A technique for replica platingCoprinus lagopus, a filamentous fungus

SUMMARYThe problem of replica plating filamentous fungi such asCoprinus lagopusis overcome by inducing micro-colonies with sodium deoxycholate and using ‘Velcro’, a hooked material, to replace velveteen in the standard replica plating technique. ‘Velcro’ is advantageous in that it has a regular pattern of closely spaced hooks which transfer small inocula from the colonies on the master plate.

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A Replica Plating Technique for the Isolation of Nutritionally Exacting Mutants of a Filamentous Fungus (Aspergillus nidulans)

Journal of General Microbiology ◽

10.1099/00221287-20-3-540 ◽

1959 ◽

Vol 20 (3) ◽

pp. 540-548 ◽

Cited By ~ 44

Author(s):

C. F. ROBERTS

Keyword(s):

Aspergillus Nidulans ◽

Filamentous Fungus ◽

Plating Technique ◽

Replica Plating

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A replica plating technique for in vitro study of susceptibility of Canadida albicans to miconazole, econazole and ketoconazole: Some data for standaridization

Annales de l Institut Pasteur Microbiologie ◽

10.1016/s0769-2609(85)80099-5 ◽

1985 ◽

Vol 136 (3) ◽

pp. 371-380 ◽

Cited By ~ 1

Author(s):

A. Sanchez-Sousa ◽

C. Gomez-Criado ◽

M.L. Jimenez ◽

A. Fernandez-Jorge ◽

F. Baquero

Keyword(s):

In Vitro Study ◽

Vitro Study ◽

Plating Technique ◽

Replica Plating

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REPLICA PLATING TECHNIQUE FOR STUDYING MICROBIAL INTERACTIONS IN SOIL

Canadian Journal of Microbiology ◽

10.1139/m65-084 ◽

1965 ◽

Vol 11 (4) ◽

pp. 629-636 ◽

Cited By ~ 28

Author(s):

G. Stotzky

Keyword(s):

Nutritional Composition ◽

Mixed Population ◽

Microbial Interactions ◽

Individual Species ◽

Selective Inhibitors ◽

Subsequent Growth ◽

Sterile Soil ◽

Plating Technique ◽

Replica Plating ◽

Plating Method

A replica plating method was developed to study ecology of microorganisms in soil. Precise placement of inocula and amendments at desired loci in sterile soil contained in petri plates were accomplished with a template. Subsequent growth and distribution of individual species, even when part of a mixed population, was measured by periodic transfer with an easily constructed replicator to agar plates of differing nutritional composition or containing selective inhibitors. The method is rapid and reproducible, and permits the study of many variables and interactions in a single soil plate; it can also be used with non-sterile soil and other suitable microbial habitats.

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Isolation of the Streptomycin Resistant Variants of E. Coli B by a Modified Replica Plating Technique

Pathobiology ◽

10.1159/000160749 ◽

1956 ◽

Vol 19 (6) ◽

pp. 792-797

Author(s):

S. Banič

Keyword(s):

E Coli ◽

Plating Technique ◽

Replica Plating

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Defining individual size in the model filamentous fungus Neurospora crassa

Proceedings of The Royal Society B Biological Sciences ◽

10.1098/rspb.2015.2470 ◽

2016 ◽

Vol 283 (1826) ◽

pp. 20152470 ◽

Cited By ~ 2

Author(s):

Linda Ma ◽

Boya Song ◽

Thomas Curran ◽

Nhu Phong ◽

Emilie Dressaire ◽

...

Keyword(s):

