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- Title
Targeted gene analysis in Ulmus americana and U. pumila tissues.
- Authors
Nasmith, C.; Jeng, R.; Hubbes, M.
- Abstract
Steady-state gene expression was compared between Dutch elm disease (DED)-susceptible Ulmus americana and DED-resistant U. pumila callus, leaf midrib, root and inner bark tissues. Stress-related cDNAs including phenylalanine ammonia-lyase (PAL), chitinase (CHT) and polygalacturonase-inhibiting protein (PGIP) were isolated and compared following RT-PCR of elm tissues. Complete CHT and partial PAL and PGIP cDNA transcripts were identified, each displaying sequence variation between elm species. These transcripts were Dig-labelled and subsequently used for northern analyses of the elm tissues. Midrib and root tissue displayed highest steady-state gene expression compared with inner bark and callus tissues. A modified nucleic acid isolation technique was necessary for downstream RNA analyses. Lithium chloride and polyvinylpyrrolidone were critical for efficient removal of polysaccharides and phenolics associated with some of the elm tissues. Steady-state gene expression is discussed in relation to the tissues investigated. The use of tissues other than in vitro callus culture more closely represents the tissues associated with the elm’s vascular response to DED.
- Subjects
CRYOBIOLOGY; PLANT diseases; AGRICULTURAL pests; CALLUS (Botany); DUTCH elm disease; ELM diseases &; pests; FUNGAL diseases of plants; NITROGEN compounds; RNA
- Publication
Forest Pathology, 2008, Vol 38, Issue 2, p90
- ISSN
1437-4781
- Publication type
Article
- DOI
10.1111/j.1439-0329.2007.00523.x