The isolation and functional characterisation of a B. napus acyl carrier protein 5' flanking region involved in the regulation of seed storage lipid synthesis.

Abstract

Acyl carrier protein (ACP) is a key component of the fatty acid biosynthetic machinery in plants. A 1.4 kb 5' flanking region of a Brassica napus ACP gene (ACP05) was transcriptionally fused to the reporter gene beta-glucuronidase (GUS), and expression of the chimeric gene monitored in transgenic tobacco. GUS activity was found to increase through seed development reaching a maximum value, coincident with the most active phase of storage lipid synthesis that was, on average, 100-fold higher than that observed in leaf. In control plants transformed with CaMV 35S-GUS constructs, GUS activity was similar in leaf and all stages of seed development. Based on average values, the level of GUS expression obtained via the ACP promoter was comparable to that obtained from the CaMV 35S promoter. We therefore conclude that the isolated 5' ACP flanking sequence represents a strong promoter element involved in the developmental regulation of storage lipid synthesis in B. napus seed tissue. Putative regulatory elements in the 5' upstream region of ACP05 were identified by dot matrix analysis and by sequence comparison with the upstream regions from a second seed-expressed rape ACP gene and from an Arabidopsis ACP gene.