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CELL BIOLOGY & MOLECULAR GENETICS

Isolation and Characterization of Two Promoters from Linseed for Genetic Engineering

^a National Research Council of Canada, Plant Biotechnology Institute, Saskatoon, SK, Canada S7N 0W9
^b Crop Development Centre, Univ. of Saskatchewan, Saskatoon, SK, Canada S7N 5A8

jravinder{at}pbi.nrc.ca

Linseed (Linum usitatissimum L.) is an important oilseed crop worldwide and is cultivated for the high level of linolenic acid (18:3) in its seed oil. Currently, there is a concerted effort to improve linseed by genetic engineering. This will require appropriate transgenes and tissue-specific or constitutive promoters. We report the isolation and characterization of two linseed promoters from a two-member gene family encoding the enzyme stearoyl-acyl carrier protein desaturase (SAD). The SAD1 and SAD2 gene promoter were each fused transcriptionally with the reporter gene for ß-glucuronidase (uidA; GUS) and were transferred to linseed to study their expression pattern. In transgenic linseed, GUS activity mediated by the SAD2 promoter appeared to be constitutive and was detected in leaves, apices, stem, roots, flower buds, flowers, and seeds. In contrast, GUS activity mediated by the SAD1 promoter appeared to be root- and seed-specific. In developing seeds, both the promoters exhibited a temporal expression pattern concomitant with protein and lipid biosyntheses. The GUS activity could be detected as early as 4 days after pollination (dap) and in mature seeds (50 dap) with the highest activities around mid-development. The first pair of linseed promoters will be useful for manipulating the expression of indigenous as well as transgenes in linseed to create value-added cultivars.

Abbreviations: dap, days after pollination • OLs, oligoribonucleotides • SAD, stearoyl-acyl carrier protein desaturase • uidA, ß-glucuronidase