Peroxidase isoenzymes as markers for the rooting ability of easy-to-root and difficult-to-root Grevillea species and cultivars of Protea obstusifolia (Proteaceae)

Abstract

One easy-to-root and one difficult-to-root species of the ornamental plant Grevillea were investigated for rooting potential in relation to peroxidase activity. In vitro-grown shoot segments of both species started to root 30 d after transplanting to rooting medium containing indole-3-butyric acid (IBA), however, fewer roots were found on fewer segments of the difficult-to-root species G. petrophioides compared to the easy-to-root species G. rondeau. Total peroxidase (POX) activity was measured during the rooting process. G. petrophioides showed higher total POX activity at the time point of adventitious root formation than G. rondeau. Isoelectric focusing electrophoresis showed that G. rondeau contained more acidic isoforms than G. petrophioides, but the basic isoforms were more prominent in the difficult-to-root species, especially at the time point of lateral root emergence. In addition, the ability of different hormones to induced POX activity in upper and lower stem segments of both species was tested. Indole-3-acetic acid (IAA), IBA and α-naphthaleneacetic acid induced POX activity in the upper stem segments of G. rondeau, whereas the same hormones led to the induction of POX activity in the lower stem segments of G. petrophioides. Similar to the results obtained with Grevillea, the difficult-to-root variety of Protea showed higher POX activity, especially in the middle stem part and the leaves. Feeding of radiolabeled IAA to the Grevillea stem segments resulted in the synthesis of three different compounds in both species. After 1h incubation no differences were found in the uptake of IAA and the appearance of other labeled compounds. However, after 2 and 4 h incubation IAA uptake was faster in the easy-to-root species and IAA was also metabolized to a higher extent in G. rondeau. Three metabolites were found, tentatively identified as IAA-aspartate, IBA, and an IBA conjugate.