Presence of multiple cDNAs encoding an isoform of ADP-glucose pyrophosphorylase large subunit from sweet potato and characterization of expression levels.

Abstract

Three cDNAs (iAGPLI-1, iAGPLI-2, and iAGPLI-3) encoding an isoform of AGPase large subunit were isolated from a sweet potato cDNA library constructed from tuberous root tissue. iAGPLI-1 was 2,161 bp in length and contained an open reading frame of 517 amino acids with a calculated molecular mass of 57,689 Da. iAGPLI-2 and iAGPLI-3 were 1,804 and 1,524 bp in length, respectively, and contained partial open reading frames of 490 and 385 amino acids. Deduced amino acid sequence comparison analysis showed that iAGPLI-1 has sequence identity with iAGPLI-2 (97.9) and iAGPLI-3 (98.7%) while iAGPLI-2 and iAGPLI-3 have 96.8% sequence identity. iAGPLI-1 had the highest sequence identity of 77.8% with potato AGPase (sAGPL1). Steady-state levels of iAGPLI-1 transcripts were expressed predominantly in the stem, and moderately in the tuberous root, but not in either the roots or leaves. However, AGPase activity was present in all tissues. The expression level in the stem declined dramatically after a 12 h incubation in the dark to nearly 3% of the value under light, although the activity under a dark condition remained at half the levels in light. The activity levels were not correlated with the transcript levels. iAGPL transcripts in leaves were induced by sucrose feeding but not by glucose or fructose. Therefore, the expression of iAGPLI-1 is regulated in stem tissue preferentially and by sucrose. Southern blot analysis showed that the sweet potato genome contained several copies of iAGPLI gene probably due to polyploidy.