoit, Isabelle; Zhou, Miaomiao; Vivas Duarte, Alexandra; Downes, Damien J.; Todd, Richard B.; Kloezen, Wendy; Post, Harm; Heck, Albert J. R.; Maarten Altelaar, A. F.; de Vries, Ronald P.

2015-01-01

Degradation of plant biomass to fermentable sugars is of critical importance for the use of plant materials for biofuels. Filamentous fungi are ubiquitous organisms and major plant biomass degraders. Single colonies of some fungal species can colonize massive areas as large as five soccer stadia. During growth, the mycelium encounters heterogeneous carbon sources. Here we assessed whether substrate heterogeneity is a major determinant of spatial gene expression in colonies of Aspergillus niger. We analyzed whole-genome gene expression in five concentric zones of 5-day-old colonies utilizing sugar beet pulp as a complex carbon source. Growth, protein production and secretion occurred throughout the colony. Genes involved in carbon catabolism were expressed uniformly from the centre to the periphery whereas genes encoding plant biomass degrading enzymes and nitrate utilization were expressed differentially across the colony. A combined adaptive response of carbon-catabolism and enzyme production to locally available monosaccharides was observed. Finally, our results demonstrate that A. niger employs different enzymatic tools to adapt its metabolism as it colonizes complex environments. PMID:26314379

Spatial differentiation of gene expression in Aspergillus niger colony grown for sugar beet pulp utilization.

PubMed

Benoit, Isabelle; Zhou, Miaomiao; Vivas Duarte, Alexandra; Downes, Damien J; Todd, Richard B; Kloezen, Wendy; Post, Harm; Heck, Albert J R; Maarten Altelaar, A F; de Vries, Ronald P

2015-08-28

Degradation of plant biomass to fermentable sugars is of critical importance for the use of plant materials for biofuels. Filamentous fungi are ubiquitous organisms and major plant biomass degraders. Single colonies of some fungal species can colonize massive areas as large as five soccer stadia. During growth, the mycelium encounters heterogeneous carbon sources. Here we assessed whether substrate heterogeneity is a major determinant of spatial gene expression in colonies of Aspergillus niger. We analyzed whole-genome gene expression in five concentric zones of 5-day-old colonies utilizing sugar beet pulp as a complex carbon source. Growth, protein production and secretion occurred throughout the colony. Genes involved in carbon catabolism were expressed uniformly from the centre to the periphery whereas genes encoding plant biomass degrading enzymes and nitrate utilization were expressed differentially across the colony. A combined adaptive response of carbon-catabolism and enzyme production to locally available monosaccharides was observed. Finally, our results demonstrate that A. niger employs different enzymatic tools to adapt its metabolism as it colonizes complex environments.

Identification of Fourier transform infrared photoacoustic spectral features for detection of Aspergillus flavus infection in corn.

PubMed

Gordon, S H; Schudy, R B; Wheeler, B C; Wicklow, D T; Greene, R V

1997-04-01

Aspergillus flavus and other pathogenic fungi display typical infrared spectra which differ significantly from spectra of substrate materials such as corn. On this basis, specific spectral features have been identified which permit detection of fungal infection on the surface of corn kernels by photoacoustic infrared spectroscopy. In a blind study, ten corn kernels showing bright greenish yellow fluorescence (BGYF) in the germ or endosperm and ten BGYF-negative kernels were correctly classified as infected or not infected by Fourier transform infrared photoacoustic spectroscopy. Earlier studies have shown that BGYF-positive kernels contain the bulk of the aflatoxin contaminating grain at harvest. Ten major spectral features, identified by visual inspection of the photoacoustic spectra of A. flavus mycelium grown in culture versus uninfected corn, were interpreted and assigned by theoretical comparisons of the relative chemical compositions of fungi and corn. The spectral features can be built into either empirical or knowledge-based computer models (expert systems) for automatic infrared detection and segregation of grains or kernels containing aflatoxin from the food and feed supply.

Re-inoculation strategies enhance the degradation of emerging pollutants in fungal bioaugmentation of sewage sludge.

PubMed

Rodríguez-Rodríguez, Carlos E; Lucas, Daniel; Barón, Enrique; Gago-Ferrero, Pablo; Molins-Delgado, Daniel; Rodríguez-Mozaz, Sara; Eljarrat, Ethel; Díaz-Cruz, M Silvia; Barceló, Damià; Caminal, Glòria; Vicent, Teresa

2014-09-01

The use of Trametes versicolor has been partially successful in the removal of some pharmaceuticals from sewage sludge in laboratory-scale biopile systems. The application of two strategies for the re-inoculation of biomass was assessed during the fungal bioaugmentation of non-sterile sludge (42-d treatment) as an approach to improve the elimination of pharmaceuticals and other groups of emerging pollutants. Globally, the re-inoculation of biopiles with blended mycelium exerted a major effect on the removal of pharmaceuticals (86%), brominated-flame-retardants (81%) and UV filters (80%) with respect to the re-inoculation with additional lignocellulosic substrate colonized by the fungus (69-67-22%). The performance was better than that of the analogous non-re-inoculated systems that were assayed previously for the removal of pharmaceuticals. The results demonstrate the ability of T. versicolor to remove a wide spectrum of emerging micropollutants under non-sterile conditions, while re-inoculation appears to be a useful step to improve the fungal treatment of sludge. Copyright © 2014 Elsevier Ltd. All rights reserved.

Pilot-scale bioconversion of rice and sunflower agro-residues into medicinal mushrooms and laccase enzymes through solid-state fermentation with Ganoderma lucidum.

PubMed

Postemsky, P D; Bidegain, M A; González-Matute, R; Figlas, N D; Cubitto, M A

2017-05-01

Solid-state fermentation was evaluated at the pilot-scale for the bioconversion and valorization of rice husks and straw (RSH), or sunflower seed hulls (SSH), into medicinal mushrooms and crude extracts, with laccase activity. The average mushroom yield was 56kg dry weight per ton of agro-residues. Laccase activity in crude aqueous extracts showed its maximum value of 10,927Ukg -1 in RSH (day 10, Exudate phase) and 16,442Ukg -1 in SSH (day 5, Full colonization phase), the activity at the Residual substrate phase being 511Ukg -1 in RSH and 803Ukg -1 in SSH, respectively. Crude extracts obtained with various protocols revealed differences in the extraction yields. Lyophilization followed by storage at 4°C allowed the preservation of laccase activity for more than one month. It is proposed that standard mushroom farms could increase their profits by obtaining laccase as a byproduct during the gaps in mycelium running. Copyright © 2017 Elsevier Ltd. All rights reserved.

[Experiment to study some suspension media for the lyophilization of actinomycetes].

PubMed

Semenov, S M

1975-09-01

Viability and cultural properties of 59 actinomycetes and 17 bacteria lyophilized in polyvinylpyrrolidone (PVP), sodium glutamate, their combinations and horse serum were studied after storage for 2 years at a temperature of 4-10 degrees. A 5 per cent solution of sodium glutamate had a high protective effect on viability of the above organisms. The solution containing 3 per cent of sodium glutamate and 3 per cent of PVP was somewhat less effective. The cultures lyophilized in 5 per cent solution of sodium glutamate had the same viability levels as those lyophilized in horse serum, while the latter had better growth rates on their plating out on nutrient media. A 5 per cent solution of PVP had no advantages over sodium glutamate or horse serum with respect to preservation of the organism viability. No significant differences in the cultural properties: colour of the aerial and substrate mycelium and pigment production were noted in the actinomycetes lyophilized in various protective media and the analogous control cultures maintained by means of passages on fresh nutrient media.

Differential Response of Extracellular Proteases of Trichoderma Harzianum Against Fungal Phytopathogens.

PubMed

Sharma, Vivek; Salwan, Richa; Sharma, Prem N

2016-09-01

In the present study, production of extracellular proteases by Trichoderma harzianum was evaluated based on the relative gene expression and spectrophotometric assay. The fungal isolates were grown in Czapek Dox Broth medium supplemented with deactivated mycelium of plant fungal pathogens such as Fusarium oxysporum, Colletotrichum capsici, Gloeocercospora sorghi, and Colletotrichum truncatum. The maximum protease activity was detected after 48 h of incubation against Colletotrichum spp. Similarly in qRT-PCR, the relative gene expression of four proteases varied from 48 to 96 h against host pathogens in a time-independent manner. Among proteases, statistically significant upregulation of asp, asp, and srp was observed against Colletotrichum spp., followed by F. oxysporum. But in the case of pepM22, maximum upregulation was observed against F. oxysporum. The variation in enzyme assay and qRT-PCR of proteases at different time intervals against various fungal phytopathogens could be due to the limitation of using casein as a substrate for all types of proteases or protease-encoding transcripts selected for qRT-PCR, which may not be true representative of total protease activity.

Nanoscale biophysical properties of the cell surface galactosaminogalactan from the fungal pathogen Aspergillus fumigatus

NASA Astrophysics Data System (ADS)

Beaussart, Audrey; El-Kirat-Chatel, Sofiane; Fontaine, Thierry; Latgé, Jean-Paul; Dufrêne, Yves F.

2015-09-01

Many fungal pathogens produce cell surface polysaccharides that play essential roles in host-pathogen interactions. In Aspergillus fumigatus, the newly discovered polysaccharide galactosaminogalactan (GAG) mediates adherence to a variety of substrates through molecular mechanisms that are poorly understood. Here we use atomic force microscopy to unravel the localization and adhesion of GAG on living fungal cells. Using single-molecule imaging with tips bearing anti-GAG antibodies, we found that GAG is massively exposed on wild-type (WT) germ tubes, consistent with the notion that this glycopolymer is secreted by the mycelium of A. fumigatus, while it is lacking on WT resting conidia and on germ tubes from a mutant (Δuge3) deficient in GAG. Imaging germ tubes with tips bearing anti-β-glucan antibodies shows that exposure of β-glucan is strongly increased in the Δuge3 mutant, indicating that this polysaccharide is masked by GAG during hyphal growth. Single-cell force measurements show that expression of GAG on germ tubes promotes specific adhesion to pneumocytes and non-specific adhesion to hydrophobic substrates. These results provide a molecular foundation for the multifunctional adhesion properties of GAG, thus suggesting it could be used as a potential target in anti-adhesion therapy and immunotherapy. Our methodology represents a powerful approach for characterizing the nanoscale organization and adhesion of cell wall polysaccharides during fungal morphogenesis, thereby contributing to increase our understanding of their role in biofilm formation and immune responses.

Root development during soil genesis: effects of root-root interactions, mycorrhizae, and substrate

NASA Astrophysics Data System (ADS)

Salinas, A.; Zaharescu, D. G.

2015-12-01

A major driver of soil formation is the colonization and transformation of rock by plants and associated microbiota. In turn, substrate chemical composition can also influence the capacity for plant colonization and development. In order to better define these relationships, a mesocosm study was set up to analyze the effect mycorrhizal fungi, plant density and rock have on root development, and to determine the effect of root morphology on weathering and soil formation. We hypothesized that plant-plant and plant-fungi interactions have a stronger influence on root architecture and rock weathering than the substrate composition alone. Buffalo grass (Bouteloua dactyloides) was grown in a controlled environment in columns filled with either granular granite, schist, rhyolite or basalt. Each substrate was given two different treatments, including grass-microbes and grass-microbes-mycorrhizae and incubated for 120, 240, and 480 days. Columns were then extracted and analyzed for root morphology, fine fraction, and pore water major element content. Preliminary results showed that plants produced more biomass in rhyolite, followed by schist, basalt, and granite, indicating that substrate composition is an important driver of root development. In support of our hypothesis, mycorrhizae was a strong driver of root development by stimulating length growth, biomass production, and branching. However, average root length and branching also appeared to decrease in response to high plant density, though this trend was only present among roots with mycorrhizal fungi. Interestingly, fine fraction production was negatively correlated with average root thickness and volume. There is also slight evidence indicating that fine fraction production is more related to substrate composition than root morphology, though this data needs to be further analyzed. Our hope is that the results of this study can one day be applied to agricultural research in order to promote the production of crops on traditionally un-arable land.

Protein phosphatase 2Cm is a critical regulator of branched-chain amino acid catabolism in mice and cultured cells.

PubMed

Lu, Gang; Sun, Haipeng; She, Pengxiang; Youn, Ji-Youn; Warburton, Sarah; Ping, Peipei; Vondriska, Thomas M; Cai, Hua; Lynch, Christopher J; Wang, Yibin

2009-06-01

The branched-chain amino acids (BCAA) are essential amino acids required for protein homeostasis, energy balance, and nutrient signaling. In individuals with deficiencies in BCAA, these amino acids can be preserved through inhibition of the branched-chain-alpha-ketoacid dehydrogenase (BCKD) complex, the rate-limiting step in their metabolism. BCKD is inhibited by phosphorylation of its E1alpha subunit at Ser293, which is catalyzed by BCKD kinase. During BCAA excess, phosphorylated Ser293 (pSer293) becomes dephosphorylated through the concerted inhibition of BCKD kinase and the activity of an unknown intramitochondrial phosphatase. Using unbiased, proteomic approaches, we have found that a mitochondrial-targeted phosphatase, PP2Cm, specifically binds the BCKD complex and induces dephosphorylation of Ser293 in the presence of BCKD substrates. Loss of PP2Cm completely abolished substrate-induced E1alpha dephosphorylation both in vitro and in vivo. PP2Cm-deficient mice exhibited BCAA catabolic defects and a metabolic phenotype similar to the intermittent or intermediate types of human maple syrup urine disease (MSUD), a hereditary disorder caused by defects in BCKD activity. These results indicate that PP2Cm is the endogenous BCKD phosphatase required for nutrient-mediated regulation of BCKD activity and suggest that defects in PP2Cm may be responsible for a subset of human MSUD.

Ecophysiology of horse chestnut (Aesculus Hippocastanum L.) in degraded and restored urban sites

Treesearch

Jacek Oleksyn; Brian D. Kloeppel; Szymon Lukasiewicz; Piotr Karolewski; Peter B. Reich

2007-01-01

We explored changes in growth, phenology, net CO2 assimilation rate, water use efficiency, secondary defense compounds, substrate and foliage nutrient concentration of a degraded urban horse chestnut (Aesculus hippocastanum L.) site restored for three years using mulching (tree branches including foliage) and fertilization (...

The annealing helicase and branch migration activities of Drosophila HARP.

PubMed

Kassavetis, George A; Kadonaga, James T

2014-01-01

HARP (SMARCAL1, MARCAL1) is an annealing helicase that functions in the repair and restart of damaged DNA replication forks through its DNA branch migration and replication fork regression activities. HARP is conserved among metazoans. HARP from invertebrates differs by the absence of one of the two HARP-specific domain repeats found in vertebrates. The annealing helicase and branch migration activity of invertebrate HARP has not been documented. We found that HARP from Drosophila melanogaster retains the annealing helicase activity of human HARP, the ability to disrupt D-loops and to branch migrate Holliday junctions, but fails to regress model DNA replication fork structures. A comparison of human and Drosophila HARP on additional substrates revealed that both HARPs are competent in branch migrating a bidirectional replication bubble composed of either DNA:DNA or RNA:DNA hybrid. Human, but not Drosophila, HARP is also capable of regressing a replication fork structure containing a highly stable poly rG:dC hybrid. Persistent RNA:DNA hybrids in vivo can lead to replication fork arrest and genome instability. The ability of HARP to strand transfer hybrids may signify a hybrid removal function for this enzyme, in vivo.

Identification of a novel nematotoxic protein by challenging the model mushroom Coprinopsis cinerea with a fungivorous nematode

DOE PAGES

Plaza, David Fernando; Schmieder, Stefanie Sofia; Lipzen, Anna; ...

2015-11-19

The dung of herbivores, the natural habitat of the model mushroom Coprinopsis cinerea, is a nutrient-rich but also very competitive environment for a saprophytic fungus. Previously we showed that C. cinerea expresses constitutive, tissue-specific armories against antagonists such as animal predators and bacterial competitors. In order to dissect the inducible armories against such antagonists, we sequenced the poly(A)-positive transcriptome of C. cinerea vegetative mycelium upon challenge with fungivorous and bacterivorous nematodes, Gram-negative and Gram-positive bacteria and mechanical damage. As a response to the fungivorous nematode Aphelenchus avenae, C. cinerea was found to specifically induce the transcription of several genes encodingmore » previously characterized nematotoxic lectins. In addition, a previously not characterized gene encoding a cytoplasmic protein with several predicted Ricin B-fold domains, was found to be strongly up-regulated under this condition. Functional analysis of the recombinant protein revealed a high toxicity towards the bacterivorous nematode Caenorhabditis elegans. Challenge of the mycelium with A. avenae also lead to the induction of several genes encoding putative antibacterial proteins. Some of these genes were also induced upon challenge of the mycelium with the bacteria Escherichia coli and Bacillus subtilis. Lastly, these results suggest that fungi have the ability to induce specific innate defense responses similar to plants and animals.« less

[Study on the detection of active ingredient contents of Paecilomyces hepiali mycelium via near infrared spectroscopy].

PubMed

Teng, Wei-Zhuo; Song, Jia; Meng, Fan-Xin; Meng, Qing-Fan; Lu, Jia-Hui; Hu, Shuang; Teng, Li-Rong; Wang, Di; Xie, Jing

2014-10-01

Partial least squares (PLS) and radial basis function neural network (RBFNN) combined with near infrared spectros- copy (NIR) were applied to develop models for cordycepic acid, polysaccharide and adenosine analysis in Paecilomyces hepialid fermentation mycelium. The developed models possess well generalization and predictive ability which can be applied for crude drugs and related productions determination. During the experiment, 214 Paecilomyces hepialid mycelium samples were obtained via chemical mutagenesis combined with submerged fermentation. The contents of cordycepic acid, polysaccharide and adenosine were determined via traditional methods and the near infrared spectroscopy data were collected. The outliers were removed and the numbers of calibration set were confirmed via Monte Carlo partial least square (MCPLS) method. Based on the values of degree of approach (Da), both moving window partial least squares (MWPLS) and moving window radial basis function neural network (MWRBFNN) were applied to optimize characteristic wavelength variables, optimum preprocessing methods and other important variables in the models. After comparison, the RBFNN, RBFNN and PLS models were developed successfully for cordycepic acid, polysaccharide and adenosine detection, and the correlation between reference values and predictive values in both calibration set (R2c) and validation set (R2p) of optimum models was 0.9417 and 0.9663, 0.9803 and 0.9850, and 0.9761 and 0.9728, respectively. All the data suggest that these models possess well fitness and predictive ability.

Biocide effects of volatile organic compounds produced by potential biocontrol rhizobacteria on Sclerotinia sclerotiorum

PubMed Central

Giorgio, Annalisa; De Stradis, Angelo; Lo Cantore, Pietro; Iacobellis, Nicola S.

2015-01-01

Six rhizobacteria isolated from common bean and able to protect bean plants from the common bacterial blight (CBB) causal agent, were in vitro evaluated for their potential antifungal effects toward different plant pathogenic fungi, mostly soil-borne. By dual culture assays, the above bacteria resulted producing diffusible and volatile metabolites which inhibited the growth of the majority of the pathogens under study. In particular, the latter substances highly affected the mycelium growth of Sclerotinia sclerotiorum strains, one of which was selected for further studies either on mycelium or sclerotia. Gas chromatographic analysis of the bacterial volatiles led to the identification of an array of volatile organic compounds (VOCs). Time course studies showed the modification of the VOCs profile along a period of 5 days. In order to evaluate the single detected VOC effects on fungal growth, some of the pure compounds were tested on S. sclerotiorum mycelium and their minimal inhibitory quantities were determined. Similarly, the minimal inhibitory quantities on sclerotia germination were also defined. Moreover, observations by light and transmission electron microscopes highlighted hyphae cytoplasm granulation and ultrastructural alterations at cell organelles, mostly membranes, mitochondria, and endoplasmic reticulum. The membranes appeared one of the primary targets of bacterial volatiles, as confirmed by hemolytic activity observed for the majority of pure VOCs. However, of interest is the alteration observed on mitochondria as well. PMID:26500617

Evaluation of the Antioxidant Activities and Tyrosinase Inhibitory Property from Mycelium Culture Extracts

PubMed Central

Park, Ki Moon; Kwon, Kyung Min; Lee, Seung Ho

2015-01-01

Since mushrooms have many bioactive components, they have been used as components in folk medicine. Because mycelium has an advantage when it comes to large-scale production, this study aimed to evaluate the antioxidant properties and anti-tyrosinase activity from 55 mycelia in culture media. Relatively high 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging capacity was detected from the ethanol extract of culture media including mycelium (EECiM) of Morchella esculenta var. esculenta (MEVE), Auricularia polytricha (APO), Tremella aurantia (TAU), Volvariella bombycina (VBO), and Oudemansiella sp. (Osp), which also showed strong reducing power and inhibitory activity in relation to the thiobarbituric acid (TBA) value. On the other hand, relatively high tyrosinase inhibitory activity was detected in Inonotus mikadoi (IMI), Coriolus versicolor (CVE), Volvariella volvacea (VVO), Panellus serotinus (PSE), Auricularia auricula (AAU), and Fomitopsis sp. (Fsp). Interestingly, the APO EECiM exhibited the highest DPPH radical scavenging rate (77.5 ± 4.3%) and reducing power (1.18 ± 0.041), while the highest inhibitory power of the TBA value and antityrosinase activity were detected in that of TAU (64.5 ± 4.1%) and IMI (46.0 ± 7.5%), respectively. Overall, our study suggested potential candidates for EECiMs that exhibited powerful antioxidant and tyrosinase inhibitory properties and might be used as natural antioxidant tyrosinase inhibitor. PMID:26345142

Comparative studies on the induction of Trichoderma harzianum mutanase by α-(1→3)-glucan-rich fruiting bodies and mycelia of Laetiporus sulphureus.

PubMed

Wiater, Adrian; Pleszczyńska, Małgorzata; Szczodrak, Janusz; Janusz, Grzegorz

2012-01-01

Mutanase (α-(1→3)-glucanase) is a little-known inductive enzyme that is potentially useful in dentistry. Here, it was shown that the cell wall preparation (CWP) obtained from the fruiting body or vegetative mycelium of polypore fungus Laetiporus sulphureus is rich in α-(1→3)-glucan and can be successfully used for mutanase induction in Trichoderma harzianum. The content of this biopolymer in the CWP depended on the age of fruiting bodies and increased along with their maturation. In the case of CWP prepared from vegetative mycelia, the amount of α-(1→3)-glucan depended on the mycelium age and also on the kind of medium used for its cultivation. All CWPs prepared from the individually harvested fruiting body specimens induced high mutanase activity (0.53-0.82 U/mL) in T. harzianum after 3 days of cultivation. As for the CWPs obtained from the hyphal mycelia of L. sulpureus, the maximal enzyme productivity (0.34 U/mL after 3 days of incubation) was recorded for CWP prepared from the 3 week-old mycelium cultivated in Sabouraud medium. Statistically, a high positive correlation was found between the total percentage content of α-(1→3)-glucan in the CWP and the mutanase activity.

Evaluation of the toxicological safety of erinacine A-enriched Hericium erinaceus in a 28-day oral feeding study in Sprague-Dawley rats.

PubMed

Li, I-Chen; Chen, Yen-Lien; Lee, Li-Ya; Chen, Wan-Ping; Tsai, Yueh-Ting; Chen, Chin-Chu; Chen, Chin-Shuh

2014-08-01

Natural products have attained great importance as they are believed to be the new alternative medicines for conventional therapy. As numerous studies have proved the tremendous medicinal values of Hericium erinaceus, it is necessary to take into account its safety as well as its risk for the recipient. However, mushroom mycelium has an identity distinct from mushrooms, as two specific classes of compounds, hericenones and erinacines, can only be extracted from both the fruit body and the cultured mycelium, respectively. Therefore, this is the first report on the evaluation of the toxicity of H.erinaceus mycelium, enriched with 5mg/g erinacine A, by a 28-day repeated oral administration study in Sprague-Dawley rats. Three doses of 1 (Low), 2 (Mid) and 3 (High) g/kg body weight/day were selected for the study while distilled water served as control. All animals survived to the end of the study. No abnormal changes were observed in clinical signs. No adverse or test article-related differences were found in urinalysis, haematology and serum biochemistry parameters, between the treatment and control groups. No gross pathological findings and histopathological differences were seen. Therefore, the no-observed-adverse-effect level of erinacine A-enriched H.erinaceus is greater than 3g/kgbody weight/day. Copyright © 2014 Elsevier Ltd. All rights reserved.

IRRADIATION METHOD OF CONVERTING ORGANIC COMPOUNDS

DOEpatents

Allen, A.O.; Caffrey, J.M. Jr.

1960-10-11

A method is given for changing the distribution of organic compounds from that produced by the irradiation of bulk alkane hydrocarbons. This method consists of depositing an alkane hydrocarbon on the surface of a substrate material and irradiating with gamma radiation at a dose rate of more than 100,000 rads. The substrate material may be a metal, metal salts, metal oxides, or carbons having a surface area in excess of 1 m/sup 2//g. The hydrocarbons are deposited in layers of from 0.1 to 10 monolayers on the surfaces of these substrates and irradiated. The product yields are found to vary from those which result from the irradiation of bulk hydrocarbons in that there is an increase in the quantity of branched hydrocarbons.

[Isolation and regeneration of the protoplasts of the streptomycete producers of actinomycins C and X].

PubMed

Orlova, T I; Masha, G G; Kliueva, N A

1986-09-01

Protoplasts of S. michiganensis, S. chrysomallus and Streptomyces sp. 26-115, organisms producing actinomycins C and X form in hypertonic salt solution under the action of 3-4,5 mg/ml of lysozyme on the mycelium suspension. For protoplasting, the streptomycetes were grown on the soybean medium in the presence of 0.2-0.8 per cent of glycine. The mycelium of the streptomycete exponential growth phase was more favourable for protoplast formation. Protoplast regeneration was studied on the medium described by Okanishi et al. The quantitative composition of this medium was not optimal for regeneration of protoplasts of the above streptomycetes. The level of their regeneration depended to various extents on concentration of phosphate, magnesium and calcium ions and sucrose in the regeneration medium.

Myxococcus xanthus induces actinorhodin overproduction and aerial mycelium formation by Streptomyces coelicolor

PubMed Central

Pérez, Juana; Muñoz‐Dorado, José; Braña, Alfredo F.; Shimkets, Lawrence J.; Sevillano, Laura; Santamaría, Ramón I.

2011-01-01

Summary Interaction of the predatory myxobacterium Myxococcus xanthus with the non‐motile, antibiotic producer Streptomyces coelicolor was examined using a variety of experimental approaches. Myxococcus xanthus cells prey on S. coelicolor, forming streams of ordered cells that lyse the S. coelicolor hyphae in the contact area between the two colonies. The interaction increases actinorhodin production by S. coelicolor up to 20‐fold and triggers aerial mycelium production. Other bacteria are also able to induce these processes in S. coelicolor though to a lesser extent. These studies offer new clues about the expression of genes that remain silent or are expressed at low level in axenic cultures and open the possibility of overproducing compounds of biotechnological interest by using potent inducers synthesized by other bacteria. PMID:21342463

Field-temperature phase diagrams of freestanding and substrate-constrained epitaxial Ni-Mn-Ga-Co films for magnetocaloric applications

NASA Astrophysics Data System (ADS)

Diestel, A.; Niemann, R.; Schleicher, B.; Schwabe, S.; Schultz, L.; Fähler, S.

2015-07-01

Ferroic cooling processes that rely on field-induced first-order transformations of solid materials are a promising step towards a more energy-efficient refrigeration technology. In particular, thin films are discussed for their fast heat transfer and possible applications in microsystems. Substrate-constrained films are not useful since their substrates act as a heat sink. In this article, we examine a substrate-constrained and a freestanding epitaxial film of magnetocaloric Ni-Mn-Ga-Co. We compare phase diagrams and entropy changes obtained by magnetic field and temperature scans, which differ. We observe an asymmetry of the hysteresis between heating and cooling branch, which vanishes at high magnetic fields. These effects are discussed with respect to the vector character of a magnetic field, which acts differently on the nucleation and growth processes compared to the scalar character of the temperature.

Highly efficient Cu(In,Ga)Se2 solar cells grown on flexible polymer films.

PubMed

Chirilă, Adrian; Buecheler, Stephan; Pianezzi, Fabian; Bloesch, Patrick; Gretener, Christina; Uhl, Alexander R; Fella, Carolin; Kranz, Lukas; Perrenoud, Julian; Seyrling, Sieghard; Verma, Rajneesh; Nishiwaki, Shiro; Romanyuk, Yaroslav E; Bilger, Gerhard; Tiwari, Ayodhya N

2011-09-18

Solar cells based on polycrystalline Cu(In,Ga)Se(2) absorber layers have yielded the highest conversion efficiency among all thin-film technologies, and the use of flexible polymer films as substrates offers several advantages in lowering manufacturing costs. However, given that conversion efficiency is crucial for cost-competitiveness, it is necessary to develop devices on flexible substrates that perform as well as those obtained on rigid substrates. Such comparable performance has not previously been achieved, primarily because polymer films require much lower substrate temperatures during absorber deposition, generally resulting in much lower efficiencies. Here we identify a strong composition gradient in the absorber layer as the main reason for inferior performance and show that, by adjusting it appropriately, very high efficiencies can be obtained. This implies that future manufacturing of highly efficient flexible solar cells could lower the cost of solar electricity and thus become a significant branch of the photovoltaic industry.

[GENETIC AND METABOLIC URGENCIES IN THE NEONATAL INTENSIVE CARE UNIT: MAPLE SYRUP URINE DISEASE].

PubMed

Páez Rojas, Paola Liliana; Suarez Obando, Fernando

2015-07-01

Maple syrup urine disease (MSUD) is a hereditary disorder of branched chain amino/keto acid metabolism, caused by a decreased activity of the branched-chain alpha- ketoacid dehydrogenase complex (BCKAD), which leads to abnormal elevated plasma concentrations of branched-chain amino acids (BCAAs) clinically manifested as a heavy burden for Central Nervous system. The toxic accumulation of substrates promotes the development of a severe and rapidly progressive neonatal encephalopathy if treatment is not immediately given. This disorder has a specific medical management in acute phase in order to minimize mortality and morbidity. For all those reasons, it is important to include the MSUD as a possible diagnosis in a encephalopathic newborn. We present a colombian newborn with classical MSUD with fatal outcome as an example of metabolic emergency and a differential diagnosis in the encephalopathic newborn. Copyright AULA MEDICA EDICIONES 2014. Published by AULA MEDICA. All rights reserved.

Conformational stability and thermodynamic characterization of the lipoic acid bearing domain of human mitochondrial branched chain α-ketoacid dehydrogenase

PubMed Central

Naik, Mandar T.; Huang, Tai-Huang

2004-01-01

The lipoic acid bearing domain (hbLBD) of human mitochondrial branched chain α-ketoacid dehydrogenase (BCKD) plays important role of substrate channeling in oxidative decarboxylation of the branched chain α-ketoacids. Recently hbLBD has been found to follow two-step folding mechanism without detectable presence of stable or kinetic intermediates. The present study describes the conformational stability underlying the folding of this small β-barrel domain. Thermal denaturation in presence of urea and isothermal urea denaturation titrations are used to evaluate various thermodynamic parameters defining the equilibrium unfolding. The linear extrapolation model successfully describes the two-step; native state ↔denatured state unfolding transition of hbLBD. The average temperature of maximum stability of hbLBD is estimated as 295.6 ± 0.9 K. Cold denaturation of hbLBD is also predicted and discussed. PMID:15322287

Effect of Aromatic Compounds on Cellular Fatty Acid Composition of Rhodococcus opacus

PubMed Central

Tsitko, Irina V.; Zaitsev, Gennadi M.; Lobanok, Anatoli G.; Salkinoja-Salonen, Mirja S.

1999-01-01

In cells of Rhodococcus opacus GM-14, GM-29, and 1CP, the contents of branched (10-methyl) fatty acids increased from 3% to 15 to 34% of the total fatty acids when the cells were grown on benzene, phenol, 4-chlorophenol, chlorobenzene, or toluene as the sole source of carbon and energy, in comparison with cells grown on fructose. In addition, the content of trans-hexadecenoic acid increased from 5% to 8 to 18% with phenol or chlorophenol as the carbon source. The 10-methyl branched fatty acid content of R. opacus GM-14 cells increased in a dose-related manner following exposure to phenol or toluene when toluene was not utilized as the growth substrate. The results suggest that 10-methyl branched fatty acids may participate in the adaptation of R. opacus to lipophilic aromatic compounds. PMID:9925629

Peripheral nervous control of cold-induced reduction in the respiratory quotient of the rat

NASA Astrophysics Data System (ADS)

Refinetti, Roberto

1990-03-01

Cold-exposed rats show a reduction in the respiratory quotient which is indicative of a relative shift from carbohydrates to lipids as substrates for oxidative metabolism. In the present study, the effects of food deprivation and cold exposure on the respiratory quotient were observed. In addition, the involvement of the three main branches of the peripheral nervous system (sympathetic, parasympathetic, and somatic) was investigated by means of synaptic blockade with propranolol, atropine, and quinine, respectively. Both propranolol and quinine blocked the cold-induced decrease in respiratory quotient and increase in heat production, whereas atropine had only minor and very brief effects. It is concluded that both the sympathetic and somatic branches are involved in the metabolic changes associated with cold-induced thermogenesis and that the increase in metabolic heat production involves a shift from carbohydrate to lipid utilization irrespective of which of the two branches is activated.

Arginine-glycine-aspartic acid functional branched semi-interpenetrating hydrogels.

PubMed

Plenderleith, Richard A; Pateman, Christopher J; Rodenburg, Cornelia; Haycock, John W; Claeyssens, Frederik; Sammon, Chris; Rimmer, Stephen

2015-10-14

For the first time a series of functional hydrogels based on semi-interpenetrating networks with both branched and crosslinked polymer components have been prepared and we show the successful use of these materials as substrates for cell culture. The materials consist of highly branched poly(N-isopropyl acrylamide)s with peptide functionalised end groups in a continuous phase of crosslinked poly(vinyl pyrrolidone). Functionalisation of the end groups of the branched polymer component with the GRGDS peptide produces a hydrogel that supports cell adhesion and proliferation. The materials provide a new synthetic functional biomaterial that has many of the features of extracellular matrix, and as such can be used to support tissue regeneration and cell culture. This class of high water content hydrogel material has important advantages over other functional hydrogels in its synthesis and does not require post-processing modifications nor are functional-monomers, which change the polymerisation process, required. Thus, the systems are amenable to large scale and bespoke manufacturing using conventional moulding or additive manufacturing techniques. Processing using additive manufacturing is exemplified by producing tubes using microstereolithography.

Enzymatic Mechanism for Arabinan Degradation and Transport in the Thermophilic Bacterium Caldanaerobius polysaccharolyticus.

PubMed

Wefers, Daniel; Dong, Jia; Abdel-Hamid, Ahmed M; Paul, Hans Müller; Pereira, Gabriel V; Han, Yejun; Dodd, Dylan; Baskaran, Ramiya; Mayer, Beth; Mackie, Roderick I; Cann, Isaac

2017-09-15

The plant cell wall polysaccharide arabinan provides an important supply of arabinose, and unraveling arabinan-degrading strategies by microbes is important for understanding its use as a source of energy. Here, we explored the arabinan-degrading enzymes in the thermophilic bacterium Caldanaerobius polysaccharolyticus and identified a gene cluster encoding two glycoside hydrolase (GH) family 51 α-l-arabinofuranosidases (CpAbf51A, CpAbf51B), a GH43 endoarabinanase (CpAbn43A), a GH27 β-l-arabinopyranosidase (CpAbp27A), and two GH127 β-l-arabinofuranosidases (CpAbf127A, CpAbf127B). The genes were expressed as recombinant proteins, and the functions of the purified proteins were determined with para -nitrophenyl ( p NP)-linked sugars and naturally occurring pectin structural elements as the substrates. The results demonstrated that CpAbn43A is an endoarabinanase while CpAbf51A and CpAbf51B are α-l-arabinofuranosidases that exhibit diverse substrate specificities, cleaving α-1,2, α-1,3, and α-1,5 linkages of purified arabinan-oligosaccharides. Furthermore, both CpAbf127A and CpAbf127B cleaved β-arabinofuranose residues in complex arabinan side chains, thus providing evidence of the function of this family of enzymes on such polysaccharides. The optimal temperatures of the enzymes ranged between 60°C and 75°C, and CpAbf43A and CpAbf51A worked synergistically to release arabinose from branched and debranched arabinan. Furthermore, the hydrolytic activity on branched arabinan oligosaccharides and degradation of pectic substrates by the endoarabinanase and l-arabinofuranosidases suggested a microbe equipped with diverse activities to degrade complex arabinan in the environment. Based on our functional analyses of the genes in the arabinan degradation cluster and the substrate-binding studies on a component of the cognate transporter system, we propose a model for arabinan degradation and transport by C. polysaccharolyticus IMPORTANCE Genomic DNA sequencing and bioinformatic analysis allowed the identification of a gene cluster encoding several proteins predicted to function in arabinan degradation and transport in C. polysaccharolyticus The analysis of the recombinant proteins yielded detailed insights into the putative arabinan metabolism of this thermophilic bacterium. The use of various branched arabinan oligosaccharides provided a detailed understanding of the substrate specificities of the enzymes and allowed assignment of two new GH127 polypeptides as β-l-arabinofuranosidases able to degrade pectic substrates, thus expanding our knowledge of this rare group of glycoside hydrolases. In addition, the enzymes showed synergistic effects for the degradation of arabinans at elevated temperatures. The enzymes characterized from the gene cluster are, therefore, of utility for arabinose production in both the biofuel and food industries. Copyright © 2017 American Society for Microbiology.

Enzymatic Mechanism for Arabinan Degradation and Transport in the Thermophilic Bacterium Caldanaerobius polysaccharolyticus

PubMed Central

Dong, Jia; Abdel-Hamid, Ahmed M.; Paul, Hans Müller; Pereira, Gabriel V.; Han, Yejun; Dodd, Dylan; Baskaran, Ramiya; Mayer, Beth; Mackie, Roderick I.

2017-01-01

ABSTRACT The plant cell wall polysaccharide arabinan provides an important supply of arabinose, and unraveling arabinan-degrading strategies by microbes is important for understanding its use as a source of energy. Here, we explored the arabinan-degrading enzymes in the thermophilic bacterium Caldanaerobius polysaccharolyticus and identified a gene cluster encoding two glycoside hydrolase (GH) family 51 α-l-arabinofuranosidases (CpAbf51A, CpAbf51B), a GH43 endoarabinanase (CpAbn43A), a GH27 β-l-arabinopyranosidase (CpAbp27A), and two GH127 β-l-arabinofuranosidases (CpAbf127A, CpAbf127B). The genes were expressed as recombinant proteins, and the functions of the purified proteins were determined with para-nitrophenyl (pNP)-linked sugars and naturally occurring pectin structural elements as the substrates. The results demonstrated that CpAbn43A is an endoarabinanase while CpAbf51A and CpAbf51B are α-l-arabinofuranosidases that exhibit diverse substrate specificities, cleaving α-1,2, α-1,3, and α-1,5 linkages of purified arabinan-oligosaccharides. Furthermore, both CpAbf127A and CpAbf127B cleaved β-arabinofuranose residues in complex arabinan side chains, thus providing evidence of the function of this family of enzymes on such polysaccharides. The optimal temperatures of the enzymes ranged between 60°C and 75°C, and CpAbf43A and CpAbf51A worked synergistically to release arabinose from branched and debranched arabinan. Furthermore, the hydrolytic activity on branched arabinan oligosaccharides and degradation of pectic substrates by the endoarabinanase and l-arabinofuranosidases suggested a microbe equipped with diverse activities to degrade complex arabinan in the environment. Based on our functional analyses of the genes in the arabinan degradation cluster and the substrate-binding studies on a component of the cognate transporter system, we propose a model for arabinan degradation and transport by C. polysaccharolyticus. IMPORTANCE Genomic DNA sequencing and bioinformatic analysis allowed the identification of a gene cluster encoding several proteins predicted to function in arabinan degradation and transport in C. polysaccharolyticus. The analysis of the recombinant proteins yielded detailed insights into the putative arabinan metabolism of this thermophilic bacterium. The use of various branched arabinan oligosaccharides provided a detailed understanding of the substrate specificities of the enzymes and allowed assignment of two new GH127 polypeptides as β-l-arabinofuranosidases able to degrade pectic substrates, thus expanding our knowledge of this rare group of glycoside hydrolases. In addition, the enzymes showed synergistic effects for the degradation of arabinans at elevated temperatures. The enzymes characterized from the gene cluster are, therefore, of utility for arabinose production in both the biofuel and food industries. PMID:28710263

Do fungi need to be included within environmental radiation protection assessment models?

