MASMC 2010

Usif M., O. S. Isikhuemhen, F. Anike and E. I. Ohimain

Department of Natural Resources & Environmental Design, NC A&T State University, Greensboro, NC 27411.

Abstract: Peanut shells are of particular interest in bioconversion because of its mass accumulation as a waste from agro-industrial activity. However, its bioconversion to bioproducts has been limited by the high lignin content (40%). Corn stalks, is also an abundant agricultural wastes reported to be amenable to bioconversion by white rot fungi.  The aim of this research is to determine the ability of Pleurotus ostreatus to convert peanut shells and in combination with corn stalks to mushrooms and macromolecule profiles of degraded substrates. Preliminary studies on PS & CS in 21 substrate combinations, with 5% and 10% rye added as a supplement to some of the combinations, showed that faster colonization of mixed  PS and CS substrates (70PS:5Rye:25CS, 85PS:5Rye:10CS, & 50PS:50CS) was favored by Pleurotus ostreatus over the non-mixed substrates. Fruit induction studies also indicated that mixed PS and CS substrates, supplemented with 10% rye, produced the highest mushroom yield. From the previous study and results, peanut shells could perform better as a mixed substrate for mushroom cultivation than non-mixed. The Present work will evaluate the bioconversion of PS and CS substrates in seven substrate combinations (SCs) incubated at five colonization periods as well as mushroom cultivation in large mass substrates of 3 SCs selected from the previous study. Analysis of lignin, hemicelluloses & cellulose, water soluble carbohydrates, Amino acids, in vitro digestibility, Loss of organic matter (LOM), Average weight of mushroom and Biological Efficiency (BE) are among key parameters that will be compared and evaluated in this study.

Meghan A. McCormick, Marc Cubeta and Larry Grand

Center for Integrated Fungal Research

Department of Plant Pathology

North Carolina State University

Abstract

The objective of this study is to compare the morphological and genetic characteristics from individuals of the wood decay fungi Fomes fasciatus, Fomes fomentarius, and Piptoporus betulinus, to each other and to other individuals collected from the geographical extremes within their known range. We propose to test the hypothesis that species from a similar geographic area are more closely related and that genetic differentiation will increase with geographic distance, for each of the species. A method to isolate DNA directly from fresh and dried herbarium basidiocarps has been developed in our laboratory and involves placing a small piece of context tissue from the fruiting body into a plastic microcentrifuge tube that contains extraction buffer and glass beads. This DNA will be subsequently amplified by polymerase chain reaction (PCR) and sequenced. The DNA sequence from the internal transcribed spacer region (ITS) of the ribosomal DNA repeat and the coding gene for RNA polymerase subunit II (RPB2) will be compared to sequences deposited in the public database GenBank to confirm species identity. Sequences will be aligned and used to assess genetic diversity and genetic differentiation within and between individuals of the same and different species from each geographic area.

Timothy J. Baroni

Department of Biological Sciences, State University of New York – College at Cortland, Cortland, NY  13045

email: tim.baroni@cortland.edu

Abstract:  A decade of field work in the Greater Antilles and Belize will be summarized. Some of the more attractive species discovered during that work will be illustrated. The rate of new discoveries for selected groups of macrofungi will be discussed and the case made for more intensive field exploration in these regions in the future.  An overview of two unique expeditions to the highest cloud forest in the Maya Mountains will also be presented to illustrate the high adventure of field mycology.

Hannah T. Reynolds¹, Michael Castellano², Kentaro Hosaka³, Rytas J. Vilgalys¹

¹Duke University, ²USDA Forest Service Corvallis OR, ³Japan National Museum of Nature and Science

Abstract: The false truffle Elaphomyces is parasitized by Elaphocordyceps, which are related to parasites of cicada nymphs. When the parasite and host are
collected together, the host often cannot be identified because it a) may not
produce spores which are often needed for identification and b) it may be
discolored and degraded. Little is known about the specificity and possible
co-evolution of the parasite on its host. Recent publications on the phylogeny
of Cordyceps will be reviewed, as will preliminary molecular data on the
phylogenetics of Elaphomyces.  We have developed primers specific to both host
and parasite in order to identify the players through molecular means. The
current molecular and morphological data on the specificity of Elaphocordyceps species will be discussed.

