Eiji Tanesaka

WOOD DEGRADING ABILITY OF BASIDIOMYCETES THAT ARE WOOD DECOMPOSERS, LITTER DECOMPOSERS, OR MYCORRHIZAL SYMBIONTS

The wood degrading ability of basidiomycetes, 41 wood decomposers, 22 litter decomposers, and 5 mycorrhizal symbionts, was measured in aseptic culture on an agar medium. The wood decomposers had greater variation in this ability than the litter decomposers. Ratios of percent lignin loss to percent weight loss of the wood specimens (the lignin loss ratio) were somewhat higher in the white-rot fungi than in the brown-rot fungi. The species collected from the L and F layers of litter deposits, such as Collybia confluens, Collybia dryophila, and Hygrocybe sp., and Marasmius maximus, decomposed wood more than the species from the H layer. These species also produced lignin loss ratios two to three times that of the white-rot species of wood decomposers. Ten litter decomposers bleached wood specimens to yellowish white, as typical white-rot species do. None of the mycorrhizal species degraded wood during 80 days of culture. The wood and litter decomposers showed highly significant variation among species within a genus for wood degrading ability.

Electrophoretic karyotype of Flammulina velutipes and its variation among monokaryotic progenies

Abstract The karyotype of Flammulina velutipes (Curt. : Fr.) Sing. was investigated using contour-clamped homogeneous electric fields (CHEF) gel electrophoresis. A parental dikaryotic stock, JA, was resolved into at least eight chromosomal DNA bands ranging from 1.4- to 4.9-megabase (Mb) pairs. Overall, little size variation was found among monokaryotic strains with a few major exceptions. Among 13 monokaryotic progenies examined, 11 strains were resolved into at least eight chromosomal DNA bands in a manner similar to the parent dikaryon, whereas the other 2 were resolved into at least seven chromosomes lacking the 2.1-Mb chromosome possessed in the former. A slightly larger size variation was found in a chromosome carrying ribosomal DNA. An estimated haploid genome size of this stock was 24.0 Mb or more.

Colonizing success of saprotrophic and ectomycorrhizal basidiomycetes on islands

The biodiversity of saprotrophic and ectomycorrhizal basidiomycetous macrofungi growing on seven islands in central Japan were compared to examine colonizing success within the context of island biogeography theory. Two hypotheses were tested: that the number of the fungal species depends on island area and that the slope of the species-area curve for saprotrophic and ectomycorrhizal macrofungi differ in response to differences in their nutritional requirements. Data for the number of species that were identified based on sporocarps closely fit the conventional species-area curve. The slopes of the species-area curve for saprotrophic fungi (0.316) and ectomycorrhizal fungi (0.469) were similar to those reported for insects and birds, and plants on other archipelagos, respectively. In addition species-area curve data showed that ectomycorrhizal fungi colonized only islands > 630 m2. While the species composition of saprotrophic fungi found on any pair of islands was positively correlated to the ratio of the areas of the island pair being compared (smaller/larger), no such relationship was observed for ectomycorrhizal fungi. Conversely similar ectomycorrhizal fungi, mostly those belonging to the genera Amanita, Inocybe, Boletellus and Russula, were found on pairs of islands with similar vegetation in the same geographic region. These results suggested that the colonizing success by ectomycorrhizal fungi is limited by host plant diversity, which is lower on smaller islands, instead of restricted immigration resulting from limited spore dispersal ability.

Nuclear selection and nuclear substitution in fully compatible di-mon matings in Pleurotus ostreatus

Nuclear behavior in fully compatible dikaryon-monokaryon matings inPleurotus ostreatus was examined with esterase isozymes as markers, which are located at the EST1 locus independent of mating type factors A and B. Monokaryotic strains mMA91 and mOW89 were obtained from the stocks MA91 from Japan and OW89 from Thailand, respectively. The mMA91 strains represented genotype EST1 (100), and the mOW89 strains represented either EST1 (86) or EST1 (93). Most of the stocks derived from mMA91×OW89 represented the genotype EST1 (93)+EST1 (100). This result suggests that the donor nucleus of OW89 carrying EST1 (93) migrated preferentially into the monokaryon mMA91 to conjugate with the recipient nucleus of the latter. Some of the stocks from mOW89 × MA91 represented the genotype EST1 (100)+EST1 (100). This seems to have resulted from substitution of a nucleus of the recipient mOW89 by a pair of nuclei of the donor MA91. The variations in mycelial products, mycelial growth rate, and wood degrading ability among the D1 populations of stocks from mMA91 × OW89 were significantly smaller than those from mOW89 × MA91, reflecting the selective nuclear migration from OW89 to mMA91.