PubMed

Guillén, J; Baeza, A; Beresford, N A; Wood, M D

2017-09-01

Fungi are used as biomonitors of forest ecosystems, having comparatively high uptakes of anthropogenic and naturally occurring radionuclides. However, whilst they are known to accumulate radionuclides they are not typically considered in radiological assessment tools for environmental (non-human biota) assessment. In this paper the total dose rate to fungi is estimated using the ERICA Tool, assuming different fruiting body geometries, a single ellipsoid and more complex geometries considering the different components of the fruit body and their differing radionuclide contents based upon measurement data. Anthropogenic and naturally occurring radionuclide concentrations from the Mediterranean ecosystem (Spain) were used in this assessment. The total estimated weighted dose rate was in the range 0.31-3.4 μGy/h (5 th -95 th percentile), similar to natural exposure rates reported for other wild groups. The total estimated dose was dominated by internal exposure, especially from 226 Ra and 210 Po. Differences in dose rate between complex geometries and a simple ellipsoid model were negligible. Therefore, the simple ellipsoid model is recommended to assess dose rates to fungal fruiting bodies. Fungal mycelium was also modelled assuming a long filament. Using these geometries, assessments for fungal fruiting bodies and mycelium under different scenarios (post-accident, planned release and existing exposure) were conducted, each being based on available monitoring data. The estimated total dose rate in each case was below the ERICA screening benchmark dose, except for the example post-accident existing exposure scenario (the Chernobyl Exclusion Zone) for which a dose rate in excess of 35 μGy/h was estimated for the fruiting body. Estimated mycelium dose rate in this post-accident existing exposure scenario was close to the 400 μGy/h benchmark for plants, although fungi are generally considered to be less radiosensitive than plants. Further research on appropriate mycelium geometries and their radionuclide content is required. Based on the assessments presented in this paper, there is no need to recommend that fungi should be added to the existing assessment tools and frameworks; if required some tools allow a geometry representing fungi to be created and used within a dose assessment. Copyright © 2017 Elsevier Ltd. All rights reserved.

pH regulation of mitochondrial branch chain alpha-keto acid transport and oxidation in rat heart mitochondria.

PubMed

Hutson, S M

1987-07-15

The kinetics of branched chain alpha-keto acid uptake and efflux were studied as a function of varied external and matrix pH. Matrix pH was determined by the distribution of 5,5'-dimethyloxazolidine-2,4-dione. When rat heart mitochondria were incubated under transport conditions at pH 7.0 with succinate as respiratory substrate, the matrix pH was significantly greater than 8.0. Matrix pH remained greater than or equal to 8.0 when the medium pH was varied from 6.3 to 8.3, and it was lowered below 8.0 by addition of 5 mM phosphate or uncoupler. No pH gradient was detectable when mitochondria were incubated in the presence of valinomycin and uncoupler. Efflux of alpha-ketoisocaproate or alpha-ketoisovalerate from rat heart mitochondria obeyed first order kinetics. Varying the external pH from 6.6 to 8.3 had no significant effect on efflux, and at an external pH of 7.0, the first order rate constant for efflux was not affected by decreasing the matrix pH. On the other hand, exchange was sensitive to changes in medium but not matrix pH. The K0.5 for external branched chain alpha-keto acid was lowered by changing the medium pH from 7.6 to 6.3. At medium pH values greater than or equal to 8.0 both K0.5 and Vmax were affected. Uptake was determined either by measuring initial rates or was calculated after measuring the first order approach to a final equilibrium value. Unlike efflux, uptake was sensitive to changes in both external and matrix pH. The rate of branched chain alpha-keto acid uptake was stimulated by decreasing the medium pH from 8.3 to 6.3 and by alkalinization of the mitochondrial matrix. The estimated external pK for proton binding was 6.9. The data indicate that the branched chain alpha-keto acid transporter is asymmetric, that is, binding sites for substrate on the inside and outside of the mitochondrial membrane are not identical. alpha-Ketoisocaproate oxidation was measured at 37 degrees C in isolated mitochondria over the pH range of 6.6 to 8.1. Changes in the rate of branched chain alpha-keto acid oxidation, particularly when ATP was added (increase delta pH), were found to parallel the pH effects observed on branched chain alpha-keto acid uptake. Therefore, transport, and by implication oxidation, can be regulated by pH changes within the physiological range. Furthermore, intracellular pH may affect the degree of compartmentation between the cytosolic and mitochondrial branched chain alpha-keto acid pools.

Interspecies variation of Kitasatospora recifensis endophytic from yam bean producing thermostable amylases in alternative media.

PubMed

Stamford, Tania Lucia Montenegro; Stamford, Thayza Christina Montenegro; Stamford, Newton Pereira; Santos, Carolina Etienne Rosália Silva; de Lyra, Maria do Carmo Catanho Pereira; Ha-Park, Yong; Bae, Jin-Won; Araújo, Janete Magali

2007-12-01

An endophytic actinomycete isolated from tubers of yam beam (Pachyrhizus erosus L. Urban) was classified as a novel species nominated Kitasatospora recifensis based in phenotypic and genotypic analysis (16S rDNA gene sequence). Monosporic culture using specific ISP2 media revealed three interspecies, which were identified by DNA southern hybridization (Wild strain 13817 W, Aerial Mycelium strain 13817 AM and Vegetative Mycelium strain 13817 VM). The strains were tested for the production of amylolitic enzymes in alternative media. Maximum yields for both enzymes were observed in starch-casein. Higher α-amylase was obtained with strain 13817 W in starch-urea, and amyloglucosidase with strain 13817 AM in starch-ammonium that are economic sources and may be important for industrial purposes. Type strain (DAUFPE 13817(T) = KCTC 9972(T )= DSM 44943(T)).

Biodegradation of kerosene by Aspergillus ochraceus NCIM-1146.

PubMed

Saratale, Ganesh; Kalme, Satish; Bhosale, Sanjyot; Govindwar, Sanjay

2007-10-01

The filamentous fungus Aspergillus ochraceus NCIM-1146 was found to degrade kerosene, when previously grown mycelium (96 h) was incubated in the broth containing kerosene. Higher levels of NADPH-DCIP reductase, aminopyrine N-demethylase and kerosene biodegradation activities were found to be present after the growth in potato dextrose broth for 96 h, when compared with the activities at different time intervals during the growth phase. NADPH was the preferred cofactor for enzyme activity, which was inhibited by CO, indicating cytochrome P450 mediated reactions. A significant increase in all the enzyme activities was observed when mycelium incubated for 18 h in mineral salts medium, containing cholesterol, camphor, naphthalene, 1,2-dimethoxybenzene, phenobarbital, n-hexane, kerosene or saffola oil as inducers. Acetaldehyde produced by alcohol dehydrogenase could be used as an indicator for the kerosene biodegradation.

Generation and Analysis of the Expressed Sequence Tags from the Mycelium of Ganoderma lucidum

PubMed Central

Huang, Yen-Hua; Wu, Hung-Yi; Wu, Keh-Ming; Liu, Tze-Tze; Liou, Ruey-Fen; Tsai, Shih-Feng; Shiao, Ming-Shi; Ho, Low-Tone; Tzean, Shean-Shong; Yang, Ueng-Cheng

2013-01-01

Ganoderma lucidum (G. lucidum) is a medicinal mushroom renowned in East Asia for its potential biological effects. To enable a systematic exploration of the genes associated with the various phenotypes of the fungus, the genome consortium of G. lucidum has carried out an expressed sequence tag (EST) sequencing project. Using a Sanger sequencing based approach, 47,285 ESTs were obtained from in vitro cultures of G. lucidum mycelium of various durations. These ESTs were further clustered and merged into 7,774 non-redundant expressed loci. The features of these expressed contigs were explored in terms of over-representation, alternative splicing, and natural antisense transcripts. Our results provide an invaluable information resource for exploring the G. lucidum transcriptome and its regulation. Many cases of the genes over-represented in fast-growing dikaryotic mycelium are closely related to growth, such as cell wall and bioactive compound synthesis. In addition, the EST-genome alignments containing putative cassette exons and retained introns were manually curated and then used to make inferences about the predominating splice-site recognition mechanism of G. lucidum. Moreover, a number of putative antisense transcripts have been pinpointed, from which we noticed that two cases are likely to reveal hitherto undiscovered biological pathways. To allow users to access the data and the initial analysis of the results of this project, a dedicated web site has been created at http://csb2.ym.edu.tw/est/. PMID:23658685

Risk assessment of fungal spoilage: A case study of Aspergillus niger on yogurt.

PubMed

Gougouli, Maria; Koutsoumanis, Konstantinos P

2017-08-01

A quantitative risk assessment model of yogurt spoilage by Aspergillus niger was developed based on a stochastic modeling approach for mycelium growth by taking into account the important sources of variability such as time-temperature conditions during the different stages of chill chain and individual spore behavior. Input parameters were fitted to the appropriate distributions and A. niger colony's diameter at each stage of the chill chain was estimated using Monte Carlo simulation. By combining the output of the growth model with the fungus prevalence, that can be estimated by the industry using challenge tests, the risk of spoilage translated to number of yogurt cups in which a visible mycelium of A. niger is being formed at the time of consumption was assessed. The risk assessment output showed that for a batch of 100,000 cups in which the percentage of contaminated cups with A. niger was 1% the predicted numbers (median (5 th , 95 th percentiles)) of the cups with a visible mycelium at consumption time were 8 (5, 14). For higher percentages of 3, 5 and 10 the predicted numbers (median (5 th , 95 th percentiles)) of the spoiled cups at consumption time were estimated to be 24 (16, 35), 39 (29, 52) and 80 (64, 94), respectively. The developed model can lead to a more effective risk-based quality management of yogurt and support the decision making in yogurt production. Copyright © 2017 Elsevier Ltd. All rights reserved.

MreB of Streptomyces coelicolor is not essential for vegetative growth but is required for the integrity of aerial hyphae and spores.

PubMed

Mazza, Paola; Noens, Elke E; Schirner, Kathrin; Grantcharova, Nina; Mommaas, A Mieke; Koerten, Henk K; Muth, Günther; Flärdh, Klas; van Wezel, Gilles P; Wohlleben, Wolfgang

2006-05-01

MreB forms a cytoskeleton in many rod-shaped bacteria which is involved in cell shape determination and chromosome segregation. PCR-based and Southern analysis of various actinomycetes, supported by analysis of genome sequences, revealed mreB homologues only in genera that form an aerial mycelium and sporulate. We analysed MreB in one such organism, Streptomyces coelicolor. Ectopic overexpression of mreB impaired growth, and caused swellings and lysis of hyphae. A null mutant with apparently normal vegetative growth was generated. However, aerial hyphae of this mutant were swelling and lysing; spores doubled their volume and lost their characteristic resistance to stress conditions. Loss of cell wall consistency was observed in MreB-depleted spores by transmission electron microscopy. An MreB-EGFP fusion was constructed to localize MreB in the mycelium. No clearly localized signal was seen in vegetative mycelium. However, strong fluorescence was observed at the septa of sporulating aerial hyphae, then as bipolar foci in young spores, and finally in a ring- or shell-like pattern inside the spores. Immunogold electron microscopy using MreB-specific antibodies revealed that MreB is located immediately underneath the internal spore wall. Thus, MreB is not essential for vegetative growth of S. coelicolor, but exerts its function in the formation of environmentally stable spores, and appears to primarily influence the assembly of the spore cell wall.

Toxicity of organic and inorganic nanoparticles to four species of white-rot fungi.

PubMed

Galindo, T P S; Pereira, R; Freitas, A C; Santos-Rocha, T A P; Rasteiro, M G; Antunes, F; Rodrigues, D; Soares, A M V M; Gonçalves, F; Duarte, A C; Lopes, I

2013-08-01

The rapid development of nanoparticles (NP) for industrial applications and large-volume manufacturing, with its subsequent release into the environment, raised the need to understand and characterize the potential effects of NP to biota. Accordingly, this work aimed to assess sublethal effects of five NP to the white-rot fungi species Trametes versicolor, Lentinus sajor caju, Pleurotus ostreatus, and Phanerochaete chrysosporium. Each species was exposed to serial dilutions of the following NP: organic-vesicles of SDS/DDAB and of Mo/NaO; gold-NP, quantum dot CdSe/ZnS, and Fe/Co. Fungi growth rate was monitored every day, and at the end of assay the mycelium from each replicate was collected to evaluate possible changes in its chemical composition. For all NP-suspensions the following parameters were characterized: hydrodynamic diameter, surface charge, aggregation index, zeta potential, and conductivity. All tested NP tended to aggregate when suspended in aqueous media. The obtained results showed that gold-NP, CdSe/ZnS, Mo/NaO, and SDS/DDAB significantly inhibited the growth of fungi with effects on the mycelium chemical composition. Among the tested NP, gold-NP and CdSe/ZnS were the ones exerting a higher effect on the four fungi. Finally to our knowledge, this is the first study reporting that different types of NP induce changes in the chemical composition of fungi mycelium. Copyright © 2013 Elsevier B.V. All rights reserved.

Fermentation optimization for the production of bioactive polysaccharides from Cordyceps sinensis fungus UM01.

PubMed

Wang, Lan-Ying; Cheong, Kit-Leong; Wu, Ding-Tao; Meng, Lan-Zhen; Zhao, Jing; Li, Shao-Ping

2015-08-01

The optimal fermentation conditions and medium for the production of bioactive polysaccharides from the mycelium of Cordyceps sinensis fungus UM01 were investigated by using orthogonal design and high performance size exclusion chromatography coupled with multi-angel laser light scattering and refractive index detector (HPSEC-MALLS-RID). Results showed that the optimal temperature, initial pH, rotation speed, medium capacity (ratio of medium volume to the volume of flask bottle) and inoculums volume for the mycelium growth were 15 °C, pH 6.0, 150 rpm, 2/5 (v/v), and 3% (v/v), respectively. Furthermore, bioactive polysaccharides from the mycelium of C. sinensis fungus UM01 were determined as polysaccharide fractions with the molecular weight above 10 kDa. The optimal fermentation medium was determined as a composition of glucose 30.0 g/L, sucrose 30.0 g/L, KH2PO4 1.0 g/L, CaCl2 0.5 g/L, yeast extract 3.0 g/L, and MgCl2 0.1g/L according to the maximum amount of the bioactive polysaccharides (486.16±19.60 mg/L) measured by HPSEC-MALLS/RID. Results are helpful to establish an efficient and controllable fermentation process for the industrial production of bioactive polysaccharides from C. sinensis UM01, and beneficial to develop a unique health and functional product in future. Copyright © 2015 Elsevier B.V. All rights reserved.

Mechanisms of Bacterial (Serratia marcescens) Attachment to, Migration along, and Killing of Fungal Hyphae.

PubMed

Hover, Tal; Maya, Tal; Ron, Sapir; Sandovsky, Hani; Shadkchan, Yana; Kijner, Nitzan; Mitiagin, Yulia; Fichtman, Boris; Harel, Amnon; Shanks, Robert M Q; Bruna, Roberto E; García-Véscovi, Eleonora; Osherov, Nir

2016-05-01

We have found a remarkable capacity for the ubiquitous Gram-negative rod bacterium Serratia marcescens to migrate along and kill the mycelia of zygomycete molds. This migration was restricted to zygomycete molds and several basidiomycete species. No migration was seen on any molds of the phylum Ascomycota. S. marcescens migration did not require fungal viability or surrounding growth medium, as bacteria migrated along aerial hyphae as well.S. marcescens did not exhibit growth tropism toward zygomycete mycelium. Bacterial migration along hyphae proceeded only when the hyphae grew into the bacterial colony. S. marcescens cells initially migrated along the hyphae, forming attached microcolonies that grew and coalesced to generate a biofilm that covered and killed the mycelium. Flagellum-defective strains of S. marcescens were able to migrate along zygomycete hyphae, although they were significantly slower than the wild-type strain and were delayed in fungal killing. Bacterial attachment to the mycelium does not necessitate type 1 fimbrial adhesion, since mutants defective in this adhesin migrated equally well as or faster than the wild-type strain. Killing does not depend on the secretion of S. marcescens chitinases, as mutants in which all three chitinase genes were deleted retained wild-type killing abilities. A better understanding of the mechanisms by which S. marcescens binds to, spreads on, and kills fungal hyphae might serve as an excellent model system for such interactions in general; fungal killing could be employed in agricultural fungal biocontrol. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

Mechanisms of Bacterial (Serratia marcescens) Attachment to, Migration along, and Killing of Fungal Hyphae

PubMed Central

Hover, Tal; Maya, Tal; Ron, Sapir; Sandovsky, Hani; Shadkchan, Yana; Kijner, Nitzan; Mitiagin, Yulia; Fichtman, Boris; Harel, Amnon; Shanks, Robert M. Q.; Bruna, Roberto E.; García-Véscovi, Eleonora

2016-01-01

We have found a remarkable capacity for the ubiquitous Gram-negative rod bacterium Serratia marcescens to migrate along and kill the mycelia of zygomycete molds. This migration was restricted to zygomycete molds and several basidiomycete species. No migration was seen on any molds of the phylum Ascomycota. S. marcescens migration did not require fungal viability or surrounding growth medium, as bacteria migrated along aerial hyphae as well. S. marcescens did not exhibit growth tropism toward zygomycete mycelium. Bacterial migration along hyphae proceeded only when the hyphae grew into the bacterial colony. S. marcescens cells initially migrated along the hyphae, forming attached microcolonies that grew and coalesced to generate a biofilm that covered and killed the mycelium. Flagellum-defective strains of S. marcescens were able to migrate along zygomycete hyphae, although they were significantly slower than the wild-type strain and were delayed in fungal killing. Bacterial attachment to the mycelium does not necessitate type 1 fimbrial adhesion, since mutants defective in this adhesin migrated equally well as or faster than the wild-type strain. Killing does not depend on the secretion of S. marcescens chitinases, as mutants in which all three chitinase genes were deleted retained wild-type killing abilities. A better understanding of the mechanisms by which S. marcescens binds to, spreads on, and kills fungal hyphae might serve as an excellent model system for such interactions in general; fungal killing could be employed in agricultural fungal biocontrol. PMID:26896140

Foraging behavior of pileated woodpeckers in partial cut and uncut bottomland hardwood forest

USGS Publications Warehouse

Newell, P.; King, Sammy L.; Kaller, Michael D.

2009-01-01

In bottomland hardwood forests, partial cutting techniques are increasingly advocated and used to create habitat for priority wildlife like Louisiana black bear (Ursus americanus luteolus), white-tailed deer (Odocoileus virginianus), and Neotropical migrants. Although partial cutting may be beneficial to some species, those that use dead wood may be negatively affected since large diameter and poor quality trees (deformed, moribund, or dead) are rare, but normally targeted for removal. On the other hand, partial cutting can create dead wood if logging slash is left on-site. We studied foraging behavior of pileated woodpeckers (Dryocopus pileatus) in one- and two-year-old partial cuts designed to benefit priority species and in uncut forest during winter, spring, and summer of 2006 and 2007 in Louisiana. Males and females did not differ in their use of tree species, dbh class, decay class, foraging height, use of foraging tactics or substrate types; however, males foraged on larger substrates than females. In both partial cut and uncut forest, standing live trees were most frequently used (83% compared to 14% for standing dead trees and 3% for coarse woody debris); however, dead trees were selected (i.e. used out of proportion to availability). Overcup oak (Quercus lyrata) and bitter pecan (Carya aquatica) were also selected and sugarberry (Celtis laevigata) avoided. Pileated woodpeckers selected trees >= 50 cm dbh and avoided trees in smaller dbh classes (10-20 cm). Density of selected foraging substrates was the same in partial cut and uncut forest. Of the foraging substrates, woodpeckers spent 54% of foraging time on live branches and boles, 37% on dead branches and boles, and 9% on vines. Of the foraging tactics, the highest proportion of foraging time was spent excavating (58%), followed by pecking (14%), gleaning (14%), scaling (7%), berry-eating (4%), and probing (3%). Woodpecker use of foraging tactics and substrates, and foraging height and substrate diameter did not differ between recent partial cut and uncut forest. Partial cutting designed to improve or maintain habitat for priority wildlife did not affect pileated woodpecker foraging behavior or availability of selected trees compared to uncut forest in the short term.

Foraging behavior of pileated woodpeckers in partial cut and uncut bottomland hardwood forest

USGS Publications Warehouse

Newell, P.; King, S.; Kaller, M.

2009-01-01

In bottomland hardwood forests, partial cutting techniques are increasingly advocated and used to create habitat for priority wildlife like Louisiana black bear (Ursus americanus luteolus), white-tailed deer (Odocoileus virginianus), and Neotropical migrants. Although partial cutting may be beneficial to some species, those that use dead wood may be negatively affected since large diameter and poor quality trees (deformed, moribund, or dead) are rare, but normally targeted for removal. On the other hand, partial cutting can create dead wood if logging slash is left on-site. We studied foraging behavior of pileated woodpeckers (Dryocopus pileatus) in one- and two-year-old partial cuts designed to benefit priority species and in uncut forest during winter, spring, and summer of 2006 and 2007 in Louisiana. Males and females did not differ in their use of tree species, dbh class, decay class, foraging height, use of foraging tactics or substrate types; however, males foraged on larger substrates than females. In both partial cut and uncut forest, standing live trees were most frequently used (83% compared to 14% for standing dead trees and 3% for coarse woody debris); however, dead trees were selected (i.e. used out of proportion to availability). Overcup oak (Quercus lyrata) and bitter pecan (Carya aquatica) were also selected and sugarberry (Celtis laevigata) avoided. Pileated woodpeckers selected trees ???50 cm dbh and avoided trees in smaller dbh classes (10-20 cm). Density of selected foraging substrates was the same in partial cut and uncut forest. Of the foraging substrates, woodpeckers spent 54% of foraging time on live branches and boles, 37% on dead branches and boles, and 9% on vines. Of the foraging tactics, the highest proportion of foraging time was spent excavating (58%), followed by pecking (14%), gleaning (14%), scaling (7%), berry-eating (4%), and probing (3%). Woodpecker use of foraging tactics and substrates, and foraging height and substrate diameter did not differ between recent partial cut and uncut forest. Partial cutting designed to improve or maintain habitat for priority wildlife did not affect pileated woodpecker foraging behavior or availability of selected trees compared to uncut forest in the short term. ?? 2009 Elsevier B.V.

Phylogenetic lineages in Entomophthoromycota

USDA-ARS?s Scientific Manuscript database

Entomophthoromycota Humber is one of five major phylogenetic lineages among the former phylum Zygomycota. These early terrestrial fungi share evolutionarily ancestral characters such as coenocytic mycelium and gametangiogamy as a sexual process resulting in zygospore formation. Previous molecular st...

Substrate Topography Induces a Crossover from 2D to 3D Behavior in Fibroblast Migration

PubMed Central

Ghibaudo, Marion; Trichet, Léa; Le Digabel, Jimmy; Richert, Alain; Hersen, Pascal; Ladoux, Benoît

2009-01-01

Abstract In a three-dimensional environment, cells migrate through complex topographical features. Using microstructured substrates, we investigate the role of substrate topography in cell adhesion and migration. To do so, fibroblasts are plated on chemically identical substrates composed of microfabricated pillars. When the dimensions of the pillars (i.e., the diameter, length, and spacing) are varied, migrating cells encounter alternating flat and rough surfaces that depend on the spacing between the pillars. Consequently, we show that substrate topography affects cell shape and migration by modifying cell-to-substrate interactions. Cells on micropillar substrates exhibit more elongated and branched shapes with fewer actin stress fibers compared with cells on flat surfaces. By analyzing the migration paths in various environments, we observe different mechanisms of cell migration, including a persistent type of migration, that depend on the organization of the topographical features. These responses can be attributed to a spatial reorganization of the actin cytoskeleton due to physical constraints and a preferential formation of focal adhesions on the micropillars, with an increased lifetime compared to that observed on flat surfaces. By changing myosin II activity, we show that actomyosin contractility is essential in the cellular response to micron-scale topographic signals. Finally, the analysis of cell movements at the frontier between flat and micropillar substrates shows that cell transmigration through the micropillar substrates depends on the spacing between the pillars. PMID:19580774

Quantification of the biocontrol agent Trichoderma harzianum with real-time TaqMan PCR and its potential extrapolation to the hyphal biomass.

PubMed

López-Mondéjar, Rubén; Antón, Anabel; Raidl, Stefan; Ros, Margarita; Pascual, José Antonio

2010-04-01

The species of the genus Trichoderma are used successfully as biocontrol agents against a wide range of phytopathogenic fungi. Among them, Trichoderma harzianum is especially effective. However, to develop more effective fungal biocontrol strategies in organic substrates and soil, tools for monitoring the control agents are required. Real-time PCR is potentially an effective tool for the quantification of fungi in environmental samples. The aim of this study consisted of the development and application of a real-time PCR-based method to the quantification of T. harzianum, and the extrapolation of these data to fungal biomass values. A set of primers and a TaqMan probe for the ITS region of the fungal genome were designed and tested, and amplification was correlated to biomass measurements obtained with optical microscopy and image analysis, of the hyphal length of the mycelium of the colony. A correlation of 0.76 between ITS copies and biomass was obtained. The extrapolation of the quantity of ITS copies, calculated based on real-time PCR data, into quantities of fungal biomass provides potentially a more accurate value of the quantity of soil fungi. Copyright 2009 Elsevier Ltd. All rights reserved.

Determination of lytic enzyme activities of indigenous Trichoderma isolates from Pakistan.

PubMed

Asad, Saeed Ahmad; Tabassum, Ayesha; Hameed, Abdul; Hassan, Fayyaz Ul; Afzal, Aftab; Khan, Sabaz Ali; Ahmed, Rafiq; Shahzad, Muhammad

2015-01-01

This study investigated lytic enzyme activities in three indigenous Trichoderma strains namely, Trichoderma asperellum, Trichoderma harzianum and Trichoderma sp. Native Trichoderma strains and a virulent strain of Rhizoctonia solani isolated from infected bean plants were also included in the study. Enzyme activities were determined by measuring sugar reduction by dinitrosalicylic acid (DNS) method using suitable substrates. The antagonists were cultured in minimal salt medium with the following modifications: medium A (1 g of glucose), medium B (0.5 g of glucose + 0.5 g of deactivated R. solani mycelia), medium C (1.0 g of deactivated respective antagonist mycelium) and medium D (1 g of deactivated R. solani mycelia). T asperellum showed presence of higher amounts of chitinases, β-1, 3-glucanases and xylanases in extracellular protein extracts from medium D as compared to medium A. While, the higher activities of glucosidases and endoglucanses were shown in medium D extracts by T. harzianum. β-glucosidase activities were lower compared with other enzymes; however, activities of the extracts of medium D were significantly different. T. asperellum exhibited maximum inhibition (97.7%). On the other hand, Trichoderma sp. did not show any effect on mycelia growth of R. solani on crude extract.

Determination of lytic enzyme activities of indigenous Trichoderma isolates from Pakistan

PubMed Central

Asad, Saeed Ahmad; Tabassum, Ayesha; Hameed, Abdul; Hassan, Fayyaz ul; Afzal, Aftab; Khan, Sabaz Ali; Ahmed, Rafiq; Shahzad, Muhammad

2015-01-01

Abstract This study investigated lytic enzyme activities in three indigenous Trichoderma strains namely, Trichoderma asperellum, Trichoderma harzianum and Trichoderma sp. Native Trichoderma strains and a virulent strain of Rhizoctonia solani isolated from infected bean plants were also included in the study. Enzyme activities were determined by measuring sugar reduction by dinitrosalicylic acid (DNS) method using suitable substrates. The antagonists were cultured in minimal salt medium with the following modifications: medium A (1 g of glucose), medium B (0.5 g of glucose + 0.5 g of deactivated R. solani mycelia), medium C (1.0 g of deactivated respective antagonist mycelium) and medium D (1 g of deactivated R. solani mycelia). T asperellum showed presence of higher amounts of chitinases, β-1, 3-glucanases and xylanases in extracellular protein extracts from medium D as compared to medium A. While, the higher activities of glucosidases and endoglucanses were shown in medium D extracts by T. harzianum. β-glucosidase activities were lower compared with other enzymes; however, activities of the extracts of medium D were significantly different. T. asperellum exhibited maximum inhibition (97.7%). On the other hand, Trichoderma sp. did not show any effect on mycelia growth of R. solani on crude extract. PMID:26691463

Agaricus bisporus genome sequence: a commentary.

PubMed

Kerrigan, Richard W; Challen, Michael P; Burton, Kerry S

2013-06-01

The genomes of two isolates of Agaricus bisporus have been sequenced recently. This soil-inhabiting fungus has a wide geographical distribution in nature and it is also cultivated in an industrialized indoor process ($4.7bn annual worldwide value) to produce edible mushrooms. Previously this lignocellulosic fungus has resisted precise econutritional classification, i.e. into white- or brown-rot decomposers. The generation of the genome sequence and transcriptomic analyses has revealed a new classification, 'humicolous', for species adapted to grow in humic-rich, partially decomposed leaf material. The Agaricus biporus genomes contain a collection of polysaccharide and lignin-degrading genes and more interestingly an expanded number of genes (relative to other lignocellulosic fungi) that enhance degradation of lignin derivatives, i.e. heme-thiolate peroxidases and β-etherases. A motif that is hypothesized to be a promoter element in the humicolous adaptation suite is present in a large number of genes specifically up-regulated when the mycelium is grown on humic-rich substrate. The genome sequence of A. bisporus offers a platform to explore fungal biology in carbon-rich soil environments and terrestrial cycling of carbon, nitrogen, phosphorus and potassium. Copyright © 2013 Elsevier Inc. All rights reserved.

Insulin receptor substrate signaling controls cardiac energy metabolism and heart failure.

PubMed

Guo, Cathy A; Guo, Shaodong

2017-06-01

The heart is an insulin-dependent and energy-consuming organ in which insulin and nutritional signaling integrates to the regulation of cardiac metabolism, growth and survival. Heart failure is highly associated with insulin resistance, and heart failure patients suffer from the cardiac energy deficiency and structural and functional dysfunction. Chronic pathological conditions, such as obesity and type 2 diabetes mellitus, involve various mechanisms in promoting heart failure by remodeling metabolic pathways, modulating cardiac energetics and impairing cardiac contractility. Recent studies demonstrated that insulin receptor substrates 1 and 2 (IRS-1,-2) are major mediators of both insulin and insulin-like growth factor-1 (IGF-1) signaling responsible for myocardial energetics, structure, function and organismal survival. Importantly, the insulin receptor substrates (IRS) play an important role in the activation of the phosphatidylinositide-3-dependent kinase (PI-3K) that controls Akt and Foxo1 signaling cascade, regulating the mitochondrial function, cardiac energy metabolism and the renin-angiotensin system. Dysregulation of this branch in signaling cascades by insulin resistance in the heart through the endocrine system promotes heart failure, providing a novel mechanism for diabetic cardiomyopathy. Therefore, targeting this branch of IRS→PI-3K→Foxo1 signaling cascade and associated pathways may provide a fundamental strategy for the therapeutic and nutritional development in control of metabolic and cardiovascular diseases. In this review, we focus on insulin signaling and resistance in the heart and the role energetics play in cardiac metabolism, structure and function. © 2017 Society for Endocrinology.

Magneto-optic evaluation of antiferromagnetic α-Fe2O3 nanoparticles coated on a quartz substrate

NASA Astrophysics Data System (ADS)

Balasubramanian, Srinath; Panmand, Rajendra; Kumar, Ganapathy; Mahajan, Satish M.; Kale, Bharat B.

2016-03-01

This paper presents a prima facie study of the magneto-optic response of antiferromagnetic α-Fe2O3 nanoparticles coated on a quartz substrate investigated by MOKE. The concentrations of the iron oxide nanoparticles in the films were varied from 8.6% to 21.5% and showed a linear increase in film thicknesses. As the concentration of the iron oxide nanoparticles were increased, the samples changed from a net-like morphology to a crystalline morphology. Magnetization reversals in the lower concentration samples were asymmetric with the reversals for the ascending and descending branch of the hysteresis loop occurring on the same side. The asymmetry in the magnetization reversal was attributed to the angle between the antiferromagnetic easy axis and the external magnetic field. With increase in concentration, an improvement in the magneto-optic response was observed with the magnetization reversal occurring via coherent rotation for both ascending and descending branches of the hysteresis loop. The changes in the magneto-optic behavior for the samples with higher concentrations is attributed to the strong exchange interactions and changes in the shape of the nanoparticles. Sensitivity studies performed on the samples showed an increased magneto-optic sensitivity to changes in magnetic field for samples of higher concentration. The high sensitivity of these samples could be exploited in magneto-optic sensors. Nanoparticles on a quartz substrate could find applications in bio-medicine due to their bio-compatibility.

Structure of an XPF endonuclease with and without DNA suggests a model for substrate recognition

PubMed Central

Newman, Matthew; Murray-Rust, Judith; Lally, John; Rudolf, Jana; Fadden, Andrew; Knowles, Philip P; White, Malcolm F; McDonald, Neil Q

2005-01-01

The XPF/Mus81 structure-specific endonucleases cleave double-stranded DNA (dsDNA) within asymmetric branched DNA substrates and play an essential role in nucleotide excision repair, recombination and genome integrity. We report the structure of an archaeal XPF homodimer alone and bound to dsDNA. Superposition of these structures reveals a large domain movement upon binding DNA, indicating how the (HhH)2 domain and the nuclease domain are coupled to allow the recognition of double-stranded/single-stranded DNA junctions. We identify two nonequivalent DNA-binding sites and propose a model in which XPF distorts the 3′ flap substrate in order to engage both binding sites and promote strand cleavage. The model rationalises published biochemical data and implies a novel role for the ERCC1 subunit of eukaryotic XPF complexes. PMID:15719018

The influence of biotic and abiotic factors on (137)Cs accumulation in higher fungi after the accident at Chernobyl NPP.

PubMed

Zarubina, N

2016-09-01

Levels of soil contamination with (137)Cs, the belonging of fungi to a certain ecological group, the localization depth of the main part of mycelium in soil are the primary factors influencing the value of (137)Cs specific activity in higher fungi after the accident at Chernobyl NPP. It has been found that the value of (137)Cs specific activity in fungi of one species could vary by more than 10 times during a vegetation period. A correlation between the changes of (137)Cs content in fungi during the vegetation period and the amount of precipitates during various periods preceding the collection of samples has not been determined. An assumption has been proposed stating dependence between peculiarities of mycelium growth during the vegetation period and the changes of (137)Cs specific activity in fungi. Copyright © 2015 Elsevier Ltd. All rights reserved.

Quantitative Trait Loci Controlling Vegetative Growth Rate in the Edible Basidiomycete Pleurotus ostreatus

PubMed Central

Larraya, Luis M.; Idareta, Eneko; Arana, Dani; Ritter, Enrique; Pisabarro, Antonio G.; Ramírez, Lucia

2002-01-01

Mycelium growth rate is a quantitative characteristic that exhibits continuous variation. This trait has applied interest, as growth rate is correlated with production yield and increased advantage against competitors. In this work, we studied growth rate variation in the edible basidiomycete Pleurotus ostreatus growing as monokaryotic or dikaryotic mycelium on Eger medium or on wheat straw. Our analysis resulted in identification of several genomic regions (quantitative trait loci [QTLs]) involved in the control of growth rate that can be mapped on the genetic linkage map of this fungus. In some cases monokaryotic and dikaryotic QTLs clustered at the same map position, indicating that there are principal genomic areas responsible for growth rate control. The availability of this linkage map of growth rate QTLs can help in the design of rational strain breeding programs based on genomic information. PMID:11872457

Naphthoquinone spiroketals and organic extracts from the endophytic fungus Edenia gomezpompae as potential herbicides.

PubMed

Macías-Rubalcava, Martha L; Ruiz-Velasco Sobrino, M Emma; Meléndez-González, Claudio; Hernández-Ortega, Simón

2014-04-23

From the fermentation mycelium of the endophytic fungus Edenia gomezpompae were obtained several phytotoxic compounds including two new members of the naphthoquinone spiroketal family, namely, palmarumycin EG1 (1) and preussomerin EG4 (4). In addition, preussomerins EG1-EG3 (7-9) and palmarumycins CP19 (2), CP17 (3), and CP2 (6), as well as ergosta-4,6,8(14),22-tetraen-3-one (5), were obtained. Compounds 2, 3, and 5 are new to this species. The structures of palmarumycins CP19 (2) and CP17 (3) were unambiguously determined by X-ray analysis. The isolates and mycelium organic extracts from four morphological variants of E. gomezpompae caused significant inhibition of seed germination, root elongation, and seedling respiration of Amaranthus hypochondriacus, Solanum lycopersicum, and Echinochloa crus-galli. The treatments also affected respiration on intact mitochondria isolated from spinach.

Polysaccharides in fungi. XXXII. Hypoglycemic activity and chemical properties of a polysaccharide from the cultural mycelium of Cordyceps sinensis.

PubMed

Kiho, T; Hui, J; Yamane, A; Ukai, S

1993-12-01

Crude polysaccharides were obtained from a hot-water extract and alkaline extracts of the cultural mycelium of Cordyceps sinensis. They showed significant activity in normal mice and streptozotocin-induced diabetic mice as a result of intraperitoneal (i.p.) injection. A crude polysaccharide (CS-OHEP) obtained from 5% sodium hydroxide extract slightly lowered the plasma glucose level in normal mice by oral (p.o.) administration. A neutral polysaccharide (CS-F30) exhibited higher hypoglycemic activity than its crude polysaccharide (CS-OHEP), exhibited by i.p. injection, and it significantly lowered the glucose level by p.o. administration (50 mg/kg). However, it hardly affected the plasma insulin level in normal mice. CS-F30 ([alpha]D + 21 degrees in water) is composed of galactose, glucose and mannose (molar percent, 62:28:10), and its molecular weight is about 45000.

Identification, Cloning, and Characterization of a Lactococcus lactis Branched-Chain α-Keto Acid Decarboxylase Involved in Flavor Formation

PubMed Central

Smit, Bart A.; van Hylckama Vlieg, Johan E. T.; Engels, Wim J. M.; Meijer, Laura; Wouters, Jan T. M.; Smit, Gerrit

2005-01-01

The biochemical pathway for formation of branched-chain aldehydes, which are important flavor compounds derived from proteins in fermented dairy products, consists of a protease, peptidases, a transaminase, and a branched-chain α-keto acid decarboxylase (KdcA). The activity of the latter enzyme has been found only in a limited number of Lactococcus lactis strains. By using a random mutagenesis approach, the gene encoding KdcA in L. lactis B1157 was identified. The gene for this enzyme is highly homologous to the gene annotated ipd, which encodes a putative indole pyruvate decarboxylase, in L. lactis IL1403. Strain IL1403 does not produce KdcA, which could be explained by a 270-nucleotide deletion at the 3′ terminus of the ipd gene encoding a truncated nonfunctional decarboxylase. The kdcA gene was overexpressed in L. lactis for further characterization of the decarboxylase enzyme. Of all of the potential substrates tested, the highest activity was observed with branched-chain α-keto acids. Moreover, the enzyme activity was hardly affected by high salinity, and optimal activity was found at pH 6.3, indicating that the enzyme might be active under cheese ripening conditions. PMID:15640202

Substrate-Driven Mapping of the Degradome by Comparison of Sequence Logos

PubMed Central

Fuchs, Julian E.; von Grafenstein, Susanne; Huber, Roland G.; Kramer, Christian; Liedl, Klaus R.

