Fungal hyphae--and in some cases, spores--are multi-nucleate. During genetic transformation of these spores or mycelia, only one nucleus generally receives the transferred T-DNA generating heterokaryotic colonies. Characterization of genetic changes, such as the effects of gene disruption in the transformants, requires purified homokaryotic lines. Hyphal tip transfer has conventionally been used to isolate homokaryons. We developed an alternative method for purification of fungal homokaryons from transformed heterokaryotic lines of Sclerotinia sclerotiorum. Ultrasound pulses were used to generate bi-septate, unicellular hyphal fragments that regenerate under selection to produce homokaryotic lines that can be easily identified using colony PCR. This technique facilitates the purification of transformed lines, which allows for routine genome manipulation, and should be adaptable for other filamentous fungi.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.2144/000113243 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Variation in allele frequencies at the bg112 locus reveals unequal inheritance of nuclei in a dikaryotic isolate of the fungus Rhizophagus irregularis.
Mycorrhiza
May 2018
Department of Ecology and Evolution, Biophore Building, University of Lausanne, 1015, Lausanne, Switzerland.
The genetic state of the arbuscular mycorrhizal fungus species Rhizophagus irregularis differs among isolates, including both homokaryotic and dikaryotic isolates. Via the production of multi-nucleate axexual spores, siblings of dikaryotic isolates may inherit unequal frequencies of nucleotypes. Using bg112, a microsatellite marker, previous studies revealed that lines deriving from single spores of the dikaryotic R.
View Article and Find Full Text PDFQuantitative trait locus mapping for bruising sensitivity and cap color of Agaricus bisporus (button mushrooms).
Fungal Genet Biol
April 2015
Plant Breeding, Wageningen University & Research Centre, Droevendaalsestaag 1, 6700 AA Wageningen, The Netherlands.
White button mushrooms discolor after mechanical damage of the cap skin. This hampers the development of a mechanical harvest system for the fresh market. To unravel the genetic basis for bruising sensitivity, two haploid populations (single spore cultures) were generated derived from crosses between parental lines differing in discoloration after mechanical damage (bruising sensitivity).
View Article and Find Full Text PDFGenetic variation and combining ability analysis of bruising sensitivity in Agaricus bisporus.
PLoS One
August 2014
Plant Breeding, Wageningen University and Research Center, Wageningen, The Netherlands ; Institute of Agricultural Resources and Regional Planning, Chinese Academy of Agricultural Sciences, Beijing, PR China.
Advanced button mushroom cultivars that are less sensitive to mechanical bruising are required by the mushroom industry, where automated harvesting still cannot be used for the fresh mushroom market. The genetic variation in bruising sensitivity (BS) of Agaricus bisporus was studied through an incomplete set of diallel crosses to get insight in the heritability of BS and the combining ability of the parental lines used and, in this way, to estimate their breeding value. To this end nineteen homokaryotic lines recovered from wild strains and cultivars were inter-crossed in a diallel scheme.
View Article and Find Full Text PDFIsolation of fungal homokaryotic lines from heterokaryotic transformants by sonic disruption of mycelia.
Biotechniques
January 2010
Agriculture and Agri-Food Canada, 107 Science Place, Saskatoon, SK, Canada.
Fungal hyphae--and in some cases, spores--are multi-nucleate. During genetic transformation of these spores or mycelia, only one nucleus generally receives the transferred T-DNA generating heterokaryotic colonies. Characterization of genetic changes, such as the effects of gene disruption in the transformants, requires purified homokaryotic lines.
View Article and Find Full Text PDFA genetic linkage map for the ectomycorrhizal fungus Laccaria bicolor and its alignment to the whole-genome sequence assemblies.
New Phytol
March 2009
UMR 1136, INRA-Nancy Université, Interactions Arbres/Microorganismes, INRA-Nancy, 54280 Champenoux, France.
A genetic linkage map for the ectomycorrhizal basidiomycete Laccaria bicolor was constructed from 45 sib-homokaryotic haploid mycelial lines derived from the parental S238N strain progeny. For map construction, 294 simple sequence repeats (SSRs), single-nucleotide polymorphisms (SNPs), amplified fragment length polymorphisms (AFLPs) and random amplified polymorphic DNA (RAPD) markers were employed to identify and assay loci that segregated in backcross configuration. Using SNP, RAPD and SSR sequences, the L.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!