Proliferating axillary shoots of the difficult-to-root apple cultivar `Jonathan' acquired an enhanced ability to form adventitious roots with increasing number of subcultures in vitro . The transition between the difficult-to-root and the easy-to-root condition occurred at the fourth subculture .Endogenous levels of free IAA and ABA in shoot tissues were analysed by gas chromatography/mass spectrometry/single ion monitoring (GC/MS/SIM) using negative ion chemical ionisation . Tissues from the mother plants grown in the glasshouse contained more IAA and ABA than those from tissue-culture material . After establishment in vitro there was no variation in the IAA content throughout the subcultures but a decrease in ABA content was observed after the fourth transfer . The IAA/ABA ratio increased from 0 .2 in difficult-to-root shoots from the initial culture up to 0 .7 in easy-to-root shoots from the long-term subculture.
1 . IntroductionThe propagation of apples from cuttings is difficult because most apple cultivars and rootstocks do not readily form roots under conventional nursery conditions [23] . However, the rooting ability of apple cuttings can be improved by `conditioning' the mother plants using treatments such as heavy pruning and blanching [11], application of growth retardants [6,9] or exposure to low light intensity [7] . Success in the striking of cuttings is determined as much by the management of the stock plants prior to severance as by the conditions under which root formation occurred [18] . Most of the research done on root formation is concerned with cuttings after excision from the stock plants .Recent reviews on adventitious root formation [10,13] have drawn attention to the importance of the physiological state of the mother plants at time of collection of the cuttings . Investigations on the hormonal balance in stock and the results are conflicting . In Dahlia [3] and in Rhododendron [24] the capacity of cuttings to form adventitious roots is not correlated with the endogenous level of IAA in the mother plants . In stock plant of Bougainvillea, Hibiscus [5] and Vitis [2], however, enhanced root formation of cuttings is associated with high IAA content . Similarly very little is known about the occurrence of ABA in mother plants . In Rhododendron the rooting ability of cuttings is positively correlated with a high ABA content in stock plants [24] but high ABA levels are found to be inhibitory in Sequoia sempervirens [8] .Difficult-and easy-to-root microcuttings of the same apple cultivar can be produced in vitro by repeated subculture under specific conditions [19] .