2013-01-01

Sequence logos are frequently used to illustrate substrate preferences and specificity of proteases. Here, we employed the compiled substrates of the MEROPS database to introduce a novel metric for comparison of protease substrate preferences. The constructed similarity matrix of 62 proteases can be used to intuitively visualize similarities in protease substrate readout via principal component analysis and construction of protease specificity trees. Since our new metric is solely based on substrate data, we can engraft the protease tree including proteolytic enzymes of different evolutionary origin. Thereby, our analyses confirm pronounced overlaps in substrate recognition not only between proteases closely related on sequence basis but also between proteolytic enzymes of different evolutionary origin and catalytic type. To illustrate the applicability of our approach we analyze the distribution of targets of small molecules from the ChEMBL database in our substrate-based protease specificity trees. We observe a striking clustering of annotated targets in tree branches even though these grouped targets do not necessarily share similarity on protein sequence level. This highlights the value and applicability of knowledge acquired from peptide substrates in drug design of small molecules, e.g., for the prediction of off-target effects or drug repurposing. Consequently, our similarity metric allows to map the degradome and its associated drug target network via comparison of known substrate peptides. The substrate-driven view of protein-protein interfaces is not limited to the field of proteases but can be applied to any target class where a sufficient amount of known substrate data is available. PMID:24244149

Structure, function and regulation of the enzymes in the starch biosynthetic pathway.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Geiger, Jim

Starch is the major reserve polysaccharide in nature and accounts for the majority of the caloric intact of humans. It is also gaining importance as a renewable and biodegradable industrial material. There is burgeoning interest in increasing the amount and altering the properties of the plant starches by plant genetic modification. A rational approach to this effort will require a detailed, atomic-level understanding of the enzymatic processes that produce the starch granule. The starch granule is a complex particle made up of alternating layers of crystalline and amorphous lamellae. It consists of two types of polymer, amylose, a polymer ofmore » relatively long chains of α-1,4-linked glucans that contain virtually no branches, and amylopectin, which is highly branched and contains much shorter chains. This complex structure is synthesized by the coordinate activities of the starch synthases (SS), which elongate the polysaccharide chain by addition of glucose units via α-1,4 linkages using ADP- glucose as a donor, and branching enzymes (BE), which branch the polysaccharide chain by cleavage of α₋1,4 linkages and subsequent re-attachment via α₋1,6 linkages. Several isoforms of both starch synthase (SS) and branching enzyme (BE) are found in plants, including SSI, SSII, SSIII and granule- bound SS (GBSS), and SBEI, SBEIIa and SBEIIb. These isoforms have different activities and substrate and product specificities and play different roles in creating the granule and determining the properties of the resulting starch. The overarching goal of this proposal is to begin to understand the regulation and specificities of these enzymes at the atomic level. High-resolution X-ray structures of these enzymes bound to substrates and products will be determined to visualize the molecular interactions responsible for the properties of the enzymes. Hypotheses regarding these issues will then be tested using mutagenesis and enzyme assays. To date, we have determined the structure of ADP- Glucose pyrophosphorylase from potato in its inhibited conformation, and bound to both ATP and ADP-glucose. In addition, we have determined the first structure of glycogen synthase in its "closed", catalytically active conformation bound to ADP-glucose. We also determined the structure of glycogen synthase bound to malto-oligosaccharides, showing for the first time that an enzyme in the starch biosynthetic pathway recognizes glucans not just in its active site but on binding sites on the surface of the enzyme ten’s of Angstroms from the active site. In addition our structure of a glycogen branching enzyme bound to malto-oligosaccharides identified seven distinct binding sites distributed about the surface of the enzyme. We will now determine the function of these sites to get a molecular-level picture of exactly how these enzymes interact with their polymeric substrates and confer specificity leading to the complex structure of the starch granule. We will extend our studies to other isoforms of the enzymes, to understand how their structures give rise to their distinct function. Our goal is to understand what accounts for the various functional differences between SS and SBE isoforms at a molecular level.« less

Chance, destiny, and the inner workings of ClpXP.

PubMed

Russell, Rick; Matouschek, Andreas

2014-07-31

AAA+ proteases are responsible for protein degradation in all branches of life. Using single-molecule and ensemble assays, Cordova et al. investigate how the bacterial protease ClpXP steps through a substrate's polypeptide chain and construct a quantitative kinetic model that recapitulates the interplay between stochastic and deterministic behaviors of ClpXP. Copyright © 2014 Elsevier Inc. All rights reserved.

Coral reef recovery in Florida and the Persian Gulf

USGS Publications Warehouse

Shinn, Eugene A.

1976-01-01

Long-term observations and study of coral reef destruction by hurricanes in the Florida Keys show, surprisingly, that although corals are devastated on a grand scale during storms, recovery is rapid. Recovery occurs because of the widespread scattering of live fragments, many of which become growth sites of new colonies. Reef recovery from death by chilling in the Persian Gulf was well under way when last observed, but it is not yet known if the recovery rate was as rapid as recovery from the storm destruction in Florida. Recovery from death by chilling requires settlement of transported coral larvae and a substrate suitable for larval attachment. Such resettlement is subject to the effects of currents, predators, pollution, and competition for substrate. A growth rate of 10 cm per year combined with geometrical progression of branch formation accounts for rapid recovery. Although calculated coral proliferation seems unusually high, it has been confirmed by serial underwater photographs spanning ten years. More precise measurements of growth and branching are needed, along with growth data for other common reef-building corals. Such data would be useful for predicting standing crop of a restocked or transplanted reef.

Functions of Calcium-Dependent Protein Kinases in Plant Innate Immunity

PubMed Central

Gao, Xiquan; Cox, Kevin L.; He, Ping

2014-01-01

An increase of cytosolic Ca2+ is generated by diverse physiological stimuli and stresses, including pathogen attack. Plants have evolved two branches of the immune system to defend against pathogen infections. The primary innate immune response is triggered by the detection of evolutionarily conserved pathogen-associated molecular pattern (PAMP), which is called PAMP-triggered immunity (PTI). The second branch of plant innate immunity is triggered by the recognition of specific pathogen effector proteins and known as effector-triggered immunity (ETI). Calcium (Ca2+) signaling is essential in both plant PTI and ETI responses. Calcium-dependent protein kinases (CDPKs) have emerged as important Ca2+ sensor proteins in transducing differential Ca2+ signatures, triggered by PAMPs or effectors and activating complex downstream responses. CDPKs directly transmit calcium signals by calcium binding to the elongation factor (EF)-hand domain at the C-terminus and substrate phosphorylation by the catalytic kinase domain at the N-terminus. Emerging evidence suggests that specific and overlapping CDPKs phosphorylate distinct substrates in PTI and ETI to regulate diverse plant immune responses, including production of reactive oxygen species, transcriptional reprogramming of immune genes, and the hypersensitive response. PMID:27135498

FY02 Final Report on Phytoremediation of Chlorinated Ethenes in Southern Sector Sediments of the Savannah River Site

DOE Office of Scientific and Technical Information (OSTI.GOV)

Brigmon, R..L.

This final report details the operations and results of a 3-year Seepline Phytoremediation Project performed adjacent to Tims Branch, which is located in the Southern Sector of the Savannah River Site (SRS) A/M Area. Phytoremediation is a process where interactions between vegetation, associated microorganisms, and the host substrate combine to effectively degrade contaminated soils, sediments, and groundwater. Phytoremediation is a rapidly developing technology that shows promise for the effective and safe cleanup of certain hazardous wastes. It has the potential to remediate numerous volatile organic compounds (VOCs). Extensive characterization work has demonstrated that two VOCs, tetrachloroethylene (PCE) and trichloroethylene (TCE)more » are the major components of the VOC-contaminated groundwater that is migrating through the Southern Sector and Tims Branch seepline area (WSRC, 1999). The PCE and TCE are chlorinated ethenes (CE), and have been detected in seepline soils and ground water adjacent to the ecologically-sensitive Tims Branch seepline area.« less

[Preliminary assessment of habitat of juvenile Collichthys lucidus in the Yangtze estuary].

PubMed

Yang, Gang; Zhang, Tao; Zhuang, Ping; Hou, Jun-Li; Wang, Yu; Song, Chao; Zhang, Long-Zhen

2014-08-01

To evaluate the choice preference of fish habitat in the Yangtze estuary, juvenile Collichthys lucidus which is the dominant species in spring was selected. The 4 indicator factors, including abundance of Pseudograpsus albus, salinity, substrate type and water depth, were selected from 19 environmental factors. Then, the indices of the habitat suitability curves of the 4 indicator factors were established, and the HSI of juvenile C. lucidus at each site was calculated. The results indicated that HSI was almost more than 0.5 in North Branch, and less than 0.2 in South Branch. It showed that the North Branch of Yangtze estuary was the main nursery area of C. lucidus. The most suitable growth sector was the area with salinity more than 14, mean grain size of substrate less than 29 μm and water depth 2 to 5 m, which was consistent with the distribution of HSI. The study demonstrated that biological factors could be characterized by the response of juvenile C. lucidus to the environment. Chemical oxygen demand, ammonium nitrogen, total phosphorus and volatile phenol did not have significant correlation with the fish abundance, with which nitrite nitrogen, nitrate nitrogen and total nitrogen had significant positive correlation. It suggested that the eutrophication of the survey area had not damaged the habitat of C. lucidus. However, copper ion and cadmium ion had significant negative correlation with the fish abundance, which indicated that the heavy metal pollution had harmed the growth and distribution of juvenile C. lucidus. It was inferred that the heavy metal pollution was the restrictive factor influencing the fish habitat in Yangtze estuary.

Chemical Composition and Medicinal Value of the New Ganoderma tsugae var. jannieae CBS-120304 Medicinal Higher Basidiomycete Mushroom.

PubMed

Chan, Jannie Siew Lee; Asatiani, Mikheil D; Sharvit, Lital E; Trabelcy, Beny; Barseghyan, Gayane S; Wasser, Solomon P

2015-01-01

In this research, the chemical composition and anticancer and antioxidant activity of the new medicinal mushroom Ganoderma tsugae var. jannieae CBS-120304 were evaluated. The chemical composition assay includes amounts of total carbohydrates and proteins, amino acids, fatty acids, micro- and macroelements, and vitamins. The investigated medicinal mushroom seemed to be a rich source of nutritional components. Mycelium accumulated more than 2-fold more total protein compared with the fruiting body and reached 37% and 16% of dry weight, respectively. Carbohydrate content in the fruiting body seemed to be conspicuously higher than in the mycelium (50% of dry weight) and reached 80% of dry weight. Quantification of the identified fatty acids indicated that, in general, palmitic acid, oleic acid, and linoleic acid were the major fatty acids. Toxic elements, such as silver, arsenic, cadmium, and mercury, were found only in trace amounts in mycelium and were not detected in the fruiting body. Furthermore, the 1,1-diphenyl-2-picrylhydrazyl free radical scavenging assay was used to evaluate antioxidant activity. The highest radical scavenging activity was 9.0 mg/mL (65.9%) by ethanol extract. In addition, mycelial extracts were tested to inhibit MCF7 breast cancer cells. Ganoderma tsugae var. jannieae ethyl acetate extract (GTEAE) extract showed high potential by inhibiting reporter activity by more than 70%. Results demonstrated that GTEAE had a strong effect on inhibitory protein κΒα level in the higher concentration used (200 gg/mL), which could be compared with the effect of parthenolide. Furthermore, GTEAE demonstrated strong inhibition of IκΒα phosphorylation.

Mycoremediation of manganese and phenanthrene by Pleurotus eryngii mycelium enhanced by Tween 80 and saponin.

PubMed

Wu, Minghui; Xu, Yongan; Ding, Wenbo; Li, Yuanyuan; Xu, Heng

2016-08-01

Bioremediation of areas co-contaminated with metals and polycyclic aromatic hydrocarbons (PAHs) by mushrooms has attracted considerable attention in recent years. In this study, Pleurotus eryngii was introduced for the removal of Mn and phenanthrene (Phe) from potato liquid medium (PDL) simultaneously. Effects of Tween 80 and saponin on P. eryngii growth together with Mn uptake as well as Phe removal were investigated. Although pollutants had a negative effect on mycelial morphology and growth, P. eryngii could still tolerate and remove Mn and Phe. Tween 80 increased removal of Mn and Phe through increase of P. eryngii growth, Phe solubility, pollutants bioavailability, and specific surface area of mycelium pellets, moreover, the activities of manganese peroxidase (MnP) and laccase, which played an important role on PAHs biodegradation. The maximal removal of Mn and Phe was achieved (92.17 and 93.85 % after 15 days incubation, respectively) with 0.6 g L(-1) Tween 80. Treatments with saponin markedly inhibited P. eryngii growth (50.17-66.32 % lower relative to control) due to its fungistatic activity. Nevertheless, saponin could slightly enhance Phe removal through increasing solubility of Phe, and Phe removal rate varied from 80.53 to 87.06 % in saponin treatments. Joint stress of Mn and Phe induced a strong antioxidative response, and superoxide dismutase (SOD) activity decreased in surfactants-treated mycelium compared with control. Generally, Tween 80 was more suitable for strengthening mycoremediation by P. eryngii than saponin, and could be a promising alternative for the remediation of heavy metals and PAHs co-contaminated sites by mushrooms.

Branching instability in expanding bacterial colonies.

PubMed

Giverso, Chiara; Verani, Marco; Ciarletta, Pasquale

2015-03-06

Self-organization in developing living organisms relies on the capability of cells to duplicate and perform a collective motion inside the surrounding environment. Chemical and mechanical interactions coordinate such a cooperative behaviour, driving the dynamical evolution of the macroscopic system. In this work, we perform an analytical and computational analysis to study pattern formation during the spreading of an initially circular bacterial colony on a Petri dish. The continuous mathematical model addresses the growth and the chemotactic migration of the living monolayer, together with the diffusion and consumption of nutrients in the agar. The governing equations contain four dimensionless parameters, accounting for the interplay among the chemotactic response, the bacteria-substrate interaction and the experimental geometry. The spreading colony is found to be always linearly unstable to perturbations of the interface, whereas branching instability arises in finite-element numerical simulations. The typical length scales of such fingers, which align in the radial direction and later undergo further branching, are controlled by the size parameters of the problem, whereas the emergence of branching is favoured if the diffusion is dominant on the chemotaxis. The model is able to predict the experimental morphologies, confirming that compact (resp. branched) patterns arise for fast (resp. slow) expanding colonies. Such results, while providing new insights into pattern selection in bacterial colonies, may finally have important applications for designing controlled patterns. © 2015 The Author(s) Published by the Royal Society. All rights reserved.

Methyl-branched lipids promote the membrane adsorption of α-synuclein by enhancing shallow lipid-packing defects.

PubMed

Garten, Matthias; Prévost, Coline; Cadart, Clotilde; Gautier, Romain; Bousset, Luc; Melki, Ronald; Bassereau, Patricia; Vanni, Stefano

2015-06-28

Alpha-synuclein (AS) is a synaptic protein that is directly involved in Parkinson's disease due to its tendency to form protein aggregates. Since AS aggregation can be dependent on the interactions between the protein and the cell plasma membrane, elucidating the membrane binding properties of AS is of crucial importance to establish the molecular basis of AS aggregation into toxic fibrils. Using a combination of in vitro reconstitution experiments based on Giant Unilamellar Vesicles (GUVs), confocal microscopy and all-atom molecular dynamics simulations, we have investigated the membrane binding properties of AS, with a focus on the relative contribution of hydrophobic versus electrostatic interactions. In contrast with previous observations, we did not observe any binding of AS to membranes containing the ganglioside GM1, even at relatively high GM1 content. AS, on the other hand, showed a stronger affinity for neutral flat membranes consisting of methyl-branched lipids. To rationalize these results, we used all-atom molecular dynamics simulations to investigate the influence of methyl-branched lipids on interfacial membrane properties. We found that methyl-branched lipids promote the membrane adsorption of AS by creating shallow lipid-packing defects to a larger extent than polyunsaturated and monounsaturated lipids. Our findings suggest that methyl-branched lipids may constitute a remarkably adhesive substrate for peripheral proteins that adsorb on membranes via hydrophobic insertions.

New secondary metabolites from bioactive extracts of the fungus Armillaria tabescens

USDA-ARS?s Scientific Manuscript database

Ethyl acetate extracts of Armillaria tabescens (strain JNB-OZ344) mycelium showed significant fungistatic and bacteristatic activities against several major human pathogens including Candida albicans, Cryptococcus neoformans, Escherichia coli and Mycobacterium intracellulare. Chemical analysis of th...

Parasitic Aspects of a Fairy Ring Fungus, Marasmius oreades

Treesearch

T. H. Filer

1965-01-01

Marasmius oreades parasitizes Poa pratensis, Festuca rubra, and Agrostis tenuis. The fungus penetrates the root directly in all three species and does not require natural openings or wounds. The mycelium ramifies in the cortical cells and destroys the cell contents.

Atomic force microscopy investigation of growth process of organic TCNQ aggregates on SiO2 and mica substrates

NASA Astrophysics Data System (ADS)

Huan, Qing; Hu, Hao; Pan, Li-Da; Xiao, Jiang; Du, Shi-Xuan; Gao, Hong-Jun

2010-08-01

Deposition patterns of tetracyanoquinodimethane (TCNQ) molecules on different surfaces are investigated by atomic force microscopy. A homemade physical vapour deposition system allows the better control of molecule deposition. Taking advantage of this system, we investigate TCNQ thin film growth on both SiO2 and mica surfaces. It is found that dense island patterns form at a high deposition rate, and a unique seahorse-like pattern forms at a low deposition rate. Growth patterns on different substrates suggest that the fractal pattern formation is dominated by molecule-molecule interaction. Finally, a phenomenal “two-branch" model is proposed to simulate the growth process of the seahorse pattern.

Substrate-controlled Rh(II)-catalyzed single-electron-transfer (SET): divergent synthesis of fused indoles.

PubMed

Chen, Kai; Zhu, Zi-Zhong; Liu, Jia-Xin; Tang, Xiang-Ying; Wei, Yin; Shi, Min

2016-01-07

Rh(II)-catalyzed diversified ring expansions controlled by single-electron-transfer (SET) have been disclosed in this communication, producing a series of indole-fused azetidines and 1H-carbazoles or related derivatives in moderate to good yields via Rh2(III,II) nitrene radical intermediates. The direction of ring expansion branches according to different ring sizes of methylenecycloalkanes.

Genome sequence of the button mushroom Agaricus bisporus reveals mechanisms governing adaptation to a humic-rich ecological niche

DOE Office of Scientific and Technical Information (OSTI.GOV)

Morin, Emmanuelle; Kohler, Annegret; Baker, Adam R.

Agaricus bisporus is the model fungus for the adaptation, persistence, and growth in the humic-rich leaf-litter environment. Aside from its ecological role, A. bisporus has been an important component of the human diet for over 200 y and worldwide cultivation of the button mushroom forms a multibillion dollar industry. We present two A. bisporus genomes, their gene repertoires and transcript profiles on compost and during mushroom formation. The genomes encode a full repertoire of polysaccharide-degrading enzymes similar to that of wood-decayers. Comparative transcriptomics of mycelium grown on defined medium, casing-soil, and compost revealed genes encoding enzymes involved in xylan, cellulose,more » pectin, and protein degradation are more highly expressed in compost. The striking expansion of heme-thiolate peroxidases and etherases is distinctive from Agaricomycotina wood-decayers and suggests a broad attack on decaying lignin and related metabolites found in humic acid-rich environment. Similarly, up-regulation of these genes together with a lignolytic manganese peroxidase, multiple copper radical oxidases, and cytochrome P450s is consistent with challenges posed by complex humic-rich substrates. The gene repertoire and expression of hydrolytic enzymes in A. bisporus is substantially different from the taxonomically related ectomycorrhizal symbiont Laccaria bicolor. A common promoter motif was also identified in genes very highly expressed in humic-rich substrates. These observations reveal genetic and enzymatic mechanisms governing adaptation to the humic-rich ecological niche formed during plant degradation, further defining the critical role such fungi contribute to soil structure and carbon sequestration in terrestrial ecosystems. Genome sequence will expedite mushroom breeding for improved agronomic characteristics.« less

Genome sequence of the button mushroom Agaricus bisporus reveals mechanisms governing adaptation to a humic-rich ecological niche

PubMed Central

Morin, Emmanuelle; Kohler, Annegret; Baker, Adam R.; Foulongne-Oriol, Marie; Lombard, Vincent; Nagye, Laszlo G.; Ohm, Robin A.; Patyshakuliyeva, Aleksandrina; Brun, Annick; Aerts, Andrea L.; Bailey, Andrew M.; Billette, Christophe; Coutinho, Pedro M.; Deakin, Greg; Doddapaneni, Harshavardhan; Floudas, Dimitrios; Grimwood, Jane; Hildén, Kristiina; Kües, Ursula; LaButti, Kurt M.; Lapidus, Alla; Lindquist, Erika A.; Lucas, Susan M.; Murat, Claude; Riley, Robert W.; Salamov, Asaf A.; Schmutz, Jeremy; Subramanian, Venkataramanan; Wösten, Han A. B.; Xu, Jianping; Eastwood, Daniel C.; Foster, Gary D.; Sonnenberg, Anton S. M.; Cullen, Dan; de Vries, Ronald P.; Lundell, Taina; Hibbett, David S.; Henrissat, Bernard; Burton, Kerry S.; Kerrigan, Richard W.; Challen, Michael P.; Grigoriev, Igor V.; Martin, Francis

2012-01-01

Agaricus bisporus is the model fungus for the adaptation, persistence, and growth in the humic-rich leaf-litter environment. Aside from its ecological role, A. bisporus has been an important component of the human diet for over 200 y and worldwide cultivation of the “button mushroom” forms a multibillion dollar industry. We present two A. bisporus genomes, their gene repertoires and transcript profiles on compost and during mushroom formation. The genomes encode a full repertoire of polysaccharide-degrading enzymes similar to that of wood-decayers. Comparative transcriptomics of mycelium grown on defined medium, casing-soil, and compost revealed genes encoding enzymes involved in xylan, cellulose, pectin, and protein degradation are more highly expressed in compost. The striking expansion of heme-thiolate peroxidases and β-etherases is distinctive from Agaricomycotina wood-decayers and suggests a broad attack on decaying lignin and related metabolites found in humic acid-rich environment. Similarly, up-regulation of these genes together with a lignolytic manganese peroxidase, multiple copper radical oxidases, and cytochrome P450s is consistent with challenges posed by complex humic-rich substrates. The gene repertoire and expression of hydrolytic enzymes in A. bisporus is substantially different from the taxonomically related ectomycorrhizal symbiont Laccaria bicolor. A common promoter motif was also identified in genes very highly expressed in humic-rich substrates. These observations reveal genetic and enzymatic mechanisms governing adaptation to the humic-rich ecological niche formed during plant degradation, further defining the critical role such fungi contribute to soil structure and carbon sequestration in terrestrial ecosystems. Genome sequence will expedite mushroom breeding for improved agronomic characteristics. PMID:23045686

Differential allergy responses to Metarhizium anisopliae fungal component extracts in BALB/c mice

EPA Science Inventory

Intratracheal aspiration (IA) exposure to Metarhizium anisopliae crude antigen (MACA), which is composed of equal protein amounts of mycelium (MYC), conidia (CON) and inducible proteases/chitinases (IND) extracts/filtrates, has resulted in responses characteristic of human allerg...

Paecilomyces tabacinus

USDA-ARS?s Scientific Manuscript database

Culture characteristics - (in the dark, 25 °C after 7 d): Colonies on MEA 30–31 mm diam, velutinous to slightly floccose, mycelium white, sporulation abundant, cream-buff [R30; Ridgway (1912)], exudate clear, small droplets dominate, soluble pigments not observed, reverse cream-buff to chamois (R30)...

Rasamsonia columbiensis

USDA-ARS?s Scientific Manuscript database

Culture characteristics - (in the dark, 25 °C after 7 d): Colonies on MEA 30–31 mm diam, velutinous to slightly floccose, mycelium white, sporulation abundant, cream-buff [R30; Ridgway (1912)], exudate clear, small droplets dominate, soluble pigments not observed, reverse cream-buff to chamois (R30)...

Morphology and mycelial growth rate of Pleurotus spp. strains from the Mexican mixtec region

PubMed Central

Guadarrama-Mendoza, P.C.; del Toro, G. Valencia; Ramírez-Carrillo, R.; Robles-Martínez, F.; Yáñez-Fernández, J.; Garín-Aguilar, M.E.; Hernández, C.G.; Bravo-Villa, G.

2014-01-01

Two native Pleurotus spp. strains (white LB-050 and pale pink LB-051) were isolated from rotten tree trunks of cazahuate (Ipomoea murucoides) from the Mexican Mixtec Region. Both strains were chemically dedikaryotized to obtain their symmetrical monokaryotic components (neohaplonts). This was achieved employing homogenization time periods from 60 to 65 s, and 3 day incubation at 28 °C in a peptone-glucose solution (PGS). Pairing of compatible neohaplonts resulted in 56 hybrid strains which were classified into the four following hybrid types: (R1-nxB1-n, R1-nxB2-1, R2-nxB1-n and R2-nxB2-1). The mycelial growth of Pleurotus spp. monokaryotic and dikaryotic strains showed differences in texture (cottony or floccose), growth (scarce, regular or abundant), density (high, regular or low), and pigmentation (off-white, white or pale pink). To determine the rate and the amount of mycelium growth in malt extract agar at 28 °C, the diameter of the colony was measured every 24 h until the Petri dish was completely colonized. A linear model had the best fit to the mycelial growth kinetics. A direct relationship between mycelial morphology and growth rate was observed. Cottony mycelium presented significantly higher growth rates (p < 0.01) in comparison with floccose mycelium. Thus, mycelial morphology can be used as criterion to select which pairs must be used for optimizing compatible-mating studies. Hybrids resulting from cottony neohaplonts maintained the characteristically high growth rates of their parental strains with the hybrid R1-nxB1-n being faster than the latter. PMID:25477920

Submerged cultivation of medicinal mushrooms: bioprocesses and products (review).

PubMed

Elisashvili, Vladimir

2012-01-01

Medicinal mushrooms belonging to higher Basidiomycetes are an immensely rich yet largely untapped resource of useful, easily accessible, natural compounds with various biological activities that may promote human well-being. The medicinal properties are found in various cellular components and secondary metabolites (polysaccharides, proteins and their complexes, phenolic compounds, polyketides, triterpenoids, steroids, alkaloids, nucleotides, etc.), which have been isolated and identified from the fruiting bodies, culture mycelium, and culture broth of mushrooms. Some of these compounds have cholesterol-lowering, anti-diabetic, antioxidant, antitumor, immunomodulating, antimicrobial, and antiviral activities ready for industrial trials and further commercialization, while others are in various stages of development. Recently, the submerged cultivation of medicinal mushrooms has received a great deal of attention as a promising and reproducible alternative for the efficient production of mushroom mycelium and metabolites. Submerged cultivation of mushrooms has significant industrial potential, but its success on a commercial scale depends on increasing product yields and development of novel production systems that address the problems associated with this technique of mushroom cultivation. In spite of many researchers' efforts for the production of bioactive metabolites by mushrooms, the physiological and engineering aspects of submerged cultures are still far from being thoroughly studied. The vast majority of studies have focused on polysaccharide and ganoderic acid production in submerged cultivation of medicinal mushrooms, and very little has been written so far on the antioxidant and hemagglutinating activity of submerged mushroom cultures. The purpose of this review is to provide an update of the present state of the art and future prospects of submerged cultivation of medicinal mushrooms to produce mycelium and bioactive metabolites, and to make a contribution for the research and development of new pharmaceutical products from mushrooms. A brief overview of the metabolic diversity and bioactive compounds of mushrooms produced by submerged cultures is also given.

Comparative Analysis of the Number and Structure of the Complexes of Microscopic Fungi in Tundra and Taiga Soils in the North of the Kola Peninsula

NASA Astrophysics Data System (ADS)

Korneikova, M. V.

2018-01-01

The number, biomass, length of fungal mycelium, and species diversity of microscopic fungi have been studied in soils of the tundra and taiga zones in the northern part of the Kola Peninsula: Al-Fe-humus podzols (Albic Podzols), podburs (Entic Podzols), dry peaty soils (Folic Histosols), low-moor peat soils (Sapric Histosols), and soils of frost bare spots (Cryosols). The number of cultivated microscopic fungi in tundra soils varied from 8 to 328 thousand CFU/g, their biomass averaged 1.81 ± 0.19 mg/g, and the length of fungal mycelium averaged 245 ± 25 m/g. The number of micromycetes in taiga soils varied from 80 to 350 thousand CFU/g, the number of fungal propagules in some years reached 600 thousand CFU/g; the fungal biomass varied from 0.23 to 6.2 mg/g, and the length of fungal mycelium varied from 32 to 3900 m/g. Overall, 36 species of fungi belonging to 16 genera, 13 families, and 8 orders were isolated from tundra soils. The species diversity of microscopic fungi in taiga soils was significantly higher: 87 species belonging to 31 genera, 21 families, and 11 orders. Fungi from the Penicillium genus predominated in both natural zones and constituted 38-50% of the total number of isolated species. The soils of tundra and taiga zones were characterized by their own complexes of micromycetes; the similarity of their species composition was about 40%. In soils of the tundra zone, Mortierella longicollis, Penicillium melinii, P. raistrickii, and P. simplicissimum predominated; dominant fungal species in soils of the taiga zone were represented by M. longicollis, P. decumbens, P. implicatum, and Umbelopsis isabellina.

Anti-Inflammatory Activity of Sanghuangporus sanghuang Mycelium.

PubMed

Lin, Wang-Ching; Deng, Jeng-Shyan; Huang, Shyh-Shyun; Wu, Sheng-Hua; Chen, Chin-Chu; Lin, Wan-Rong; Lin, Hui-Yi; Huang, Guan-Jhong

2017-02-07

Acute lung injury (ALI) is characterized by inflammation of the lung tissue and oxidative injury caused by excessive accumulation of reactive oxygen species. Studies have suggested that anti-inflammatory or antioxidant agents could be used for the treatment of ALI with a good outcome. Therefore, our study aimed to test whether the mycelium extract of Sanghuangporus sanghuang (SS-1), believed to exhibit antioxidant and anti-inflammatory properties, could be used against the excessive inflammatory response associated with lipopolysaccharides (LPS)-induced ALI in mice and to investigate its possible mechanism of action. The experimental results showed that the administration of SS-1 could inhibit LPS-induced inflammation. SS-1 could reduce the number of inflammatory cells, inhibit myeloperoxidase (MPO) activity, regulate the TLR4/PI3K/Akt/mTOR pathway and the signal transduction of NF-κB and MAPK pathways in the lung tissue, and inhibit high mobility group box-1 protein 1 (HNGB1) activity in BALF. In addition, SS-1 could affect the synthesis of antioxidant enzymes Heme oxygenase 1 (HO-1) and Thioredoxin-1 (Trx-1) in the lung tissue and regulate signal transduction in the KRAB-associated protein-1 (KAP1)/nuclear factor erythroid-2-related factor Nrf2/Kelch Like ECH associated Protein 1 (Keap1) pathway. Histological results showed that administration of SS-1 prior to induction could inhibit the large-scale LPS-induced neutrophil infiltration of the lung tissue. Therefore, based on all experimental results, we propose that SS-1 exhibits a protective effect against LPS-induced ALI in mice. The mycelium of S. sanghuang can potentially be used for the treatment or prevention of inflammation-related diseases.

[Influence of antitumor preparations on the concentration of free radicals in cells of Fusarium bulbigenum var. blasticola fungus during primary and tumour-like secondary growth].

PubMed

Riabikin, Iu A; Nikitina, E T; Balgimbatva, A S; Zashkvara, O V; Shakiev, S Sh

2007-01-01

The fungus Fusarium bulbigenum var. blasticola in which secondary tumor-like formations appear under certain conditions in aging was used as a new test system to examine the action of antitumor preparations. Free radicals in the primary mycelium and tumor-like formations without introduction of preparations (control samples) and after the introduction of preparation into the cultivation medium of the fungus have been studied by EPR spectroscopy. The EPR spectra of the fungus represent single, somewhat asymmetrical lines with a width of deltaH = 0.4 divided by 0.6 mT and g = 2.0036 +/- 0.006, which enabled one to assign the paramagnetic centers observed to melanine radicals. It was found that the concentration of free radicals in tumor-like formations is always higher than in the primary mycelium, which may be related to intensive metabolism in tumor-like formations. It has been established that several antitumor preparations (fluorouracil, hydrea, methotrexat, and vepezide) completely inhibit the growth of tumor-like formations. Another group of preparations (cyclophosphanum, dacarbazin, adriablastin, and vinblastin), on the contrary, stimulate their growth, which is accompanied by an increase in the concentration of free radicals in cells of the primary mycelium and tumor-like formations. The preparations of the third group (mercaptopurine, lanvis, and farmorubicin), despite the increased level of free radicals in cells, have a weak inhibitory effect. It has been shown that, in the concentration range studied, vitamins B2, B12, C, and PP stimulate the growth of tumor-like formations, and, when used in combination with antitumor preparations, enhance or reduce the inhibitory properties of these preparations.

Environmental Conditions Affect Botrytis cinerea Infection of Mature Grape Berries More Than the Strain or Transposon Genotype.

PubMed

Ciliberti, Nicola; Fermaud, Marc; Roudet, Jean; Rossi, Vittorio

2015-08-01

Effects of environment, Botrytis cinerea strain, and their interaction on the infection of mature grape berries were investigated. The combined effect of temperature (T) of 15, 20, 25, and 30°C and relative humidity (RH) of 65, 80, 90, and 100% was studied by inoculating berries with mycelium plugs. Regardless of the T, no disease occurred at 65% RH, and both disease incidence and severity increased with increasing RH. The combined effect of T (5 to 30°C) and wetness duration (WD) of 3, 6, 12, 24, and 36 h was studied by inoculating berries with conidia. At WD of 36 h, disease incidence was approximately 75% of affected berries at 20 or 25°C, 50% at 15°C, and 30 to 20% at 30 and 10°C; no infection occurred at 5°C. Under favorable conditions (100% RH or 36 h of WD) and unfavorable conditions (65% RH or 3 h of WD), berry wounding did not significantly affect disease incidence; under moderately favorable conditions (80% RH or 6 to 12 h of WD), disease incidence was approximately 1.5 to 5 times higher in wounded than in intact berries. Our data collectively showed that (i) T and RH or WD were more important than strain for mature berry infection by either mycelium or conidia and (ii) the effect of the environment on the different strains was similar. Two equations were developed describing the combined effect of T and RH, or T and WD, on disease incidence following inoculation by mycelium (R2=0.99) or conidia (R2=0.96), respectively. These equations may be useful in the development of models used to predict and control Botrytis bunch rot during berry ripening.

P. brasiliensis Virulence Is Affected by SconC, the Negative Regulator of Inorganic Sulfur Assimilation

PubMed Central

Menino, João Filipe; Saraiva, Margarida; Gomes-Rezende, Jéssica; Sturme, Mark; Pedrosa, Jorge; Castro, António Gil; Ludovico, Paula; Goldman, Gustavo H.; Rodrigues, Fernando

2013-01-01

Conidia/mycelium-to-yeast transition of Paracoccidioides brasiliensis is a critical step for the establishment of paracoccidioidomycosis, a systemic mycosis endemic in Latin America. Thus, knowledge of the factors that mediate this transition is of major importance for the design of intervention strategies. So far, the only known pre-requisites for the accomplishment of the morphological transition are the temperature shift to 37°C and the availability of organic sulfur compounds. In this study, we investigated the auxotrophic nature to organic sulfur of the yeast phase of Paracoccidioides , with special attention to P. brasiliensis species. For this, we addressed the role of SconCp, the negative regulator of the inorganic sulfur assimilation pathway, in the dimorphism and virulence of this pathogen. We show that down-regulation of SCONC allows initial steps of mycelium-to-yeast transition in the absence of organic sulfur compounds, contrarily to the wild-type fungus that cannot undergo mycelium-to-yeast transition under such conditions. However, SCONC down-regulated transformants were unable to sustain yeast growth using inorganic sulfur compounds only. Moreover, pulses with inorganic sulfur in SCONC down-regulated transformants triggered an increase of the inorganic sulfur metabolism, which culminated in a drastic reduction of the ATP and NADPH cellular levels and in higher oxidative stress. Importantly, the down-regulation of SCONC resulted in a decreased virulence of P. brasiliensis, as validated in an in vivo model of infection. Overall, our findings shed light on the inability of P. brasiliensis yeast to rely on inorganic sulfur compounds, correlating its metabolism with cellular energy and redox imbalances. Furthermore, the data herein presented reveal SconCp as a novel virulence determinant of P. brasiliensis. PMID:24066151

Effects of physical and nutritional stress conditions during mycelial growth on conidial germination speed, adhesion to host cuticle, and virulence of Metarhizium anisopliae, an entomopathogenic fungus.

PubMed

Rangel, Drauzio E N; Alston, Diane G; Roberts, Donald W

2008-11-01

Growth under stress may influence pathogen virulence and other phenotypic traits. Conidia of the entomopathogenic fungus Metarhizium anisopliae var. anisopliae (isolate ARSEF 2575) were produced under different stress conditions and then examined for influences on in vitro conidial germination speed, adhesion to the insect cuticle, and virulence to an insect host, Tenebrio molitor. Conidia were produced under non-stress conditions [on potato-dextrose agar plus 1gl(-1) yeast extract (PDAY; control)], or under the following stress conditions: osmotic (PDAY+sodium chloride or potassium chloride, 0.6 or 0.8m); oxidative [(PDAY+hydrogen peroxide, 5mm) or UV-A (irradiation of mycelium on PDAY)]; heat shock (heat treatment of mycelium on PDAY at 45 degrees C, 40min); and nutritive [minimal medium (MM) with no carbon source, or on MM plus 3gl(-1) lactose (MML)]. Conidia were most virulent (based on mortality at 3d) and had the fastest germination rates when produced on MML, followed by MM. In addition, conidial adhesion to host cuticle was greatest when the conidia were produced on MML. Media with high osmolarity (0.8m) produced conidia with slightly elevated virulence and faster germination rates than conidia produced on the control medium (PDAY), but this trend did not hold for media with the lower osmolarity, (0.6m). Conidia produced from mycelium irradiated with UV-A while growing on PDAY had somewhat elevated virulence levels similar to that of conidia produced on MM, but their germination rate was not increased. Hydrogen peroxide and heat shock treatments did not alter virulence. These results demonstrate that the germination, adhesion and virulence of M. anisopliae conidia can be strongly influenced by culture conditions (including stresses) during production of the conidia.

Circulating metabolite predictors of glycemia in middle-aged men and women.

PubMed

Würtz, Peter; Tiainen, Mika; Mäkinen, Ville-Petteri; Kangas, Antti J; Soininen, Pasi; Saltevo, Juha; Keinänen-Kiukaanniemi, Sirkka; Mäntyselkä, Pekka; Lehtimäki, Terho; Laakso, Markku; Jula, Antti; Kähönen, Mika; Vanhala, Mauno; Ala-Korpela, Mika

2012-08-01

Metabolite predictors of deteriorating glucose tolerance may elucidate the pathogenesis of type 2 diabetes. We investigated associations of circulating metabolites from high-throughput profiling with fasting and postload glycemia cross-sectionally and prospectively on the population level. Oral glucose tolerance was assessed in two Finnish, population-based studies consisting of 1,873 individuals (mean age 52 years, 58% women) and reexamined after 6.5 years for 618 individuals in one of the cohorts. Metabolites were quantified by nuclear magnetic resonance spectroscopy from fasting serum samples. Associations were studied by linear regression models adjusted for established risk factors. Nineteen circulating metabolites, including amino acids, gluconeogenic substrates, and fatty acid measures, were cross-sectionally associated with fasting and/or postload glucose (P < 0.001). Among these metabolic intermediates, branched-chain amino acids, phenylalanine, and α1-acid glycoprotein were predictors of both fasting and 2-h glucose at 6.5-year follow-up (P < 0.05), whereas alanine, lactate, pyruvate, and tyrosine were uniquely associated with 6.5-year postload glucose (P = 0.003-0.04). None of the fatty acid measures were prospectively associated with glycemia. Changes in fatty acid concentrations were associated with changes in fasting and postload glycemia during follow-up; however, changes in branched-chain amino acids did not follow glucose dynamics, and gluconeogenic substrates only paralleled changes in fasting glucose. Alterations in branched-chain and aromatic amino acid metabolism precede hyperglycemia in the general population. Further, alanine, lactate, and pyruvate were predictive of postchallenge glucose exclusively. These gluconeogenic precursors are potential markers of long-term impaired insulin sensitivity that may relate to attenuated glucose tolerance later in life.

Polymer thermal optical switch for a flexible photonic circuit.

PubMed

Sun, Yue; Cao, Yue; Wang, Qi; Yi, Yunji; Sun, Xiaoqiang; Wu, Yuanda; Wang, Fei; Zhang, Daming

2018-01-01

Flexible and wearable optoelectronic devices are the new trend for an active lifestyle. These devices are polymer-based for flexibility. We demonstrated flexible polymer waveguide optical switches for a flexible photonic integrated circuit. The optical switches are composed of a single-mode inverted waveguide with dimensions of 5 μm waveguide width, 3 μm ridge height, and 3 μm slab height. A Mach-Zehnder structure was used in the device, with the Y-branch horizontal length of 0.1 cm, the distance between two heating branches of 30 μm, and the heating branch length of 1 cm. The optical field of the device was simulated by beam propagation to optimize the electrode position. The switching properties of the flexible optical switch with different working conditions, such as contact to the polymer, silicon, and skin, were simulated. The device was prepared based on the photo curved polymer and lithography method. The end faces of the flexible film device were processed using an excimer laser with optimized parameters of 28  mJ/cm 2 and 15 Hz. The response rise time and fall time on the PMMA substrate were measured as 1.98 ms and 2.71 ms, respectively. The power consumption was 16 mW and the extinction ratio was 11 dB. The response rise and fall times on the Si substrate were measured as 1.08 ms and 1.62 ms, respectively. The power consumption was 17 mW and the extinction ratio was 11 dB. The demonstrated properties indicate that this flexible optical waveguide structure can be used in the light control area of a wearable device.

