Canadian Forest Service Publications

Characterization of a fall protein of sugar pine and detection of its homologue associated with frost hardiness of western white pine needles. 1995. Ekramoddoullah, A.K.M.; Taylor, D.; Hawkins, B.J. Canadian Journal of Forest Research 25: 1137-1147.

Year: 1995

Issued by: Pacific Forestry Centre

Catalog ID: 4164

Language: English

Availability: Order paper copy (free)

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Abstract

A 19-kDa protein (named Pin l I) of sugar pine was detected in increasing amounts in the fall. This protein was separated by SDS-PAGE and also by two-dimensional gel electrophoresis. Pin l I was composed of two acidic isoforms. This protein was subjected to N-terminal amino acid sequence analysis. The two isoforms had an identical N-terminal amino acid sequence. The N-terminal peptide was synthesized and purified, and the purity of the synthetic peptide was greater than 95%. The peptide was conjugated to a carrier protein, Kehole limphet hemocyanin (KLH). Rabbits were immunized with peptide-KLH and the anti-peptide antibody was purified from the crude antisera by immuno-affinity chromatography. The antibody was shown to bind specifically to Pin l I. This anti-Pin l I antibody was used in a Western immunoblot to detect the homologues of Pin l I in two other five-needled white pines: western (named Pin m III) and eastern white pine. The antibody was also used to monitor the seasonal variation of Pin m III in western white pine needles. Pin m III was shown to be associated with overwintering of western white pine seedlings. A significant correlation was observed between the frost hardiness of western white pine foliage and the content of Pin m III.