Cloning and analysis of a cDNA encoding an endo-polygalacturonase expressed during the desiccation period of the silique-valves of turnip-tops (Brassica rapa L. cv. Rapa).
During zygotic embryogenesis of turnip-tops (Brassica rapa L. cv. Rapa), the polygalacturonase activity (PG; EC 3.2.1.15), measured as a decrease in viscosity of polygalacturonic acid, reached a high when the desiccation process in the seeded silique was triggered and the valves had lost more than 70-75% of their moisture (45-50 DPA). The PG activity was not detected in any phases of developing seeds. This work also characterizes a cDNA with an open reading frame of 1303 bp and that codes for a putative PG called BrPG1. This falls into the category of clade-B, which includes PG related to shattering and abscission processes. The deduced BrPG1 sequence predicted a 434-residue-long precursor protein (46.7kDa) with a transit peptide sequence 23 amino acids long. A molecular mass of 44.3 kDa was calculated for the mature form of BrPG1, which showed high sequence similarity to PGA1 (97%) of B. napus (X98373) and ADPG1 (87%) of Arabidopsis thaliana (AJ002532). All conserved amino acids at the catalytic site of PGs belonging to clade-B were preserved on BrPG1. This BrPG1 gene was specifically expressed in the silique valves of turnip-tops and was temporally expressed at the beginning of its desiccation.
Rodríguez-Gacio
María del Carmen
Mdel C
Departamento de Fisiología Vegetal, Facultad de Farmacia, Universidad de Santiago de Compostela, Campus Sur s/n. 15782-Santiago de Compostela, Spain.
Nicolás
Carlos
C
Matilla
Angel J
AJ
D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics; D001483:Base Sequence; D001937:Brassica / Q000201:enzymology / Q000235:genetics; D029741:Brassica rapa / Q000145:classification / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics*; D003890:Desiccation; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics; D018506:Gene Expression Regulation, Plant / Q000235:genetics*; D018525:Germination; D007700:Kinetics; D008969:Molecular Sequence Data; D010802:Phylogeny; D035261:Plant Components, Aerial / Q000201:enzymology / Q000235:genetics / Q000254:growth & development*; D011096:Polygalacturonase / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012639:Seeds; D016415:Sequence Alignment
D018076:DNA, Complementary; D011096:Polygalacturonase