Neurospora Crassa ◽

Filamentous Fungi ◽

Filamentous Fungus ◽

Reproductive Potential ◽

Fungal Mycelium ◽

Dependent Property ◽

Mycelial Network ◽

And Function ◽

Individual Size ◽

Over Time

It is challenging to apply the tenets of individuality to filamentous fungi: a fungal mycelium can contain millions of genetically diverse but totipotent nuclei, each capable of founding new mycelia. Moreover, a single mycelium can potentially stretch over kilometres, and it is unlikely that its distant parts share resources or have the same fitness. Here, we directly measure how a single mycelium of the model ascomycete Neurospora crassa is patterned into reproductive units (RUs), meaning subpopulations of nuclei that propagate together as spores, and function as reproductive individuals. The density of RUs is sensitive to the geometry of growth; we detected 50-fold smaller RUs when mycelia had expanding frontiers than when they were constrained to grow in one direction only. RUs fragmented further when the mycelial network was perturbed. In mycelia with expanding frontiers, RU composition was strongly influenced by the distribution of genotypes early in development. Our results provide a concept of fungal individuality that is directly connected to reproductive potential, and therefore to theories of how fungal individuals adapt and evolve over time. Our data show that the size of reproductive individuals is a dynamic and environment-dependent property, even within apparently totally connected fungal mycelia.

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Isolation of sugar and antibiotic mutants in Chlorella vulgaris with a new replica plating technique

Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis ◽

10.1016/0027-5107(73)90231-5 ◽

1973 ◽

Vol 19 (3) ◽

pp. 313-318 ◽

Cited By ~ 1

Author(s):

Sipra Guha-Mukherjee ◽

E.C. Keller

Keyword(s):

Chlorella Vulgaris ◽

Plating Technique ◽

Replica Plating

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Modifying the growth habit of the filamentous fungus Fusarium solani to facilitate replica plating procedures

Canadian Journal of Botany ◽

10.1139/b77-101 ◽

1977 ◽

Vol 55 (7) ◽

pp. 848-851 ◽

Cited By ~ 9

Author(s):

Hans D. VanEtten ◽

H. Gunnar Kølmark

Keyword(s):

Filamentous Fungus ◽

Fusarium Solani ◽

Radial Growth ◽

Growth Habit ◽

Fast Growing ◽

Replica Plating ◽

Morphological Mutants ◽

Triton X

The surfactants Triton X-100 and X-171 were found to facilitate the replica plating of Fusarium solani by markedly inhibiting the radial growth of this fast-growing fungus. In addition, specific types of morphological mutants were isolated which assisted the replica plating of F. solani.

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Transient Silencing of DNA Repair Genes Improves Targeted Gene Integration in the Filamentous Fungus Trichoderma reesei

Applied and Environmental Microbiology ◽

10.1128/aem.00535-17 ◽

2017 ◽

Vol 83 (15) ◽

Cited By ~ 6

Author(s):

Pak Yang Chum ◽

Georg Schmidt ◽

Markku Saloheimo ◽

Christopher P. Landowski

Keyword(s):

Dna Repair ◽

Trichoderma Reesei ◽

Filamentous Fungi ◽

Filamentous Fungus ◽

Unintended Consequences ◽

Dna Repair Genes ◽

Content Type ◽

Targeted Integration ◽

Gene Integration ◽

Repair Genes

ABSTRACT Trichoderma reesei is a filamentous fungus that is used worldwide to produce industrial enzymes. Industrial strains have traditionally been created though systematic strain improvement using mutagenesis and screening approaches. It is also desirable to specifically manipulate the genes of the organism to further improve and to modify the strain. Targeted integration in filamentous fungi is typically hampered by very low frequencies of homologous recombination. To address this limitation, we have developed a simple transient method for silencing genes in T. reesei. Using gene-specific small interfering RNAs (siRNAs) targeted to mus53, we could achieve up to 90% knockdown of mus53 mRNA. As a practical example, we demonstrated that transient silencing of DNA repair genes significantly improved homologous integration of DNA at a specific locus in a standard protoplast transformation. The best transient silencing of mus53 with siRNAs in protoplasts could achieve up to 59% marker gene integration. IMPORTANCE The previous solution for improving targeted integration efficiency has been deleting nonhomologous end joining (NHEJ) DNA repair genes. However, deleting these important repair genes may lead to unintended consequences for genomic stability and could lead to the accumulation of spontaneous mutations. Our method of transiently silencing NHEJ repair pathway genes allows recovery of their important repair functions. Here we report a silencing approach for improving targeted DNA integration in filamentous fungi. Furthermore, our transient silencing method is a truly flexible approach that is capable of knocking down the expression of a target gene in growing mycelial cultures, which could facilitate the broad study of gene functions in T. reesei.

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