Characterization of a New β(1–3)-Glucan Branching Activity of Aspergillus fumigatus

PubMed Central

Gastebois, Amandine; Mouyna, Isabelle; Simenel, Catherine; Clavaud, Cécile; Coddeville, Bernadette; Delepierre, Muriel; Latgé, Jean-Paul; Fontaine, Thierry

2010-01-01

A new HPLC method was developed to separate linear from β(1–6)-branched β(1–3)-glucooligosaccharides. This methodology has permitted the isolation of the first fungal β(1–6)/β(1–3)-glucan branching transglycosidase using a cell wall autolysate of Aspergillus fumigatus (Af). The encoding gene, AfBGT2 is an ortholog of AfBGT1, another transglycosidase of A. fumigatus previously analyzed (Mouyna, I., Hartland, R. P., Fontaine, T., Diaquin, M., Simenel, C., Delepierre, M., Henrissat, B., and Latgé, J. P. (1998) Microbiology 144, 3171–3180). Both enzymes release laminaribiose from the reducing end of a β(1–3)-linked oligosaccharide and transfer the remaining chain to another molecule of the original substrate. The AfBgt1p transfer occurs at C-6 of the non-reducing end group of the acceptor, creating a kinked β(1–3;1–6) linear molecule. The AfBgt2p transfer takes place at the C-6 of an internal group of the acceptor, resulting in a β(1–3)-linked product with a β(1–6)-linked side branch. The single Afbgt2 mutant and the double Afbgt1/Afbgt2 mutant in A. fumigatus did not display any cell wall phenotype showing that these activities were not responsible for the construction of the branched β(1–3)-glucans of the cell wall. PMID:19948732

Existence of bound states of a polaron with a breather in soft potentials

NASA Astrophysics Data System (ADS)

Cuevas, J.; Kevrekidis, P. G.; Frantzeskakis, D. J.; Bishop, A. R.

2006-08-01

We consider polarons in models of coupled electronic and vibrational degrees of freedom, in the presence of a soft nonlinear substrate potential (Morse potential). In particular, we focus on a bound state of a polaron with a breather, a so-called “polarobreather.” We analyze the existence of these states based on frequency resonance conditions and illustrate their stability using Floquet spectrum techniques. Multisite solutions of this type are also obtained both in the stationary case (bond-centered and twisted polarons) and in the breathing case (bond-centered and twisted polarobreathers). For all the branches examined, the dynamical evolution of instabilities pertinent to the corresponding solutions are also briefly discussed. Finally, a different branch of so-called phantom polarobreathers is also demonstrated.

DIFFERENTIAL ALLERGIC AND NEUROTROPHIN RESPONSES TO FUNGAL COMPONENT EXTRACTS IN BALB/C MICE

EPA Science Inventory

Metarhizium anisopliae mycelium (MYC), conidia (CON) and inducible protease (IND) extracts were combined to produce the antigen MACA to screen for allergenic potential. Involuntary aspiration (IA) exposure to MACA in BALB/c mice has caused immune, inflammatory and physiological ...

Packaging produced from natural fibers and mycelium: Optimizing biomass blends to meet performance specifications

USDA-ARS?s Scientific Manuscript database

Polystyrene is non-biodegradable and has environmental issues associated with its disposal. A replacement for polystyrene that is biodegradable, environmentally friendly, and has similar performance characteristics is of interest to many ecologically minded companies and consumers. This study utiliz...

Degradation and mineralization of DDT by the ectomycorrhizal fungi, Xerocomus chrysenteron.

PubMed

Huang, Yi; Wang, Jie

2013-08-01

One strain of ectomycorrhizal fungi, Xerocomus chrysenteron, had been investigated for its ability to degrade 1,1,1-trichloro-2,2-bis(4-chlorophe-nyl) ethane (DDT) by measuring unlabeled DDT and identifying its metabolites, and determining the mineralization of [(13)C]DDT in pure cultures. After 45d incubation, about 55% of the added DDT disappeared from the culture system, less than 5% remained in the nutrient solution, and about 44% was retained in the mycelium. Inoculation with mycelium enhanced the degradation of DDT in soil, and alleviated enrichment of DDT in plants. The metabolites identified by gas chromatography-mass spectrometry were 1,1-dichloro-2,2-bis(4-chlorophenyl) ethane (DDD), 1,1-dichloro-2,2-bis(4-chlorophenyl)ethylene (DDE), and 4,4'-dichlorobenzophenone (DBP). There were significant differences in the δ(13)C of released CO2 between [(13)C]DDT and DDT cultures, which indicated X. chrysenteron was able to mineralize DDT to CO2. Copyright © 2013 Elsevier Ltd. All rights reserved.

Biomineralization of gold by Mucor plumbeus: The progress in understanding the mechanism of nanoparticles' formation.

PubMed

Maliszewska, Irena; Tylus, Włodzimierz; Chęcmanowski, Jacek; Szczygieł, Bogdan; Pawlaczyk-Graja, Izabela; Pusz, Wojciech; Baturo-Cieśniewska, Anna

2017-09-01

This contribution describes the deposition of gold nanoparticles by microbial reduction of Au(III) ions using the mycelium of Mucor plumbeus. Biosorption as the major mechanism of Au(III) ions binding by the fungal cells and the reduction of them to the form of Au(0) on/in the cell wall, followed by the transportation of the synthesized gold nanoparticles to the cytoplasm, is postulated. The probable mechanism behind the reduction of Au(III) ions is discussed, leading to the conclusion that this process is nonenzymatic one. Chitosan of the fungal cell wall is most likely to be the major molecule involved in biomineralization of gold by the mycelium of M. plumbeus. Separation of gold nanoparticles from the cells has been carried out by the ultrasonic disintegration and the obtained nanostructures were characterized by UV-vis spectroscopy and transmission electron micrograph analysis. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:1381-1392, 2017. © 2017 American Institute of Chemical Engineers.

Mutation-Screening of Pleurotus Ferulae with High Temperature Tolerance by Nitrogen Ion Implantation

NASA Astrophysics Data System (ADS)

Chen, Henglei; Wan, Honggui; Zhang, Jun; Zeng, Xianxian

2008-08-01

In order to obtain Pleurotus ferulae with high temperature tolerance, conidiophores of wild type strain ACK were implanted with nitrogen ions in energy of 5 ~15 keV and dose of 1.5 × 1015 ~ 1.5 × 1016 cm-2, and a mutant CGMCC1763 was isolated subsequently through thermotolerant screening method. It was found that during riper period the surface layer mycelium of the mutant in mushroom bag wasn't aging neither grew tegument even above 30° C. The mycelium endurable temperature of the mutant was increased by 5°C compared to that of the wild type strain. The fruiting bodies growth temperature of the mutant was 18 ~22°C in daytime and 8~14°C at night. The highest growth temperature of fruiting bodies of the mutant was increased about 7°C w.r.t. that of original strain. Through three generations investigations, it was found that the mutant CGMCC1763 was stable with high temperature tolerance.

Development of nutraceutical formulations based on the mycelium of Pleurotus ostreatus and Agaricus bisporus.

PubMed

Cardoso, Rossana V C; Fernandes, Ângela; Oliveira, M Beatriz P P; Calhelha, Ricardo C; Barros, Lillian; Martins, Anabela; Ferreira, Isabel C F R

2017-06-21

The present work is aimed at developing nutraceutical formulations based on the mycelium of Agaricus bisporus and Pleurotus ostreatus, highlighting the potential of in vitro culture as a tool to improve the production of bioactive compounds, namely phenolic acids and ergosterol. The mycelia of both species were cultured in different solid and liquid media in order to compare the growth rate and yielded biomass. Fruiting bodies, mycelia and culture media were compared regarding the antioxidant activity, anti-inflammatory effects in RAW264.7 cells and cytotoxicity in human tumor cell lines and non-tumor porcine liver cells. P. ostreatus mycelia showed higher contents of ergosterol and phenolic compounds, and stronger antioxidant activity than the corresponding fruiting body. P. ostreatus and A. bisporus did not show anti-inflammatory activity, and P. ostreatus was the only one showing cytotoxicity in tumor cell lines. The results show that these mushrooms provide compounds with antioxidant and cytotoxic capacities, with variations among species.

Production of Mushroom Mycelium as a Protein and Fat Source in Submerged Culture in Medium of Vinasse

PubMed Central

Falanghe, H.

1962-01-01

Of ten mushroom cultures investigated, only Agaricus campestris, Boletus indecisus, and Tricholoma nudum were capable of growing in submerged culture in medium of vinasse with added salts. Higher fermentative efficiencies were found under these conditions than in medium containing molasses or waste sulfite liquor. A. campestris showed a better capacity to produce protein but, since B. indecisus is capable of developing greater mycelium weight, its fermentative efficiencies are comparable. Both microorganisms could be grown in medium of vinasse with greatly varied amounts, producing higher mycelial weight in media with greater vinasse. The capacity of B. indecisus and A. campestris to utilize the noncarbohydrate fraction in total solids, instead of the total carbohydrates when they are in smaller amount, was observed in medium containing vinasse. B. indecisus and A. campestris were easily separated by filtration from the medium, although T. nudum was difficult to separate by this procedure. In experiments with A. campestris, the adaptative capacity of the organism to vinasse was demonstrated. PMID:13962715

A toolbox to measure changes in the cell wall glycopolymer composition during differentiation of Streptomyces coelicolor A3(2).

PubMed

Sigle, Steffen; Steblau, Nadja; Wohlleben, Wolfgang; Muth, Günther

2016-09-01

Cell wall glycopolymers (CWG) represent an important component of the Gram-positive cell envelope with many biological functions. The mycelial soil bacterium Streptomyces coelicolor A3(2) incorporates two distinct CWGs, polydiglycosylphosphate (PDP) and teichulosonic acid, into the cell wall of its vegetative mycelium but only little is known about their role in the complex life cycle of this microorganism. In this study we established assays to measure the total amount of CWGs in mycelial cell walls and spore walls, to quantify the individual CWGs and to determine the length of PDP. By applying these assays, we discovered that the relative amount of CWGs, especially of PDP, is reduced in spores compared to vegetative mycelium. Furthermore we found that PDP extracted from mycelial cell walls consisted of at least 19 repeating units, whereas spore walls contained substantially longer PDP polymers. Copyright © 2016 Elsevier B.V. All rights reserved.

Degradation and decolorization of monosodium glutamate wastewater with Coriolus versicolor.

PubMed

Jia, Cuiying; Kang, Ruijuan; Zhang, Yuhui; Zhang, Yong; Cong, Wei

2007-10-01

Degradation and decolorization of monosodium glutamate wastewater (MSGW) with Coriolus versicolor were firstly carried out. The effects of various operation parameters namely wastewater concentrations, pH, culture time and incidence of sterilization on maximum percentage of degradation and decolorization of wastewater were investigated. Studies of mycelium and enzyme for C. versicolor degradation and decolorization were estimated in this study. Ten percentage of wastewater concentration and pH = 5.0 were found to be the most suitable ones among the other experiments. The highest degradation and decolorization efficiency of wastewater was obtained at the fifth day of cultivation, which was displayed with more than 70% chemical oxygen demand removal, 83% total sugar removal and 55% color removal, respectively. Sterile operation had no remarkable effect on the degradation and decolorization efficiency for C. versicolor. Mycelium and the extra cellular fungal enzyme were both necessary for the degradation and decolorization of MSGW. C. versicolor possesses great potential and economic advantages in MSGW treatment.

Structural data and immunomodulatory properties of a water-soluble heteroglycan extracted from the mycelium of an Italian isolate of Ganoderma lucidum.

PubMed

Carrieri, Raffaele; Manco, Rosanna; Sapio, Daniela; Iannaccone, Marco; Fulgione, Andrea; Papaianni, Marina; de Falco, Bruna; Grauso, Laura; Tarantino, Paola; Ianniello, Flora; Lanzotti, Virginia; Lahoz, Ernesto; Capparelli, Rosanna

2017-09-01

Mushrooms produce a wide range of bioactive polysaccharides, different from each other in chemical structure and biological effects. In the last years, the idea to develop functional foods or drugs containing fungal polysaccharides is attracting great attention. Fruiting bodies of Basidiomycetes Ganoderma lucidum are commonly used in Oriental medicine to treat several disorders. G. lucidum polysaccharides - mainly β-glucans and heteroglycans - have numerous biological properties such as antitumour and immunomodulatory activities. This report shows, by gene expression analyses and bioenergetic assays, immunomodulatory properties and capacity to improve glucose metabolism of a water-soluble heteroglycan extracted from mycelium of an Italian isolate of G. lucidum. The findings suggest the use of the heteroglycan as probiotic or ingredient in functional foods, being easy to produce and disperse in a food matrix thanks to its water-solubility. Heteroglycan could exert protective effects in pro-inflammatory conditions and benefits for people characterised by suppressed immune response.

Selection of Lecanicillium Strain with High Virulence against Developmental Stages of Bemisia tabaci

PubMed Central

Park, Heeyong

2010-01-01

Selection of fungal strains with high virulence against the developmental stages of Bemisia tabaci was performed using internal transcribed spacer regions. The growth rate of hyphae was measured and bioassay of each developmental stage of B. tabaci was conducted for seven days. All of the fungal strains tested were identified as Lecanicillium spp., with strain 4078 showing the fastest mycelium growth rate (colony diameter, 16.3 ± 0.9 mm) among the strains. Compared to strain 4075, which showed the slowest growth rate, the growth rate of strain 4078 was increased almost 2-fold after seven days. Strains 4078 and Btab01 were most virulent against the egg and larva stages, respectively. The virulence of fungal strains against the adult stage was high, except for strains 41185 and 3387. Based on the growth rate of mycelium and level of virulence, strains 4078 and Btab01 were selected as the best fungal strains for application to B. tabaci, regardless of developmental stage. PMID:23956657

137Cs in the fungal compartment of Swedish forest soils.

PubMed

Vinichuk, Mykhaylo M; Johanson, Karl J; Taylor, Andy F S

2004-05-05

The (137)Cs activities in soil profiles and in the mycelia of four ectomycorrhizal fungi were studied in a Swedish forest in an attempt to understand the mechanisms governing the transfer and retention of (137)Cs in forest soil. The biomass of four species of fungi was determined and estimated to be 16 g m(-2) in a peat soil and 47-189 g m(-2) in non-peat soil to the depth of 10 cm. The vertical distribution was rather homogeneous for two species (Tylospora spp. and Piloderma fallax) and very superficial for Hydnellum peckii. Most of the (137)Cs activity in mycelium of non-peat soils was found in the upper 5 cm. Transfer factors were quite high even for those species producing resupinate sporocarps. In the peat soil only approximately 0.3% of the total (137)Cs inventory in soil was found in the fungal mycelium. The corresponding values for non-peat soil were 1.3, 1.8 and 1.9%.

Influence of 2,4,6-trinitrotoluene (TNT) concentration on the degradation of TNT in explosive-contaminated soils by the white rot fungus Phanerochaete chrysosporium.

PubMed Central

Spiker, J K; Crawford, D L; Crawford, R L

1992-01-01

The ability of Phanerochaete chrysosporium to bioremediate TNT (2,4,6-trinitrotoluene) in a soil containing 12,000 ppm of TNT and the explosives RDX (hexahydro-1,3,5-trinitro-1,3,5- triazine; 3,000 ppm) and HMX (octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine; 300 ppm) was investigated. The fungus did not grow in malt extract broth containing more than 0.02% (wt/vol; 24 ppm of TNT) soil. Pure TNT or explosives extracted from the soil were degraded by P. chrysosporium spore-inoculated cultures at TNT concentrations of up to 20 ppm. Mycelium-inoculated cultures degraded 100 ppm of TNT, but further growth was inhibited above 20 ppm. In malt extract broth, spore-inoculated cultures mineralized 10% of added [14C]TNT (5 ppm) in 27 days at 37 degrees C. No mineralization occurred during [14C]TNT biotransformation by mycelium-inoculated cultures, although the TNT was transformed. PMID:1444437

Effect of water stress on in vitro mycelium cultures of two mycorrhizal desert truffles.

PubMed

Navarro-Ródenas, Alfonso; Lozano-Carrillo, M Cecilia; Pérez-Gilabert, Manuela; Morte, Asunción

2011-05-01

The ability of two species of desert truffle, Terfezia claveryi strain TcS2 and Picoa lefebvrei strain OL2, to tolerate water stress in pure culture has been investigated. Both T. claveryi and P. lefebvrei strains exhibited a mycelium growth pattern characteristic of drought tolerant species. However, they were only tolerant to moderate water stress, below -1.07 MPa, with the P. lefebvrei isolate being slightly more drought tolerant than the T. claveryi isolate. The increased alkaline phosphatase (ALP) activity observed in both fungi at moderate water stress with respect to the control indicated the functional adaptation of these mycelia to these drought conditions. ALP activity can be used as an indicator of the metabolic activity of these fungi. Slight water stress (-0.45 MPa) could improve mycelial inoculum production of these desert truffles. Moreover, P. lefebvrei could be a good candidate for further desert truffle mycorrhizal plant cultivation programmes in semiarid Mediterranean areas.

JPRS Report Science & Technology Japan

DTIC Science & Technology

1989-10-20

plant callus o Adaptation of protozoans to microgravity o Fertilization and embryogeny of newt in space o Fertilization and embryogeny of sea urchin ...for by the mining and manufacturing branch. Moreover, about 50 percent of the energy consumed is exhausted to air, river, sea , etc., as waste heat...Cross section of substrate Figure 6. Photo of Cross Section of Coating Film Taken by Scanning Electron Microscope ( SEM ) It was heated for 60 hours

Oviposition traps to survey eggs of Lambdina fiscellaria (Lepidoptera: Geometridae).

PubMed

Hébert, Christian; Jobin, Luc; Auger, Michel; Dupont, Alain

2003-06-01

Outbreaks of the hemlock looper, Lambdina fiscellaria (Gueneé), are characterized by rapid increase and patchy distribution over widespread areas, which make it difficult to detect impending outbreaks. This is a major problem with this insect. Population forecasting is based on tedious and expensive egg surveys in which eggs are extracted from 1-m branches; careful observation is needed to avoid counting old unhatched eggs of previous year populations. The efficacy of artificial substrates as oviposition traps to sample hemlock looper eggs was tested as a means of improving outbreak detection and population forecasting. A white polyurethane foam substrate (1,095 lb/ft3) used with the Luminoc insect trap, a portable light trap, was highly efficient in sampling eggs of the hemlock looper. Foam strips placed on tree trunks at breast height were less efficient but easier and less expensive to use for the establishment of extensive survey networks. Estimates based on oviposition traps were highly correlated with those obtained from the 1-m branch extraction method. The oviposition trap is a standard, inexpensive, easy, and robust method that can be used by nonspecialists. This technique makes it possible to sample higher numbers of plots in widespread monitoring networks, which is crucial for improving the management of hemlock looper populations.

The Dual Activity Responsible for the Elongation and Branching of β-(1,3)-Glucan in the Fungal Cell Wall.

PubMed

Aimanianda, Vishukumar; Simenel, Catherine; Garnaud, Cecile; Clavaud, Cecile; Tada, Rui; Barbin, Lise; Mouyna, Isabelle; Heddergott, Christoph; Popolo, Laura; Ohya, Yoshikazu; Delepierre, Muriel; Latge, Jean-Paul

2017-06-20

β-(1,3)-Glucan, the major fungal cell wall component, ramifies through β-(1,6)-glycosidic linkages, which facilitates its binding with other cell wall components contributing to proper cell wall assembly. Using Saccharomyces cerevisiae as a model, we developed a protocol to quantify β-(1,6)-branching on β-(1,3)-glucan. Permeabilized S. cerevisiae and radiolabeled substrate UDP-( 14 C)glucose allowed us to determine branching kinetics. A screening aimed at identifying deletion mutants with reduced branching among them revealed only two, the bgl2 Δ and gas1 Δ mutants, showing 15% and 70% reductions in the branching, respectively, compared to the wild-type strain. Interestingly, a recombinant Gas1p introduced β-(1,6)-branching on the β-(1,3)-oligomers following its β-(1,3)-elongase activity. Sequential elongation and branching activity of Gas1p occurred on linear β-(1,3)-oligomers as well as Bgl2p-catalyzed products [short β-(1,3)-oligomers linked by a linear β-(1,6)-linkage]. The double S. cerevisiae gas1 Δ bgl2 Δ mutant showed a drastically sick phenotype. An Sc Gas1p ortholog, Gel4p from Aspergillus fumigatus , also showed dual β-(1,3)-glucan elongating and branching activity. Both Sc Gas1p and A. fumigatus Gel4p sequences are endowed with a carbohydrate binding module (CBM), CBM43, which was required for the dual β-(1,3)-glucan elongating and branching activity. Our report unravels the β-(1,3)-glucan branching mechanism, a phenomenon occurring during construction of the cell wall which is essential for fungal life. IMPORTANCE The fungal cell wall is essential for growth, morphogenesis, protection, and survival. In spite of being essential, cell wall biogenesis, especially the core β-(1,3)-glucan ramification, is poorly understood; the ramified β-(1,3)-glucan interconnects other cell wall components. Once linear β-(1,3)-glucan is synthesized by plasma membrane-bound glucan synthase, the subsequent event is its branching event in the cell wall space. Using Saccharomyces cerevisiae as a model, we identified GH72 and GH17 family glycosyltransferases, Gas1p and Bgl2p, respectively, involved in the β-(1,3)-glucan branching. The sick phenotype of the double Scgas1 Δ bgl2 Δ mutant suggested that β-(1,3)-glucan branching is essential. In addition to Sc Gas1p, GH72 family Sc Gas2p and Aspergillus fumigatus Gel4p, having CBM43 in their sequences, showed dual β-(1,3)-glucan elongating and branching activity. Our report identifies the fungal cell wall β-(1,3)-glucan branching mechanism. The essentiality of β-(1,3)-glucan branching suggests that enzymes involved in the glucan branching could be exploited as antifungal targets. Copyright © 2017 Aimanianda et al.

Expanding the chemical diversity of natural esters by engineering a polyketide-derived pathway into Escherichia coli.

PubMed

Menendez-Bravo, Simón; Comba, Santiago; Sabatini, Martín; Arabolaza, Ana; Gramajo, Hugo

2014-07-01

Microbial fatty acid (FA)-derived molecules have emerged as promising alternatives to petroleum-based chemicals for reducing dependence on fossil hydrocarbons. However, native FA biosynthetic pathways often yield limited structural diversity, and therefore restricted physicochemical properties, of the end products by providing only a limited variety of usually linear hydrocarbons. Here we have engineered into Escherichia coli a mycocerosic polyketide synthase-based biosynthetic pathway from Mycobacterium tuberculosis and redefined its biological role towards the production of multi-methyl-branched-esters (MBEs) with novel chemical structures. Expression of FadD28, Mas and PapA5 enzymes enabled the biosynthesis of multi-methyl-branched-FA and their further esterification to an alcohol. The high substrate tolerance of these enzymes towards different FA and alcohol moieties resulted in the biosynthesis of a broad range of MBE. Further metabolic engineering of the MBE producer strain coupled this system to long-chain-alcohol biosynthetic pathways resulting in de novo production of branched wax esters following addition of only propionate. Copyright © 2014 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

Tobacco leaf spot and root rot caused by Rhizoctonia solani Kühn.

PubMed

Gonzalez, Marleny; Pujol, Merardo; Metraux, Jean-Pierre; Gonzalez-Garcia, Vicente; Bolton, Melvin D; Borrás-Hidalgo, Orlando

2011-04-01

Rhizoctonia solani Kühn is a soil-borne fungal pathogen that causes disease in a wide range of plants worldwide. Strains of the fungus are traditionally grouped into genetically isolated anastomosis groups (AGs) based on hyphal anastomosis reactions. This article summarizes aspects related to the infection process, colonization of the host and molecular mechanisms employed by tobacco plants in resistance against R. solani diseases. Teleomorph: Thanatephorus cucumeris (Frank) Donk; anamorph: Rhizoctonia solani Kühn; Kingdom Fungi; Phylum Basidiomycota; Class Agaricomycetes; Order Cantharellales; Family Ceratobasidiaceae; genus Thanatephorus. Somatic hyphae in culture and hyphae colonizing a substrate or host are first hyaline, then buff to dark brown in colour when aging. Hyphae tend to form at right angles at branching points that are usually constricted. Cells lack clamp connections, but possess a complex dolipore septum with continuous parenthesomes and are multinucleate. Hyphae are variable in size, ranging from 3 to 17 µm in diameter. Although the fungus does not produce any conidial structure, ellipsoid to globose, barrel-shaped cells, named monilioid cells, 10-20 µm wide, can be produced in chains and can give rise to sclerotia. Sclerotia are irregularly shaped, up to 8-10 mm in diameter and light to dark brown in colour. Symptoms in tobacco depend on AG as well as on the tissue being colonized. Rhizoctonia solani AG-2-2 and AG-3 infect tobacco seedlings and cause damping off and stem rot. Rhizoctonia solani AG-3 causes 'sore shin' and 'target spot' in mature tobacco plants. In general, water-soaked lesions start on leaves and extend up the stem. Stem lesions vary in colour from brown to black. During late stages, diseased leaves are easily separated from the plant because of severe wilting. In seed beds, disease areas are typically in the form of circular to irregular patches of poorly growing, yellowish and/or stunted seedlings. Knowledge is scarce regarding the mechanisms associated with resistance to R. solani in tobacco. However, recent evidence suggests a complex response that involves several constitutive factors, as well as induced barriers controlled by multiple defence pathways. This fungus can survive for many years in soil as mycelium, and also by producing sclerotia, which makes the management of the disease using conventional means very difficult. Integrated pest management has been most successful; it includes timely fungicide applications, crop rotation and attention to soil moisture levels. Recent developments in biocontrol may provide other tools to control R. solani in tobacco. © 2010 CIGB. MOLECULAR PLANT PATHOLOGY © 2010 BSPP AND BLACKWELL PUBLISHING LTD.

Optimizing biomass blends for manufacturing molded packaging materials using mycelium

USDA-ARS?s Scientific Manuscript database

Polystyrene is one of the most widely used plastics and is commonly produced in three forms: 1) Extruded polystyrene – disposable utensils, CD/DVD cases, yogurt containers, smoke alarm housing, etc.; 2) Expanded polystyrene foam – molded packaging materials and packaging "peanuts"; 3) Extruded polys...

Fungal mycelia in soils - a new method for quantification of their biomass

NASA Astrophysics Data System (ADS)

Drabløs Eldhuset, Toril; Lange, Holger; Svetlik, Jan; Børja, Isabella

2013-04-01

All plant-bearing soils are interwoven with fungal hyphae. Their structure and function are affected by environmental factors like drought, which might be a stress factor of increasing importance in many world regions due to climate change. The fungal mycelium in soil is important both for mycorrhizal symbiosis with plant roots and for litter decomposition, and thereby also for carbon turnover in soils. However, the mycelium biomass has been difficult to assess. Here we describe a simple and feasible method to quantify the biomass of fungal mycelium. We report on a manipulation study in the field where drought stress has been induced. The experiment was performed in a Norway spruce (Picea abies) 20 years old stand planted on former agricultural land, with a control plot and a roofed plot where precipitation was excluded. To investigate the fungal mycelium, nylon nets (mesh size 1 mm, width 7 cm and length 25 cm), were inserted vertically into the soil down to 20 cm depth. The nets were left in the soil from October to June, removed and replaced by new nets that were left in the soil from June to October. After removal, by cutting a block of soil around each net, the nets were cleaned from residual soil and scanned using the image scanner CanoScan 9000F. The resulting images were analyzed using the image processing software ImageJ. The image analysis was based on the distribution of grey values in the individual pixels which characterize the different components in the image (voids, hyphae, the nylon net, and soil). Based on the repeated visual evaluation of hyphal coverage in the net segments, we obtained an exponential equation allowing us to determine automatically the coverage of net windows by hyphae in percentage for each net scanned. In this way we can compare the hyphal coverage in the control and the drought-exposed plots. Based on the hyphal coverage scans together with hyphal dry weight on clean nets, we account for the soil particles adhering to the nets. Using this analysis method, the hyphal mat coverage in mm2 on any net is quantified and the hyphal biomass on the net can be calculated and compared between treatments. Also, the hyphal biomass per cm3 soil at the spot where the net has been inserted can be assessed. In addition, DNA from net-bound hyphae may be extracted to determine the identity of fungal species at different soil depths for the individual treatments.

Analysis of some metallic elements and metalloids composition and relationships in parasol mushroom Macrolepiota procera.

PubMed

Falandysz, Jerzy; Sapkota, Atindra; Dryżałowska, Anna; Mędyk, Małgorzata; Feng, Xinbin

2017-06-01

The aim of the study was to characterise the multi-elemental composition and associations between a group of 32 elements and 16 rare earth elements collected by mycelium from growing substrates and accumulated in fruiting bodies of Macrolepiota procera from 16 sites from the lowland areas of Poland. The elements were quantified by inductively coupled plasma quadrupole mass spectrometry using validated method. The correlation matrix obtained from a possible 48 × 16 data matrix has been used to examine if any association exits between 48 elements in mushrooms foraged from 16 sampling localizations by multivariate approach using principal component (PC) analysis. The model could explain up to 93% variability by eight factors for which an eigenvalue value was ≥1. Absolute values of the correlation coefficient were above 0.72 (significance at p < 0.05) for 43 elements. From a point of view by consumer, the absolute content of Cd, Hg, Pb in caps of M. procera collected from background (unpolluted) areas could be considered elevated while sporadic/occasional ingestion of this mushroom is considered safe. The multivariate functional analysis revealed on associated accumulation of many elements in this mushroom. M. procera seem to possess some features of a bio-indicative species for anthropogenic Pb but also for some geogenic metals.

Differential response of oyster shell powder on enzyme profile and nutritional value of oyster mushroom Pleurotus florida PF05.

PubMed

Naraian, Ram; Narayan, Om Prakash; Srivastava, Jatin

2014-01-01

Oyster mushroom Pleurotus florida was cultivated on different combinations of wheat straw (WS) as basal substrate and oyster shell powder (OSP) supplement. The OSP supplementation considerably responded to different cultivation phases. The mycelium grew fast and showed rapid growth rate (8.91 mmd(-1)) in WS + OSP (97 + 3) combination while WS + OSP (92 + 8) showed maximum laccase (3.133 U/g) and Mn peroxidase (MnP) activities (0.091 U/g). The climax level of laccase (5.433 U/g) and MnP (0.097 U/g) was recorded during fruit body initiation in WS + OSP (97 + 3) and WS + OSP (98 + 2) combinations, respectively. The WS + OSP (97 + 3) combination represented the best condition for mushroom cultivation and produced the highest biological efficiency (147%). In addition, protein and lipid contents in fruit bodies were slightly improved in response to OSP. The carbohydrate was significantly increased by raising concentration of OSP. The highest values of protein, carbohydrate, and lipid noted were 31.3 μg/g, 0.0639 (g/g), and 0.373 (g/g) correspondingly. Conclusively it was evident that lower concentrations of OSP acted positively and relatively to higher concentrations and improved nutritional content which may suitably be used to enhance both yield and nutritional values of mushroom.

Boric Acid Inhibition of Trichophyton rubrum Growth and Conidia Formation.

PubMed

Schmidt, Martin

2017-12-01

Trichophyton rubrum is a common human dermatophyte that is the causative agent of 80-93% of fungal infections of the skin and nails. While dermatophyte infections in healthy people are easily treatable with over-the-counter medications, such infections pose a higher risk for patients with compromised immune function and impaired regenerative potential. The efficacy of boric acid (BA) for the treatment of vaginal yeast infections prompted an investigation of the effect of BA on growth and morphology of T. rubrum. This is of particular interest since BA facilitates wound healing, raising the possibility that treating athlete's foot with BA, either alone or in combination with other antifungal drugs, would combine the benefits of antimicrobial activity and tissue regeneration to accelerate healing of infected skin. The data presented here show that BA represses T. rubrum growth at a concentration reported to be beneficial for host tissue regeneration. Oxygen exposure increases BA toxicity, and mycelia growing under BA stress avoid colonizing the surface of the growth surface, which leads to a suppression of aerial mycelium growth and surface conidia formation. BA penetrates into solid agar matrices, but the relative lack of oxygen below the substrate surface limits the effectiveness of BA in suppressing growth of embedded T. rubrum cells.

Actinomycetes benefaction role in soil and plant health.

PubMed

Bhatti, Asma Absar; Haq, Shamsul; Bhat, Rouf Ahmad

2017-10-01

Actinomycetes are aerobic, spore forming gram-positive bacteria, belonging to the order actinomycetales characterized with substrate and aerial mycelium growth. They are the most abundant organisms that form thread-like filaments in the soil and are responsible for characteristically "earthy" smell of freshly turned healthy soil. They play major roles in the cycling of organic matter; inhibit the growth of several plant pathogens in the rhizosphere and decompose complex mixtures of polymer in dead plant, animal and fungal material results in production of many extracellular enzymes which are conductive to crop production. The major contribution in biological buffering of soils, biological control of soil environments by nitrogen fixation and degradation of high molecular weight compounds like hydrocarbons in the polluted soils are remarkable characteristics of actinomycetes. Besides this, they are known to improve the availability of nutrients, minerals, enhance the production of metabolites and promote plant growth regulators. Furthermore, actinobacteria do not contaminate the environment instead, they help sustainably in improving soil health by formation and stabilization of compost piles, formation of stable humus and combine with other soil microorganisms in breaking down the tough plant residues such as cellulose and animal residues to maintain the biotic equilibrium of soil by cooperating with nutrient cycling. Copyright © 2017 Elsevier Ltd. All rights reserved.

Characterization, Molecular Cloning, and Differential Expression Analysis of Laccase Genes from the Edible Mushroom Lentinula edodes

PubMed Central

Zhao, J.; Kwan, H. S.

1999-01-01

The effect of different substrates and various developmental stages (mycelium growth, primordium appearance, and fruiting-body formation) on laccase production in the edible mushroom Lentinula edodes was studied. The cap of the mature mushroom showed the highest laccase activity, and laccase activity was not stimulated by some well-known laccase inducers or sawdust. For our molecular studies, two genomic DNA sequences, representing allelic variants of the L. edodes lac1 gene, were isolated, and DNA sequence analysis demonstrated that lac1 encodes a putative polypeptide of 526 amino acids which is interrupted by 13 introns. The two allelic genes differ at 95 nucleotides, which results in seven amino acid differences in the encoded protein. The copper-binding domains found in other laccase enzymes are conserved in the L. edodes Lac1 proteins. A fragment of a second laccase gene (lac2) was also isolated, and competitive PCR showed that expression of lac1 and lac2 genes was different under various conditions. Our results suggest that laccases may play a role in the morphogenesis of the mushroom. To our knowledge, this is the first report on the cloning of genes involved in lignocellulose degradation in this economically important edible fungus. PMID:10543802

Biosynthetic Potential of Bioactive Streptomycetes Isolated From Arid Region of the Thar Desert, Rajasthan (India)

PubMed Central

Masand, Meeta; Sivakala, Kunjukrishnan Kamalakshi; Menghani, Ekta; Thinesh, Thangathurai; Anandham, Rangasamy; Sharma, Gaurav; Sivakumar, Natesan; Jebakumar, Solomon R. D.; Jose, Polpass Arul

2018-01-01

Acquisition of Actinobacteria, especially Streptomyces from previously underexplored habitats and the exploration of their biosynthetic potential have gained much attention in the rejuvenated antibiotics search programs. Herein, we isolated some Streptomyces strains, from an arid region of the Great Indian Thar Desert, which possess an ability to produce novel bioactive compounds. Twenty-one morphologically distinctive strains differing in their aerial and substrate mycelium were isolated by employing a stamping method. Among them, 12 strains were identified by a two-level antimicrobial screening method, exerting antimicrobial effects against a panel of indicator strains including methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococcus species. Based on their potent antimicrobial activity, four isolates were further explored by 16S rRNA gene-based identification, genetic screening, and metabolomic analysis; and it was found that these strains belong to the genus Streptomyces. The selected strains were found to have polyketide synthase and non-ribosomal peptide synthetase systems. In addition, extracellular metabolomic screening revealed that the isolates produced analogs of doxorubicinol, pyrromycin, erythromycin, and 6-13 other putative novel metabolites. These results demonstrate the significance of Streptomyces inhabiting the arid region of Thar Desert, suggesting that similar arid environments can be considered as the reservoirs of novel Streptomyces strains that could have biotechnological significance. PMID:29720968

In Vivo Study of Trichoderma-Pathogen-Plant Interactions, Using Constitutive and Inducible Green Fluorescent Protein Reporter Systems

PubMed Central

Lu, Zexun; Tombolini, Riccardo; Woo, Sheridan; Zeilinger, Susanne; Lorito, Matteo; Jansson, Janet K.

2004-01-01

Plant tissue colonization by Trichoderma atroviride plays a critical role in the reduction of diseases caused by phytopathogenic fungi, but this process has not been thoroughly studied in situ. We monitored in situ interactions between gfp-tagged biocontrol strains of T. atroviride and soilborne plant pathogens that were grown in cocultures and on cucumber seeds by confocal scanning laser microscopy and fluorescence stereomicroscopy. Spores of T. atroviride adhered to Pythium ultimum mycelia in coculture experiments. In mycoparasitic interactions of T. atroviride with P. ultimum or Rhizoctonia solani, the mycoparasitic hyphae grew alongside the pathogen mycelia, and this was followed by coiling and formation of specialized structures similar to hooks, appressoria, and papillae. The morphological changes observed depended on the pathogen tested. Branching of T. atroviride mycelium appeared to be an active response to the presence of the pathogenic host. Mycoparasitism of P. ultimum by T. atroviride occurred on cucumber seed surfaces while the seeds were germinating. The interaction of these fungi on the cucumber seeds was similar to the interaction observed in coculture experiments. Green fluorescent protein expression under the control of host-inducible promoters was also studied. The induction of specific Trichoderma genes was monitored visually in cocultures, on plant surfaces, and in soil in the presence of colloidal chitin or Rhizoctonia by confocal microscopy and fluorescence stereomicroscopy. These tools allowed initiation of the mycoparasitic gene expression cascade to be monitored in vivo. PMID:15128569

Basis for substrate recognition and distinction by matrix metalloproteinases

PubMed Central

Ratnikov, Boris I.; Cieplak, Piotr; Gramatikoff, Kosi; Pierce, James; Eroshkin, Alexey; Igarashi, Yoshinobu; Kazanov, Marat; Sun, Qing; Godzik, Adam; Osterman, Andrei; Stec, Boguslaw; Strongin, Alex; Smith, Jeffrey W.

2014-01-01

Genomic sequencing and structural genomics produced a vast amount of sequence and structural data, creating an opportunity for structure–function analysis in silico [Radivojac P, et al. (2013) Nat Methods 10(3):221–227]. Unfortunately, only a few large experimental datasets exist to serve as benchmarks for function-related predictions. Furthermore, currently there are no reliable means to predict the extent of functional similarity among proteins. Here, we quantify structure–function relationships among three phylogenetic branches of the matrix metalloproteinase (MMP) family by comparing their cleavage efficiencies toward an extended set of phage peptide substrates that were selected from ∼64 million peptide sequences (i.e., a large unbiased representation of substrate space). The observed second-order rate constants [k(obs)] across the substrate space provide a distance measure of functional similarity among the MMPs. These functional distances directly correlate with MMP phylogenetic distance. There is also a remarkable and near-perfect correlation between the MMP substrate preference and sequence identity of 50–57 discontinuous residues surrounding the catalytic groove. We conclude that these residues represent the specificity-determining positions (SDPs) that allowed for the expansion of MMP proteolytic function during evolution. A transmutation of only a few selected SDPs proximal to the bound substrate peptide, and contributing the most to selectivity among the MMPs, is sufficient to enact a global change in the substrate preference of one MMP to that of another, indicating the potential for the rational and focused redesign of cleavage specificity in MMPs. PMID:25246591

Imaging complex nutrient dynamics in mycelial networks.

