Preparation and Isolation of Oligogalacturonic Acids and Their Biological Effects in Cockscomb (Celosia argentea L.) Seedlings

Abstract

A mixture of oligogalacturonic acids, the partial degradation substances of polygalacturonic acid, promoted shoot growth in cockscomb (Celosia argentea L.) seedlings, which generally had a high sensitivity for growth-promoting substances. The effect of the mixture of oligogalacturonic acids on shoot growth of cockscomb was higher than that of the polygalacturonic acid at concentrations above 30 ppm. These oligomers were loaded onto an anion exchange column, DEAE Sephadex A-25, and separated into individual oligomer sizes using the NH4HCO3 eluent system. This separation method has the advantage of using NH4HCO3 as the eluent solution; NH4HCO3 in the sample solution is effectively removed by lyophilization. Each of the isolated oligogalacturonic acids gave a single band on a fluorophore-assisted carbohydrate electrophoresis (FACE), and they showed the m/z value which corresponded to their molecular ion peaks [M-H]- on a fast atom bombardment mass spectrometry (FAB-MS) analysis. These results showed that the successive chromatography method used in this study is well suited for the preparation of oligogalacturonic acid for the plant growth test. Furthermore, we showed that the effective degree of polymerization (DP) of oligogalacturonic acid was around 8 on shoot growth of cockscomb seedlings, and the effects of both smaller and larger oligogalacturonic acids were slightly lower than that of octa-galacturonide. Octa-galacturonide promoted shoot growth of cockscomb at concentrations above 10 mM, and showed a 66% promotion at the most effective concentration of 300 mM. Root growth was slightly inhibited at concentrations above 300 mM. These results suggest that DP around 8 of oligogalacturonic acids has the function to control shoot growth in cockscomb as a growth-promoting substance.