Effect ofin vitro storage at 4°C on survival and proliferation of two apple rootstocks
One cm long shoot explants of dwarf apple rootstocks P 2 and M.9 taken from 2 year-old cultures were stored at 4°C in the dark in three media differing in concentration of growth regulators. Every 6 weeks, some explants were transferred into proliferation medium and multiplication rate was observed during three or four consecutive passages. In a second experiment, the influence of explant type (1 cm long shoot tips, 1 cm long middle part of shoots or three-shoot tufts smaller than 1 cm) and transfer time to the cold room (immediately, 10 days, or 20 days after subculture) on explant survival and proliferation were analysed.
Survival of explants was influenced by composition of the storage media. On medium without 6-benzylaminopurine, 70% of P 2 and 17% of M.9 explants became necrotic during 18 weeks of storage. P 2 rootstock proliferated better in three passages after storage than did unstored controls. Storage of M.9 rootstock reduced proliferation in the first and second passages if stored in media containing 6-benzylaminopurine in comparison with unstored controls. Explants stored as tufts and transferred to the cold room directly after subculture produced more shoots during two passages than cultures stored as single shoots.
Key wordsapple rootstocks in vitro storage shoot proliferation
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