Enzymatic Staining for Detection of Phenol-Oxidizing Isozymes Involved in Lignin- Degradation by Lentinula edodes on Native-PAGE

Lignocellulose is the most abundant organic compound in the terrestrial environment. Nonetheless, with the exception of basidiomycetous fungi, most organisms are either unable to degrade lignocellulose, or if they can, they do so with difficulty (Kirk & Fenn, 1982). Wood-decomposing basidiomycetes can be grouped into two categories: white-rot and brown-rot fungi. White-rot fungi have cellulases and lignin-degrading enzymes that decompose most cell wall components, whereas brown-rot fungi have enzymatic systems that selectively degrade cellulose and hemicelluloses, leaving brown shrunken lumps of tissue composed mainly of a loose lignin matrix (Enoki et al., 1988; Highley et al., 1985; Highley & Murmanis, 1987; Kirk & Highley, 1973). The name ‘white-rot’ is derived from the bleaching effect that this fungus has when degrading wood; the lignin-degrading enzymes that they secrete have the effect of promoting lignin loss and exposing the white cellulose fibrils. White-rot fungi are known to produce polyphenol oxidases (phenoloxidases), which, when the fungi are plated on agar media containing gallic or tannic acids, change the color of the agar to a dark reddish-brown in what is referred to as Bavendamm’s polyphenol oxidase test or Bavendamm reaction (Bavendamm, 1928, as cited in JØrgensen & Vejlby, 1953). Based on this reaction, phenoloxidases are considered to be one of putative lignindegrading enzymes (Higuchi 1990). Laccase (Lcc, EC 1.10.3.2), catechol oxidase (EC 1.10.3.1) and tyrosinase (monophenol monooxygenase, EC 1.14.18.1) are phenoloxidases with considerable overlap in their substrate affinities (Burke & Cairney, 2002). Lcc catalyze the reduction of O2 to H2O using a range of phenolics, aromatic amines, and other electron-rich substances as hydrogen donors (Thurston, 1994). Similar phenol-oxidizing activities are also observed in peroxidases (EC 1.11.1.x), which use H2O2 as an electron donor. Lignin peroxidase (ligninase, LiP, EC 1.11.1.14) was first discovered in Phanerochaete chrysosporium in which the H2O2-dependent C┙-C┚ cleavage of non-phenolic lignin model compounds was first described (Tien & Kirk, 1983, 1984). Manganese peroxidase (MnP, EC 1.11.1.13) also strongly degrades lignin model compounds and the reaction is mediated by H2O2 and Mn2+ (Glenn et al., 1983; Glenn & Gold, 1985; Kuwahara et al., 1984). Whereas lignin can effectively be oxidized by LiP directly, as reviewed previously (Cullen & Kersten, 2004;

Functional analysis of novel genes expressed specifically in the early maturation of sesame seeds

