Optimization of an efficient micropropagation protocol and assessment of plant genetic fidelity by RAPD markers in pistachio (Pistacia vera L.).

摘要

An efficient protocol for the micropropagation of pistachio (P. vera cv. Kirmizi) was developed. Various concentrations and combinations of 6-benzyladenine [benzyladenine] (BA), kinetin (KIN), gibberellic acid (GA3) and (NA) were evaluated to optimize axillary bud development and multiple shoot formation from uni-nodal explants excised from in vitro-grown seedlings. The best shoot proliferation was obtained in Murashige and Skoog's (MS) medium supplemented with 8.0 micro M BA and 1.0 micro M GA3. Elongated shoots were rooted in the MS medium containing 12.0 micro m IBA, and successfully acclimatized under greenhouse conditions. Microshoots, which underwent at least 10 cycles of in vitro subculturing, were used in random amplified polymorphic DNA (RAPD) analysis to assess the maintenance of genetic fidelity to the donor plant. A total of 624 amplified reproducible bands were produced from 10 random decamer primers out of 16; 76 were found to be polymorphic (producing polymorphism ranging between 5.3 and 48.5%), and the average number of bands produced per primer was 6.2. The similarity matrix established through the analysis of the band patterns ranged from 0.83 to 0.99 between the donor and micropropagated plants, and from 0.79 to 0.96 among the cloned plantlets. The results suggested that somaclonal variations may be present in microshoots of pistachio which came from many cycles of axillary bud proliferation..