Demonstration that the Neurospora crassa mutation un-4 is a single nucleotide change in the tim16 gene encoding a subunit of the mitochondrial inner membrane translocase

The Neurospora crassa temperature sensitive mutation known as un-4 has been shown by a map-based complementation approach to be a single nucleotide change in the open reading frame of the mitochondrial inner membrane translocase subunit tim16 (NCU05515)

Fantastic growth as the FGSC turns 50

Comparative Medicine - OneHealth and Comparative Medicine Poster SessionFounded in 1960, the Fungal Genetics Stock Center enters it's fiftieth year of operation during a period of tremendous growth. The collection has more than doubled since moving to UMKC in 2004 and has added new materials that reach out beyond it's traditional constituency. Among these are deletion sets for Neurospora, Cryptococcus and Candida as well as molecular genetic tools for working with industrial fungi, model organisms, and plant and human pathogens. With distribution growing every year, the FGSC sends materials to scientists in over 35 countries every year; approximately half of our orders are from within the US. In addition to being part of an NIH funded multi-institution Functional Genomics Program for Neurospora, the FGSC is involved in cutting edge genomics research with collaborators at the US DOE Joint Genome Institute. The FGSC and its staff are actively involved in national and international societies and ad hoc working groups fostering the development of collection resources in the US and around the world

The effect of repeated freeze-thaw cycles on cryopreserved Neurospora crassa samples.

To better characterize handling parameters for the arrayed mutants prepared for the Neurospora functional genomics program, we have put 7 day old conidia from strain FGSC 2489 through a series of cycles of freezing at -80 C in 25% glycerol and 3.5% reconstituted non-fat dry milk

Analysis of the DNA sequence of the putative ABC transporter NCU09975 in Neurospora crassa strains carrying acriflavin resistance markers.

Genomic DNA sequence was determined for the putative Neurospora crassa ABC transporter NCU09975 from several different classical mutant strains including several acriflavin resistant mutants. The sensitivity of these strains to acriflavin was tested. While the open reading frame NCU09975 has multiple polymorphisms in strains sequenced for other purposes, none of the acriflavin resistant classical mutants tested had polymorphisms in the NCU09975 coding region or in the 195 bases upstream of the translation start site

Identification of the Neurospora crassa mutation un-10 as a point mutation in a gene encoding eukaryotic translation initiation factor 3, subunit B.

The Neurospora crassa temperature-sensitive mutant known as un-10 has been shown by a map-based complementation approach to be a single nucleotide change in the open reading frame of the eukaryotic translation initiation factor 3b (NCU02208.3)

Role of gliotoxin in the symbiotic and pathogenic interactions of Trichoderma virens

Using a gene disruption strategy, we generated mutants in the gliP locus of the plant-beneficial fungus Trichoderma virens that were no longer capable of producing gliotoxin. Phenotypic assays demonstrated that the gliP-disrupted mutants grew faster, were more sensitive to oxidative stress and exhibited a sparse colony edge compared with the WT strain. In a plate confrontation assay, the mutants deficient in gliotoxin production were ineffective as mycoparasites against the oomycete, Pythium ultimum, and the necrotrophic fungal pathogen, Sclerotinia sclerotiorum, but retained mycoparasitic ability against Rhizoctonia solani. Biocontrol assays in soil showed that the mutants were incapable of protecting cotton seedlings from attack by P. ultimum, against which the WT strain was highly effective. The mutants, however, were as effective as the WT strain in protecting cotton seedlings against R. solani. Loss of gliotoxin production also resulted in a reduced ability of the mutants to attack the sclerotia of S. sclerotiorum compared with the WT. The addition of exogenous gliotoxin to the sclerotia colonized by the mutants partially restored their degradative abilities. Interestingly, as in Aspergillus fumigatus, an opportunistic human pathogen, gliotoxin was found to be involved in pathogenicity of T. virens against larvae of the wax moth, Galleria mellonella. The loss of gliotoxin production in T. virens was restored by complementation with the gliP gene from A. fumigatus. We have, thus, demonstrated that the putative gliP cluster of T. virens is responsible for the biosynthesis of gliotoxin, and gliotoxin is involved in mycoparasitism and biocontrol properties of this plant-beneficial fungusFil: Vargas, Walter Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Centro de Estudios Fotosintéticos y Bioquímicos (i); ArgentinaFil: Mukherjee, Prasun K.. Bhabha Atomic Research Centre; IndiaFil: Laughlin, David. Texas A&M University; Estados UnidosFil: Wiest, Aric. University Of Missouri; Estados UnidosFil: Moran Diez, Maria E.. Texas A&M University; Estados UnidosFil: Kenerley, Charles M.. Texas A; Estados Unido

