Characterisation of Agaricus bisporus response genes to Verticillium fungicola infection
Fecha
2006Versión
Acceso abierto / Sarbide irekia
Tipo
Contribución a congreso / Biltzarrerako ekarpena
Versión
Versión publicada / Argitaratu den bertsioa
Impacto
|
nodoi-noplumx
|
Resumen
The mycoparasite Verticillium fungicola is a persistent threat to the cultivation
of the mushroom Agaricus bisporus. Mushroom “dry bubble” is characterised
by an undifferentiated mass of cells and can result in major crop losses. During
the establishment of “dry bubble” substantial changes occur in the biochemistry
and physiology of both partners. To enable new insights to be
made into the m ...
[++]
The mycoparasite Verticillium fungicola is a persistent threat to the cultivation
of the mushroom Agaricus bisporus. Mushroom “dry bubble” is characterised
by an undifferentiated mass of cells and can result in major crop losses. During
the establishment of “dry bubble” substantial changes occur in the biochemistry
and physiology of both partners. To enable new insights to be
made into the molecular events underlying the disease, work is in progress to
identify genes expressed during pathogen infection. Subtractive Suppressive
Hybridisation (SSH) has enabled recovery of 65 expressed sequenced tags
(ESTs) differentially expressed during infection. After database searches 27
of the genes were identified as most likely from V. fungicola, 25 from A. bisporus
and 13 unknown. Bioinformatic analysis suggested that the response
genes identified were involved in a range of biological functions that included
stress, signalling, protein synthesis and cell wall structure and function.
Specific full-length genes will be recovered using cDNA library constructed
from lesions of A. bisporus infected with V. fungicola, enabling silencing
approaches to be used to further investigate the role of the identified
genes in disease. An alternative higher-throughput method of gene function
analysis, RNA interference (RNAi) using A. bisporus model genes (URA3,
CBX), is also being developed. Silencing constructs expressing RNAi hairpin
were transformed into A. bisporus using Agrobacterium tumefaciens and hygromycin
resistance. Screening of the transformants by PCR confirmed integration
of the silencing construct in 24 transformants. RT-PCR is being
used to confirm transcription of the RNAi hairpin. Quantitative PCR will be
used to analyse levels of target gene transcripts post RNAi transformation.
The role of A. bisporus genes identified, in the infection process, will be determined
through infection trails with A. bisporus silenced lines. [--]
Materias
Verticillum fungicola,
Agaricus bisporus
Editor
Universidad Pública de Navarra / Nafarroako Unibertsitate Publikoa
Publicado en
Antonio G. Pisabarro and Lucía Ramírez (eds.): VI Meeting on Genetics and Cellular Biology of Basidiomycetes (GCBB-VI). Pamplona: Universidad Pública de Navarra / Nafarroako Unibertsitate Publikoa, 2006.
Notas
Resumen de la conferencia presentada al VI Meeting on Genetics and Cellular Biology of Basidiomycetes (GCBB-VI), organizado por y celebrado en la Universidad Pública de Navarra el 3-6 de junio de 2005.