Evaluation and characterization of an endosperm-specific sbeⅡa promoter in wheat

Starch,the main component of the wheat grain,is the product of a complex biochemical pathway. The sbeⅡα gene plays a key role in controlling the synthesis of starch, in particular, the biosynthesis of amylopectin,in maturing wheat grain.To investigate its regulatory mechanisms and endosperm-specific expression pattern, the sbeⅡα promoter (3094 bp in length) was cloned using APCR and sequenced.The effect of a series of deletions was studied using a GUS transient assay system. Results showed that the 3094 bp sequence (sbe.g construct) exhibited full stable promoting activity and that the activities of 5′ or 3′ deletions reduced levels of GUS expression. Some constructs with internal deletions showed only weak activity, however,sbe.e, with a deletion from -1579—--1210 bp resulted in higher levels of expression than the full-length promoter sequence, sbe.g. This indicates that motifs such as the -300 bp element, G-box and/or P-box act as positive elements and are necessary in determining the promoter‘s endosperm-specific pattern and that negative repressor elements or motifs may also be present within the -1579—-1210 bp sequence. The age of wheate ndosperm tissue used in the GUS-transient assay system is shown to be of significant importance.

Evaluation and characterization of an endosperm-specific sbeIIa promoter in wheat

Starch, the main component of the wheat grain, is the product of a complex biochemical pathway. The sbeIIa gene plays a key role in controlling the synthesis of starch, in particular, the biosynthesis of amylopectin, in maturing wheat grain. To investigate its regulatory mechanisms and endosperm-specific expression pattern, the sbeIIa promoter (3094 bp in length) was cloned using APCR and sequenced. The effect of a series of deletions was studied using a GUS transient assay system. Results showed that the 3094 bp sequence (sbe.g construct) exhibited full stable promoting activity and that the activities of 5′ or 3′ deletions reduced levels of GUS expression. Some constructs with internal deletions showed only weak activity, however, sbe.e, with a deletion from −1579 – −1210 bp resulted in higher levels of expression than the full-length promoter sequence, sbe.g. This indicates that motifs such as the −300 bp element, G-box and/or P-box act as positive elements and are necessary in determining the promoter’s endosperm-specific pattern and that negative repressor elements or motifs may also be present within the −1579 – −1210 bp sequence. The age of wheat endosperm tissue used in the GUS-transient assay system is shown to be of significant importance.