PubMed

Fricker, M D; Lee, J A; Bebber, D P; Tlalka, M; Hynes, J; Darrah, P R; Watkinson, S C; Boddy, L

2008-08-01

Transport networks are vital components of multi-cellular organisms, distributing nutrients and removing waste products. Animal cardiovascular and respiratory systems, and plant vasculature, are branching trees whose architecture is thought to determine universal scaling laws in these organisms. In contrast, the transport systems of many multi-cellular fungi do not fit into this conceptual framework, as they have evolved to explore a patchy environment in search of new resources, rather than ramify through a three-dimensional organism. These fungi grow as a foraging mycelium, formed by the branching and fusion of threadlike hyphae, that gives rise to a complex network. To function efficiently, the mycelial network must both transport nutrients between spatially separated source and sink regions and also maintain its integrity in the face of continuous attack by mycophagous insects or random damage. Here we review the development of novel imaging approaches and software tools that we have used to characterise nutrient transport and network formation in foraging mycelia over a range of spatial scales. On a millimetre scale, we have used a combination of time-lapse confocal imaging and fluorescence recovery after photobleaching to quantify the rate of diffusive transport through the unique vacuole system in individual hyphae. These data then form the basis of a simulation model to predict the impact of such diffusion-based movement on a scale of several millimetres. On a centimetre scale, we have used novel photon-counting scintillation imaging techniques to visualize radiolabel movement in small microcosms. This approach has revealed novel N-transport phenomena, including rapid, preferential N-resource allocation to C-rich sinks, induction of simultaneous bi-directional transport, abrupt switching between different pre-existing transport routes, and a strong pulsatile component to transport in some species. Analysis of the pulsatile transport component using Fourier techniques shows that as the colony forms, it self-organizes into well demarcated domains that are identifiable by differences in the phase relationship of the pulses. On the centimetre to metre scale, we have begun to use techniques borrowed from graph theory to characterize the development and dynamics of the network, and used these abstracted network models to predict the transport characteristics, resilience, and cost of the network.

Protein farnesyltransferase isoprenoid substrate discrimination is dependent on isoprene double bonds and branched methyl groups.

PubMed

Micali, E; Chehade, K A; Isaacs, R J; Andres, D A; Spielmann, H P

2001-10-16

Farnesylation is a posttranslational lipid modification in which a 15-carbon farnesyl isoprenoid is linked via a thioether bond to specific cysteine residues of proteins in a reaction catalyzed by protein farnesyltransferase (FTase). We synthesized the benzyloxyisoprenyl pyrophosphate (BnPP) series of transferable farnesyl pyrophosphate (FPP) analogues (1a-e) to test the length dependence of the isoprenoid substrate on the FTase-catalyzed transfer of lipid to protein substrate. Kinetic analyses show that pyrophosphates 1a-e and geranyl pyrophosphate (GPP) transfer with a lower efficiency than FPP whereas geranylgeranyl pyrophosphate (GGPP) does not transfer at all. While a correlation was found between K(m) and analogue hydrophobicity and length, there was no correlation between k(cat) and these properties. Potential binding geometries of FPP, GPP, GGPP, and analogues 1a-e were examined by modeling the molecules into the active site of the FTase crystal structure. We found that analogue 1d displaces approximately the same volume of the active site as does FPP, whereas GPP and analogues 1a-c occupy lesser volumes and 1e occupies a slightly larger volume. Modeling also indicated that GGPP adopts a different conformation than the farnesyl chain of FPP, partially occluding the space occupied by the Ca(1)a(2)X peptide in the ternary X-ray crystal structure. Within the confines of the FTase pocket, the double bonds and branched methyl groups of the geranylgeranyl chain significantly restrict the number of possible conformations relative to the more flexible lipid chain of analogues 1a-e. The modeling results also provide a molecular explanation for the observation that an aromatic ring is a good isostere for the terminal isoprene of FPP.

Temporal and Spatial Regulation of Gene Expression during Asexual Development of Neurospora crassa

USDA-ARS?s Scientific Manuscript database

In this study we profiled spatial and temporal transcriptional changes during asexual sporulation in the filamentous fungus Neurospora crassa. Aerial tissue was separated from the mycelium to allow detection of genes specific to each tissue. We identified 2641 genes that were differentially expresse...

Evaluation of physico-mechanical properties of mycelium reinforced green biocomposites made from cellulosic fibers

USDA-ARS?s Scientific Manuscript database

Biodegradable and sustainable feedstock’s are increasingly replacing petroleum based materials commonly used for single or multi use packaging applications. A study was conducted to evaluate the potential of an innovative biocomposite material patented by Ecovative Design LLC for use in commercial p...

Evaluation of mycelium reinforced agricultural fiber biocomposites for diverse applications

USDA-ARS?s Scientific Manuscript database

There is a genuine need for new biodegradable alternatives to the inert plastics and expanded foams that are common in both manufacturing processes and device componentry. The material emphasized in this report is a bio-composite patented by Ecovative Design, LLC. The bio-composite utilizes fungus m...

Effects of varying levels of forest thinning on Tricholoma magnivelare (American matsutake)

Treesearch

Joyce L. Eberhart; Daniel L. Luoma

2013-01-01

Commercial matsutake mushroom (Tricholoma magnivelare) harvest in the Pacific Northwest is common in Lodgepole Pine (Pinus contorta) forests. Insuffi cient ecological knowledge of the mycorrhizae and mycelium has challenged monitoring of this resource. Among many management issues are concerns over logging practices, a lack of...

Germination of Phytophthora ramorum chlamydospores: a comparison of separation method and chlamydospore age

Treesearch

Justin P. Shaffer; Jennifer L. Parke

2013-01-01

Phytophthora ramorum characteristically produces large amounts of chlamydospores in vitro, but the role of these propagules in the disease cycle remains unclear. Germination is difficult to observe and quantify if chlamydospores are not free of mycelium, and the low frequency of germination commonly reported suggests that...

Laser Journal (Selected Articles),

DTIC Science & Technology

1982-09-10

temperature CO2 branch selection laser with a lifetime already exceeding 6500 hours which may be even longer. HIGH POWER LONG LIFE HeCd LASER Qu Shipu...method of plating single crystal gold film in a vacuum with the foreign material extension method. First mica is used as the substrate. Then a special...Hospital) Chen Zhasping Zhou Yiping et al (Eye, Ear, Nose, Throat Hospital, Examination Department, Shanghai Medical School Number 1.) Qu Zhipu et al

Solid State Technology Branch of NASA Lewis Research Center Second Annual Digest, June 1989 - June 1990

NASA Technical Reports Server (NTRS)

1990-01-01

A collection of papers and presentations authored by the branch between June 1989 and June 1990 is presented. The papers are organized into four sections. Section 1 deals with research in microwave circuits and includes full integrated circuits, the demonstration of optical/RF interfaces, and the evaluation of some hybrid circuitry. Section 2 indicates developments in coplanar waveguides and their use in breadboard circuits. Section 3 addresses high temperature superconductivity and includes: thin film deposition, transport measurement of film characteristics, RF surface resistant measurements, substrate permittivity measurements, measurements of microstrip line characteristics at cryogenic temperatures, patterning of superconducting films, and evaluation of simple passive microstrip circuitry based on YBaCuO films. Section 4 deals with carbon films, silicon carbide, GaAs/AlGaAs, HgCdTe, and other materials.

Early-branching Gut Fungi Possess A Large, And Comprehensive Array Of Biomass-Degrading Enzymes

DOE Office of Scientific and Technical Information (OSTI.GOV)

Solomon, Kevin V.; Haitjema, Charles; Henske, John K.

The fungal kingdom is the source of almost all industrial enzymes in use for lignocellulose bioprocessing. Its more primitive members, however, remain relatively unexploited. We developed a systems-level approach that integrates RNA-Seq, proteomics, phenotype and biochemical studies of relatively unexplored early-branching free-living fungi. Anaerobic gut fungi isolated from herbivores produce a large array of biomass-degrading enzymes that synergistically degrade crude, unpretreated plant biomass, and are competitive with optimized commercial preparations from Aspergillus and Trichoderma. Compared to these model platforms, gut fungal enzymes are unbiased in substrate preference due to a wealth of xylan-degrading enzymes. These enzymes are universally catabolite repressed,more » and are further regulated by a rich landscape of noncoding regulatory RNAs. Furthermore, we identified several promising sequence divergent enzyme candidates for lignocellulosic bioprocessing.« less

Asymmetric Fluorination of α-Branched Cyclohexanones Enabled by a Combination of Chiral Anion Phase-Transfer Catalysis and Enamine Catalysis using Protected Amino Acids

PubMed Central

2015-01-01

We report a study involving the successful merger of two separate chiral catalytic cycles: a chiral anion phase-transfer catalysis cycle to activate Selectfluor and an enamine activation cycle, using a protected amino acid as organocatalyst. We have demonstrated the viability of this approach with the direct asymmetric fluorination of α-substituted cyclohexanones to generate quaternary fluorine-containing stereocenters. With these two chiral catalytic cycles operating together in a matched sense, high enantioselectivites can be achieved, and we envisage that this dual catalysis method has the potential to be more broadly applicable, given the breadth of enamine catalysis. It also represents a rare example of chiral enamine catalysis operating successfully on α-branched ketones, substrates commonly inert to this activation mode. PMID:24684209

The pentose phosphate pathway leads to enhanced succinic acid flux in biofilms of wild-type Actinobacillus succinogenes.

PubMed

Bradfield, Michael F A; Nicol, Willie

2016-11-01

Increased pentose phosphate pathway flux, relative to total substrate uptake flux, is shown to enhance succinic acid (SA) yields under continuous, non-growth conditions of Actinobacillus succinogenes biofilms. Separate fermentations of glucose and xylose were conducted in a custom, continuous biofilm reactor at four different dilution rates. Glucose-6-phosphate dehydrogenase assays were performed on cell extracts derived from in situ removal of biofilm at each steady state. The results of the assays were coupled to a kinetic model that revealed an increase in oxidative pentose phosphate pathway (OPPP) flux relative to total substrate flux with increasing SA titre, for both substrates. Furthermore, applying metabolite concentration data to metabolic flux models that include the OPPP revealed similar flux relationships to those observed in the experimental kinetic analysis. A relative increase in OPPP flux produces additional reduction power that enables increased flux through the reductive branch of the TCA cycle, leading to increased SA yields, reduced by-product formation and complete closure of the overall redox balance.

Redirection of pyruvate flux toward desired metabolic pathways through substrate channeling between pyruvate kinase and pyruvate-converting enzymes in Saccharomyces cerevisiae.

PubMed

Kim, Sujin; Bae, Sang-Jeong; Hahn, Ji-Sook

2016-04-07

Spatial organization of metabolic enzymes allows substrate channeling, which accelerates processing of intermediates. Here, we investigated the effect of substrate channeling on the flux partitioning at a metabolic branch point, focusing on pyruvate metabolism in Saccharomyces cerevisiae. As a platform strain for the channeling of pyruvate flux, PYK1-Coh-Myc strain was constructed in which PYK1 gene encoding pyruvate kinase is tagged with cohesin domain. By using high-affinity cohesin-dockerin interaction, the pyruvate-forming enzyme Pyk1 was tethered to heterologous pyruvate-converting enzymes, lactate dehydrogenase and α-acetolactate synthase, to produce lactic acid and 2,3-butanediol, respectively. Pyruvate flux was successfully redirected toward desired pathways, with a concomitant decrease in ethanol production even without genetic attenuation of the ethanol-producing pathway. This pyruvate channeling strategy led to an improvement of 2,3-butanediol production by 38%, while showing a limitation in improving lactic acid production due to a reduced activity of lactate dehydrogenase by dockerin tagging.

Integrating metabolomics and transcriptomics data to discover a biocatalyst that can generate the amine precursors for alkamide biosynthesis

PubMed Central

Rizhsky, Ludmila; Jin, Huanan; Shepard, Michael R.; Scott, Harry W.; Teitgen, Alicen M.; Perera, M. Ann; Mhaske, Vandana; Jose, Adarsh; Zheng, Xiaobin; Crispin, Matt; Wurtele, Eve S.; Jones, Dallas; Hur, Manhoi; Góngora-Castillo, Elsa; Buell, C. Robin; Minto, Robert E.; Nikolau, Basil J.

2016-01-01

Summary The Echinacea genus is exemplary of over 30 plant families that produce a set of bioactive amides, called alkamides. The Echinacea alkamides may be assembled from two distinct moieties, a branched-chain amine that is acylated with a novel polyunsaturated fatty acid. In this study we identified the potential enzymological source of the amine moiety as a pyridoxal phosphate dependent decarboxylating enzyme that uses branched chain amino acids as substrate. This identification was based on a correlative analysis of the transcriptomes and metabolomes of 36 different E. purpurea tissues and organs, which expressed distinct alkamide profiles. Although no correlation was found between the accumulation patterns of the alkamides and their putative metabolic precursors (i.e., fatty acids and branched chain amino acids), isotope-labeling analyses supported the transformation of valine and isoleucine to isobutylamine and 2-methylbutylamine as reactions of alkamide biosynthesis. Sequence homology identified the pyridoxal phosphate dependent decarboxylase-like proteins in the translated proteome of E. purpurea. These sequences were prioritized for direct characterization by correlating their transcript levels with alkamide accumulation patterns in different organs and tissues, and this multi-pronged approach led to the identification and characterization of a branched-chain amino acid decarboxylase, which would appear to be responsible for generating the amine moieties of naturally occurring alkamides. PMID:27497272

Cellular response of preosteoblasts to nanograined/ultrafine-grained structures.

PubMed

Misra, R D K; Thein-Han, W W; Pesacreta, T C; Hasenstein, K H; Somani, M C; Karjalainen, L P

2009-06-01

Metallic materials with submicron- to nanometer-sized grains provide surfaces that are different from conventional polycrystalline materials because of the large proportion of grain boundaries with high free energy. In the study described here, the combination of cellular and molecular biology, materials science and engineering advances our understanding of cell-substrate interactions, especially the cellular activity between preosteoblasts and nanostructured metallic surfaces. Experiments on the effect of nano-/ultrafine grains have shown that cell attachment, proliferation, viability, morphology and spread are favorably modulated and significantly different from conventional coarse-grained structures. Additionally, immunofluorescence studies demonstrated stronger vinculin signals associated with actin stress fibers in the outer regions of the cells and cellular extensions on nanograined/ultrafine-grained substrate. These observations suggest enhanced cell-substrate interaction and activity. The differences in the cellular response on nanograined/ultrafine-grained and coarse-grained substrates are attributed to grain size and degree of hydrophilicity. The outcomes of the study are expected to reduce challenges to engineer bulk nanostructured materials with specific physical and surface properties for medical devices with improved cellular attachment and response. The data lay the foundation for a new branch of nanostructured materials for biomedical applications.

The activity of pyruvate carrier in a reconstituted system: substrate specificity and inhibitor sensitivity.

PubMed

Nałecz, K A; Kamińska, J; Nałecz, M J; Azzi, A

1992-08-15

The pyruvate carrier, of molecular mass 34 kDa, was purified from mitochondria isolated from rat liver, rat brain, and bovine heart, by affinity chromatography on immobilized 2-cyano-4-hydroxycinnamate. Its activity after reconstitution in phosphatidylcholine vesicles was measured either as uptake of [1-14C]pyruvate or as exchange with different 2-oxoacids. All preparations exhibited similar apparent Km values for pyruvate, but somewhat different V(max) values. The ability to exchange different anions of physiological significance, including branched-chain 2-oxoacids, confirmed the known substrate specificity described for the pyruvate carrier in mitochondria. The sensitivity of pyruvate transport toward phenylglyoxal suggested an important role of arginyl residues in the transport activity, while a role of lysyl and histidyl residues was not confirmed.

THE SYNTHESIS OF THE STARCH GRANULE.

PubMed

Smith, A. M.; Denyer, K.; Martin, C.

1997-06-01

This review describes and discusses the implications of recent discoveries about how starch polymers are synthesized and organized to form a starch granule. Three issues are highlighted. 1. The role and importance of ADPglucose pyrophosphorylase in the generation of ADPglucose as the substrate for polymer synthesis. 2. The contributions of isoforms of starch-branching enzyme, starch synthase, and debranching enzyme to the synthesis and ordered packing of amylopectin molecules. 3. The requirements for and regulation of the synthesis of amylose.

Diastereoselective sp2-sp3 coupling of sugar enol ethers with unactivated cycloalkenes: new entries to C-branched sugars.

PubMed

Hussain, Nazar; Tatina, Madhu Babu; Rasool, Faheem; Mukherjee, Debaraj

2016-10-25

Sugar enol ethers undergo efficient coupling at C-2 with unactivated cycloalkenes under a low Pd loading affording allylic substitution products. High diastereoselectivity was observed at the allylic centre with sterically hindered substrates. Generation of a π-allyl complex by the Pd(ii) catalyst via cleavage of the allylic C-H bond of the cycloalkene may be responsible for the formation of sp 2 -sp 3 coupling products.

Disturbance of proteasomal and autophagic protein degradation pathways by amyotrophic lateral sclerosis-linked mutations in ubiquilin 2.

PubMed

Osaka, Mayuko; Ito, Daisuke; Suzuki, Norihiro

2016-04-01

Ubiquilin (UBQLN), a member of the ubiquitin-like (UBL)-ubiquitin-associated (UBA) family, is a dual regulator of both the proteasomal and autophagic branches of the cellular protein degradation system. Mutations in the UBQLN2 gene encoding ubiquilin 2 cause X-linked amyotrophic lateral sclerosis (ALS)/frontotemporal dementia (FTD), and UBQLN2-positive inclusions have been identified in ALS patients with UBQLN2 mutations as well as in cases of both familial and sporadic ALS without UBQLN2 mutations. Compelling evidence links UBQLN2 to disturbance of the protein quality control network in neurons, but the pathomechanisms remain obscure. This study aimed to clarify how ALS-linked mutations in UBQLN2 affect the protein degradation system. Overexpression of a UBQLN2 with ALS-associated mutations resulted in the accumulation of polyubiquitinated proteins in neuronal cells, including the ALS-associated protein TDP-43. This effect was dependent on the UBA domain but not on inclusion formation. Immunocytochemistry and protein fractionation analysis of IVm-UBQLN2 cellular distribution indicated that it sequesters ubiquitinated substrates from both the proteasomal and autophagic branches of the protein degradation system, resulting in accumulation of polyubiquitinated substrates. These findings provide a molecular basis for the development of ALS/FTD-associated proteinopathy and establish novel therapeutic targets for ALS. Copyright © 2016. Published by Elsevier Inc.

Absolute configuration of acremoxanthone C, a potent calmodulin inhibitor from Purpureocillium lilacinum

USDA-ARS?s Scientific Manuscript database

Bioassay-guided fractionation of an extract prepared from the culture medium and mycelium of Purpureocillium lilacinum allowed the isolation of two calmodulin (CaM) inhibitors, namely, acremoxanthone C (1) and acremonidin A (2). The absolute configuration of 1 was established as 2R, 3R, 1'S, 11'S, ...

Survey and Detection of Endophytic Fungi in Lolium Germ Plasm by Direct Staining and Aphid Assays

Treesearch

A. Dan Wilson; Stephen L. Clement; Walter J. Kaiser

1991-01-01

Clavicipitaceous anamorphic endophytes were detected in 28 of 85 accessions from five of eight species in a collection of Lolium germ plasm. Comparative descriptions of endophytic mycelium in seeds of L. multiflorum, L. perenne, L. persicum, L. rigidum, and L. temulentum revealed morphological characteristics...

Optimization of biomass blends in the manufacture of molded packaging materials produced using fungal mycelium

USDA-ARS?s Scientific Manuscript database

Polystyrene is one of the most widely used plastics and is commonly produced in three forms: 1) Extruded polystyrene – disposable utensils, CD/DVD cases, yogurt containers, smoke alarm housing, etc.; 2) Expanded polystyrene foam – molded packaging materials and packaging "peanuts"; 3) Extruded polys...

Stawamycin analog, JBIR-11 from Streptomyces viridochromogenes subsp. sulfomycini NBRC 13830.

PubMed

Izumikawa, Miho; Komaki, Hisayuki; Hashimoto, Junko; Takagi, Motoki; Shin-ya, Kazuo

2008-05-01

A stawamycin analog, JBIR-11 (1) was isolated from mycelium of Streptomyces viridochromogenes subsp. sulfomycini NBRC 13830. The structure was determined on the basis of the spectroscopic data. Compound 1 exhibited growth inhibitory effect against human fibrosarcoma HT1080 cells with an IC50 value of 25 microM.

Differential expression in Phanerochaete chrysosporium of membrane- associated proteins relevant to lignin degradation

Treesearch

Semarjit Shary; Alexander N. Kapich; Ellen A. Panisko; Jon K. Magnuson; Daniel Cullen; Kenneth E. Hammel

2008-01-01

Fungal lignin-degrading systems likely include membrane-associated proteins that participate in diverse processes such as uptake and oxidation of lignin fragments, production of ligninolytic secondary metabolites, and defense of the mycelium against ligninolytic oxidants. Little is known about the nature or regulation of these membrane-associated components. We grew...

Use of mycelium and detached leaves in bioassays for assessing resistance to boxwood blight

USDA-ARS?s Scientific Manuscript database

Boxwood blight caused by Calonectria pseudonaviculata is a newly emergent disease of boxwood (Buxus L.) in the United States that causes leaf drop, stem lesions, and plant death. A rapid and reliable laboratory assay that enables screening hundreds of boxwood genotypes for resistance to boxwood blig...

pH-Signaling Transcription Factor AopacC Regulates Ochratoxin A Biosynthesis in Aspergillus ochraceus.

PubMed

Wang, Yan; Liu, Fei; Wang, Liuqing; Wang, Qi; Selvaraj, Jonathan Nimal; Zhao, Yueju; Wang, Yun; Xing, Fuguo; Liu, Yang

2018-05-02

In Aspergillus and Penicillium species, an essential pH-response transcription factor pacC is involved in growth, pathogenicity, and toxigenicity. To investigate the connection between ochratoxin A (OTA) biosynthesis and ambient pH, the AopacC in Aspergillus ochraceus was functionally characterized using a loss-of-function mutant. The mycelium growth was inhibited under pH 4.5 and 10.0, while the sporulation increased under alkaline condition. A reduction of mycelium growth and an elevation of sporulation was observed in Δ AopacC mutant. Compared to neutral condition, OTA contents were respectively reduced by 71.6 and 79.8% under acidic and alkaline conditions. The expression of AopacC increased with the elevated pH, and deleting AopacC dramatically decreased OTA production and biosynthetic genes Aopks expression. Additionally, the Δ AopacC mutant exhibited attenuated infection ability toward pear fruits. These results suggest that AopacC is an alkaline-induced regulator responsible for growth and OTA biosynthesis in A. ochraceus and this regulatory mechanism might be pH-dependent.

[Degradation of the herbicide atrazine by the soil mycelial fungus INBI 2-26(-)--a producer of cellobiose dehydrogenase].

PubMed

Khromonygina, V V; Saltykova, A I; Vasil'chenko, L G; Kozlov, Iu P; Rabinovich, M L

2004-01-01

Nonsporulating mycelial fungi producing cellobiose dehydrogenase (CDH) and isolated from soils of South Vietnam with high residual content of dioxins are capable of growing on a solid medium in the presence of high atrazine concentrations (to 500 mg/l). At 20 and 50 mg/l atrazine, the area of fungal colonies was 1.5-1.2-fold larger, respectively, compared with control colonies of the same age, whereas development of the colonies at 500 mg/l atrazine was delayed by 5 days, compared with controls grown in the absence of atrazine. Surface cultivation of the fungus on a minimal medium with glucose as a sole source of carbon and energy decreased the initial concentration of atrazine (20 mg/l) 50 times in 40 days; in addition, no pronounced sorption of atrazine by mycelium was detected. This was paralleled by accumulation in the culture medium of extracellular CDH; atrazine increased the synthesis of this enzyme two- to threefold. Accumulation of beta-glucosidase (a mycelium-associated enzyme) and cellulases preceded the formation of CDH.

Investigation of lead bioimmobilization and transformation by Penicillium oxalicum SL2.

PubMed

Ye, Binhui; Luo, Yating; He, Junyu; Sun, Lijuan; Long, Bibo; Liu, Qinglin; Yuan, Xiaofeng; Dai, Peibin; Shi, Jiyan

2018-05-18

Fungi Penicillium oxalicum SL2 was applied for Pb 2+ bioremediation in aqueous solution in this study. After 7 days of incubation at different initial concentrations of Pb 2+ (0, 100, 500 and 2500 mg L -1 ), most of Pb 2+ were removed (90, 98.3, and 86.2%), the maximum Pb content in mycelium reached about 155.6 mg g -1  dw. Meanwhile, the formation of extracellular secondary minerals and intracellular Pb-complex were observed and identified, the speciation of Pb in mycelium was also detected by X-ray absorption near-edge structure (XANES) spectroscopy, i.e., Pb-oxalate, Pb-citrate, Pb-hydrogen phosphate and Pb-glutathione analogues. In addition, content of glutathione and oxidized glutathione was increased under the exposure of Pb 2+ , which implied that glutathione might play a key role in Pb immobilization and detoxification in P. oxalicum SL2. This study elucidated partial mechanisms of Pb immobilization and speciation transformation of this strain, providing an alternative biomaterial in the bioremediation of Pb-contaminated wastewater. Copyright © 2018 Elsevier Ltd. All rights reserved.

Biological Modification of Trichothecene Mycotoxins: Acetylation and Deacetylation of Deoxynivalenols by Fusarium spp

PubMed Central

Yoshizawa, Takumi; Morooka, Nobuichi

1975-01-01

Attempts were made to elucidate the acetyl transformation of novel trichothecene mycotoxins, 3α,7α,15-trihydroxy-12,13-epoxytrichothec-9-en-8-one (deoxynivalenol) and its derivatives, by trichothecene-producing strains of Fusarium nivale, F. roseum, and F. solani. In the peptone-supplemented Czapek-Dox medium, F. roseum converted 3α-acetoxy-7α,15-dihydroxy-12,13-epoxytrichothec-9-en-8-one (3-acetyldeoxynivalenol) to deoxynivalenol. 3-Acetyldeoxynivalenol was also deacetylated by intact mycelia of the three strains in sugar-free Czapek-Dox medium. The growing F. nivale acetylated deoxynivalenol to afford a small amount of 3-acetyldeoxynivalenol. 3α,7α,15-Triacetoxy-12,13-epoxytrichothec-9-en-8-one (deoxynivalenol triacetate) was transformed by the intact mycelium of F. solani into 7α,15-diacetoxy-3α-hydroxy-12,13-epoxytrichothec-9-en-8-one (7,15-diacetyl-deoxynivalenol), which was then deacetylated to give 7α-acetoxy-3α,15-dihydroxy-12,13-epoxytrichothec-9-en-8-one (7-acetyldeoxynivalenol). It was noted that the ester at C-7 was not hydrolyzed by the fungal mycelium. PMID:234156

Four novel antibacterial sesquiterpene-α-amino acid quaternary ammonium hybrids from the mycelium of mushroom Stereum hirsutum.

PubMed

Duan, Yuan-Chang; Feng, Jun; Bai, Na; Li, Guo-Hong; Zhang, Ke-Qin; Zhao, Pei-Ji

2018-05-21

The mushroom Stereum hirsutum is parasitized by Tremella aurantia to form a heterogeneous basidiocarp Jin'er, which has been used as food and folk medicine in Chinese society. In present work, the S. hirsutum was fermented in YMG broth, and four novel mixed terpenes, stereumamides A-D (1-4), which are sesquiterpenes combined with α-amino acids to form quaternary ammonium hybrids, were isolated from the Stereum hirsutum FP-91666 and their structures were elucidated by spectroscopic data analysis. Stereumamides A and D showed antibacterial activity against Escherichia coli, Staphylococcus aureus, and Salmonella typhimurium, with the minimum inhibitory concentration (MIC) values of 12.5-25.0 μg/mL. The stereumamides A-D should be apparently the first example of naturally occurring a quaternary ammonium compound (QAC) conjugated by sesquiterpene with an α-amino acid. QAC is a common antibacterial agent in food industry, which is found in the mycelium of Stereum hirsutum would suggest that the complex basidiocarp is a functional food and veritable folk medicine. Copyright © 2017. Published by Elsevier B.V.

Laccase production by free and immobilized mycelia of Peniophora cinerea and Trametes versicolor: a comparative study.

PubMed

Silvério, Sara C; Moreira, Sérgio; Milagres, Adriane M F; Macedo, Eugénia A; Teixeira, José A; Mussatto, Solange I

2013-03-01

The production of laccase by immobilized mycelia of Peniophora cinerea and Trametes versicolor was studied. In an initial stage, experimental assays were performed in Erlenmeyer flasks using free and immobilized mycelium, and the performance of the fungal strains to produce the enzyme was compared. Both fungi adhered into the support material (a synthetic fiber), growing not only on the surface but also in the interspaces of the fibers. Immobilization of P. cinerea provided a 35-fold increase in laccase production when compared to the production obtained by using free mycelium. On the other hand, immobilization of T. versicolor caused a decrease in laccase activity. A comparison between the strains revealed that immobilized P. cinerea (3,500 U/L) surpassed the enzyme production by free T. versicolor (800 U/L). When the conditions that gave the best laccase production to each fungus were employed in a stirred tank bioreactor, very low laccase production was observed for both the cases, suggesting that shear stress and mycelia damage caused by the agitation impellers negatively affected the enzyme production.

Transcriptomic Analysis of Calonectria pseudoreteaudii during Various Stages of Eucalyptus Infection

PubMed Central

Ye, Xiaozhen; Liu, Hongyi; Jin, Yajie; Guo, Mengmeng; Huang, Aizhen; Chen, Quanzhu; Guo, Wenshuo; Zhang, Feiping; Feng, Lizhen

2017-01-01

Eucalyptus leaf blight caused by Calonectria spp. is a serious disease in Eucalyptus seedling and plantations. However, the molecular mechanisms of the infection process and pathogenesis of Calonectria to Eucalyptus is not well-studied. In this study, we analyzed the transcriptomes of C. pseudoreteaudii at three stages of Eucalyptus leaf infection, and in mycelium grown in potato dextrose broth using Illumina RNA-Seq technology. We identified 161 differentially expressed genes between C. pseudoreteaudii from leaf and mycelium grown in potato dextrose broth. GO and KEGG enrichment analyses of these genes suggested that they were mainly involved in oxidoreductase activity, hydrolase activity, and transmembrane transporter activity. Most of the differentially expressed genes at the early infection stage were upregulated. These upregulated genes were mainly involved in cell wall hydrolysis and toxin synthesis, suggesting a role for toxin and cell wall hydrolases in the establishment of Calonectria leaf blight. Genes related to detoxification of phytoalexins were continually upregulated during infection. The candidate effectors and putative pathogenicity determinants identified in this study will help in the functional analysis of C. pseudoreteaudii virulence and pathogenicity. PMID:28072879

Transcriptomic Analysis of Calonectria pseudoreteaudii during Various Stages of Eucalyptus Infection.

PubMed

Ye, Xiaozhen; Liu, Hongyi; Jin, Yajie; Guo, Mengmeng; Huang, Aizhen; Chen, Quanzhu; Guo, Wenshuo; Zhang, Feiping; Feng, Lizhen

2017-01-01

Eucalyptus leaf blight caused by Calonectria spp. is a serious disease in Eucalyptus seedling and plantations. However, the molecular mechanisms of the infection process and pathogenesis of Calonectria to Eucalyptus is not well-studied. In this study, we analyzed the transcriptomes of C. pseudoreteaudii at three stages of Eucalyptus leaf infection, and in mycelium grown in potato dextrose broth using Illumina RNA-Seq technology. We identified 161 differentially expressed genes between C. pseudoreteaudii from leaf and mycelium grown in potato dextrose broth. GO and KEGG enrichment analyses of these genes suggested that they were mainly involved in oxidoreductase activity, hydrolase activity, and transmembrane transporter activity. Most of the differentially expressed genes at the early infection stage were upregulated. These upregulated genes were mainly involved in cell wall hydrolysis and toxin synthesis, suggesting a role for toxin and cell wall hydrolases in the establishment of Calonectria leaf blight. Genes related to detoxification of phytoalexins were continually upregulated during infection. The candidate effectors and putative pathogenicity determinants identified in this study will help in the functional analysis of C. pseudoreteaudii virulence and pathogenicity.

Emergence of an Out-of-Plane Optical Phonon (ZO) Kohn Anomaly in Quasifreestanding Epitaxial Graphene.

PubMed

Politano, Antonio; de Juan, Fernando; Chiarello, Gennaro; Fertig, Herbert A

2015-08-14

In neutral graphene, two prominent cusps known as Kohn anomalies are found in the phonon dispersion of the highest optical phonon at q=Γ (LO branch) and q=K (TO branch), reflecting a significant electron-phonon coupling (EPC) to undoped Dirac electrons. In this work, high-resolution electron energy loss spectroscopy is used to measure the phonon dispersion around the Γ point in quasifreestanding graphene epitaxially grown on Pt(111). The Kohn anomaly for the LO phonon is observed at finite momentum q~2k_{F} from Γ, with a shape in excellent agreement with the theory and consistent with known values of the EPC and the Fermi level. More strikingly, we also observe a Kohn anomaly at the same momentum for the out-of-plane optical phonon (ZO) branch. This observation is the first direct evidence of the coupling of the ZO mode with Dirac electrons, which is forbidden for freestanding graphene but becomes allowed in the presence of a substrate. Moreover, we estimate the EPC to be even greater than that of the LO mode, making graphene on Pt(111) an optimal system to explore the effects of this new coupling in the electronic properties.

Correlation between polymer architecture, mesoscale structure and photovoltaic performance in side-chain-modified PAE-PAV:fullerene bulk-heterojunction solar cells

NASA Astrophysics Data System (ADS)

Rathgeber, S.; Kuehnlenz, F.; Hoppe, H.; Egbe, D. A. M.; Tuerk, S.; Perlich, J.; Gehrke, R.

2012-02-01

A poly(arylene-ethynylene)-alt-poly(arylene-vinylene) statistical copolymer carrying linear and branched alkoxy side chains along the conjugated backbone in a random manner, yields, compared to its regular substituted counterparts, an improved performance in polymer:fullerene bulk-heterojunction solar cells. Results obtained from GiWAXS experiments show that the improved performance of the statistical copolymer may be attributed to the following structural characteristics: 1) Well, ordered stacked domains that promote backbone planarization and thus improve the ππ-overlap. 2) Partly face-on alignment of domains relative to the electrodes for an improved active layer electrode charge transfer. Branched side chains seem to promote face-on domain orientation. Most likely they can minimize their unfavorable contact with the interface by just bringing the CH3 groups of the branches into direct contact with the surface so that favorable phenylene-substrate interaction can promote face-on orientation. 3) A more isotropic domain orientation throughout the active layer to ensure that the backbone alignment direction has components perpendicular and parallel to the electrodes in order to compromise between light absorption and efficient intra-chain charge transport.

Fire adaptation in Neblinaria celiae (Theaceae), a high-elevation rosette shrub endemic to a wet equatorial tepui

USGS Publications Warehouse

Givnish, T.J.; McDiarmid, R.W.; Buck, W.R.

1986-01-01

Neblinaria celiae (Theaceae), a rosette shrub endemic to the exceedingly rainy summit of remote Cerro de la Neblina in southern Venezuela, has a previously undescribed set of adaptations to fire. Its growth form entails sparse branching, massive terminal leaf rosettes, and thick bark. It is highly fire-tolerant, with a survival rate of 93% in a stand recently ignited by lightning, vs. 0% in seven co-occurring woody species. Survival increases sharply with rosette height, favoring a sparsely branched habit that would maximize the rate of upward growth through the sparse fuel layer supported by a sterile substrate. Thick bark and massive rosettes help protect cambial and foliar meristems from brief exposure to high temperatures. Rosettes on shorter plants are exposed to greater damage from fire near the ground and, as expected, are bigger and impound more rainwater; the greater number of leaves nearly balances the greater leaf mortality caused by fire. We relate Neblinaria's growth form to its dominance atop Neblina, to a general model for the evolution of sparse branching, and to the evolution of growth form in other tepui plants.

Substrate specificity of platypus venom L-to-D-peptide isomerase.

PubMed

Bansal, Paramjit S; Torres, Allan M; Crossett, Ben; Wong, Karen K Y; Koh, Jennifer M S; Geraghty, Dominic P; Vandenberg, Jamie I; Kuchel, Philip W

2008-04-04

The L-to-D-peptide isomerase from the venom of the platypus (Ornithorhyncus anatinus) is the first such enzyme to be reported for a mammal. In delineating its catalytic mechanism and broader roles in the animal, its substrate specificity was explored. We used N-terminal segments of defensin-like peptides DLP-2 and DLP-4 and natriuretic peptide OvCNP from the venom as substrates. The DLP analogues IMFsrs and ImFsrs (srs is a solubilizing chain; lowercase letters denote D-amino acid) were effective substrates for the isomerase; it appears to recognize the N-terminal tripeptide sequence Ile-Xaa-Phe-. A suite of 26 mutants of these hexapeptides was synthesized by replacing the second residue (Met) with another amino acid, viz. Ala, alpha-aminobutyric acid, Ile, Leu, Lys, norleucine, Phe, Tyr, and Val. It was shown that mutant peptides incorporating norleucine and Phe are substrates and exhibit L- or D-amino acid isomerization, but mutant peptides that contain residues with shorter, beta-branched or long side chains with polar terminal groups, viz. Ala, alpha-aminobutyric acid, Ile, Val, Leu, Lys, and Tyr, respectively, are not substrates. It was demonstrated that at least three N-terminal amino acid residues are absolutely essential for L-to-D-isomerization; furthermore, the third amino acid must be a Phe residue. None of the hexapeptides based on LLH, the first three residues of OvCNP, were substrates. A consistent 2-base mechanism is proposed for the isomerization; abstraction of a proton by 1 base is concomitant with delivery of a proton by the conjugate acid of a second base.

Understanding Which Residues of the Active Site and Loop Structure of a Tyrosine Aminomutase Define Its Mutase and Lyase Activities.

PubMed

Attanayake, Gayanthi; Walter, Tyler; Walker, Kevin D

2018-05-30

Site-directed mutations and substrate analogues were used to gain insights into the branch-point reaction of the 3,5-dihydro-5-methylidene-4 H-imidazol-4-one (MIO)-tyrosine aminomutase from Oryza sativa ( OsTAM). Exchanging the active residues of OsTAM (Y125C/N446K) for those in a phenylalanine aminomutase TcPAM altered its substrate specificity from tyrosine to phenylalanine. The aminomutase mechanism of OsTAM surprisingly changed almost exclusively to that of an ammonia lyase making cinnamic acid (>95%) over β-phenylalanine [Walter, T., et al. (2016) Biochemistry 55, 3497-3503]. We hypothesized that the missing electronics or sterics on the aryl ring of the phenylalanine substrate, compared with the sizable electron-donating hydroxyl of the natural tyrosine substrate, influenced the unexpected lyase reactivity of the OsTAM mutant. The double mutant was incubated with 16 α-phenylalanine substituent analogues of varying electronic strengths and sterics. The mutant converted each analogue principally to its acrylate with ∼50% conversion of the p-Br substrate, making only a small amount of the β-amino acid. The inner loop structure over the entrance to the active site was also mutated to assess how the lyase and mutase activities are affected. An OsTAM loop mutant, matching the loop residues of TcPAM, still chiefly made >95% of the acrylate from each substrate. A combined active site:loop mutant was most reactive but remained a lyase, making 10-fold more acrylates than other mutants did. While mutations within the active site changed the substrate specificity of OsTAM, continued exploration is needed to fully understand the interplay among the inner loop, the substrate, and the active site in defining the mutase and lyase activities.

Predicting and preventing mold spoilage of food products.

PubMed

Dagnas, Stéphane; Membré, Jeanne-Marie

2013-03-01

This article is a review of how to quantify mold spoilage and consequently shelf life of a food product. Mold spoilage results from having a product contaminated with fungal spores that germinate and form a visible mycelium before the end of the shelf life. The spoilage can be then expressed as the combination of the probability of having a product contaminated and the probability of mold growth (germination and proliferation) up to a visible mycelium before the end of the shelf life. For products packed before being distributed to the retailers, the probability of having a product contaminated is a function of factors strictly linked to the factory design, process, and environment. The in-factory fungal contamination of a product might be controlled by good manufacturing hygiene practices and reduced by particular processing practices such as an adequate air-renewal system. To determine the probability of mold growth, both germination and mycelium proliferation can be mathematically described by primary models. When mold contamination on the product is scarce, the spores are spread on the product and more than a few spores are unlikely to be found at the same spot. In such a case, models applicable for a single spore should be used. Secondary models can be used to describe the effect of intrinsic and extrinsic factors on either the germination or proliferation of molds. Several polynomial models and gamma-type models quantifying the effect of water activity and temperature on mold growth are available. To a lesser extent, the effect of pH, ethanol, heat treatment, addition of preservatives, and modified atmospheres on mold growth also have been quantified. However, mold species variability has not yet been properly addressed, and only a few secondary models have been validated for food products. Once the probability of having mold spoilage is calculated for various shelf lives and product formulations, the model can be implemented as part of a risk management decision tool.

Innervation of taste buds revealed with Brainbow-labeling in mouse.

