Leaf number and position effects on the survival and performance of grape microcuttings in vitro, and the sensitivity of the cut nodal region to the medium

The presence of leaf in microcuttings of grape cvs. Arka Neelamani and Thompson Seedless promoted rooting in vitro (MS, 1 micromolar IAA, 0.1 micromolar GA3, 3% sucrose) but the effect varied depending on the number of leaves and position of the leaf on the cutting. Single node cuttings with a full-length lower internode and a lamina at top (LAT) showed earlier rooting and more root and shoot growth than cuttings with lamina positioned at the middle (LAM), while cuttings with a leaf at the base (LAB) of the cutting and full-length upper internode exhibited a lower percent rooting and sprouting, poor root and shoot growth, and low survival. Partial or complete removal of the upper internodal segment in LAB cuttings improved rooting and sprouting suggesting the possible operation of an inhibitory effect by the upper internode. Retaining an upper leaf in LAB cuttings (LAB+UL) resulted in necrosis of the upper leaf often followed by the lower one. The extent of necrotic damage was influenced by the leaf area and position or age of the cutting on the stock shoot. Retaining the lower internode in LAB and LAB+UL cuttings which held the node leaf junction away from the medium, or reducing the concentration of MS medium helped significantly in improving the survival and performance of these cuttings. The difference in reaction between LAB and LAT cuttings was attributable mainly to the difference in the sensitivity of the stem part that came in contact with the medium. Removal of the leaf in LAB cuttings reduced this sensitivity. The majority of the LAB and LAB+UL cuttings, as well as non-rooting or delayed rooting LAT and LAM cuttings, exhibited high purple pigmentation of leaf, petiole and stem. Two-leafed cuttings in vitro showed poor survival, less rooting and low plantlet output compared to single-leafed cuttings.