Abstract
Amyβ is a single gene that encodes β-amylase, a starch-hydrolyzing enzyme that confers sweetness to sweet potato roots upon cooking. To clarify the genetic basis underlying β-amylase null activity, we sequenced a highly conserved region of Amyβ, including the Amyβ active center, from normal and null cultivars. Comparison of the sequences revealed the presence of the In-Del sequences in null cultivars, which was thought to be a loss-of-function mutation causing a change from sweet to non-sweet. The In-Del frequency revealed using a multiple sequencing approach, and its association with β-amylase activity, showed that an inactive allele of Amyβ gene (Amyβ-I) exists. Amyβ expression analysis showed that Amyβ-I is transcribed normally, suggesting that translational control of β-amylase activity occurs in null mutants. The insertion in Amyβ-I caused a sequence frameshift resulting in an amino acid substitution with the incorporation of a stop codon in Amyβ-I. The Amyβ-I amino acid substitution and premature termination affect the substrate binding and catalytic site of the enzyme, which may result in an inactive protein that is responsible for the β-amylase inactivity in non-sweet cultivars. These findings are the basis of selection markers for non-sweetness in sweet potato breeding.
