Cloning symbiosis‐related cDNAs from eucalypt ectomycorrhiza by PCR‐assisted differential screening
Open Access
- 1 October 1993
- journal article
- Published by Wiley in New Phytologist
- Vol. 125 (2) , 339-343
- https://doi.org/10.1111/j.1469-8137.1993.tb03884.x
Abstract
As part of a project to identify symbiosis‐related genes, we report here a simple differential screening procedure for isolating up‐ and down‐regulated fungal transcripts from a cDNA library of the developing Eucalyptus globulus‐Pisolithus tinctorius mycorrhiza. cDNA inserts of randomly selected λZAP plaques were amplified by PCR and separated by agarose gel electrophoresis. The PCR‐amplified cDNA samples were then screened by Southern blotting, using radiolabelled‐cDNA probes of high specific activity. We have applied this method to fungal transcripts that are differentially expressed in ectomycorrhizas during the early stages of development. We estimate that about 50 % of the fungal mRNA population is regulated by development of the symbiosis; several up‐ and down‐regulated cDNAs have been isolated for further analysis.Keywords
This publication has 19 references indexed in Scilit:
- Symbiosis‐related polypeptides associated with the early stages of ectomycorrhiza organogenesis in birch (Betula pendula Roth)New Phytologist, 1993
- Cold‐plaque screening: a simple technique for the isolation of low abundance, differentially expressed transcripts from conventional cDNA librariesThe Plant Journal, 1992
- Ectomycorrhizins ? symbiosis-specific or artifactual polypeptides from ectomycorrhizas?Planta, 1992
- Ectomycorrhizin Synthesis and Polypeptide Changes during the Early Stage of Eucalypt Mycorrhiza DevelopmentPlant Physiology, 1991
- Morphological, biochemical and molecular changes during ectomycorrhiza developmentCellular and Molecular Life Sciences, 1991
- Infectivity of pine and eucalypt isolates of Pisolithus tinctorius (Pers.) Coker & Couch on roots of Eucalyptus urophylla S. T. Blake in vitro. II. Ultrastructural and biochemical changes at the early stage of mycorrhiza formationNew Phytologist, 1990
- Regulation of gene expression in ectomycorrhizas. I. Protein changes and the presence of ectomycorrhiza-specific polypeptides in the Pisolithus-Eucalyptus symbiosisNew Phytologist, 1988
- Time sequence of the infection process eucalypt ectomycorrhizasNew Phytologist, 1988
- Primer-Directed Enzymatic Amplification of DNA with a Thermostable DNA PolymeraseScience, 1988
- Ontogeny of Eucalyptus pilularis – Pisolithus tinctorius ectomycorrhizae. I. Light microscopy and scanning electron microscopyCanadian Journal of Botany, 1987