Development of an adventitious shoot regeneration system in cherries Public Deposited

http://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/qv33s098h

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  • Shoot regeneration in vitro is a difficult problem for cherry species. This study addressed three approaches to the problem, regeneration of adventitious shoots from roots and leaves of mature cherry clones, and from cotyledons of open-pollinated 'Royal Ann' cotyledons. Regeneration on roots occurred at a very low frequency. Adventitious shoots regenerated from only 17.5% of the cotyledons of open pollinated 'Royal Ann' cherries on Murashige and Skoog (MS) medium (Murashige et al., 1962) supplemented with 10 or 20 μM thidiazuron (TDZ) and 0 to 10 μM indolebutyric acid (IBA). Therefore, a two-stage process was developed for regeneration from leaves that resulted in high rates of adventitious shoot regeneration in some genotypes. Initially, leaves of several clones previously established in tissue culture were tested for shoot regeneration. The inter-specific Prunus hybrids Giessen 154-4 and Giessen 154-7, two of 15 clones tested, regenerated a small number of shoots when placed on MS medium and a combination of TDZ (5 to 10 μM) and naphthaleneacetic acid (NAA) or IBA (0.5 to 2.5 μM). Using these two clones, a two-stage process was developed to initiate and then promote elongation of adventitious shoots. In the first stage, increasing the agar concentration from 8 to 12 g/l and optimizing the TDZ and NAA concentrations to 10 μM and 1.25 μM respectively, markedly improved shoot initiation. After 20 days the cultures were transferred to a medium containing 5 μM BAP and 0.5 μM NAA. This second stage retarded callus growth and allowed meristems to elongate and form visible shoots. Applying this procedure to seven of the original 15 clones and one additional clone resulted in seven of the eight clones regenerating adventitious shoots, with four clones, 154-4, 154-7, 173-1 and 195-2, regenerating shoots on 50- 100% of the leaves. Changing the basal medium from Murashige and Skoog to Driver Kuniyuki walnut (DKW) medium (McGranahan et al., 1987) further increased the number of regenerating sites per leaf and meristem per leaf.
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