High expression of GUS activities in sweet potato storage roots by sucrose-inducible minimal promoter.

KEY MESSAGE: We developed transgenic sweet potato with Spomin (sucrose-inducible minimal promoter)-GUS gene-fused constructs. Induced GUS activities by Spomin were higher than those by CaMV 35S promoter. We developed transgenic sweet potato (Ipomoea batatas L. Lam. cv. Kokei no. 14) plants with Spomin (sucrose-inducible minimal promoter)-GUS gene-fused constructs with signal peptides for sorting to cytosol, apoplast and ER, and we analyzed the GUS expression pattern of cut tissue after sucrose treatment. Induced GUS activities by Spomin were several hundred times higher than those by the CaMV 35S promoter. Also, GUS activities in storage roots induced with a Spomin-cytosol-GUS construct were higher than those with either Spomin-apoplast or -ER-GUS constructs. The induced GUS activities by Spomin were higher in storage roots without sucrose treatment than those with sucrose treatment. Chilling (4 °C) storage roots with Spomin constructs for 4 weeks produced higher GUS activities than in storage roots stored at 25 °C for 4 weeks. The calculated maximum GUS content in the storage roots was up to about 224.2 µg/g fresh weight. The chilling treatment increased the free sucrose content in the storage roots, and this increase in endogenous sugar levels induced increased GUS activities in the storage roots. Therefore, Spomin appears to be a useful promoter to develop protein production systems using sweet potato variety Kokei no. 14 storage roots by postharvest treatment.

The alteration of plant morphology by small peptides released from the proteolytic processing of the bacterial peptide TENGU.

Phytoplasmas are insect-borne plant pathogenic bacteria that alter host morphology. TENGU, a small peptide of 38 residues, is a virulence factor secreted by phytoplasmas that induces dwarfism and witches' broom in the host plant. In this study, we demonstrate that plants process TENGU in order to generate small functional peptides. First, virus vector-mediated transient expression demonstrated that the amino-terminal 11 amino acids of TENGU are capable of causing symptom development in Nicotiana benthamiana plants. The deletion of the 11th residue significantly diminished the symptom-inducing activity of TENGU, suggesting that these 11 amino acids constitute a functional domain. Second, we found that TENGU undergoes proteolytic processing in vitro, generating peptides of 19 and 21 residues including the functional domain. Third, we observed similar processing of TENGU in planta, and an alanine substitution mutant of TENGU, for which processing was compromised, showed reduced symptom induction activity. All TENGU homologs from several phytoplasma strains possessed similar symptom induction activity and went through processing, which suggests that the processing of TENGU might be related to its function.