K also called as metastin, is the cognate ligand GPR-54 (G-coupled receptor), which was previously an orphan receptor. Kisspeptin consists of 54 amino acids and its biological activity can be localized to the C-terminal segment which is cleaved into C-10, C-13, and C-14 segment. Kisspeptin-GPR-54 interaction stimulates GnRH secretion and has been shown to be essential for the initiation of the pubertal LH surge. A 14 amino acid derivative of kisspeptin (Asp-58-Val59-Ser60-Ala61-Tyr62Asn63-Trp64-Asn65-Ser66-Phe67-Gly68-Leu69-Arg70-Tyr71NH2) was synthesized by solid phase peptide synthesis using F-moc (9-fluorenyl methoxy carbonyl) strategy. Kisspeptin-14 was synthesized on Rinkamide resin after swelling it for 2h. Amino acid coupling steps were carried out by the treatment of deprotected resin with a 3 fold molar excess of F-moc protected amino acid, HBTU (O-Benzotriazole-N,N,N’,N’-tetramethyl-uronium-hexafluoro-phosphate) and HOBt (N-Hydroxybenzotriazole) for 2h at room temperature under constant shaking. Deprotection was carried out using 20% piperidine for 20 min under constant shaking. Between coupling and deprotection, the resin was washed twice with di-methyl formamide followed by three washing of di-chloromethane. Peptide was cleaved out of the resin using the cleavage mixture (Trifluoro acetic acid-82.5%, Thioanisole-5%, Ethanedithiol-5%, Water-10%, Phenol-5%). Kisspeptin peptide was purified by RP-HPLC using a gradient of 65-90% using C-18 column in binary gradient module consisting of 5% acetonitrile in water and a limiting organic solvent. The peptide was recovered directly after lyophilization. Peptide was evaluated for its secondary conformation by performing CD spectroscopy which showed primarily a random coil structure in water which can be induced to adopt more ordered conformation using solvent like tri-fluoroethanol.L hydrolysis of castor oil yields a high value product free from odour, colour and undesired side products, associated with conventional hydrolytic methods. However, use of immobilized lipase is limited by diffusional constraints of conventional two phase reaction media. Smart design of reaction media allowing oil homogenization was carried out in this study for hydrolysis of castor oil using indigenously immobilized lipases. Optimization of various parameters towards complete hydrolysis was carried out in batch experiments. Maximum hydrolysis of 85% was obtained for the reusable immobilized lipase preparation under packed bed conditions. The activity of immobilized enzyme for the said reaction under optimised conditions was observed to be >85% with no loss of activity for upto 10 cycles tested. The product so formed can be further converted to sugar alcohols, estolides, conjugated linolenic acids or lactones either by enzymatic/microbial methods. The advantage of biotransformation allows production of clean materials that find wider applicability and also higher commercial value.A are an important class of Pharmacological agents used for treating infections, which are a major cause of human morbidity and mortality. Although antibiotics were first isolated from fungi and bacteria (natural Source), but over the years more and more synthetic antibiotics are flowing in market. During the last two decades, the development of drug resistance as well as the appearance of undesirable side effects of certain antibiotics has lead to the search of new antimicrobial agent. There is ample of documented evidence which show antibiotic properties in plant extracts. The present study is based on antibacterial and antifungal activity of wildly growing indigenous plants. Aqueous and ethanol extract of different part ( leaves, flower, fruit, root) of Oxalis amara, Argemone maxicana, Datura inoxia, Calatropis procera , Amranthus, Pithecellobium dulce, Ziziphus mauritiana, Croton bonplandianum, Cannibus sativa, Leucaena leucophela, Andographis Peniculata were taken for study, against pathogenic bacteria Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and fungus Candida albicans. Results were observed/ calculated by their zone of inhibition after 24 hours by agar plate gel diffusion method. These plants are showing their antibacterial and antifungal activities, differently for different microorganism. This laboratory experiments has shown that plant extracts are effective against a broad spectrum of microbes (Both Gram positive and Gram negative bacteria and fungus) & antibiotics from Plant extracts (Herbal antibiotics) are promising agents for developing newer antibioticsT effect of increased industrialization has adverse impact on the environment thus leading to air pollution and water pollution. The major impact of industrial systems on the environment is the emissions of gaseous, liquid and particulate materials in the atmosphere which leads to air pollution. Air pollution is aggravated now-a-days because of economic development of societies across the world. The category of air emissions include the criteria pollutants given by the Environmental Protection Agency (EPA), USA which include sulfur dioxide (SO2), nitrogen dioxide (NO2), carbon