PubMed

Zaidi, Faisal N; Cicchini, Vanessa; Kaufman, Daniel; Ko, Elizabeth; Ko, Abraham; Van Tassel, Heather; Whitehead, Mark C

2016-12-01

Nerve fibers that surround and innervate the taste bud were visualized with inherent fluorescence using Brainbow transgenic mice that were generated by mating the founder line L with nestin-cre mice. Multicolor fluorescence revealed perigemmal fibers as branched within the non-taste epithelium and ending in clusters of multiple rounded swellings surrounding the taste pore. Brainbow-labeling also revealed the morphology and branching pattern of single intragemmal fibers. These taste bud fibers frequently innervated both the peripheral bud, where immature gemmal cells are located, and the central bud, where mature, differentiated cells are located. The fibers typically bore preterminal and terminal swellings, growth cones with filopodia, swellings, and rounded retraction bulbs. These results establish an anatomical substrate for taste nerve fibers to contact and remodel among receptor cells at all stages of their differentiation, an interpretation that was supported by staining with GAP-43, a marker for growing fibers and growth cones. © 2016 Anatomical Society.

Dynamics of Gram-negative bacteria population density in a soil in the course of the succession initiated by chitin and cellulose

NASA Astrophysics Data System (ADS)

Konstantin, Ivanov; Lubov, Polyanskaya

2014-05-01

The functions of actinomycetes in polymer destruction in soil traditionally considered as the dominant, compare to another groups of bacteria. Gram-positive bacteria also have ecological functions in destruction of soil organic matter. The role of Gram-negative bacteria has been researched in the microbial succession in terms of polymers destruction, which are widely spreads in soils: chitin and cellulose. The method with nalidixic acid as an inhibitor of DNA division of Gram-negative bacteria was modified. By modified method microbial succession of Gram-negative bacteria in the different horizons of a chernozem under aerobic and anaerobic conditions was researched. Chitin and cellulose as the source of nutrients with moistening was used in experiments. The introduction of chitin had no positive effect on the population density of Gram-negative bacteria in a chernozem, but it advanced the date of their appearance in microbial succession: the maximum of Gram-negative bacteria population density was registered on the 3rd- 7th day of the experiment with adding chitin. Compare to the control, which one was without any nutrient adding this dynamics registered much earlier. Consequently, the introduction of chitin as an additional source of nutrition promoted revealing of the Gram-negative bacteria in soil already at the early stages of the succession. In the course of the succession, when the fungal mycelium begins to die off, the actinomycetic mycelium increases in length, i.e., Gram-negative bacteria are replaced at this stage with Gram-positive ones, the leading role among which belongs to actinomycetes. The growth rate of Gram-negative bacteria is higher than that of actinomycetes, so they start chitin utilization at the early stages of the succession, whereas actinomycetes dominate at the late stages. The population density of Gram-negative bacteria was lower under the anaerobic conditions as compared with that in the aerobic ones. The population density of Gram-negative bacteria in the lower layer of the A horizon of the chernozem and in the B horizon was slightly higher only in the case of the chitin introduction. When cellulose was introduced into the soil under aerobic conditions, the population density of Gram-negative bacteria in all the layers of the A horizon of the chernozem was maximal from the 14th to the 22nd day of the experiment. Simultaneously, an increase in the length of the actinomycetal mycelium was observed, as these organisms also perform cellulose hydrolysis in soils. The Gram-negative bacteria began to develop at the stage of the fungal mycelium destruction, which indirectly confirmed the chitinolytic activity of these bacteria.

Alteration of the fatty acid profile of Streptomyces coelicolor by replacement of the initiation enzyme 3-ketoacyl acyl carrier protein synthase III (FabH).

PubMed

Li, Yongli; Florova, Galina; Reynolds, Kevin A

2005-06-01

The first elongation step of fatty acid biosynthesis by a type II dissociated fatty acid synthases is catalyzed by 3-ketoacyl-acyl carrier protein (ACP) synthase III (KASIII, FabH). This enzyme, encoded by the fabH gene, catalyzes a decarboxylative condensation between an acyl coenzyme A (CoA) primer and malonyl-ACP. In organisms such as Escherichia coli, which generate only straight-chain fatty acids (SCFAs), FabH has a substrate preference for acetyl-CoA. In streptomycetes and other organisms which produce a mixture of both SCFAs and branched-chain fatty acids (BCFAs), FabH has been shown to utilize straight- and branched-chain acyl-CoA substrates. We report herein the generation of a Streptomyces coelicolor mutant (YL/ecFabH) in which the chromosomal copy of the fabH gene has been replaced and the essential process of fatty acid biosynthesis is initiated by plasmid-based expression of the E. coli FabH (bearing only 35% amino acid identity to the Streptomyces enzyme). The YL/ecFabH mutant produces predominantly SCFAs (86%). In contrast, BCFAs predominate (approximately 70%) in both the S. coelicolor parental strain and S. coelicolor YL/sgFabH (a deltafabH mutant carrying a plasmid expressing the Streptomyces glaucescens FabH). These results provide the first unequivocal evidence that the substrate specificity of FabH observed in vitro is a determinant of the fatty acid made in an organism. The YL/ecFabH strain grows significantly slower on both solid and liquid media. The levels of FabH activity in cell extracts of YL/ecFabH were also significantly lower than those in cell extracts of YL/sgFabH, suggesting that a decreased rate of fatty acid synthesis may account for the observed decreased growth rate. The production of low levels of BCFAs in YL/ecFabH suggests either that the E. coli FabH is more tolerant of different acyl-CoAs substrates than previously thought or that there is an additional pathway for initiation of BCFA biosynthesis in Streptomyces coelicolor.

Diagnosis of Annosus Root Disease in Mixed Conifer Forests in the Northwestern United States

Treesearch

Craig L. Schmitt

1989-01-01

Recognizing annosus root disease affecting conifers in northwestern United States forests is discussed. Field diagnosis can bemade by observing characteristic stand patterns, wood stain and decay, ectotrophic mycelium, and sporophores. Most seriously affected trees include hemlocks, grand fir, white fir and Pacific silver fir. Ponderosa pine and other true firs may...

First report of clavicipitaceous anamorphic endophytes in hordeum species

Treesearch

A.D. Wilson; S.L. Clement; W.J. Kaiser; D.G. Lester

1991-01-01

Clavicipitaceous endophytes systemically infect many grass species and produce alkaloids that confer resistance to insects (2) and toxicity to mammals (1). The mutualistic anamorphic forms (e.g., Acremonium spp.) do not sporulate or cause symptoms, but they produce distinctive mycelium in their hosts. The incidence of anamorphic endophytes in a portion of the U.S....

Fungal mycelium and cotton plant materials in the manufacture of biodegradable molded packaging material: Evaluation study of select blends of cotton byproducts

USDA-ARS?s Scientific Manuscript database

The primary material used by the packaging industry is extruded polystyrene foam, which is commonly marketed as Styrofoam™. In its original formulation, Styrofoam™ is resistant to photolysis and effectively does not decompose. The light weight of Styrofoam™ packaging materials reduces the likelihood...

Influence of Stenocarpella maydis infected corn on the composition of corn kernel and its conversion into ethanol

USDA-ARS?s Scientific Manuscript database

Stenocarpella ear rot (formerly Diplodia ear rot) is resurfacing as a concern in the central United States Corn Belt. There are reports of some fields containing more than 50% mummified ears. Ears infected within two weeks of silking may be completely mummified with white to grayish brown mycelium c...

The Ubiquitin Code in the Ubiquitin-Proteasome System and Autophagy.

PubMed

Kwon, Yong Tae; Ciechanover, Aaron

2017-11-01

The conjugation of the 76 amino acid protein ubiquitin to other proteins can alter the metabolic stability or non-proteolytic functions of the substrate. Once attached to a substrate (monoubiquitination), ubiquitin can itself be ubiquitinated on any of its seven lysine (Lys) residues or its N-terminal methionine (Met1). A single ubiquitin polymer may contain mixed linkages and/or two or more branches. In addition, ubiquitin can be conjugated with ubiquitin-like modifiers such as SUMO or small molecules such as phosphate. The diverse ways to assemble ubiquitin chains provide countless means to modulate biological processes. We overview here the complexity of the ubiquitin code, with an emphasis on the emerging role of linkage-specific degradation signals (degrons) in the ubiquitin-proteasome system (UPS) and the autophagy-lysosome system (hereafter autophagy). Copyright © 2017 Elsevier Ltd. All rights reserved.

Endocytosis in the Shiitake Mushroom Lentinula edodes and Involvement of GTPase LeRAB7▿

PubMed Central

Lee, Ming Tsung; Szeto, Carol Ying Ying; Ng, Tak Pan; Kwan, Hoi Shan

2007-01-01

Endocytosis is the process by which substrates enter a cell without passing through the plasma membrane but rather invaginate the cell membrane and form intracellular vesicles. Rab7 regulates endocytic trafficking between early and late endosomes and between late endosomes and lysosomes. LeRab7 in Lentinula edodes is strongly homologous to Rab7 in Homo sapiens. Receptors for activated C kinase-1 (LeRACK1) and Rab5 GTPase (LeRAB5) were isolated as interacting partners of LeRab7, and the interactions were confirmed by in vivo and in vitro protein interaction assays. The three genes showed differential expression in the various developmental stages of the mushroom. In situ hybridization showed that the three transcripts were localized in regions of active growth, such as the outer region of trama cells, and the subhymenium of the hymenophore of mature fruiting bodies and the prehymenophore of young fruiting bodies. The existence of endocytosis in the mycelium and hymenophores was confirmed by the internalization of FM4-64. LeRAB7 was partially colocalized with the AM4-64 and was located in the late endocytic pathway. This is the first report of the presence of endocytosis in homobasidiomycetes. LeRAB7, LeRAB5, and LeRACK1 may contribute to the growth of L. edodes and cell differentiation in hymenophores. PMID:17921351

Systematic search for cultivatable fungi that best deconstruct cell walls of Miscanthus and sugarcane in the field.

PubMed

Shrestha, Prachand; Szaro, Timothy M; Bruns, Thomas D; Taylor, John W

2011-08-01

The goals of our project were to document the diversity and distributions of cultivable fungi associated with decaying Miscanthus and sugarcane plants in nature and to further assess biodegradation of host plant cell walls by these fungi in pure cultures. Late in 2008 and early in 2009 we collected decaying Miscanthus and Saccharum from 8 sites in Illinois and 11 sites in Louisiana, respectively. To recover fungi that truly decay plants and to recover slow-growing fungi, we washed the plant material repeatedly to remove spores and cultivated fungi from plant fragments small enough to harbor at most one mycelium. We randomly selected 950 fungal colonies out of 4,560 microwell colonies and used molecular identification to discover that the most frequently recovered fungal species resided in Hypocreales (Sordariomycetes), Pleosporales (Dothideomycetes), and Chaetothryiales (Eurotiomycetes) and that only a few weedy species were recovered. We were particularly interested in Pleosporales and Chaetothyriales, groups that have not been mined for plant decay fungi. To confirm that we had truly recovered fungi that deconstruct plant cell walls, we assayed the capacity of the fungi to consume whole, alkali-pretreated, ground Miscanthus. Solid substrate cultures of the nine most commonly encountered Ascomycota resulted in Miscanthus weight loss of 8 to 13% over 4 weeks. This is the first systematic, high-throughput, isolation and biodegradation assessment of fungi isolated from decaying bioenergy grasses.

Whole genome and global gene expression analyses of the model mushroom Flammulina velutipes reveal a high capacity for lignocellulose degradation.

PubMed

Park, Young-Jin; Baek, Jeong Hun; Lee, Seonwook; Kim, Changhoon; Rhee, Hwanseok; Kim, Hyungtae; Seo, Jeong-Sun; Park, Hae-Ran; Yoon, Dae-Eun; Nam, Jae-Young; Kim, Hong-Il; Kim, Jong-Guk; Yoon, Hyeokjun; Kang, Hee-Wan; Cho, Jae-Yong; Song, Eun-Sung; Sung, Gi-Ho; Yoo, Young-Bok; Lee, Chang-Soo; Lee, Byoung-Moo; Kong, Won-Sik

2014-01-01

Flammulina velutipes is a fungus with health and medicinal benefits that has been used for consumption and cultivation in East Asia. F. velutipes is also known to degrade lignocellulose and produce ethanol. The overlapping interests of mushroom production and wood bioconversion make F. velutipes an attractive new model for fungal wood related studies. Here, we present the complete sequence of the F. velutipes genome. This is the first sequenced genome for a commercially produced edible mushroom that also degrades wood. The 35.6-Mb genome contained 12,218 predicted protein-encoding genes and 287 tRNA genes assembled into 11 scaffolds corresponding with the 11 chromosomes of strain KACC42780. The 88.4-kb mitochondrial genome contained 35 genes. Well-developed wood degrading machinery with strong potential for lignin degradation (69 auxiliary activities, formerly FOLymes) and carbohydrate degradation (392 CAZymes), along with 58 alcohol dehydrogenase genes were highly expressed in the mycelium, demonstrating the potential application of this organism to bioethanol production. Thus, the newly uncovered wood degrading capacity and sequential nature of this process in F. velutipes, offer interesting possibilities for more detailed studies on either lignin or (hemi-) cellulose degradation in complex wood substrates. The mutual interest in wood degradation by the mushroom industry and (ligno-)cellulose biomass related industries further increase the significance of F. velutipes as a new model.

Whole Genome and Global Gene Expression Analyses of the Model Mushroom Flammulina velutipes Reveal a High Capacity for Lignocellulose Degradation

PubMed Central

Park, Young-Jin; Baek, Jeong Hun; Lee, Seonwook; Kim, Changhoon; Rhee, Hwanseok; Kim, Hyungtae; Seo, Jeong-Sun; Park, Hae-Ran; Yoon, Dae-Eun; Nam, Jae-Young; Kim, Hong-Il; Kim, Jong-Guk; Yoon, Hyeokjun; Kang, Hee-Wan; Cho, Jae-Yong; Song, Eun-Sung; Sung, Gi-Ho; Yoo, Young-Bok; Lee, Chang-Soo; Lee, Byoung-Moo; Kong, Won-Sik

2014-01-01

Flammulina velutipes is a fungus with health and medicinal benefits that has been used for consumption and cultivation in East Asia. F. velutipes is also known to degrade lignocellulose and produce ethanol. The overlapping interests of mushroom production and wood bioconversion make F. velutipes an attractive new model for fungal wood related studies. Here, we present the complete sequence of the F. velutipes genome. This is the first sequenced genome for a commercially produced edible mushroom that also degrades wood. The 35.6-Mb genome contained 12,218 predicted protein-encoding genes and 287 tRNA genes assembled into 11 scaffolds corresponding with the 11 chromosomes of strain KACC42780. The 88.4-kb mitochondrial genome contained 35 genes. Well-developed wood degrading machinery with strong potential for lignin degradation (69 auxiliary activities, formerly FOLymes) and carbohydrate degradation (392 CAZymes), along with 58 alcohol dehydrogenase genes were highly expressed in the mycelium, demonstrating the potential application of this organism to bioethanol production. Thus, the newly uncovered wood degrading capacity and sequential nature of this process in F. velutipes, offer interesting possibilities for more detailed studies on either lignin or (hemi-) cellulose degradation in complex wood substrates. The mutual interest in wood degradation by the mushroom industry and (ligno-)cellulose biomass related industries further increase the significance of F. velutipes as a new model. PMID:24714189

Can organic matter hide from decomposers in the labyrinth of soil aggregates? Micro-engineered Soil Chips challenging foraging fungi

NASA Astrophysics Data System (ADS)

Hammer, Edith C.; Aleklett, Kristin; Arellano Caicedo, Carlos G.; Bengtsson, Martin; Micaela Mafla Endara, Paola; Ohlsson, Pelle

2017-04-01

From the point of view of microorganisms, the soil environment is an enormously complex labyrinth with paths and dead-end streets, where resources and shelters are unevenly distributed. We study foraging strategies of soil organisms, especially fungi, and the possibility of physio-spatial stabilization of organic matter by "hiding" in occluded soil spaces. We manipulate growth habitat microstructure with lab-on-a-chip techniques, where we designed complex environments with channels and obstacle at dimensions of the size of hyphae, and construct them in the transparent, gas-permeable polymer PDMS. We fill those with different nutrient solutions or combine with mineral nutrient gradients, and inoculate them with soil organisms. We analyze organisms and substrates with microscopy, fluorescence microscopy and analytical chemistry. We compared different soil litter decomposers and an arbuscular mycorrhizal fungus for their ability to forage through complex air-gap structures and attempt to classify them into functional traits concerning their mycelium directionality, space-exploring approach and ability to grow through acute angles and narrow constrictions. We identified structures which are very difficult to penetrate for most species, and compounds located behind such features may thus be spatially unavailable for decomposers. We discuss our approach in comparison to soil pore space tomographic analyses and findings we made in the pore space of colonized wood biochar.

Use of Green Fluorescent Protein-Transgenic Strains to Study Pathogenic and Nonpathogenic Lifestyles in Colletotrichum acutatum.

PubMed

Horowitz, Sigal; Freeman, Stanley; Sharon, Amir

2002-07-01

ABSTRACT Colletotrichum acutatum, which causes anthracnose disease on strawberry, can also persist on several other plant species without causing disease symptoms. The genetic and molecular bases that determine pathogenic and nonpathogenic lifestyles in C. acutatum are unclear. We developed a transformation system for C. acutatum by electroporation of germinating conidia, and transgenic isolates that express the green fluorescent protein (GFP) were produced. Details of the pathogenic and nonpathogenic lifestyles of C. acutatum were determined by using GFP-transgenic isolates. Major differences between colonization-mediating processes of strawberry and of other plants were observed. On the main host, strawberry, the germinating conidia formed branched, thick hyphae, and large numbers of appressoria were produced that were essential for plant penetration. In strawberry, the fungus developed rapidly, filling the mesophyll with dense mycelium that invaded the cells and caused necrosis of the tissue. In nonpathogenic interactions on pepper, eggplant, and tomato, the conidia germinated, producing thin, straight germ tubes. Appressoria were produced but failed to germinate and penetrate leaf tissue, resulting in epiphytic growth without invasion of the plant. Penetration of the plant occurred only several days after inoculation and was restricted to the intercellular spaces of the first cell layers of infected tissue without causing any visible damage. Much of the new fungal biomass continued to develop on the surface of inoculated organs in the nonpathogenic interaction. The differences in fungal development on strawberry compared with the other plant species suggest that signal molecules, which may be present only in strawberry, trigger appressorial germination and penetration of the primary host.

Pahoehoe toe dimensions, morphology, and branching relationships at Mauna Ulu, Kilauea Volcano, Hawai'i

NASA Astrophysics Data System (ADS)

Crown, David A.; Baloga, Stephen M.

Pahoehoe toe dimensions, morphology, and branching relationships were analyzed in flows emplaced during 1972 at Mauna Ulu, a satellitic shield on the east rift zone of Kilauea Volcano, Hawai'i. In order to characterize regions within flow fields dominated by networks of pahoehoe toes, measurements of toe length, width, thickness, and orientation were completed for 445 toes at 13 sites. Variations in site characteristics, including slope, substrate, and position in the flow field allow an evaluation of the effects of such parameters on toe dimensions. Toe surface morphology (ropy or smooth), local flow lobe position (interior or margin), and connective relationships between toes were documented in the form of detailed toe maps. These maps show the number of branches connecting a given toe to other toes in its local pahoehoe network and illustrate branching patterns. Statistical analyses of toe dimensions and comparisons of pahoehoe toe study sites and sub-populations combined with field observations, evaluation of toe maps, and qualitative examination of toe dimension size distributions show the following: (a) Although there are significant variations at a given site, toes typically have mean lengths (101cm) greater than mean widths (74cm) and mean widths greater than mean thicknesses (19cm) sites that have mean widths greater than mean lengths are those with lower slopes. (b) Where significant site-to-site variations in mean values of a given toe dimension were apparent, these differences could not be directly related to site characteristics. (c) Ropy toes have significantly larger mean values of length, width, and number of branches than smooth toes, and toes with three or more branches have greater lengths, widths, and thicknesses than toes with two or fewer branches, suggesting concentration of flow in these toe types. (d) The skewness of all size distributions of toe length and width to larger values suggests that toes are transitional to larger sheets and channels, consistent with field observations; and (e) Two distinct types of branching patterns (called monolayer and centrally ridged) were observed in preserved pahoehoe flow lobes. The significant variability in measured toe dimensions at Mauna Ulu suggests that toe dimensions are influenced by numerous locally defined, random factors, and that an approach based on stochastic methods can be used to model pahoehoe flow emplacement.

Study of anisotropy, magnetization reversal and damping in ultrathin Co films on MgO (0 0 1) substrate

NASA Astrophysics Data System (ADS)

Mallik, Srijani; Bedanta, Subhankar

2018-01-01

Ultrathin Co films of 3 nm thickness have been prepared on MgO (0 0 1) substrate in presence or absence of substrate pre-annealing. Uniaxial anisotropy is induced in the samples due to the deposition under oblique angle of incidence. Along with the oblique deposition induced anisotropy, another uniaxial anisotropy contribution has been observed due to pre-annealing. However, no cubic anisotropy has been observed here as compared to the thicker films. Angle dependent ferromagnetic resonance (FMR) measurement confirms the presence of two anisotropies in the pre-annealed sample with ∼18° misalignment with each other. The two anisotropy constants were calculated from both superconducting quantum interference device (SQUID) magnetometry and FMR spectroscopy. The magnetization reversal is governed by nucleation dominated aftereffect followed by domain wall motion for the pre-annealed sample. Branched domains are observed for the sample prepared without pre-annealing which indicates grain disorientation of Co. However, in the thicker (25 nm) Co films ripple domains were observed in contrary to ultrathin (3 nm) films.

Phonon dynamics of graphene on metals

NASA Astrophysics Data System (ADS)

Taleb, Amjad Al; Farías, Daniel

2016-03-01

The study of surface phonon dispersion curves is motivated by the quest for a detailed understanding of the forces between the atoms at the surface and in the bulk. In the case of graphene, additional motivation comes from the fact that thermal conductivity is dominated by contributions from acoustic phonons, while optical phonon properties are essential to understand Raman spectra. In this article, we review recent progress made in the experimental determination of phonon dispersion curves of graphene grown on several single-crystal metal surfaces. The two main experimental techniques usually employed are high-resolution electron energy loss spectroscopy (HREELS) and inelastic helium atom scattering (HAS). The different dispersion branches provide a detailed insight into the graphene-substrate interaction. Softening of optical modes and signatures of the substrate‧s Rayleigh wave are observed for strong graphene-substrate interactions, while acoustic phonon modes resemble those of free-standing graphene for weakly interacting systems. The latter allows determining the bending rigidity and the graphene-substrate coupling strength. A comparison between theory and experiment is discussed for several illustrative examples. Perspectives for future experiments are discussed.

Excavated substrate modulates growth instability during nest building in ants

PubMed Central

Toffin, Etienne; Kindekens, Jonathan; Deneubourg, Jean-Louis

2010-01-01

In social insects, the nests of the same species can show a large difference in size and shape. Despite these large variations, the nests share the same substructures, some appearing during nest growth. In ants, the interplay between nest size and digging activity leads to two successive morphological transitions from circular to branched shapes (budding along the perimeter of the circular cavity and tunnelling of the galleries). Like several other self-organized collective behaviours, this phenomenon, as well as the entire nest-digging process, is thought to be modulated by environmental properties. The present study investigates the effect of excavated substrate on the nest morphogenesis and the morphological transitions by using two materials with different cohesions. Here, we show that the two morphological transitions occur more frequently with a cohesive substrate than with a granular one: 96 per cent of cohesive experiments showed both transitions, whereas only 50 per cent did in granular experiments. We found that transitions and excavation cessation follow area–response thresholds: the shape transitions take place and the digging activity stops when the dug area reaches the corresponding threshold values. The shape transition thresholds are lower with the cohesive substrate and that of stopping digging is independent of nest shape and material. According to simulations, the experimental frequencies of transitions found their origin in the competition between transitions and activity cessation and in the difference between the transition threshold values of each substrate. Our results demonstrate how the substrate properties modulate the collective response and lead to various patterns. Considering the non-specific mechanisms at work, such effects of substrate coarseness have their counterparts in various collective behaviours, generating alternative patterns to colonize and exploit the environment. PMID:20410036

Stationary nanoliter droplet array with a substrate of choice for single adherent/nonadherent cell incubation and analysis.

PubMed

Shemesh, Jonathan; Ben Arye, Tom; Avesar, Jonathan; Kang, Joo H; Fine, Amir; Super, Michael; Meller, Amit; Ingber, Donald E; Levenberg, Shulamit

2014-08-05

Microfluidic water-in-oil droplets that serve as separate, chemically isolated compartments can be applied for single-cell analysis; however, to investigate encapsulated cells effectively over prolonged time periods, an array of droplets must remain stationary on a versatile substrate for optimal cell compatibility. We present here a platform of unique geometry and substrate versatility that generates a stationary nanodroplet array by using wells branching off a main microfluidic channel. These droplets are confined by multiple sides of a nanowell and are in direct contact with a biocompatible substrate of choice. The device is operated by a unique and reversed loading procedure that eliminates the need for fine pressure control or external tubing. Fluorocarbon oil isolates the droplets and provides soluble oxygen for the cells. By using this approach, the metabolic activity of single adherent cells was monitored continuously over time, and the concentration of viable pathogens in blood-derived samples was determined directly by measuring the number of colony-formed droplets. The method is simple to operate, requires a few microliters of reagent volume, is portable, is reusable, and allows for cell retrieval. This technology may be particularly useful for multiplexed assays for which prolonged and simultaneous visual inspection of many isolated single adherent or nonadherent cells is required.

Two Tropinone Reductases with Distinct Stereospecificities from Cultured Roots of Hyoscyamus niger1

PubMed Central

Hashimoto, Takashi; Nakajima, Keiji; Ongena, Godelieve; Yamada, Yasuyuki

1992-01-01

Tropinone is an alkamine intermediate at the branch point of biosynthetic pathways leading to various tropane alkaloids. Two stereospecifically distinct NADPH-dependent oxidoreductases, TR-I and TR-II, which, respectively, reduce tropinone to 3α-hydroxytropane (tropine) and 3β-hydroxytropane (ψ-tropine), were detected mainly in the root of tropane alkaloid-producing plants but not in nonproducing cultured root. Both reductases were purified to near homogeneity from cultured root of Hyoscyamus niger and characterized. The TR-I reaction was reversible, whereas the TR-II reaction was essentially irreversible, reduction of the ketone being highly favored over oxidation of the alcohol ψ-tropine. Marked differences were found between the two reductase in their affinities for tropinone substrate and in the effects of amino acid modification reagents. Some differences in substrate specificity were apparent. For example, N-propyl-4-piperidone was reduced by TR-II but not by TR-I. Conversely, 3-quinuclidinone and 8-thiabicyclo[3,2,1]octane-3-one were accepted as substrates by TR-I but hardly at all by TR-II. Both enzymes were shown to be class B oxidoreductases, which transfer the pro-S hydrogen of NAD(P)H to their substrates. Possible roles of these tropinone reductases in alkaloid biosynthesis are discussed. Images Figure 6 PMID:16653065

Stationary nanoliter droplet array with a substrate of choice for single adherent/nonadherent cell incubation and analysis

PubMed Central

Shemesh, Jonathan; Ben Arye, Tom; Avesar, Jonathan; Kang, Joo H.; Fine, Amir; Super, Michael; Meller, Amit; Ingber, Donald E.; Levenberg, Shulamit

2014-01-01

Microfluidic water-in-oil droplets that serve as separate, chemically isolated compartments can be applied for single-cell analysis; however, to investigate encapsulated cells effectively over prolonged time periods, an array of droplets must remain stationary on a versatile substrate for optimal cell compatibility. We present here a platform of unique geometry and substrate versatility that generates a stationary nanodroplet array by using wells branching off a main microfluidic channel. These droplets are confined by multiple sides of a nanowell and are in direct contact with a biocompatible substrate of choice. The device is operated by a unique and reversed loading procedure that eliminates the need for fine pressure control or external tubing. Fluorocarbon oil isolates the droplets and provides soluble oxygen for the cells. By using this approach, the metabolic activity of single adherent cells was monitored continuously over time, and the concentration of viable pathogens in blood-derived samples was determined directly by measuring the number of colony-formed droplets. The method is simple to operate, requires a few microliters of reagent volume, is portable, is reusable, and allows for cell retrieval. This technology may be particularly useful for multiplexed assays for which prolonged and simultaneous visual inspection of many isolated single adherent or nonadherent cells is required. PMID:25053808

Metabolic remodeling of substrate utilization during heart failure progression.

PubMed

Chen, Liang; Song, Jiangping; Hu, Shengshou

2018-05-23

Heart failure (HF) is a clinical syndrome caused by a decline in cardiac systolic or diastolic function, which leaves the heart unable to pump enough blood to meet the normal physiological requirements of the human body. It is a serious disease burden worldwide affecting nearly 23 million patients. The concept that heart failure is "an engine out of fuel" has been generally accepted and metabolic remodeling has been recognized as an important aspect of this condition; it is characterized by defects in energy production and changes in metabolic pathways involved in the regulation of essential cellular functions such as the process of substrate utilization, the tricarboxylic acid cycle, oxidative phosphorylation, and high-energy phosphate metabolism. Advances in second-generation sequencing, proteomics, and metabolomics have made it possible to perform comprehensive tests on genes and metabolites that are crucial in the process of HF, thereby providing a clearer and comprehensive understanding of metabolic remodeling during HF. In recent years, new metabolic changes such as ketone bodies and branched-chain amino acids were demonstrated as alternative substrates in end-stage HF. This systematic review focuses on changes in metabolic substrate utilization during the progression of HF and the underlying regulatory mechanisms. Accordingly, the conventional concepts of metabolic remodeling characteristics are reviewed, and the latest developments, particularly multi-omics studies, are compiled.

An Unusual Fatty Acyl:Adenylate Ligase (FAAL)-Acyl Carrier Protein (ACP) Didomain in Ambruticin Biosynthesis.

PubMed

Hemmerling, Franziska; Lebe, Karen E; Wunderlich, Johannes; Hahn, Frank

2018-03-08

The divinylcyclopropane (DVC) fragment of the ambruticins is proposed to be formed by a unique polyene cyclisation mechanism, in which the unusual didomain AmbG plays a key role. It is proposed to activate the branched thioester carboxylic acid resulting from polyene cyclisation and to transfer it to its associated acyl carrier protein (ACP). After oxidative decarboxylation, the intermediate is channelled back into polyketide synthase (PKS) processing. AmbG was previously annotated as an adenylation-thiolation didomain with a very unusual substrate selectivity code but has not yet been biochemically studied. On the basis of sequence and homology model analysis, we reannotate AmbG as a fatty acyl:adenylate ligase (FAAL)-acyl carrier protein didomain with unusual substrate specificity. The expected adenylate-forming activity on fatty acids was confirmed by in vitro studies. AmbG also adenylates a number of structurally diverse carboxylic acids, including functionalised fatty acids and unsaturated and aromatic carboxylic acids. HPLC-MS analysis and competition experiments show that AmbG preferentially acylates its ACP with long-chain hydrophobic acids and tolerates a π system and a branch near the carboxylic acid. AmbG is the first characterised example of a FAAL-ACP didomain that is centrally located in a PKS and apparently activates a polyketidic intermediate. This is an important step towards deeper biosynthetic studies such as partial reconstitution of the ambruticin pathway to elucidate DVC formation. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Ubiquitin-dependent Protein Degradation at the Yeast Endoplasmic Reticulum and Nuclear Envelope

PubMed Central

Zattas, Dimitrios; Hochstrasser, Mark

2014-01-01

The endoplasmic reticulum (ER) is the primary organelle in eukaryotic cells where membrane and secreted proteins are inserted into or across cell membranes. Its membrane bilayer and luminal compartments provide a favorable environment for the folding and assembly of thousands of newly synthesized proteins. However, protein folding is intrinsically error-prone, and various stress conditions can further increase levels of protein misfolding and damage, particularly in the ER, which can lead to cellular dysfunction and disease. The ubiquitin-proteasome system (UPS) is responsible for the selective destruction of a vast array of protein substrates, either for protein quality control or to allow rapid changes in the levels of specific regulatory proteins. In this review, we will focus on the components and mechanisms of ER-associated protein degradation (ERAD), an important branch of the UPS. ER membranes extend from subcortical regions of the cell to the nuclear envelope, with its continuous outer and inner membranes; the nuclear envelope is a specialized subdomain of the ER. ERAD presents additional challenges to the UPS beyond those faced with soluble substrates of the cytoplasm and nucleus. These include recognition of sugar modifications that occur in the ER, retrotranslocation of proteins across the membrane bilayer, and transfer of substrates from the ER extraction machinery to the proteasome. Here we review characteristics of ERAD substrate degradation signals (degrons), mechanisms underlying substrate recognition and processing by the ERAD machinery, and ideas on the still unresolved problem of how substrate proteins are moved across and extracted from the ER membrane. PMID:25231236

Observation and identification of wood decay fungi from the heartwood of peach tree limbs in central Georgia, USA

USDA-ARS?s Scientific Manuscript database

Peach tree health, longevity, and limb strength can be affected by wood decay fungi, but the extent of the colonization and their identities have not been characterized in Georgia, the United States of America. In an incubation experiment, dense white fungal mycelium grew on the cross-cut faces of a...

Effect on Grass and Cereal Seedlings of Hydrogen Cyanide Produced by Mycelium and Sporophores of Marasmius oreades

Treesearch

T. H. Filer

1966-01-01

Hydrogen cyanide caused root damage to Poa pratensis, Festuca rubra, and Agrostis tenuis when the grasses were suspended 7 days above a 28-day-old viable culture Marasmius oreades (Bolt. ) Fr. The amount of HCN produced by this culture (Washington isolate 1) averaged 49 ppm per 24 hours collected in 5 ml of...

Resources and testing of endophyte-infected germplasm in national grass repository collections

Treesearch

A. D. Wilson

1996-01-01

Clavicipitaceous endophytes have been known to exist in grasses since the discovery of an endophyte in seeds of damel (Lolium temulentum L.) by Vogl in 1898 (26). The oldest known specimens of damel with endophytic mycelium were seeds retrieved from a pharoah's tomb in an Egyptian pyramid dating back to 3400 B.C. (16). Subsequent work by...

Relation of soil nutrients and light to prevalence of mycorrhizae on pine seedlings

Treesearch

Edward Hacskaylo; Albert G., Jr. Snow

1959-01-01

Odd relationships abound among the wonders of Nature. Just as men keep cows, some ants herd aphids. The bee pays Nature for its food by pollinating the flower. One symbiotic relationship that concerns foresters is the coupling of tree roots with certain fungi to form compound structures, part tree root and part fungus mycelium, which we call mycorrhizae.

A Lectin Purified from Blood Red Bracket Mushroom, Pycnoporus sanguineus (Agaricomycetidae), Mycelium Displayed Affinity Toward Bovine Transferrin.

PubMed

Albores, Silvana; Moros, Maria; Cerdeiras, Maria Pia; de la Fuente, Jesus Martinez; Grazu, Valeria; Fraguas, Laura Franco

2016-01-01

Fungal lectins constitute excellent ligands for development of affinity adsorbents useful in affinity chromatography. In this work, a lectin was purified from Pycnoporus sanguineus (PSL) mycelium using 3 procedures: by affinity chromatography, using magnetic galactosyl-nanoparticles or galactose coupled to Sepharose, and by ionic exchange chromatography (IEC). The highest lectin yield was achieved by IEC (55%); SDS-PAGE of PSL showed 2 bands with molecular mass of 68.7 and 55.2 kDa and IEC displayed 2 bands at pi 5.5 and 5.2. The lectin agglutinates rat erythrocytes, exhibiting broad specificity toward several monosaccharides, including galactose. The agglutination was also inhibited by the glycoproteins fetal calf fetuin, bovine lactoferrin, bovine transferrin, and horseradish peroxidase. The lectin was then used to synthesize an affinity adsorbent (PSL-Sepharose) and the interaction with glycoproteins was evaluated by analyzing their chromatographic behaviors. The strongest interaction with the PSL-derivative was observed with transferrin, although lower interactions were also displayed toward fetuin and lactoferrin. These results indicate that the purified PSL constitutes an interesting ligand for the design of affinity adsorbents to be used (i.e., in glycoprotein purification).

Transcriptomic analysis of the dimorphic transition of Ustilago maydis induced in vitro by a change in pH.

PubMed

Martínez-Soto, Domingo; Ruiz-Herrera, José

2013-01-01

Dimorphism is the property of fungi to grow as budding yeasts or mycelium, depending on the environmental conditions. This phenomenon is important as a model of differentiation in eukaryotic organisms, and since a large number of fungal diseases are caused by dimorphic fungi, its study is important for practical reasons. In this work, we examined the transcriptome during the dimorphic transition of the basidiomycota phytopathogenic fungus Ustilago maydis using microarrays, utilizing yeast and mycelium monomorphic mutants as controls. This way, we thereby identified 154 genes of the fungus that are specifically involved in the dimorphic transition induced by a pH change. Of these, 82 genes were up-regulated, and 72 were down-regulated. Differential categorization of these genes revealed that they mostly belonged to the classes of metabolism, cell cycle and DNA processing, transcription and protein fate, transport and cellular communication, stress, cell differentiation and biogenesis of cellular components, while a significant number of them corresponded to unclassified proteins. The data reported in this work are important for our understanding of the molecular bases of dimorphism in U. maydis, and possibly of other fungi. Copyright © 2013. Published by Elsevier Inc.

Selected phytotoxins and organic extracts from endophytic fungus Edenia gomezpompae as light reaction of photosynthesis inhibitors.

PubMed

Macías-Rubalcava, Martha Lydia; Ruiz-Velasco Sobrino, María Emma; Meléndez-González, Claudio; King-Díaz, Beatriz; Lotina-Hennsen, Blas

2014-09-05

In a search for natural herbicides, we investigated the action mechanism of the naphthoquinone spiroketals, isolated from the endophytic fungus Edenia gomezpompae: preussomerins EG1 (1) and EG4 (2), and palmarumycins CP17 (3), and CP2 (4) on the photosynthesis light reactions. The naphthoquinone spiroketals 1-4 inhibited the ATP synthesis in freshly lysed spinach thylakoids from water to MV, and they also inhibited the non-cyclic electron transport in the basal, phosphorylating and uncoupled conditions from water to MV. Therefore, they act as Hill reaction inhibitors. The results suggested that naphthoquinone spiroketals 1-4 have two interactions and inhibition site on the PSII electron transport chain. The first one involves the water splitting enzyme inhibition; and, the second on the acceptor site of PSII in a similar way that herbicide Diuron, studied by polaroghaphy and corroborated by fluorescence of the chlorophyll a of PSII. The culture medium and mycelium organic extracts from four morphological variants of E. gomezpompae were phytotoxic, and the culture medium extracts were more potent than mycelium extracts. They also act as Hill reaction inhibitors. Copyright © 2014 Elsevier B.V. All rights reserved.

X-ray absorption near-edge structure micro-spectroscopy study of vanadium speciation in Phycomyces blakesleeanus mycelium.

PubMed

Žižić, Milan; Dučić, Tanja; Grolimund, Daniel; Bajuk-Bogdanović, Danica; Nikolic, Miroslav; Stanić, Marina; Križak, Strahinja; Zakrzewska, Joanna

2015-09-01

Vanadium speciation in the fungus Phycomyces blakesleeanus was examined by X-ray absorption near-edge structure (XANES) spectroscopy, enabling assessment of oxidation states and related molecular symmetries of this transition element in the fungus. The exposure of P. blakesleeanus to two physiologically important vanadium species (V(5+) and V(4+)) resulted in the accumulation of this metal in central compartments of 24 h old mycelia, most probably in vacuoles. Tetrahedral V(5+), octahedral V(4+), and proposed intracellular complexes of V(5+) were detected simultaneously after addition of a physiologically relevant concentration of V(5+) to the mycelium. A substantial fraction of the externally added V(4+) remained mostly in its original form. However, observable variations in the pre-edge-peak intensities in the XANES spectra indicated intracellular complexation and corresponding changes in the molecular coordination symmetry. Vanadate complexation was confirmed by (51)V NMR and Raman spectroscopy, and potential binding compounds including cell-wall constituents (chitosan and/or chitin), (poly)phosphates, DNA, and proteins are proposed. The evidenced vanadate complexation and reduction could also explain the resistance of P. blakesleeanus to high extracellular concentrations of vanadium.

Sequential Formation and Accumulation of Primary and Secondary Shunt Metabolic Products in Claviceps purpurea1

PubMed Central

Taber, W. A.

1964-01-01

The fungus Claviceps purpurea was grown on a rich and a limited nutrient medium such that alkaloid was produced after 8 days on the former medium and after 3 days on the latter medium. Cultures grown on both were assayed for the primary shunt metabolic products, polyols, trehalose, lipids, ribonucleic acid, and polyphosphate, and the secondary metabolic product, ergot alkaloid. Although differing considerably in composition, the two media nevertheless allowed formation of both primary and secondary shunt products. In both instances, however, the secondary product, ergot alkaloid, did not form until formation and accumulation of the primary products had ceased and the mycelial content of these products was actually decreasing. In both instances, alkaloid formation took place after the total dry weight of the mycelium had begun to decrease but while the dry weight of the residual, or structural portion of the mycelium, was either constant or increasing. The dilution of labeling in mannitol isolated from mycelia grown on rich medium containing 1,6-C14-labeled mannitol was 2.2. Thus, about half of the mycelial mannitol was actually mannitol which had been taken up directly from the medium. PMID:14199021

Currency notes and coins as a possible source of transmitting fungal pathogens of man and plants.

