The contribution of extensin network formation to rapid, hydrogen peroxide-mediated increases in grapevine callus wall resistance to fungal lytic enzymes

Grapevine (Vitis vinifera cv. Touriga) callus cell walls contain a high level of the monomeric extensin, GvP1. Hydrogen peroxide stimulus of these cultures causes the rapid loss of monomeric GvP1, concomitant with marked increases in insoluble GvP1 amino acids and wall resistance to digestion by fungal lytic enzymes. JIM11 immunolocalization studies indicated that monomeric and network GvP1 were evenly distributed in the callus cell wall. These primary cell walls were used to investigate the specific contribution of extensin and other ionically bound cell-wall proteins to hydrogen peroxide-mediated increases in resistance to fungal lytic enzymes. This was performed by removing ionically-bound proteins and assaying for hydrogen peroxide-enhanced resistance after the addition of selected protein fractions. The results indicate that hydrogen peroxide-induced increases in resistance to digestion by fungal lytic enzymes require a co-operative action between network extensin formation and the electrostatic interaction of additional wall proteins with the extracellular matrix.