monoxide (CO), ozone (O3), suspended particulate matter (SPM) and lead (Pb) which are significant contributors in the deterioration of public health (USEPA, 1993). Another important category of pollutants is hazardous pollutants such as Volatile Organic Compounds (VOCs), ammonia (NH3), hydrogen sulfide (H2S), etc. which are responsible for major air and water pollution. Out of all listed hazardous pollutants, VOCs are the large group of organic compounds emitted into the atmosphere by a wide range of industries. In fact, VOCs are one of the major pollutants released by the industries which contaminate the atmospheric air and the fresh water resources. This technique is widely used for the removal of VOCs from and air and metal ions from water. The potential of microorganisms in consuming the VOCs as carbon source makes biodegradation and biofiltration an attractive option for the removal of pollutants from the waste air and waste water streams. The work essentially discusses the biodegradation and biofiltration studies using the mixed culture. The work also discusses the essential factors for the development of the laboratory scale biofilter column and the packing material used in the biofiltration studies. The work also details in the various parameters such as effect of time, effect of flow rate, effect of shock loads and effect of bed height to check the performance of biofiltration. The work also incorporates the various applications of biofilters.F enzyme like Superoxide Dismutase (SOD: 1.15.1.1), has wide applications in the industry of pharmaceuticals, cosmetics, food and/or biotechnology. SOD is a primary antioxidant enzyme catalyzing the dismutation of superoxide free radical (O2-) (a disease causing agent). An important requirement for SODs intended for industrial applications is their thermal stability as thermal denaturation is a common cause of enzyme inactivation. In the present study, SOD has been isolated and characterized from Juglans regia (Walnut) kernels. The SOD enzyme obtained has been partially purified into two fractions based on 0-40% and 40-80% saturation level of ammonium sulphate; 0-40% fraction (72.46 Units/mg) having higher specific activity. The temperature optima of the partially purified SOD lied between 35oC to 40oC with thermo stability up to 70oC whereas the optimum value for pH being 5.0 with stability range of 4.0-7.0. All the tested detergents viz; Sodium Lauryl Sulphate (SLS), Cween-20 and Tween-80 and Tween-20 were found to inhibit the activity of enzyme, Cween-20 being most effective. Mg2+ and Ba2+ ions in the form of their respective salts, acted as potential inhibitors with about 50% reduction in enzyme activity. The results obtained herein indicate that J. regia (Walnut) kernel constitute an excellent source of SOD enzyme, which is an effective natural dietery antioxidant. It can be inferred from the present study that J. regia, a functional food, is a promising source of natural antioxidant (SOD) enzyme with potential application in oxidative stress-related diseases. The thermostability of the enzyme further enhances its importance making it industry friendly with high economical feasibility.H is routinely used in the treatment of pulmonary insufficiency and respiratory distress in preterm and term infants and in adults with acute respiratory disease (ARDS). However, in infants, hyperoxia contributes to the development of chronic lung disease (CLD), which is termed bronchopulmonary dysplasia (BPD). The molecular mechanisms of oxygen-mediated lung injury are not understood, but reactive oxygen species (ROS) are the most likely candidates. ROSare also responsible for many other lung diseases such as acute respiratory distress syndrome (ARDS), asthma, emphysema, chronic obstructive pulmonary disease (COPD), and lung cancer. Results from our laboratory demonstrate a novel role for cytochrome P450 (CYP)1A enzymes in the detoxification of ROS-mediated lipid peroxidation products, e.g., F2-isoprostanes. Our major observations are that mice lacking the genes for CYP1A1 or 1A2 are more susceptible to hyperoxic lung injury than wild type mice, with Cyp1a2-null mice being the most sensitive. On the other hand, mice lacking the gene for CYP1B1, are less susceptible to lung injury, suggesting a pro-oxidant role for CYP1B1. Mice pre-treated with the CYP1A inducer (β-napthoflavone (BNF), followed by exposure to hyperoxia leads to protection against lung injury. We also found formation of bulky oxidative lesions (oxidative DNA adducts) in tracheal aspirates of premature infants and adults who received supplemental oxygen, and this was associated with BPD and ARDS, thereby suggesting that these adducts could serve as novel biomarkers of these diseases. Future studies could lead to the development of rational strategies for the prevention/treatment of lung diseases associated with hyperoxia.S seeds of Sesamum indicum contain abundant oil, in particular, large amounts of unsaturated fatty acid oleic and linoleic acids. These unsaturated fatty acids lower the cholesterol level in the body. Because of their effects and antioxidant