PubMed

Wanule, Dinesh; Jalander, Vaghmare; Gachande, B D; Sirsikar, A N

2011-10-01

Currency (notes and coins) handling by people during transaction is one of the most mobile objects within the community, which has a potential of transmitting pathogens. A survey carried out recently in Nanded city (Maharashtra) revealed heavy contamination of currency notes and coins by important fungal pathogens of plants and man, i.e. Aspergillus niger (60.37%), A. flavus (3.98%), A.nidulans (0.2%), Penicillium citrinum (17.80%), Alternaria tenuis (0.20%), Curvularia pallescens (0.20%), Cladosporium cladosporioides (10.69%), Rhizopus stolonifer (1.04%), an unidentified Aspergillus species .1 (0.20%) and another unidentified Aspergillus species.2 (3.14%), Fusarium sp. (0.20%), Trichoderma viride (0.20%),white sterile mycelium (0.62%) and brown sterile mycelium (0.62%). The study highlights the importance of preventing and controlling fungal contamination of currency notes and coins in public health and plant protection. Currency notes or coins are rarely suspected as infection sources and often not quarantined at airport or seaport terminal. Possible transmission of pathogens or "alien", invasive species through currency across borders or across countries needs to be taken into consideration especially under circumstances of serious outbreak of important disease or when there is a threat of biological warfare.

Cloning and Expression Analysis of Phenylalanine Ammonia-Lyase Gene in the Mycelium and Fruit Body of the Edible Mushroom Flammulina velutipes

PubMed Central

Yun, Yeo Hong; Koo, Ja Sun

2015-01-01

Phenylalanine ammonia-lyase (PAL) gene is known to be expressed in plants, and is involved in the differentiation, growth and synthesis of secondary metabolites. However, its expression in fungi remains to be explored. To understand its expression in mushroom fungi, the PAL gene of the edible mushroom Flammulina velutipes (Fvpal) was cloned and characterized. The cloned Fvpal consists of 2,175 bp, coding for a polypeptide containing 724 amino acids and having 11 introns. The translated amino acid sequence of Fvpal shares a high identity (66%) with that of ectomycorrhizal fungus Tricholoma matsutake. Distinctively, the Fvpal expression in the mycelium was higher in minimal medium supplemented with L-tyrosine than with other aromatic amino acids. During cultivation of the mushroom on sawdust medium, Fvpal expression in the fruit body correspondingly increased as the mushroom grew. In the fruiting body, Fvpal was expressed more in the stipe than in the pileus. These results suggest that F. velutipes PAL activity differs in the different organs of the mushroom. Overall, this is first report to show that the PAL gene expression is associated with mushroom growth in fungi. PMID:26539050

Isolation and Characterization of Lectins Formed by Cerrena unicolor (Higher Basidiomycetes) in Solid-State Fermentation of Sorghum and Wheat Straw.

PubMed

Davitashvili, Elene; Kapanadze, Ekaterine; Kachlishvili, Eva; Mikiashvili, Nona A; Elisashvili, Vladimir

2015-01-01

The capability of Cerrena unicolor to produce fruiting bodies and lectins was studied in solid-state fermentation of a sorghum and wheat straw mixture. The first primordia appeared on day 48 and reached 6-10 mm; however, no formation of fruiting bodies occurred and these rudiments were harvested on day 55. The protein content in the rudiment extracts was significantly higher, whereas the specific hemagglutinating activity (HA) was sixfold lower as compared with those in extracts from mycelial biomass. Moreover, the specific HA of the 80-day mycelium increased to 16,667 U/mg, exceeding by sixfold that of 55-day-old mycelium. Four protein fractions (160, 105, 67, and 8 kDa) were detected by gel-chromatography of mycelial biomass crude extract; the highest specific HA was revealed in fraction III (26336 U HA/mg). Among sugars tested, galactose was the most potent inhibitor of HA of all protein fractions, with minimal inhibition concentrations of 0.095-0.780 mM. The galactose-specific lectins isolated from the fractions II and III by affinity chromatography ranged from 15 to 116 kDa and differed with kinetic parameters.

Photoelectrocatalytic activity of a hydrothermally grown branched Zno nanorod-array electrode for paracetamol degradation.

PubMed

Lin, Chin Jung; Liao, Shu-Jun; Kao, Li-Cheng; Liou, Sofia Ya Hsuan

2015-06-30

Hierarchical branched ZnO nanorod (B-ZnR) arrays as an electrode for efficient photoelectrocatalytic degradation of paracetamol were grown on fluorine-doped tin oxide substrates using a solution route. The morphologic and structural studies show the ZnO trunks are single-crystalline hexagonal wurtzite ZnO with a [0001] growth direction and are densely covered by c-axis-oriented ZnO branches. The obvious enhancement in photocurrent response of the B-ZnR electrode was obtained than that in the ZnO nanoparticle (ZnO NP) electrode. For the photoelectrocatalytic degradation of paracetamol in 20 h, the conversion fraction of the drug increased from 32% over ZnO NP electrode to 62% over B-ZnR arrays with about 3-fold increase in initial reaction rate. The light intensity-dependent photoelectrocatalytic experiment indicated that the superior performance over the B-ZnR electrode was mainly ascribed to the increased specific surface area without significantly sacrificing the charge transport and pollutant diffusion efficiencies. Two aromatic intermediate compounds were observed and eventually converted into harmless carboxylic acids and ammonia. Hierarchical tree-like ZnO arrays can be considered effective alternatives to improve photoelectro degradation rates without the need for expensive additives. Copyright © 2015 Elsevier B.V. All rights reserved.

Interactive effects of substrate, hydroperiod, and nutrients on seedling growth of Salix nigra and Taxodium distichum

USGS Publications Warehouse

Day, Richard H.; Doyle, T.W.; Draugelis-Dale, R. O.

2006-01-01

The large river swamps of Louisiana have complex topography and hydrology, characterized by black willow (Salix nigra) dominance on accreting alluvial sediments and vast areas of baldcypress (Taxodium distichum) deepwater swamps with highly organic substrates. Seedling survival of these two wetland tree species is influenced by their growth rate in relation to the height and duration of annual flooding in riverine environments. This study examines the interactive effects of substrate, hydroperiod, and nutrients on growth rates of black willow and baldcypress seedlings. In a greenhouse experiment with a split-split-plot design, 1-year seedlings of black willow and baldcypress were subjected to two nutrient treatments (unfertilized versus fertilized), two hydroperiods (continuously flooded versus twice daily flooding/draining), and two substrates (sand versus commercial peat mix). Response variables included height, diameter, lateral branch count, biomass, and root:stem ratio. Black willow growth in height and diameter, as well as all biomass components, were significantly greater in peat substrate than in sand. Black willow showed a significant hydroperiod-nutrient interaction wherein fertilizer increased stem and root biomass under drained conditions, but flooded plants did not respond to fertilization. Baldcypress diameter and root biomass were higher in peat than in sand, and the same two variables increased with fertilization in flooded as well as drained treatments. These results can be used in Louisiana wetland forest models as inputs of seedling growth and survival, regeneration potential, and biomass accumulation rates of black willow and baldcypress.

Isolation and growth characteristics of an EDTA-degrading member of the alpha-subclass of Proteobacteria.

PubMed

Weilenmann, Hans-Ueli; Engeli, Barbara; Bucheli-Witschel, Margarete; Egli, Thomas

2004-10-01

A Gram-negative, ethylenediaminetetraacetic acid (EDTA)-degrading bacterium (deposited at the German Culture Collection as strain DSM 9103) utilising EDTA as the only source of carbon, energy and nitrogen was isolated from a mixed EDTA-degrading population that was originally enriched in a column system from a mixture of activated sludge and soil. Chemotaxonomic analysis of quinones, polar lipids and fatty acids allowed allocation of the isolate to the alpha-subclass of Proteobacteria. 16S rDNA sequencing and phylogenetic analysis revealed highest similarity to the Mesorhizobium genus followed by the Aminobacter genus. However, the EDTA-degrading strain apparently forms a new branch within the Phyllobacteriaceae/Mesorhizobia family. Growth of the strain was rather slow not only on EDTA (micro(max) = 0.05 h(-1)) but also on other substrates. Classical substrate utilisation testing in batch culture suggested a quite restricted carbon source spectrum with only lactate, glutamate, and complexing agents chemically related to EDTA (nitrilotriacetate, iminodiacetate and ethylenediaminedisuccinate) supporting growth. However, when EDTA-limited continuous cultures of strain DSM 9103 were pulsed with fumarate, succinate, glucose or acetate, these substrates were assimilated immediately. Apparently, the strain can use a broader spectrum than indicated by traditional substrate testing techniques. The EDTA species CaEDTA and MgEDTA served as growth substrates of the strain because in the mineral medium employed EDTA was predicted to be mainly present in the form of these two complexes. The bacterium was not able to degrade Fe3+-complexed EDTA.

The role of the mitochondrial pyruvate carrier in substrate regulation

PubMed Central

Vacanti, Nathaniel M.; Divakaruni, Ajit S.; Green, Courtney R.; Parker, Seth J.; Henry, Robert R.; Ciaraldi, Theodore P.; Murphy, Anne N.; Metallo, Christian M.

2014-01-01

SUMMARY Pyruvate lies at a central biochemical node connecting carbohydrate, amino acid, and fatty acid metabolism, and the regulation of pyruvate flux into mitochondria represents a critical step in intermediary metabolism impacting numerous diseases. To characterize changes in mitochondrial substrate utilization in the context of compromised mitochondrial pyruvate transport, we applied 13C metabolic flux analysis (MFA) to cells after transcriptional or pharmacological inhibition of the mitochondrial pyruvate carrier (MPC). Despite profound suppression of both glucose and pyruvate oxidation, cell growth, oxygen consumption, and tricarboxylic acid (TCA) metabolism were surprisingly maintained. Oxidative TCA flux was achieved through enhanced reliance on glutaminolysis through malic enzyme and pyruvate dehydrogenase (PDH) as well as fatty acid and branched chain amino acid oxidation. Thus, in contrast to inhibition of complex I or PDH, suppression of pyruvate transport induces a form of metabolic flexibility associated with use of lipids and amino acids as catabolic and anabolic fuels. PMID:25458843

Initial reactions involved in the dissimilation of mandelate by Rhodotorula graminis.

PubMed Central

Durham, D R

1984-01-01

Rhodotorula graminis utilized DL-mandelate, L(+)-mandelate, and D(-)-mandelate as sole sources of carbon and energy. Growth on these aromatic substrates resulted in the induction of an NAD-dependent D(-)-mandelate dehydrogenase and a dye-linked L(+)-mandelate dehydrogenase, each catalyzing the stereospecific conversion of its respective enantiomer of mandelate to benzoylformate. Benzoylformate was oxidized to benzaldehyde, which was dehydrogenated to benzoate by an NAD-dependent benzaldehyde dehydrogenase. Benzoate was further metabolized through p-hydroxybenzoate and the protocatechuate branch of the beta-ketoadipate pathway. PMID:6389497

Simultaneous fluorescent detection of multiple metal ions based on the DNAzymes and graphene oxide.

PubMed

Yun, Wen; Wu, Hong; Liu, Xingyan; Fu, Min; Jiang, Jiaolai; Du, Yunfeng; Yang, Lizhu; Huang, Yu

2017-09-15

A novel fluorescent detection strategy for simultaneous detection of Cu 2+ , Pb 2+ and Mg 2+ based on DNAzyme branched junction structure with three kinds of DNAzymes and graphene oxide (GO) was presented. Three fluorophores labeled DNA sequences consisted with enzyme-strand (E-DNA) and substrate strand (S-DNA) were annealed to form DNAzyme branched junction structure. In the presence of target metal ion, the DNAzyme was activated to cleave the fluorophore labeled S-DNA. The S-DNA fragments were released and adsorbed onto GO surface to quench the fluorescent signal. The detection limit was calculated to be 1 nM for Cu 2+ , 200 nM for Mg 2+ , and 0.3 nM for Pb 2+ , respectively. This strategy was successfully used for simultaneous detection of Cu 2+ , Mg 2+ and Pb 2+ in human serum. Moreover, it had potential application for simultaneous detection of multiple metal ions in environmental and biological samples. Copyright © 2017 Elsevier B.V. All rights reserved.

Branched Chain Amino Acids: Beyond Nutrition Metabolism.

PubMed

Nie, Cunxi; He, Ting; Zhang, Wenju; Zhang, Guolong; Ma, Xi

2018-03-23

Branched chain amino acids (BCAAs), including leucine (Leu), isoleucine (Ile), and valine (Val), play critical roles in the regulation of energy homeostasis, nutrition metabolism, gut health, immunity and disease in humans and animals. As the most abundant of essential amino acids (EAAs), BCAAs are not only the substrates for synthesis of nitrogenous compounds, they also serve as signaling molecules regulating metabolism of glucose, lipid, and protein synthesis, intestinal health, and immunity via special signaling network, especially phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/AKT/mTOR) signal pathway. Current evidence supports BCAAs and their derivatives as the potential biomarkers of diseases such as insulin resistance (IR), type 2 diabetes mellitus (T2DM), cancer, and cardiovascular diseases (CVDs). These diseases are closely associated with catabolism and balance of BCAAs. Hence, optimizing dietary BCAA levels should have a positive effect on the parameters associated with health and diseases. This review focuses on recent findings of BCAAs in metabolic pathways and regulation, and underlying the relationship of BCAAs to related disease processes.

Ubiquitination dynamics in the early-branching eukaryote Giardia intestinalis

PubMed Central

Niño, Carlos A; Chaparro, Jenny; Soffientini, Paolo; Polo, Simona; Wasserman, Moises

2013-01-01

Ubiquitination is a highly dynamic and versatile posttranslational modification that regulates protein function, stability, and interactions. To investigate the roles of ubiquitination in a primitive eukaryotic lineage, we utilized the early-branching eukaryote Giardia intestinalis. Using a combination of biochemical, immunofluorescence-based, and proteomics approaches, we assessed the ubiquitination status during the process of differentiation in Giardia. We observed that different types of ubiquitin modifications present specific cellular and temporal distribution throughout the Giardia life cycle from trophozoites to cyst maturation. Ubiquitin signal was detected in the wall of mature cysts, and enzymes implicated in cyst wall biogenesis were identified as substrates for ubiquitination. Interestingly, inhibition of proteasome activity did not affect trophozoite replication and differentiation, while it caused a decrease in cyst viability, arguing for proteasome involvement in cyst wall maturation. Using a proteomics approach, we identified around 200 high-confidence ubiquitinated candidates that vary their ubiquitination status during differentiation. Our results indicate that ubiquitination is critical for several cellular processes in this primitive eukaryote. PMID:23613346

Global gene expression profiles of Phytophthora ramorum strain pr102 in response to plant host and tissue differentiation

Treesearch

Caroline M. Press; Niklaus J. Grunwald

2008-01-01

The release of the draft genome sequence of P. ramorum strain Pr102, enabled the construction of an oligonucleotide microarray of the entire genome of Pr102. The array contains 344,680 features (oligos) that represent the transcriptome of Pr102. P. ramorum RNA was extracted from mycelium and sporangia and used to compare gene...

A novel, multiplexed, probe-based quantitative PCR assay for the soybean root- and stem-rot pathogen, Phytophthora sojae, utilizes its transposable element

USDA-ARS?s Scientific Manuscript database

Phytophthora root rot of soybean (Glycine max Merr.) is caused by the oomycete Phytophthora sojae (Kaufm. and Gerd.). P. sojae has a narrow host range, consisting primarily of soybean, and it is a serious pathogen worldwide. It exists in root and stem tissues as mycelium, wherein it can form oospo...

5'-Serial Analysis of Gene Expression studies reveal a transcriptomic switch during fruiting body development in Coprinopsis cinerea

PubMed Central

2013-01-01

Background The transition from the vegetative mycelium to the primordium during fruiting body development is the most complex and critical developmental event in the life cycle of many basidiomycete fungi. Understanding the molecular mechanisms underlying this process has long been a goal of research on basidiomycetes. Large scale assessment of the expressed transcriptomes of these developmental stages will facilitate the generation of a more comprehensive picture of the mushroom fruiting process. In this study, we coupled 5'-Serial Analysis of Gene Expression (5'-SAGE) to high-throughput pyrosequencing from 454 Life Sciences to analyze the transcriptomes and identify up-regulated genes among vegetative mycelium (Myc) and stage 1 primordium (S1-Pri) of Coprinopsis cinerea during fruiting body development. Results We evaluated the expression of >3,000 genes in the two respective growth stages and discovered that almost one-third of these genes were preferentially expressed in either stage. This identified a significant turnover of the transcriptome during the course of fruiting body development. Additionally, we annotated more than 79,000 transcription start sites (TSSs) based on the transcriptomes of the mycelium and stage 1 primoridum stages. Patterns of enrichment based on gene annotations from the GO and KEGG databases indicated that various structural and functional protein families were uniquely employed in either stage and that during primordial growth, cellular metabolism is highly up-regulated. Various signaling pathways such as the cAMP-PKA, MAPK and TOR pathways were also identified as up-regulated, consistent with the model that sensing of nutrient levels and the environment are important in this developmental transition. More than 100 up-regulated genes were also found to be unique to mushroom forming basidiomycetes, highlighting the novelty of fruiting body development in the fungal kingdom. Conclusions We implicated a wealth of new candidate genes important to early stages of mushroom fruiting development, though their precise molecular functions and biological roles are not yet fully known. This study serves to advance our understanding of the molecular mechanisms of fruiting body development in the model mushroom C. cinerea. PMID:23514374

Phytotoxic and antimicrobial activity of volatile and semi-volatile organic compounds from the endophyte Hypoxylon anthochroum strain Blaci isolated from Bursera lancifolia (Burseraceae).

PubMed

Ulloa-Benítez, Á; Medina-Romero, Y M; Sánchez-Fernández, R E; Lappe-Oliveras, P; Roque-Flores, G; Duarte Lisci, G; Herrera Suárez, T; Macías-Rubalcava, M L

2016-08-01

To evaluate the phytotoxic, antifungal and antioomycete activity; and, determine the chemical composition of the volatile organic compounds (VOCs) and semi-volatile metabolites produced by the endophyte Hypoxylon anthochroum strain Blaci isolated from Bursera lancifolia. Based on its macro- and micro-morphological features, the strain Blaci was identified as Nodulisporium sp.; partial analysis of its ITS1-5.8-ITS2 ribosomal gene sequence revealed the identity of the teleomorphic stage of the fungus as H. anthochroum. Phytotoxic and antimicrobial activities of VOCs, and culture medium and mycelium organic extracts from H. anthochroum Blaci were determined by simple and multiple antagonism bioassays, and gas phase and agar dilution bioassays respectively. The volatile and semi-volatile metabolites were identified by gas chromatography-mass spectrometry. VOCs from a 5-day H. anthochroum strain Blaci culture caused the inhibition of seed germination, root elongation and seedling respiration on Amaranthus hypochondriacus, Panicum miliaceum, Trifolium pratense and Medicago sativa. In addition, extracts, phenylethyl alcohol and eucalyptol main compounds present in the VOCs and extract displayed a high phytotoxic activity, inhibiting the three physiological processes on the four test plants in a concentration-dependent manner. The results revealed that H. anthochroum strain Blaci produces a mixture of VOCs. These VOCs showed a strong phytotoxic activity on seed germination, root elongation, and seedling respiration of four plants and slightly affected the growth of phytopathogenic fungi and oomycetes. Also, the culture medium and mycelium extracts of H. anthochroum showed a high phytotoxic activity on the four test plants and, generally, the culture medium extract was more phytotoxic than the mycelium extracts. This work firstly reports the phytotoxic activity of volatile and semi-volatile compounds produced by the endophyte H. anthochroum strain Blaci on seed germination, root elongation, and seedling respiration of four different plants; consequently, these compounds could be useful in biocontrol of weeds and plant pathogens. Journal of Applied Microbiology © 2016 The Society for Applied Microbiology.

Modeling soil organic matter reallocation in soil enhanced by fungal growth

NASA Astrophysics Data System (ADS)

Battaïa, G.; Falconer, R. E.; Otten, W.

2012-04-01

Soil, as a huge carbon reservoir having a large interface with the atmosphere, has a major role in understanding global carbon cycle. Yet, its structure gives rise to an extremely complex ecosystem in which chemical fluxes are difficult to describe. Amongst microbial organisms that inhabit soil, fungi represent an entire kingdom of life that has developed its own strategy to adapt its environment. They are thus known to have a particular importance for the reallocation of carbon (and other elements) as they are able to build a mycelium structure that can spread over several meters and through which nutrients can be translocated. This study, based on simulations, is dedicated to enlighten the role of fungal colonization to generate an ecosystem in which coexists disperse biological hotspots. The simulation environment is reconstructed from thresholded computed tomography images of soil samples. Soil organic matter acting as a resource for fungi is assumed to occur first in a particulate solid state (POM). It is degraded into dissolved organic carbon (DOC) through enzymatic activity of fungi. Fungal uptake converts DOC into an internal resource that diffuses through the mycelium and helps it for further colonization. The fungal model is an adaptation of a previously developed model. In addition to internal resource, it accounts for two states of biomass: non-insulated and insulated. One is converted into the other by insulation which is the analog of an ageing process. Being insulated, the interaction rates of the biomass with the environment (degradation and uptake) become slower and the ability to diffuse in the pore space is lost. This aims at producing a more stable state of the mycelium when all resource has been consumed. Spatially simulations reveal a transient state in POM-fungi interaction characterized by a large spread of DOC in the pore space. It is then followed by an enhanced fungal growth toward these areas. Finally a steady state occurs in which DOC is produced and consumed in a close vicinity of the POM reducing its availability for other micro-organisms.

Matrix stiffness modulates formation and activity of neuronal networks of controlled architectures.

PubMed

Lantoine, Joséphine; Grevesse, Thomas; Villers, Agnès; Delhaye, Geoffrey; Mestdagh, Camille; Versaevel, Marie; Mohammed, Danahe; Bruyère, Céline; Alaimo, Laura; Lacour, Stéphanie P; Ris, Laurence; Gabriele, Sylvain

2016-05-01

The ability to construct easily in vitro networks of primary neurons organized with imposed topologies is required for neural tissue engineering as well as for the development of neuronal interfaces with desirable characteristics. However, accumulating evidence suggests that the mechanical properties of the culture matrix can modulate important neuronal functions such as growth, extension, branching and activity. Here we designed robust and reproducible laminin-polylysine grid micropatterns on cell culture substrates that have similar biochemical properties but a 100-fold difference in Young's modulus to investigate the role of the matrix rigidity on the formation and activity of cortical neuronal networks. We found that cell bodies of primary cortical neurons gradually accumulate in circular islands, whereas axonal extensions spread on linear tracks to connect circular islands. Our findings indicate that migration of cortical neurons is enhanced on soft substrates, leading to a faster formation of neuronal networks. Furthermore, the pre-synaptic density was two times higher on stiff substrates and consistently the number of action potentials and miniature synaptic currents was enhanced on stiff substrates. Taken together, our results provide compelling evidence to indicate that matrix stiffness is a key parameter to modulate the growth dynamics, synaptic density and electrophysiological activity of cortical neuronal networks, thus providing useful information on scaffold design for neural tissue engineering. Copyright © 2016 Elsevier Ltd. All rights reserved.

Cholinephosphotransferase and Diacylglycerol Acyltransferase (Substrate Specificities at a Key Branch Point in Seed Lipid Metabolism).

PubMed

Vogel, G.; Browse, J.

1996-03-01

Many oilseed plants accumulate triacylglycerols that contain unusual fatty acyl structures rather than the common 16- and 18-carbon fatty acids found in membrane lipids of these plants. In vitro experiments demonstrate that triacylglycerols are synthesized via diacylglycerols in microsomal preparations and that this same sub-cellular fraction is the site for the synthesis of phosphatidylcholine, which in seeds is synthesized from diacylglycerol by CDP-choline: diacylglycerol cholinephosphotransferase. In microsomes from Cuphea lanceolata, a plant that accumulates fatty acids with 10 carbons and no double bonds (10:0) in its oil, the diacylglycerol acyltransferase exhibited 4-fold higher activity with 10:0/10:0 molecular species of diacylglycerol than with molecular species containing 18-carbon fatty acids. In castor bean (Ricinus communis), which accumulates oil containing ricinoleic acid, diricinoleoyldiacylglycerol was the favored substrate for triacylglycerol synthesis. In contrast to these modest specificities of the diacylglycerol acyltransferases, the cholinephosphotransferases from these plants and from safflower (Carthamus tinctorius) and rapeseed (Brassica napus) showed little or no specificity across a range of different diacylglycerol substrates. Consideration of these results and other data suggests that the targeting of unusual fatty acids to triacylglycerol synthesis and their exclusion from membrane lipids are not achieved on the basis of the diacylglycerol substrate specificities of the enzymes involved and may instead require the spatial separation of two different diacylglycerol pools.

Cholinephosphotransferase and Diacylglycerol Acyltransferase (Substrate Specificities at a Key Branch Point in Seed Lipid Metabolism).

PubMed Central

Vogel, G.; Browse, J.

1996-01-01

Many oilseed plants accumulate triacylglycerols that contain unusual fatty acyl structures rather than the common 16- and 18-carbon fatty acids found in membrane lipids of these plants. In vitro experiments demonstrate that triacylglycerols are synthesized via diacylglycerols in microsomal preparations and that this same sub-cellular fraction is the site for the synthesis of phosphatidylcholine, which in seeds is synthesized from diacylglycerol by CDP-choline: diacylglycerol cholinephosphotransferase. In microsomes from Cuphea lanceolata, a plant that accumulates fatty acids with 10 carbons and no double bonds (10:0) in its oil, the diacylglycerol acyltransferase exhibited 4-fold higher activity with 10:0/10:0 molecular species of diacylglycerol than with molecular species containing 18-carbon fatty acids. In castor bean (Ricinus communis), which accumulates oil containing ricinoleic acid, diricinoleoyldiacylglycerol was the favored substrate for triacylglycerol synthesis. In contrast to these modest specificities of the diacylglycerol acyltransferases, the cholinephosphotransferases from these plants and from safflower (Carthamus tinctorius) and rapeseed (Brassica napus) showed little or no specificity across a range of different diacylglycerol substrates. Consideration of these results and other data suggests that the targeting of unusual fatty acids to triacylglycerol synthesis and their exclusion from membrane lipids are not achieved on the basis of the diacylglycerol substrate specificities of the enzymes involved and may instead require the spatial separation of two different diacylglycerol pools. PMID:12226231

Metabolic effects of low cortisol during exercise in humans.

PubMed

Del Corral, P; Howley, E T; Hartsell, M; Ashraf, M; Younger, M S

1998-03-01

This study examined the physiological effect of reduced plasma cortisol (C) during prolonged exercise in humans. The effects of normal C (NC) were compared with metyrapone-induced low C (LC) on plasma substrate availability and the respiratory exchange ratio during 2 h of exercise at approximately 60% peak O2 consumption in nine subjects. The C responses were compared with preexercise (Pre) levels and with a rest day (Con). At rest, C was attenuated by approximately 70% for LC compared with NC. At rest, plasma glucose, lactate, glycerol, beta-hydroxybutyrate, alanine, branched-chain amino acids, insulin, glucagon, growth hormone, epinephrine, and norepinephrine were similar under LC and NC (P > 0.05). During exercise under NC, plasma C increased compared with Pre, whereas it remained unchanged during LC. During NC, plasma C was elevated at 90 min (compared with Con) and at 120 min (compared with Con and Pre). During exercise, plasma glucose decreased to the same extent and lactate was similar under both conditions, whereas plasma glycerol, beta-hydroxybutyrate, alanine, and branched-chain amino acids were higher (P < 0.01) under NC. Plasma insulin declined (P = 0.01) to a greater extent under LC, whereas growth hormone, epinephrine, and norepinephrine tended to be higher (0.05

Diacyltransferase Activity and Chain Length Specificity of Mycobacterium tuberculosis PapA5 in the Synthesis of Alkyl β-Diol Lipids

DOE Office of Scientific and Technical Information (OSTI.GOV)

Touchette, Megan H.; Bommineni, Gopal R.; Delle Bovi, Richard J.

Although classified as Gram-positive bacteria, Corynebacterineae possess an asymmetric outer membrane that imparts structural and thereby physiological similarity to more distantly related Gram-negative bacteria. Like lipopolysaccharide in Gram-negative bacteria, lipids in the outer membrane of Corynebacterineae have been associated with the virulence of pathogenic species such as Mycobacterium tuberculosis (Mtb). For example, Mtb strains that lack long, branched-chain alkyl esters known as dimycocerosates (DIMs) are significantly attenuated in model infections. The resultant interest in the biosynthetic pathway of these unusual virulence factors has led to the elucidation of many of the steps leading to the final esterification of the alkylmore » beta-diol, phthiocerol, with branched-chain fatty acids know as mycocerosates. PapA5 is an acyltransferase implicated in these final reactions. We here show that PapA5 is indeed the terminal enzyme in DIM biosynthesis by demonstrating its dual esterification activity and chain-length preference using synthetic alkyl beta-diol substrate analogues. Applying these analogues to a series of PapA5 mutants, we also revise a model for the substrate binding within PapA5. Finally, we demonstrate that the Mtb Ser/Thr kinase PknB modifies PapA5 on three Thr residues, including two (T196, T198) located on an unresolved loop. These results clarify the DIM biosynthetic pathway and suggest possible mechanisms by which DIM biosynthesis may be regulated by the post-translational modification of PapA5.« less

Type II Fatty Acid Synthesis Is Essential for the Replication of Chlamydia trachomatis*

PubMed Central

Yao, Jiangwei; Abdelrahman, Yasser M.; Robertson, Rosanna M.; Cox, John V.; Belland, Robert J.; White, Stephen W.; Rock, Charles O.

2014-01-01

The major phospholipid classes of the obligate intracellular bacterial parasite Chlamydia trachomatis are the same as its eukaryotic host except that they also contain chlamydia-made branched-chain fatty acids in the 2-position. Genomic analysis predicts that C. trachomatis is capable of type II fatty acid synthesis (FASII). AFN-1252 was deployed as a chemical tool to specifically inhibit the enoyl-acyl carrier protein reductase (FabI) of C. trachomatis to determine whether chlamydial FASII is essential for replication within the host. The C. trachomatis FabI (CtFabI) is a homotetramer and exhibited typical FabI kinetics, and its expression complemented an Escherichia coli fabI(Ts) strain. AFN-1252 inhibited CtFabI by binding to the FabI·NADH complex with an IC50 of 0.9 μm at saturating substrate concentration. The x-ray crystal structure of the CtFabI·NADH·AFN-1252 ternary complex revealed the specific interactions between the drug, protein, and cofactor within the substrate binding site. AFN-1252 treatment of C. trachomatis-infected HeLa cells at any point in the infectious cycle caused a decrease in infectious titers that correlated with a decrease in branched-chain fatty acid biosynthesis. AFN-1252 treatment at the time of infection prevented the first cell division of C. trachomatis, although the cell morphology suggested differentiation into a metabolically active reticulate body. These results demonstrate that FASII activity is essential for C. trachomatis proliferation within its eukaryotic host and validate CtFabI as a therapeutic target against C. trachomatis. PMID:24958721

Functional analysis of truncated and site-directed mutagenesis dextransucrases to produce different type dextrans.

PubMed

Wang, Chao; Zhang, Hong-Bin; Li, Meng-Qi; Hu, Xue-Qin; Li, Yao

2017-07-01

Dextrans with distinct molecular size and structure are increasingly being used in the food and pharmaceutical industries. Dextran is produced by dextransucrase (DSR, EC2.4.5.1), which is produced by Leuconostoc mesenteroides. DSR belongs to glycosyl hydrolase family (GH70) and synthesizes branched α-glucan (dextran) with both 5% α(1-3) and 95% α(1-6) glycosidic linkages. The DSR gene dex-YG (Genebank, Accession No. DQ345760) was cloned from the wild strain Leuconostoc mesenteroides 0326. This study generated a series of C-terminally truncated variants of dextransucrase and substituting the amino-acid residues in the active site of DSR. With shorter length of DSR, its polysaccharide-synthesizing capability was impaired heavily, whereas oligosaccharide (acting as prebiotics)-synthesizing capability increased significantly, efficiently producing special sizes of dextran. All truncated mutant enzymes were active. Results demonstrated that the catalytic domain dextransucrase was likely in 800 aa or less. Based on the three-dimensional structure model of dextransucrase built through homology modeling methods, the DSR and its mutants with the acceptor substrate of maltose and donor substrate of sucrose were studied by molecular-docking method. Substituting these amino-acid residues significantly affected enzyme activities. Compared with the wild-type dextran, mutant enzymes catalyzed the synthesis of a-glucan with 1-9% α(1-3) and 90-98% α(1-6) branching linkages. Some mutants introduced a small amount of α(1-4) linkages and α(1-2) linkages. This strategy can be effectively used for the rational protein design of dextransucrase. Copyright © 2017 Elsevier Inc. All rights reserved.

Current advancements and challenges in soil-root interactions modelling

NASA Astrophysics Data System (ADS)

Schnepf, Andrea; Huber, Katrin; Abesha, Betiglu; Meunier, Felicien; Leitner, Daniel; Roose, Tiina; Javaux, Mathieu; Vanderborght, Jan; Vereecken, Harry

2015-04-01

Roots change their surrounding soil chemically, physically and biologically. This includes changes in soil moisture and solute concentration, the exudation of organic substances into the rhizosphere, increased growth of soil microorganisms, or changes in soil structure. The fate of water and solutes in the root zone is highly determined by these root-soil interactions. Mathematical models of soil-root systems in combination with non-invasive techniques able to characterize root systems are a promising tool to understand and predict the behaviour of water and solutes in the root zone. With respect to different fields of applications, predictive mathematical models can contribute to the solution of optimal control problems in plant recourse efficiency. This may result in significant gains in productivity, efficiency and environmental sustainability in various land use activities. Major challenges include the coupling of model parameters of the relevant processes with the surrounding environment such as temperature, nutrient concentration or soil water content. A further challenge is the mathematical description of the different spatial and temporal scales involved. This includes in particular the branched structures formed by root systems or the external mycelium of mycorrhizal fungi. Here, reducing complexity as well as bridging between spatial scales is required. Furthermore, the combination of experimental and mathematical techniques may advance the field enormously. Here, the use of root system, soil and rhizosphere models is presented through a number of modelling case studies, including image based modelling of phosphate uptake by a root with hairs, model-based optimization of root architecture for phosphate uptake from soil, upscaling of rhizosphere models, modelling root growth in structured soil, and the effect of root hydraulic architecture on plant water uptake efficiency and drought resistance.

A mycosis-like granuloma of fish

USGS Publications Warehouse

Wood, E.M.; Yasutake, W.T.; Lehman, W.L.

1955-01-01

Mycoses of systemic distribution are rarely observed in fresh-water fish in this country. In a recent review of atypical cell growths in fishes, Nigrelli cited the only known instance of a mycetoma in a North American fresh-water fish which occurred in the head of fingerling landlocked salmon from an Idaho hatchery. The fungus associated with this granuloma was characterized by a branching septate mycelium. Rucker reported a streptomycete which was pathogenic to blueblack salmon. This organism produced internal nodules containing masses of hyphae but no inflammatory response. A pathogenic fungus has been observed frequently in marine fish, however, and in both marine and fresh-water fish in Europe. This organism was tentatively classified as a Phycomycete in or near the order Chytridiales and was assigned to the genus and species Ichthyophonus hoferi, later reclassified as Ichthyosporidium hoferi. The graduloma described in this report occurs in fresh-water trout and is apparently caused by a budding, yeast-like form with no hyphae which evokes a tremendous inflammatory reaction. Morphologically, the organism does not resemble the previously described Ichthyosporidium. The lesions were first seen accidentally in sections prepared from a diplobacillus infection of brook trout termed “kidney disease.” Subsequently, the granuloma was observed in three widely separated infections involving the diplobacillus in each instance.The histological material was received in a fixed condition; thus, no cultural data was available and the nomenclature and classification of the mycotic organism were not attempted. The present distribution of the disease, however, with its potential threat to domestic fish populations, seemed to warrant a description and discussion of the disease. Efforts are in progress to culture the organism.

Current Advancements and Challenges in Soil-Root Interactions Modelling

NASA Astrophysics Data System (ADS)

Schnepf, A.; Huber, K.; Abesha, B.; Meunier, F.; Leitner, D.; Roose, T.; Javaux, M.; Vanderborght, J.; Vereecken, H.

2014-12-01

Roots change their surrounding soil chemically, physically and biologically. This includes changes in soil moisture and solute concentration, the exudation of organic substances into the rhizosphere, increased growth of soil microorganisms, or changes in soil structure. The fate of water and solutes in the root zone is highly determined by these root-soil interactions. Mathematical models of soil-root systems in combination with non-invasive techniques able to characterize root systems are a promising tool to understand and predict the behaviour of water and solutes in the root zone. With respect to different fields of applications, predictive mathematical models can contribute to the solution of optimal control problems in plant recourse efficiency. This may result in significant gains in productivity, efficiency and environmental sustainability in various land use activities. Major challenges include the coupling of model parameters of the relevant processes with the surrounding environment such as temperature, nutrient concentration or soil water content. A further challenge is the mathematical description of the different spatial and temporal scales involved. This includes in particular the branched structures formed by root systems or the external mycelium of mycorrhizal fungi. Here, reducing complexity as well as bridging between spatial scales is required. Furthermore, the combination of experimental and mathematical techniques may advance the field enormously. Here, the use of root system, soil and rhizosphere models is presented through a number of modelling case studies, including image based modelling of phosphate uptake by a root with hairs, model-based optimization of root architecture for phosphate uptake from soil, upscaling of rhizosphere models, modelling root growth in structured soil, and the effect of root hydraulic architecture on plant water uptake efficiency and drought resistance.

Streptosporangium minutum sp. nov., isolated from garden soil exposed to microwave radiation.

PubMed

Le Roes-Hill, Marilize; Durrell, Kim; Prins, Alaric; Meyers, Paul R

2018-06-01

The actinobacterium, strain M26 T , was isolated from garden soil that was pre-treated with microwave radiation. The soil sample was collected in Roodepoort, Gauteng Province, South Africa as part of an antibiotic-screening programme. The isolate produced branched vegetative mycelium with sporangiophores bearing small sporangia ranging from 3 to 6 μm in diameter. Rapid genus identification revealed that the isolate belongs to the genus Streptosporangium. To confirm this result, the strain was subjected to polyphasic taxonomic characterisation. Chemotaxonomic characteristics were as follows: meso-DAP in the peptidoglycan, the whole-cell hydrolysate yielded madurose, predominant menaquinones were MK9 (21%), MK9(H 2 ) (40%), MK9(H 4 ) (31%) and MK9(H 6 ) (3%); the polar lipid profile included an aminolipid, phosphoglycolipids, phosphatidylethanolamine, and phosphatidylmonomethylethanolamine. In addition, the fatty acid profile showed the presence of C 16:0 (12.8%), C 17:1 ω8c (14.2%), and 10-methyl-C 17:0 (15.8%). Furthermore, 16S rRNA gene sequence phylogenetic analysis showed that the strain is closely related to members of the genus Streptosporangium, which supports its classification within the family Streptosporangiaceae. Strain M26 T exhibited antibiosis against a range of pathogenic bacteria, including, but not limited to Acinetobacter baumannii ATCC 19606 T , Enterobacter cloacae subsp. cloacae ATCC BAA-1143, Enterococcus faecalis ATCC 51299 (vancomycin resistant), Escherichia coli ATCC 25922, Listeria monocytogenes ATCC 19111, Mycobacterium tuberculosis H37Rv T , Pseudomonas aeruginosa ATCC 27853, Salmonella enterica subsp. arizonae ATCC 13314 T , and the methicillin-resistant Staphylococcus aureus subsp. aureus ATCC 33591 (MRSA). The name Streptosporangium minutum is proposed with the type strain M26 T (=LMG 28850 T =NRRL B-65295 T ).