Key Words: Elaphomyces, Cordyceps, coevolution, parasitism, ITS2

W.L. Willis*, O.S. Isikhuemhen, R. Minor, S. Hurley and  E.I. Ohimain

North Carolina Agricultural and Technical State University, Greensboro, NC

An experiment was conducted to evaluate the dietary inclusion of fungus myceliated grain (FMG) on Eimeria infected broiler performance responses.  A total of 144 broiler chicks were assigned to 8 treatments groups as follows: 1) Control-no challenge, 2) 5%-FMG no challenge, 3) Inovo-coccivac (IC) no challenge, 4) Spray-coccivac (SC)-no challenge, 5) Control-challenge, 6) 5% FMG challenged, 7) Inovo-coccivac-challenged and Spray-coccivac (SC) challenged.  Broilers were challenged with an Eimeria sp. mixture of E. acervelina, E. maxma and E. tenella at 28 d of age.  Body weight, mortality, fecal oocyst egg count, bursa and spleen weight, and some hematological parameters were measured.  The population of Eimeria counts at 35 days post challenge were significantly higher for challenged chickens in trts 5-8 when compared to the uninoculated controls trts 1-4.  Weights of challenged broilers were significantly affected by the Eimeria sp. mixture.  Eimeria sp. egg counts at 49 d in trt 5 with no protection was significantly (P > 0.05) different from the others with higher mortalities.  The result showed that significant P ≤ 0.05 differences were found in control vs challenged broiler body wts but not among challenged broilers.  The bursa/body weight percent and spleen and spleen/body weight were heavier in the unchallenged vs the challenged broilers.  The result from this study suggest that feeding a diet supplemented with 5 % FMG grain as an alternative to the use of other means of coccidiosis control can result in at least equivalent performance in the presence of Eimeria challenge during  grow out.

Key Words: Broilers, fungus myceliated grain, coccidiosis, performance.

Marianela Rodriguez-Carres^{1, 2},  Rytas Vilgalys¹, Francois Lutzoni¹ and Marc Cubeta². ¹ North Carolina State University, Department of Plant Pathology, Raleigh, NC, 27695. ² Duke University, Department of Biology, Durham, North Carolina, 27708.

The Rhizoctonia species complex is composed of genetically diverse fungi in the Ceratobasidiales that include soil saprobes, pathogens of plants in more than 180 families, and symbionts of orchids, liverworts, and mosses. Identification and classification of Rhizoctonia fungi (teleomorphs Ceratobasidium, Thanatephorus, Botryobasidium, and Tulasnella) is challenging due to the variability and limited number of morphological characters available, the lack of information about their sexual mode of reproduction, and conflicts among single gene phylogenies. Rhizoctonia fungi associated with the teleomorphs Ceratobasidium and Thanatephorus are often classified into anastomosis groups (AG) based on hyphal vegetative compatibility assays using predetermined tester strains. Phylogenetic analysis of ITS region of nuclear rDNA supported these anastomosis groups as monophyletic. However, these studies also revealed conflicts in the taxonomic delimitation of species due to variable evolutionary rates found in Rhizoctonia and their close relatives in the Cantharelloid clade. In the current study, we evaluated the phylogenetic relationships of Rhizoctonia by employing a multi-locus sequencing approach. Results from phylogenetic analyses of four loci (ITS, LSU, RPB2, EF1∝) from 29 taxa indicate that the family Ceratobasidiaceae forms a supported monophyletic clade that includes Ceratobasidium, Uthatobasidium, and Thanatephorus. Furthermore, the Ceratobasidiaceae appears to be the sister group to the Cantharelloid clade which includes Botryobasidium and Tulasnella. The genus Thanatehporus also forms a supported monophyletic group while Ceratobasidium appears to be paraphyletic, as a few species of Ceratobasidium cluster within Thanatephorus.  Therefore, the genus Ceratobasidium and the binucleate hyphal condition associated with it appear to be an ancestral condition, while the multinucleate hyphaal condition is associated with the more derived genus, Thanatephorus. Given the diverse habitat and economic importance of many of these fungal species, this study provides valuable insight into the genetic diversity, evolution, and species boundaries of Rhizoctonia, and the corticoid/resupinate fungi in the Cantharelloid clade.

Kirthi Kiran Yadagiri and Julia Kerrigan, Dept. Entomology, Soils, and Plant Sciences, Clemson University, Clemson, SC

Labyrinthula terrestris belongs to a group of organisms commonly referred to as net slime molds, which have primarily been isolated from marine environments. This species causes a disease called rapid blight on turfgrass. Symptoms include water-soaked lesions and browning or bronzing of foliage that lead to death of the infected turf, causing unsightly spots on golf course greens. Because little is known about this pathogen, its life cycle is being investigated with light and electron microscopy. L. terrestris can infect hosts indirectly through stomata, trichome bases, cut grass blades, and wounds, and through direct penetration of the host tissue. Once inside the host it has been observed in all cell types and is able to divide. Outside of the host, cells slowly aggregate and form clusters that appear to serve as resistant structures. L. terrestris is the only terrestrial species in the genus and one of only two pathogenic species; therefore, investigations on its basic biology are revealing novel findings.

