PCR-BASED CLONING OF AN ALPHA-GALACTOSIDASE GENE FROM DEACCLIMATED PETUNIA (PETUNIA x HYBRIDA)

Previous studies of plant cold hardiness have focused mainly on the cold acclimation response, while studies on the deacclimation process have been limited.
Alpha-galactosidase a key catabolic enzyme of the raffinose family oligosaccharides (RFO), cleaves the terminal-linked moiety from galactose-containing oligosaccharides.
A cDNA clone petgal, was isolated from Petunia x hybrida ‘Mitchell’ leaf RNA by RT-PCR using degenerate oligosaccharide primers designed to amplify the mature -galactosidase protein.
The putative -galactosidase cDNA sequence has high nucleotide sequence homology (<80%) to other known plant -galactosidases.
Southern blot analysis suggests that -galactosidase represents a single gene family.
Based on the amino acid alignment of the petunia -galactosidase enzyme with other known -galactosidases, it appears that the protein is conserved among species.
Up-regulation of the -galactosidase gene in response to deacclimation duration and temperature, suggests that warm temperature may regulate RFO catabolism by increasing the transcription of the -galactosidase gene.

XXVI International Horticultural Congress: Environmental Stress and Horticulture Crops

J.C. Pennycooke, M.L. Jones, R. Vepachedu, C. Stushnoff

alpha-galactosidase, cloning, deharden, petunia, RFO catabolism, RT-PCR

https://doi.org/10.17660/ActaHortic.2003.618.10