Characterization of the Temperature-Sensitive Mutations un-7 and png-1 in Neurospora crassa

The model filamentous fungus Neurospora crassa has been studied for over fifty years and many temperature-sensitive mutants have been generated. While most of these have been mapped genetically, many remain anonymous. The mutation in the N. crassa temperature-sensitive lethal mutant un-7 was identified by a complementation based approach as being in the open reading frame designated NCU00651 on linkage group I. Other mutations in this gene have been identified that lead to a temperature-sensitive morphological phenotype called png-1. The mutations underlying un-7 result in a serine to phenylalanine change at position 273 and an isoleucine to valine change at position 390, while the mutation in png-1 was found to result in a serine to leucine change at position 279 although there were other conservative changes in this allele. The overall morphology of the strain carrying the un-7 mutation is compared to strains carrying the png-1 mutation and these mutations are evaluated in the context of other temperature-sensitive mutants in Neurospora

The isolation and characterization of a peptide synthetase gene from Trichoderma virens

Due to the character of the original source materials and the nature of batch digitization, quality control issues may be present in this document. Please report any quality issues you encounter to [email protected], referencing the URI of the item.Includes bibliographical references (leaves 47-53).Issued also on microfiche from Lange Micrographics.The filamentous fungus Trichoderma has a long history of biological control, protecting numerous plant species against a broad spectrum of soilborne pathogens. Additionally, many species are significant sources of industrial enzymes. Trichoderma isolates have been shown to produce a number of antibiotic compounds, including peptaibols, which are deleterious to both fungi and bacteria. Peptaibols are known for their high α-aminoisobutyric acid content and their production as a mixture of isoforms ranging from 7-20 amino acids in length. The antibiotic activity they exhibit is probably due to an ability to form pores in lipid membranes. These pores conduct ions, leading to loss of osmotic balance and cell death. Because of the potential importance of peptaibols in the biological control of plant diseases, we sought to clone the gene(s) responsible for their synthesis. With their unusual amino acid content, we expected peptaibols to be the product of nonribosomal peptide synthetases (NRPSs). NRPSs have a modular structure in which each module is a semiautonomous unit that recognizes, activates, and incorporates a single residue into the final peptide. We have identified novel peptaibols from Trichoderma virens and cloned the NRPS gene, tex1, responsible for their biosynthesis. Disruption of tex1 in T. virens eliminates peptaibol production, as determined by HPLC analysis. This gene is the first complete peptide synthetase gene cloned from the genus Trichoderma. The 62.8 kb continuous open reading frame represents not only the first peptaibol synthetase gene cloned, but also the largest known open reading frame of any gene in any organism. Tex1 is expressed under all examined conditions, including ungerminated conidia. Since modular order is co-linear with that of the product, we were able to assign substrate amino acid(s) to each module. Comparison of protein sequences between modules is consistent with the module/substrate assignments. Here we report the first genetic evidence showing that peptaibols from T. virens are produced by an enormous peptide synthetase. Manipulation of the abundance or sequence of peptaibols or the expression of tex1 in other fungi or plants could lead to increased suppression of plant pathogens through biological control

Towards the Biological Synthesis of fullerene molecules [abstract]

Comparative Medicine - OneHealth and Comparative Medicine Poster SessionThis invention describes the synthesis of carbon fullerene molecules and nanotubes using biological or enzymatic means. Presently carbon nanotubes are synthesized by either chemical vapor deposition or by electrical discharge in a controlled atmosphere. These approaches are very technically demanding and difficult to control giving heterogeneous populations of products. The present invention describes approaches to de novo synthesis of carbon nanotubes and related fullerenes as well as the stepwise addition of carbon to growing carbon nanotubes. In the latter case, the dimensions of the growing nanotube, including both diameter and length, are customizable and limited only by the input of raw materials and extent to which the reactions are allowed to progress.

Neurospora TS lethal genes involved in protein production, transport, or quality control.

<p>Neurospora TS lethal genes involved in protein production, transport, or quality control.</p