Cesar Herrera¹, Gary Samuels², Amy Y. Rossman², Priscila Chaverri¹

1 – University of Maryland, College Park, Department of Plant Sciences and Landscape Architecture, 2102 Plant Sciences Building, University of Maryland, College Park, MD 20742
2 – United States Department of Agriculture, Systematic Mycology and Microbiology Lab, Rm. 304, B-011A, 10300 Baltimore Ave., Beltsville, MD 20705

Cosmospora vilior (Starbäck) Rossman & Samuels is a nectroid fungus that has been characterized by: 1) its tuberculate ascospores, 2) its occurrence on the surface of the stromata of xylariaceous fungi, and 3) its associated anamorph, Acremonium berkeleyanum, which produces green coloration in culture.  Cosmospora vilior has been reported to occur on other substrata including polyporaceous fungi, bark, and a new record of substrate, bone. The objective of this research is to determine the phylogenic relationship of the novel C. cf. vilior occurring on bone, and ultimately to revise C. vilior based on molecular analysis and morphological characters. Phylogenetic relationships were determined based on the analyses of sequences from six genes (internal transcribed spacer, large subunit nuclear rDNA,  translation-elongation factor 1-α, β-tubulin, α-actin, RNA polymerase II subunit 1; 3,472 total base pairs). Preliminary phylogenetic analyses suggest that C. vilior is paraphyletic. Phylogenetic analyses indicate a core clade consisting of several lineages occurring on xylariaceous fungi. The clades falling outside the core contain species occurring on non-xylariaceous fungi (e.g. polypores, bark, and bone). Morphological characters are currently being studied.

John Holliday, Aloha Medicinals Inc., 2300 Arrowhead Dr., Carson City, NV  89706

Abstract:

This paper demonstrates that many microorganisms are able to survive across geological time scales within veins of cellulosic material accumulated in the interior of incompletely carbonized organic deposits and low grade coal. Previous work from Pokorny, et al. alerts that viable fungal and bacterial spores amenable to reanimation and cultivation may well be present in such deposits. The availability of these organisms, previously thought to be extinct, presents development capabilities for novel antibiotics and other unique secondary metabolites derived from these archaic organisms. In this paper we have taken the previous work to a whole new level, showing  the isolation methodology used in reanimating  more than a hundred different species of bacteria and fungi through an assortment of growth media in an effort to determine the full organismal variety of the coal. Many of the fungal species isolated thus far were typified via genetic sequencing of the 28S-D2 ribosomal and internal transcriptional subunits (ITS), while the 5s gene sequence and Fatty Acid Methyl Ester methodology was used in identifying the bacterial species thus derived.  This work gives us a whole new genetic bank from which to work in the development of novel biologically active compounds, and indicates further research into this area of archaic organisms is needed.

Fabiano B. Rocha¹, Robert W. Barreto¹

¹Departamento de Fitopatologia, Universidade Federal de Viçosa, Viçosa, MG, 36570-000, Brasil

Despite common perceptions, not all pathogenic fungi are undesirable. Some pathogens are used to control invasive species (i.e. weeds), others are sources of pharmaceuticals, and others play an important role as biological regulators of populations of natural plants, thereby contributing to the maintenance of biological diversity. As Charles Darwin hinted, coextinction is the disappearance of one species as a consequence of its dependence on other species, which becomes extinct. Some documented examples of coextinction include pigeon-lice parasites, some parasites of primates (including Pneumocystis, nematodes and lice), pollinator wasps and herbivorous insects. Although the recognition that groups of plant pathogens such as rust and smut fungi that have coevolved with endangered plant species should be included in red lists is not new, examples of experimental demonstration that prove such claims for threatened pathogens are few if they exist at all. It is difficult to demonstrate empirically that any fungus is endangered; for this reason only three species of fungi (two lichens and one mushroom) are listed in official IUCN Red List of Threatened Species. No Brazilian fungi are listed as being endangered, but an initiative has been developed by Brazilian mycological team based in Universidade Federal de Viçosa (Minas Gerais state). This initiative started with the study of the mycobiota of two endangered tree species, Coussapoa floccosa (Cecropiaceae) and Euterpe edulis (Arecaceae), both natives of the Atlantic rainforest. Six new species and one new genus were associated with C flocossa, and four as yet unidentified species from E. edulis. The next phase of the project is to determine which, if any, of these species are strictly host-dependent. This study involves: 1- Search for those fungi in the same plant species in different locations; 2- Intensive survey of the mycobiota of plants occurring in the vicinity of C. floccosa and E. edulis, with particular reference to members of the Cecropiaceae and Arecaceae; 3- centrifugal-phylogenetic host-range testing of each fungal species by inoculation of test plants under controlled conditions; 4- complementary host-range testing by leaving selected test-plants as sentinels under the canopies of infected plants in the field (only done for C. floccosa) for one year. Preliminary results indicate that all fungi associated with C floccosa are strictly dependent, three species from E. edulis are host-specific and one is a polyphagous species of Bipolaris. This work is presently being extended to include the mycobiota of Dimorphandra wilsonii, a highly endangered tree from the Brazilian “cerrado” (South American savannah-like formation).