Inoculation effects on root-colonizing arbuscular mycorrhizal fungal communities spread beyond directly inoculated plants

PubMed Central

Krak, Karol; Vosátka, Miroslav; Püschel, David; Štorchová, Helena

2017-01-01

Inoculation with arbuscular mycorrhizal fungi (AMF) may improve plant performance at disturbed sites, but inoculation may also suppress root colonization by native AMF and decrease the diversity of the root-colonizing AMF community. This has been shown for the roots of directly inoculated plants, but little is known about the stability of inoculation effects, and to which degree the inoculant and the inoculation-induced changes in AMF community composition spread into newly emerging seedlings that were not in direct contact with the introduced propagules. We addressed this topic in a greenhouse experiment based on the soil and native AMF community of a post-mining site. Plants were cultivated in compartmented pots with substrate containing the native AMF community, where AMF extraradical mycelium radiating from directly inoculated plants was allowed to inoculate neighboring plants. The abundances of the inoculated isolate and of native AMF taxa were monitored in the roots of the directly inoculated plants and the neighboring plants by quantitative real-time PCR. As expected, inoculation suppressed root colonization of the directly inoculated plants by other AMF taxa of the native AMF community and also by native genotypes of the same species as used for inoculation. In the neighboring plants, high abundance of the inoculant and the suppression of native AMF were maintained. Thus, we demonstrate that inoculation effects on native AMF propagate into plants that were not in direct contact with the introduced inoculum, and are therefore likely to persist at the site of inoculation. PMID:28738069

Systematic Search for Cultivatable Fungi That Best Deconstruct Cell Walls of Miscanthus and Sugarcane in the Field ▿ †

PubMed Central

Shrestha, Prachand; Szaro, Timothy M.; Bruns, Thomas D.; Taylor, John W.

2011-01-01

The goals of our project were to document the diversity and distributions of cultivable fungi associated with decaying Miscanthus and sugarcane plants in nature and to further assess biodegradation of host plant cell walls by these fungi in pure cultures. Late in 2008 and early in 2009 we collected decaying Miscanthus and Saccharum from 8 sites in Illinois and 11 sites in Louisiana, respectively. To recover fungi that truly decay plants and to recover slow-growing fungi, we washed the plant material repeatedly to remove spores and cultivated fungi from plant fragments small enough to harbor at most one mycelium. We randomly selected 950 fungal colonies out of 4,560 microwell colonies and used molecular identification to discover that the most frequently recovered fungal species resided in Hypocreales (Sordariomycetes), Pleosporales (Dothideomycetes), and Chaetothryiales (Eurotiomycetes) and that only a few weedy species were recovered. We were particularly interested in Pleosporales and Chaetothyriales, groups that have not been mined for plant decay fungi. To confirm that we had truly recovered fungi that deconstruct plant cell walls, we assayed the capacity of the fungi to consume whole, alkali-pretreated, ground Miscanthus. Solid substrate cultures of the nine most commonly encountered Ascomycota resulted in Miscanthus weight loss of 8 to 13% over 4 weeks. This is the first systematic, high-throughput, isolation and biodegradation assessment of fungi isolated from decaying bioenergy grasses. PMID:21685162

Inoculation effects on root-colonizing arbuscular mycorrhizal fungal communities spread beyond directly inoculated plants.

PubMed

Janoušková, Martina; Krak, Karol; Vosátka, Miroslav; Püschel, David; Štorchová, Helena

2017-01-01

Inoculation with arbuscular mycorrhizal fungi (AMF) may improve plant performance at disturbed sites, but inoculation may also suppress root colonization by native AMF and decrease the diversity of the root-colonizing AMF community. This has been shown for the roots of directly inoculated plants, but little is known about the stability of inoculation effects, and to which degree the inoculant and the inoculation-induced changes in AMF community composition spread into newly emerging seedlings that were not in direct contact with the introduced propagules. We addressed this topic in a greenhouse experiment based on the soil and native AMF community of a post-mining site. Plants were cultivated in compartmented pots with substrate containing the native AMF community, where AMF extraradical mycelium radiating from directly inoculated plants was allowed to inoculate neighboring plants. The abundances of the inoculated isolate and of native AMF taxa were monitored in the roots of the directly inoculated plants and the neighboring plants by quantitative real-time PCR. As expected, inoculation suppressed root colonization of the directly inoculated plants by other AMF taxa of the native AMF community and also by native genotypes of the same species as used for inoculation. In the neighboring plants, high abundance of the inoculant and the suppression of native AMF were maintained. Thus, we demonstrate that inoculation effects on native AMF propagate into plants that were not in direct contact with the introduced inoculum, and are therefore likely to persist at the site of inoculation.

The status of the fungi-grower ants (Hymenoptera: Formicidae) in Puerto Rico and adjacent islands

Treesearch

J.A. Torres

1989-01-01

Ants of the tribe Attini (fungus grower) collect different organic materials that are used to grow a fungus. It was thought that the fungus mycelium was the only source of nutrition for these ants, but Quinlan and Cherrett found that Atta cephalotes (L.) squeezes oils from fresh leaves and uses them as food.  These oils supplement the fungus material eaten by this...

Surveys for Pathogens of Monoecious Hydrilla 2014

DTIC Science & Technology

2016-06-01

lateritium are all soil borne pathogens. When a host is present, the spores germinate and the mycelium penetrates plant roots and then enters the...biotypes are very different. Compared to the monoecious biotype, dioecious plants tend to have growth that is more vigorous. Dioecious plants grow...containing numerous axillary propagules (i.e., turions) drift in the water currents dispersing the plant (Steward and Van 1987). Madeira et al. (1997

A new compound from Stereum insigne CGMCC5.57.

PubMed

Tian, Meng-Qing; Wu, Qing-Lan; Wang, Xin; Zhang, Ke-Qin; Li, Guo-Hong

2017-04-01

Studies of the EtOAc extract of the culture broth and methanol extract of the mycelium of Stereum insigne CGMCC5.57 led to the isolation of one new dihydrobenzofuran (1) and six known compounds (2-7). The structures of compounds were elucidated mainly by HRESIMS experiments, and 1D, 2D NMR spectroscopy analysis. This is the first report about the chemical constitutes of the fungus S. insigne.

Hirsutella sinensis mycelium attenuates bleomycin-induced pulmonary inflammation and fibrosis in vivo

PubMed Central

Huang, Tsung-Teng; Lai, Hsin-Chih; Ko, Yun-Fei; Ojcius, David M.; Lan, Ying-Wei; Martel, Jan; Young, John D.; Chong, Kowit-Yu

2015-01-01

Hirsutella sinensis mycelium (HSM), the anamorph of Cordyceps sinensis, is a traditional Chinese medicine that has been shown to possess various pharmacological properties. We previously reported that this fungus suppresses interleukin-1β and IL-18 secretion by inhibiting both canonical and non-canonical inflammasomes in human macrophages. However, whether HSM may be used to prevent lung fibrosis and the mechanism underlying this activity remain unclear. Our results show that pretreatment with HSM inhibits TGF-β1–induced expression of fibronectin and α-SMA in lung fibroblasts. HSM also restores superoxide dismutase expression in TGF-β1–treated lung fibroblasts and inhibits reactive oxygen species production in lung epithelial cells. Furthermore, HSM pretreatment markedly reduces bleomycin–induced lung injury and fibrosis in mice. Accordingly, HSM reduces inflammatory cell accumulation in bronchoalveolar lavage fluid and proinflammatory cytokines levels in lung tissues. The HSM extract also significantly reduces TGF-β1 in lung tissues, and this effect is accompanied by decreased collagen 3α1 and α-SMA levels. Moreover, HSM reduces expression of the NLRP3 inflammasome and P2X7R in lung tissues, whereas it enhances expression of superoxide dismutase. These findings suggest that HSM may be used for the treatment of pulmonary inflammation and fibrosis. PMID:26497260

A highly sensitive NADH sensor based on a mycelium-like nanocomposite using graphene oxide and multi-walled carbon nanotubes to co-immobilize poly(luminol) and poly(neutral red) hybrid films.

PubMed

Chiang Lin, Kuo; Yu Lai, Szu; Ming Chen, Shen

2014-08-21

Hybridization of poly(luminol) (PLM) and poly(neutral red) (PNR) has been successfully performed and further enhanced by a conductive and steric hybrid nanotemplate using graphene oxide (GO) and multi-walled carbon nanotubes (MWCNTs). The morphology of the PLM-PNR-MWCNT-GO mycelium-like nanocomposite is studied by SEM and AFM and it is found to be electroactive, pH-dependent, and stable in the electrochemical system. It shows electrocatalytic activity towards NADH with a high current response and low overpotential. Using amperometry, it has been shown to have a high sensitivity of 288.9 μA mM(-1) cm(-2) to NADH (Eapp. = +0.1 V). Linearity is estimated in a concentration range of 1.33 × 10(-8) to 1.95 × 10(-4) M with a detection limit of 1.33 × 10(-8) M (S/N = 3). Particularly, it also shows another linear range of 2.08 × 10(-4) to 5.81 × 10(-4) M with a sensitivity of 151.3 μA mM(-1) cm(-2). The hybridization and activity of PLM and PNR can be effectively enhanced by MWCNTs and GO, resulting in an active hybrid nanocomposite for determination of NADH.

The carbon starvation response of the ectomycorrhizal fungus Paxillus involutus

DOE PAGES

Ellstrom, Magnus; Shah, Firoz; Johansson, Tomas; ...

2015-03-16

The amounts of carbon allocated to the fungal partner in ectomycorrhizal associations can vary substantially depending on the plant growth and the soil nutrient conditions, and the fungus may frequently be confronted with limitations in carbon. We used chemical analysis and transcriptome profiling to examine the physiological response of the ectomycorrhizal fungus Paxillus involutus to carbon starvation during axenic cultivation. Carbon starvation induced a decrease in the biomass. Concomitantly, ammonium, cell wall material (chitin) and proteolytic enzymes were released into the medium, which suggest autolysis. Compared with the transcriptome of actively growing hyphae, about 45% of the transcripts analyzed weremore » differentially regulated during C-starvation. Induced during starvation were transcripts encoding extracellular enzymes such as peptidases, chitinases and laccases. In parallel, transcripts of N-transporters were upregulated, which suggest that some of the released nitrogen compounds were re-assimilated by the mycelium. The observed changes suggest that the carbon starvation response in P. involutus is associated with complex cellular changes that involves autolysis, recycling of intracellular compounds by autophagy and reabsorption of the extracellular released material. The study provides molecular markers that can be used to examine the role of autolysis for the turnover and survival of the ectomycorrhizal mycelium in soils.« less

Haematopoiesis radioprotection in Balb/c mice by an aqueous mycelium extract from the Basidiomycete Pleurotus ostreatus mushroom.

PubMed

Llauradó, G; Morris, H J; Tamayo, V; Lebeque, Y; Beltrán, Y; Marcos, J; Moukha, S; Creppy, E E; Bermúdez, R C

2015-01-01

The study examined the radioprotective activity of an aqueous extract from Pleurotus ostreatus mycelium administered to Balb/c mice. Male mice were whole-body irradiated on day 0 ((60)Co, at 0.43 Gy/min) and divided into two groups. The extract was administered intraperitoneally to one group (100 mg/kg) on days - 10 to - 6 and - 2 to +1 with respect to the irradiation. The irradiated-control group was injected with saline solution; non-irradiated mice were used as negative controls. The radioprotective effect was evident by increases in bone marrow cellularity (5.1 × 10(6)/femur vs. 1.1 × 10(6)/femur in saline-control mice, p < 0.05), leucocyte counts (10.5 × 10(9)/L vs. 4.5 × 10(9)/L, p < 0.05), and spleen cellularity (11.2 × 10(7)/spleen vs. 6.2 × 10(7)/spleen, p < 0.05). The extract stimulated macrophage phagocytic activity as judged by a faster rate of carbon clearance in terms of absorbance ratios (1.62 vs. 2.01, p < 0.05). Therefore, this extract may be a candidate therapeutic agent with radioprotective activity for haematopoiesis damage, particularly to cells involved in immune function.

Molecular Identification of Ectomycorrhizal Mycelium in Soil Horizons

PubMed Central

Landeweert, Renske; Leeflang, Paula; Kuyper, Thom W.; Hoffland, Ellis; Rosling, Anna; Wernars, Karel; Smit, Eric

2003-01-01

Molecular identification techniques based on total DNA extraction provide a unique tool for identification of mycelium in soil. Using molecular identification techniques, the ectomycorrhizal (EM) fungal community under coniferous vegetation was analyzed. Soil samples were taken at different depths from four horizons of a podzol profile. A basidiomycete-specific primer pair (ITS1F-ITS4B) was used to amplify fungal internal transcribed spacer (ITS) sequences from total DNA extracts of the soil horizons. Amplified basidiomycete DNA was cloned and sequenced, and a selection of the obtained clones was analyzed phylogenetically. Based on sequence similarity, the fungal clone sequences were sorted into 25 different fungal groups, or operational taxonomic units (OTUs). Out of 25 basidiomycete OTUs, 7 OTUs showed high nucleotide homology (≥99%) with known EM fungal sequences and 16 were found exclusively in the mineral soil. The taxonomic positions of six OTUs remained unclear. OTU sequences were compared to sequences from morphotyped EM root tips collected from the same sites. Of the 25 OTUs, 10 OTUs had ≥98% sequence similarity with these EM root tip sequences. The present study demonstrates the use of molecular techniques to identify EM hyphae in various soil types. This approach differs from the conventional method of EM root tip identification and provides a novel approach to examine EM fungal communities in soil. PMID:12514012

The carbon starvation response of the ectomycorrhizal fungus Paxillus involutus

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ellstrom, Magnus; Shah, Firoz; Johansson, Tomas

The amounts of carbon allocated to the fungal partner in ectomycorrhizal associations can vary substantially depending on the plant growth and the soil nutrient conditions, and the fungus may frequently be confronted with limitations in carbon. We used chemical analysis and transcriptome profiling to examine the physiological response of the ectomycorrhizal fungus Paxillus involutus to carbon starvation during axenic cultivation. Carbon starvation induced a decrease in the biomass. Concomitantly, ammonium, cell wall material (chitin) and proteolytic enzymes were released into the medium, which suggest autolysis. Compared with the transcriptome of actively growing hyphae, about 45% of the transcripts analyzed weremore » differentially regulated during C-starvation. Induced during starvation were transcripts encoding extracellular enzymes such as peptidases, chitinases and laccases. In parallel, transcripts of N-transporters were upregulated, which suggest that some of the released nitrogen compounds were re-assimilated by the mycelium. The observed changes suggest that the carbon starvation response in P. involutus is associated with complex cellular changes that involves autolysis, recycling of intracellular compounds by autophagy and reabsorption of the extracellular released material. The study provides molecular markers that can be used to examine the role of autolysis for the turnover and survival of the ectomycorrhizal mycelium in soils.« less

Physicochemical and Microbiological Characteristics of Tundra Soils on the Rybachii Peninsula

NASA Astrophysics Data System (ADS)

Evdokimova, G. A.; Mozgova, N. P.; Myazin, V. A.

2018-01-01

The Rybachii Peninsula is composed of Proterozoic sedimentary rocks and differs sharply from the rest of the Kola Peninsula in its geological structure, topographic forms, and parent rocks. It is dominated by Al-Fe-humus soils formed on moraines with an admixture of local rock fragments, including slates. Organic horizons of tundra soils in the peninsula are less acid than those on granitoids of adjacent mainland of the Kola Peninsula. The content of exchangeable calcium in the organic horizons varies from 17.4 to 68.0 cmolc/kg, and the content of water-soluble carbon reaches 400 mg/100 g amounting to 1-2% of the total soil organic matter content. The total number of bacteria in the organic horizons of tundra soils varies from 3.5 × 109 to 4.8 × 109 cells/g; and bacterial biomass varies from 0.14 to 0.19 mg/g. The length of fungal mycelium and its biomass in the organic horizons are significant (>1000 m/g soil). The biomass of fungal mycelium in the organic horizons exceeds the bacterial biomass by seven times in podzols (Albic Podzols) and by ten times in podbur (Entic Podzol), dry-peat soil (Folic Histosol), and low-moor peat soil (Sapric Histosol).

Production of bio-fertilizer from microwave vacuum pyrolysis of palm kernel shell for cultivation of Oyster mushroom (Pleurotus ostreatus).

PubMed

Nam, Wai Lun; Phang, Xue Yee; Su, Man Huan; Liew, Rock Keey; Ma, Nyuk Ling; Rosli, Mohd Haqqi Nazilli Bin; Lam, Su Shiung

2018-05-15

Microwave vacuum pyrolysis of palm kernel shell (PKS) was performed to produce biochar, which was then tested as bio-fertilizer in growing Oyster mushroom (Pleurotus ostreatus). The pyrolysis approach produced biochar containing a highly porous structure with a high BET surface area of up to 270m 2 /g and low moisture content (≤10wt%), exhibiting desirable adsorption properties to be used as bio-fertilizer since it can act as a housing that provides many sites on which living microorganisms (mycelium or plant-growth promoting bacteria) and organic nutrients can be attached or adsorbed onto. This could in turn stimulate plant growth by increasing the availability and supply of nutrients to the targeted host plant. The results from growing Oyster mushroom using the biochar recorded an impressive growth rate and a monthly production of up to about 550g of mushroom. A shorter time for mycelium growth on one whole baglog (21days) and the highest yield of Oyster mushroom (550g) were obtained from cultivation medium added with 20g of biochar. Our results demonstrate that the biochar-based bio-fertilizer produced from microwave vacuum pyrolysis of PKS shows exceptional promise as growth promoting material for mushroom cultivation. Copyright © 2017 Elsevier B.V. All rights reserved.

Comparative Proteomics Analyses of Two Races of Fusarium oxysporum f. sp. conglutinans that Differ in Pathogenicity

PubMed Central

Li, Erfeng; Ling, Jian; Wang, Gang; Xiao, Jiling; Yang, Yuhong; Mao, Zhenchuan; Wang, Xuchu; Xie, Bingyan

2015-01-01

Fusarium oxysporum is a soil-inhabiting fungus that induces vascular wilt and root rot in a variety of plants. F. oxysporum f. sp. conglutinans (Foc), which comprises two races, can cause wilt disease in cabbage. Compared with race 1 (52557−TM, R1), race 2 (58385−TM, R2) exhibits much stronger pathogenicity. Here, we provide the first proteome reference maps for Foc mycelium and conidia and identify 145 proteins with different abundances among the two races. Of these proteins, most of the high-abundance proteins in the R2 mycelium and conidia are involved in carbohydrate, amino acid and ion metabolism, which indicates that these proteins may play important roles in isolate R2’s stronger pathogenicity. The expression levels of 20 typical genes demonstrate similarly altered patterns compared to the proteomic analysis. The protein glucanosyltransferase, which is involved in carbohydrate metabolism, was selected for research. We knocked out the corresponding gene (gas1) and found that Foc-∆gas1 significantly reduced growth rate and virulence compared with wild type isolates. These results deepened our understanding of the proteins related to F. oxysporum pathogenicity in cabbage Fusarium wilt and provided new opportunities to control this disease. PMID:26333982

Submerged Culture of Grifola gargal and G. sordulenta (Higher Basidiomycetes) from Argentina as a Source of Mycelia with Antioxidant Activity.

PubMed

Postemsky, Pablo Daniel; Curvetto, Néstor Raúl

2015-01-01

Submerged culture is an alternative mycelium source for Grifola gargal and G. sordulenta, two rare edible mushrooms related to Grifola frondosa. This work studies their mycelia as a source of antioxidants. The efficient concentrations of methanolic extracts in both radical scavenging (RS) and reducing power (RP) abilities in G. gargal and in G. sordulenta showed a high antioxidant activity. In the experimental design used, the antioxidant activity mainly depended on the culture conditions rather than on the media composition. Irrespective of the basal culture medium, mycelium methanolic extracts of G. sordulenta obtained from culture in Erlenmeyer flasks showed equivalents to ascorbic acid (EQ(AA)) RS-EQ(AA) and RP-EQ(AA) contents higher than the corresponding values obtained with jar cultures. Under stationary cultivation, G. sordulenta produced approximately 50% higher content in both RS-EQ(AA) and RP-EQ(AA) than the medicinal mushroom G. frondosa. Phenolics correlated with RS-EQ(AA) and RP-EQ(AA) in G. gargal and with RP-EQ(AA) in G. sordulenta; besides, thin-layer chromatography showed these compounds to be at least in part related to the RS capacity. It is concluded that G. gargal and G. sordulenta mycelia are excellent sources of antioxidant metabolites.

Modeling the Growth of Filamentous Fungi at the Particle Scale in Solid-State Fermentation Systems.

PubMed

Sugai-Guérios, Maura Harumi; Balmant, Wellington; Furigo, Agenor; Krieger, Nadia; Mitchell, David Alexander

2015-01-01

Solid-state fermentation (SSF) with filamentous fungi is a promising technique for the production of a range of biotechnological products and has the potential to play an important role in future biorefineries. The performance of such processes is intimately linked with the mycelial mode of growth of these fungi: Not only is the production of extracellular enzymes related to morphological characteristics, but also the mycelium can affect bed properties and, consequently, the efficiency of heat and mass transfer within the bed. A mathematical model that describes the development of the fungal mycelium in SSF systems at the particle scale would be a useful tool for investigating these phenomena, but, as yet, a sufficiently complete model has not been proposed. This review presents the biological and mass transfer phenomena that should be included in such a model and then evaluates how these phenomena have been modeled previously in the SSF and related literature. We conclude that a discrete lattice-based model that uses differential equations to describe the mass balances of the components within the system would be most appropriate and that mathematical expressions for describing the individual phenomena are available in the literature. It remains for these phenomena to be integrated into a complete model describing the development of fungal mycelia in SSF systems.

Impact of tricyclazole and azoxystrobin on growth, sporulation and secondary infection of the rice blast fungus, Magnaporthe oryzae.

PubMed

Kunova, Andrea; Pizzatti, Cristina; Cortesi, Paolo

2013-02-01

Rice blast, caused by Magnaporthe oryzae B. Couch sp. nov., is one of the most destructive rice diseases worldwide, causing substantial yield losses every year. In Italy, its management is based mainly on the use of two fungicides, azoxystrobin and tricyclazole, that restrain the disease progress. The aim of this study was to investigate and compare the inhibitory effects of the two fungicides on the growth, sporulation and secondary infection of M. oryzae. Magnaporthe oryzae mycelium growth was inhibited at low concentrations of azoxystrobin and relatively high concentrations of tricyclazole, while sporulation was more sensitive to both fungicides and was affected at similarly low doses. Furthermore, infection efficiency of conidia obtained from mycelia exposed to tricyclazole was affected to a higher extent than for conidia produced on azoxystrobin-amended media, even though germination of such conidia was reduced after azoxystrobin treatment. This study presents for the first time detailed azoxystrobin and tricyclazole growth-response curves for M. oryzae mycelium growth and sporulation. Furthermore, high efficacy of tricyclazole towards inhibition of sporulation and secondary infection indicates an additional possible mode of action of this fungicide that is different from inhibition of melanin biosynthesis. Copyright © 2012 Society of Chemical Industry.

Interaction of Pseudostellaria heterophylla with Fusarium oxysporum f.sp. heterophylla mediated by its root exudates in a consecutive monoculture system.

PubMed

Zhao, Yongpo; Wu, Linkun; Chu, Leixia; Yang, Yanqiu; Li, Zhenfang; Azeem, Saadia; Zhang, Zhixing; Fang, Changxun; Lin, Wenxiong

2015-02-03

In this study, quantitative real-time PCR (qPCR) was used to determine the amount of Fusarium oxysporum, an important replant disease pathogen in Pseudostellaria heterophylla rhizospheric soil. Moreover, HPLC was used to identify phenolic acids in root exudates then it was further to explore the effects of the phenolic acid allelochemicals on the growth of F. oxysporum f.sp. heterophylla. The amount of F. oxysporum increased significantly in P. heterophylla rhizosphere soil under a consecutive replant system as monitored through qPCR analysis. Furthermore, the growth of F. oxysporum f.sp. heterophylla mycelium was enhanced by root exudates with a maximum increase of 23.8%. In addition, the number of spores increased to a maximum of 12.5-fold. Some phenolic acids promoted the growth of F. oxysporum f.sp. heterophylla mycelium and spore production. Our study revealed that phenolic acids in the root secretion of P. heterophylla increased long with its development, which was closely related to changes in rhizospheric microorganisms. The population of pathogenic microorganisms such as F. oxysporum in the rhizosphere soil of P. heterophylla also sharply increased. Our results on plant-microbe communication will help to better clarify the cause of problems associated with P. heterophylla under consecutive monoculture treatment.

AMP-forming acetyl-CoA synthetases in Archaea show unexpected diversity in substrate utilization

PubMed Central

Ingram-Smith, Cheryl; Smith, Kerry S.

2007-01-01

Adenosine monophosphate (AMP)-forming acetyl-CoA synthetase (ACS; acetate:CoA ligase (AMP-forming), EC 6.2.1.1) is a key enzyme for conversion of acetate to acetyl-CoA, an essential intermediate at the junction of anabolic and catabolic pathways. Phylogenetic analysis of putative short and medium chain acyl-CoA synthetase sequences indicates that the ACSs form a distinct clade from other acyl-CoA synthetases. Within this clade, the archaeal ACSs are not monophyletic and fall into three groups composed of both bacterial and archaeal sequences. Kinetic analysis of two archaeal enzymes, an ACS from Methanothermobacter thermautotrophicus (designated as MT-ACS1) and an ACS from Archaeoglobus fulgidus (designated as AF-ACS2), revealed that these enzymes have very different properties. MT-ACS1 has nearly 11-fold higher affinity and 14-fold higher catalytic efficiency with acetate than with propionate, a property shared by most ACSs. However, AF-ACS2 has only 2.3-fold higher affinity and catalytic efficiency with acetate than with propionate. This enzyme has an affinity for propionate that is almost identical to that of MT-ACS1 for acetate and nearly tenfold higher than the affinity of MT-ACS1 for propionate. Furthermore, MT-ACS1 is limited to acetate and propionate as acyl substrates, whereas AF-ACS2 can also utilize longer straight and branched chain acyl substrates. Phylogenetic analysis, sequence alignment and structural modeling suggest a molecular basis for the altered substrate preference and expanded substrate range of AF-ACS2 versus MT-ACS1. PMID:17350930

Genoprotective Capacity of Alternatively Cultivated Lingzhi or Reishi Medicinal Mushroom, Ganoderma lucidum (Agaricomycetes), Basidiocarps.

PubMed

Cilerdzic, Jasmina; Stajic, Mirjana; Zivkovic, Lada; Vukojevic, Jelena; Bajic, Vladan; Spremo-Potparevic, Biljana

2016-01-01

Ganoderma lucidum is traditionally used in Eastern medicine to preserve vitality, promote longevity, and treat disease. It possesses immunomodulatory, antitumor, antimicrobial, and antiaging activities, among others, but one of the most important is its antioxidant property, which is the basis for other effects, because free radicals trigger many diseases. The substrate commonly used for commercial cultivation of G. lucidum is not environmentally friendly nor economically justified, so there is a need to find new alternative substrates. The aim of this study was to analyze the effect of substrate composition on the bioactivity of G. lucidum basidiocarps. G. lucidum was cultivated on 2 different substrates: (1) a mixture of wheat straw, grapevine branches, and wheat bran, and (2) wheat straw. Commercial fruiting bodies, cultivated on oak sawdust, were used as the control. 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging ability, total phenols, and flavonoid content were determined spectrophotometrically to define the antioxidative potential of basidiocarp extracts. The comet test was performed to detect the degree of DNA damage in the cells that were exposed to G. lucidum extracts before and after the effect of oxidants. Higher antioxidative potential was observed for the extract of G. lucidum basidiocarps cultivated on wheat straw compared with that from the mixed substrate and especially with commercial ones. The alternatively cultivated basidiocarps also showed stronger antigenotoxic potential compared with commercial ones. The study showed that fruiting bodies produced on wheat straw, one of the most accessible and cheapest crop residues, are more potent antioxidant and antigenotoxic agents than commercially cultivated ones.

Synthesis of customized petroleum-replica fuel molecules by targeted modification of free fatty acid pools in Escherichia coli

PubMed Central

Howard, Thomas P.; Middelhaufe, Sabine; Moore, Karen; Edner, Christoph; Kolak, Dagmara M.; Taylor, George N.; Parker, David A.; Lee, Rob; Smirnoff, Nicholas; Aves, Stephen J.; Love, John

2013-01-01

Biofuels are the most immediate, practical solution for mitigating dependence on fossil hydrocarbons, but current biofuels (alcohols and biodiesels) require significant downstream processing and are not fully compatible with modern, mass-market internal combustion engines. Rather, the ideal biofuels are structurally and chemically identical to the fossil fuels they seek to replace (i.e., aliphatic n- and iso-alkanes and -alkenes of various chain lengths). Here we report on production of such petroleum-replica hydrocarbons in Escherichia coli. The activity of the fatty acid (FA) reductase complex from Photorhabdus luminescens was coupled with aldehyde decarbonylase from Nostoc punctiforme to use free FAs as substrates for alkane biosynthesis. This combination of genes enabled rational alterations to hydrocarbon chain length (Cn) and the production of branched alkanes through upstream genetic and exogenous manipulations of the FA pool. Genetic components for targeted manipulation of the FA pool included expression of a thioesterase from Cinnamomum camphora (camphor) to alter alkane Cn and expression of the branched-chain α-keto acid dehydrogenase complex and β-keto acyl-acyl carrier protein synthase III from Bacillus subtilis to synthesize branched (iso-) alkanes. Rather than simply reconstituting existing metabolic routes to alkane production found in nature, these results demonstrate the ability to design and implement artificial molecular pathways for the production of renewable, industrially relevant fuel molecules. PMID:23610415

Cloning and Functional Characterization of Three Branch Point Oxidosqualene Cyclases from Withania somnifera (L.) Dunal*

PubMed Central

Dhar, Niha; Rana, Satiander; Razdan, Sumeer; Bhat, Wajid Waheed; Hussain, Aashiq; Dhar, Rekha S.; Vaishnavi, Samantha; Hamid, Abid; Vishwakarma, Ram; Lattoo, Surrinder K.

2014-01-01

Oxidosqualene cyclases (OSCs) positioned at a key metabolic subdividing junction execute indispensable enzymatic cyclization of 2,3-oxidosqualene for varied triterpenoid biosynthesis. Such branch points present favorable gene targets for redirecting metabolic flux toward specific secondary metabolites. However, detailed information regarding the candidate OSCs covering different branches and their regulation is necessary for the desired genetic manipulation. The aim of the present study, therefore, was to characterize members of OSC superfamily from Withania somnifera (Ws), a medicinal plant of immense repute known to synthesize a large array of biologically active steroidal lactone triterpenoids called withanolides. Three full-length OSC cDNAs, β-amyrin synthase (WsOSC/BS), lupeol synthase (WsOSC/LS), and cycloartenol synthase (WsOSC/CS), having open reading frames of 2289, 2268, and 2277 bp, were isolated. Heterologous expression in Schizosaccharomyces pombe, LC-MS analyses, and kinetic studies confirmed their monofunctionality. The three WsOSCs were found to be spatially regulated at transcriptional level with WsOSC/CS being maximally expressed in leaf tissue. Promoter analysis of three WsOSCs genes resulted in identification of distinct cis-regulatory elements. Further, transcript profiling under methyl jasmonate, gibberellic acid, and yeast extract elicitations displayed differential transcriptional regulation of each of the OSCs. Changes were also observed in mRNA levels under elicitations and further substantiated with protein expression levels by Western blotting. Negative regulation by yeast extract resulted in significant increase in withanolide content. Empirical evidence suggests that repression of competitive branch OSCs like WsOSC/BS and WsOSC/LS possibly leads to diversion of substrate pool toward WsOSC/CS for increased withanolide production. PMID:24770414

Fermentation of animal components in strict carnivores: a comparative study with cheetah fecal inoculum.

PubMed

Depauw, S; Bosch, G; Hesta, M; Whitehouse-Tedd, K; Hendriks, W H; Kaandorp, J; Janssens, G P J

2012-08-01

The natural diet of felids contains highly digestible animal tissues but also fractions resistant to small intestinal digestion, which enter the large intestine where they may be fermented by the resident microbial population. Little information exists on the microbial degradability of animal tissues in the large intestine of felids consuming a natural diet. This study aimed to rank animal substrates in their microbial degradability by means of an in vitro study using captive cheetahs fed a strict carnivorous diet as fecal donors. Fresh cheetah fecal samples were collected, pooled, and incubated with various raw animal substrates (chicken cartilage, collagen, glucosamine-chondroitin, glucosamine, rabbit bone, rabbit hair, and rabbit skin; 4 replicates per substrate) for cumulative gas production measurement in a batch culture technique. Negative (cellulose) and positive (casein and fructo-oligosaccharides; FOS) controls were incorporated in the study. Additionally, after 72 h of incubation, short-chain fatty acids (SCFA), including branched-chain fatty acids (BCFA), and ammonia concentrations were determined for each substrate. Glucosamine and glucosamine-chondroitin yielded the greatest organic matter cumulative gas volume (OMCV) among animal substrates (P < 0.05), whereas total SCFA production was greatest for collagen (P < 0.05). Collagen induced an acetate production comparable with FOS and a markedly high acetate-to-propionate ratio (8.41:1) compared with all other substrates (1.67:1 to 2.97:1). Chicken cartilage was rapidly fermentable, indicated by a greater maximal rate of gas production (R(max)) compared with all other substrates (P < 0.05). In general, animal substrates showed an earlier occurrence for maximal gas production rate compared with FOS. Rabbit hair, skin, and bone were poorly fermentable substrates, indicated by the least amount of OMCV and total SCFA among animal substrates (P < 0.05). The greatest amount of ammonia production among animal substrates was measured after incubation of collagen and rabbit bone (P < 0.05). This study provides the first insight into the potential of animal tissues to influence large intestinal fermentation in a strict carnivore, and indicates that animal tissues have potentially similar functions as soluble or insoluble plant fibers in vitro. Further research is warranted to assess the impact of fermentation of each type of animal tissue on gastro-intestinal function and health in the cheetah and other felid species.

Epitaxial growth of aligned AlGalnN nanowires by metal-organic chemical vapor deposition

DOEpatents

Han, Jung; Su, Jie

2008-08-05

Highly ordered and aligned epitaxy of III-Nitride nanowires is demonstrated in this work. <1010> M-axis is identified as a preferential nanowire growth direction through a detailed study of GaN/AlN trunk/branch nanostructures by transmission electron microscopy. Crystallographic selectivity can be used to achieve spatial and orientational control of nanowire growth. Vertically aligned (Al)GaN nanowires are prepared on M-plane AlN substrates. Horizontally ordered nanowires, extending from the M-plane sidewalls of GaN hexagonal mesas or islands demonstrate new opportunities for self-aligned nanowire devices, interconnects, and networks.

Water-Soluble Epitaxial NaCl Thin Film for Fabrication of Flexible Devices.

PubMed

Lee, Dong Kyu; Kim, Sungjoo; Oh, Sein; Choi, Jae-Young; Lee, Jong-Lam; Yu, Hak Ki

2017-08-18

We studied growth mechanisms of water-soluble NaCl thin films on single crystal substrates. Epitaxial growth of NaCl(100) on Si(100) and domain-matched growth of NaCl(111) on c-sapphire were obtained at thicknesses below 100 nm even at room temperature from low lattice mismatches in both cases. NaCl thin film, which demonstrates high solubility selectivity for water, was successfully applied as a water-soluble sacrificial layer for fabrication of several functional materials, such as WO 3 nano-helix and Sn doped In 2 O 3 nano-branches.

JPRS Report Science & Technology USSR: Life Sciences

DTIC Science & Technology

1990-06-18

water-soluble low-molecular-mass ß-l,3-ß-l,6- glucanes , suppressors present in the mycelium and zoospores of the fungus, and also in its excretions...This article studies the participation of the glucane suppressors of phytoph- thora infestans (Mont) de Bary in the suppression of various types of...potato immune response. The interac- tion of the glucanes with specific receptors on the plas- malemma of the potato cells prevents recognition by

International Conference on Separations for Biotechnology: Reading, UK,

DTIC Science & Technology

1988-02-19

tests carried hydrodynamic diameter (from diffusion out usually Involve either microcopIc coefficent measurement) and molecular examination or assay of...similar percent ard zero-percent Span 80 and compounds . The first stage of tha re- Lubrizol, respectively, after 10 min- covery of these products usually...Peni- cillin C. This compound (pK-2.75) can z: " from cell-free medium a * acidic orily be extracted from the fermentation M a from mycelium a

Lhx6 delineates a pathway mediating innate reproductive behaviors from the amygdala to the hypothalamus.

PubMed

Choi, Gloria B; Dong, Hong-Wei; Murphy, Andrew J; Valenzuela, David M; Yancopoulos, George D; Swanson, Larry W; Anderson, David J

2005-05-19

In mammals, innate reproductive and defensive behaviors are mediated by anatomically segregated connections between the amygdala and hypothalamus. This anatomic segregation poses the problem of how the brain integrates activity in these circuits when faced with conflicting stimuli eliciting such mutually exclusive behaviors. Using genetically encoded and conventional axonal tracers, we have found that the transcription factor Lhx6 delineates the reproductive branch of this pathway. Other Lhx proteins mark neurons in amygdalar nuclei implicated in defense. We have traced parallel projections from the posterior medial amygdala, activated by reproductive or defensive olfactory stimuli, respectively, to a point of convergence in the ventromedial hypothalamus. The opposite neurotransmitter phenotypes of these convergent projections suggest a "gate control" mechanism for the inhibition of reproductive behaviors by threatening stimuli. Our data therefore identify a potential neural substrate for integrating the influences of conflicting behavioral cues and a transcription factor family that may contribute to the development of this substrate.

Large-scale Topographical Screen for Investigation of Physical Neural-Guidance Cues

NASA Astrophysics Data System (ADS)

Li, Wei; Tang, Qing Yuan; Jadhav, Amol D.; Narang, Ankit; Qian, Wei Xian; Shi, Peng; Pang, Stella W.

2015-03-01

A combinatorial approach was used to present primary neurons with a large library of topographical features in the form of micropatterned substrate for high-throughput screening of physical neural-guidance cues that can effectively promote different aspects of neuronal development, including axon and dendritic outgrowth. Notably, the neuronal-guidance capability of specific features was automatically identified using a customized image processing software, thus significantly increasing the screening throughput with minimal subjective bias. Our results indicate that the anisotropic topographies promote axonal and in some cases dendritic extension relative to the isotropic topographies, while dendritic branching showed preference to plain substrates over the microscale features. The results from this work can be readily applied towards engineering novel biomaterials with precise surface topography that can serve as guidance conduits for neuro-regenerative applications. This novel topographical screening strategy combined with the automated processing capability can also be used for high-throughput screening of chemical or genetic regulatory factors in primary neurons.

The Effect of Substrate Topography on Direct Reprogramming of Fibroblasts to Induced Neurons

PubMed Central

Kulangara, Karina; Adler, Andrew F.; Wang, Hong; Chellappan, Malathi; Hammett, Ellen; Yasuda, Ryohei; Leong, Kam W.

2014-01-01

Cellular reprogramming holds tremendous potential for cell therapy and regenerative medicine. Recently, fibroblasts have been directly converted into induced neurons (iNs) by overexpression of the neuronal transcription factors Ascl1, Brn2 and Myt1L. Hypothesizing that cell-topography interactions could influence the fibroblast-to-neuron reprogramming process, we investigated the effects of various topographies on iNs produced by direct reprogramming. Final iN purity and conversion efficiency were increased on micrograting substrates. Neurite branching was increased on microposts and decreased on microgratings, with a simplified dendritic arbor characterized by the reduction of MAP2+ neurites. Neurite outgrowth increased significantly on various topographies. DNA microarray analysis detected 20 differentially expressed genes in iNs reprogrammed on smooth versus microgratings, and quantitative PCR (qPCR) confirmed the upregulation of Vip and downregulation of Thy1 and Bmp5 on microgratings. Electrophysiology and calcium imaging verified the functionality of these iNs. This study demonstrates the potential of applying topographical cues to optimize cellular reprogramming. PMID:24709523

A New Thermal Treatment Process of Low Value Volcanic Glass towards the Production of Expanded Material and its Use on CNTs’ Synthesis as Substrate Material

NASA Astrophysics Data System (ADS)

Angelopoulos, Panagiotis M.; Samouhos, Michail; Taxiarchou, Maria; Tsakiridis, P.; Haggman, John; Joyce, Paul

2018-05-01

Pitchstone is a naturally occurring volcanic glass that contains considerable amount of chemically bound water (> 6 % wt). Due to its high water content, its direct thermal processing in conventional expansion furnaces towards the production of lightweight material, similar to expanded perlite, is practically impossible. In the current research paper a sophisticated 2 stage process is presented that consists of a partial dehydration and an expansion stage towards the production of high quality expanded material. After proper treatment, low-value volcanic glass is transformed to frothy, lightweight material of closed external surface and apparent density of 52 kg·m-3 that can be used in various branches of the industry. The material produced is used as substrate for the development of multiwall CNTs through CVD method. Dense multiwall CNT clusters were identified on expanded pitchstone surface, thus rendering the material suitable for such application.