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ISHS Acta Horticulturae 736: III International Date Palm Conference

THE USE OF RAPDS FOR THE DETECTION OF GENETIC STABILITY OF REGENERATED PLANTLETS OF BARHEE PALM IN IRAQ

Authors:   T.A. Ali, J.M. Jubrail, A.M. Jasim
DOI:   10.17660/ActaHortic.2007.736.11
Abstract:
The aim of this study was the use of RAPD-PCR technique to ensure the genetic stability of date palm plantlets produced by tissue culture. To achieve this, shoot tip explants (0.51cm) were excised from date palm offshoots (Barhee) and divided into four equal parts. These parts were cultured on MS media containing NAA (25 mg/l), 2-ip (2 mg/1) and activated charcoal (3 gm/l). Nodular callus was produced from shoot tip explants after four subcultures. Somatic embryos developed and formed shoots and roots. At the end of the propagation process young date palm plantlets were produced. RAPD-PCR analysis using universal primers was performed on DNA extracted from the samples taken randomly from the stalk at the process stages: somatic embryos and plantlets, and also from the fresh healthy leaves of the mother offshoot (Barhee). PCR conditions were optimized. Reproducible RAPD patterns were obtained using 30 primers. Three primers (OPC.16, OPG.O8 and OPN.16) of the 30 produced polymorphic bands in some of the samples tested when compared to the DNA fingerprint of the mother offshoot. After mass propagation of date palm via somatic embryogenesis, genetic variation has been reported for several cultivars and has been affected by the plant genotype, number of in vitro subcultures (age of cultures), the nature of the explants and the composition of culture media. Here we report for the first time in Iraq, the efficiency of RAPD fingerprints as a control technique and a simple fast molecular marker for the detection of genetic variations in regenerated plantlets so that tissue culture can be successfully applied for large scale propagation of elite varieties.

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