substances, attention has been paid to sesame seeds as health food. The regulation of fatty acid synthesis in oilseed crops is quite unknown. It is necessary to clarify these points to breed novel varieties with high contents of unsaturated fatty acids. Full-length cDNA libraries prepared from sesame seed of 1 to 3 weeks DAF, were subtracted with cDNAs from plantlets of 4 weeks after seeding. Each of 1,545 cDNA clones was sequenced. The function of novel genes expressed during the early maturation of sesame seeds was studied by the transformation of Arabidopsis thaliana. Thirteen genes for a transcription factor were identified; four were involved in ethylene signaling. Nine genes including aquaporin-like protein, putative uncharacterized novel protein and ethylene response factor were analyzed by overexpression of A. thaliana. A. thaliana overexpression strain for novel protein and aquaporin-like protein genes, respectively showed the increase of unsaturated fatty acids. The localization of these products was investigated by the induction of the expression vectors for GFP fusion protein into onion cells and sesame seeds with a particle gun.E pollution is considered as a side effect of modern industrial society. The presence of man-made (anthropogenic) organic compounds in the environment is a very serious public health problem. Phenol, an organic compound is toxic even at low concentrations and the toxicity of phenols for microbial cells has been investigated. Owing to the toxic nature and consequent health hazard of phenol, the need to remove it from waste waters. Harnessing the potential of microbes to degrade phenol has been an area of considerable study to develop bioremediation approaches, which is considered as “Green Option” for treatment of environmental contaminants. The optimum conditions for phenol degradation by Pseudomonas putida (NCIM 2102) were at inoculum size (6%v/v), pH (7), temperature (300C), agitation speed (140 rpm), glucose (0.8g/l), ammonium sulphate (1.5g/l), peptone (0.5g/l), and concentration of metal ion Mn2+(0.02 g/l).Central Composite Design (CCD) was employed combining with Response Surface Methodology (RSM) to optimize the physical, chemical parameters for the degradation of phenol by P. putida (NCIM 2102). Response Surface method was using three-levels of physico-chemical parameters like pH, temperature, agitation speed, carbon source (glucose), inorganic nitrogen source (ammonium sulfate) and metal ion (Mn2+) concentration which also enabled the identification of significant effects of interactions for the batch studies. The experimental values are in good agreement with predicted values and the correlation coefficient of physico-chemical parameters was found to be 0.9871, 0.9028 respectively.T Transport Phenomena in Food Materials laboratory at Purdue University seeks to establish the fundamental relationships between the structure of food materials and their mechanical and functional properties as influenced by processing, composition, and environmental conditions. A current research thrust area of this laboratory: Interfacial Phenomena in Foods seeks to establish a detailed understanding of interfacial mechanisms that affect the transport of mass and heat in food matrices, particularly in terms of developing enhanced food materials and processes. Two specific examples of research within this thrust area are: a) precise control of particle size during processing (e.g. emulsification, spray drying, microencapsulation, and nanomaterials) and storage (e.g. microsintering and coalescence) via modulation of dynamic interfacial conditions; and b) identifying novel capillary and interfacial mechanisms for enhancing drying rates in porous food matrices. Advances in this area enabled by insights achieved through combinations of experimental tools (such as high-speed visualization and laser Doppler velocimetry) and recent computational modeling tools (such as first-principles approaches and direct numerical simulations) will be presented.

Dictyostelium Genes Dysregulated in an O-Glycosylation Mutant Identified by mRNA Differential Display

Seven differentially-expressed cDNA clones were isolated by using an mRNA differential display between a Dictyostelium wild-type AX2 and a mutant HG794 defective in O-glycosylation. Transcript levels for the seven clones were reduced or not detectable in the mutant HG794. Homology search showed that the four cDNA clones, DD-3 and DD-7~9 are novel and that three cDNA clones, DD-4 and DD-5, -6 encode an actin-bundling protein and phosphodiesterase inhibitors, respectively. Full-length cDNAs for DD-3 and -8 were isolated and labeled DD3-3 and DD8-14, respectively. DD3-3 consists of 2,166 bp and DD8-14 of 2,084 bp. DD3-3 was preliminarily reported in a previous paper [1]. SSL850 was named a clone by the “Dictyostelium cDNA Project in Japan”, containing a fulllength cDNA for DD-7 and was labeled DD7-1 of 902 bp. It has 60% homology with discoidin Ia. DD8-14 most likely has no direct role in glycosylation, while DD3-3 and DD7-1 very likely are involved in some aspect of recognition of glycosylation.