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Title: REGENERATION OF ALMOND (PRUNUS DULCIS MILL.) FROM IMMATURE SEED COTYLEDONS

Author
item AINSLEY, PHILLIP - ADELAIDE UNIVERSITY
item Hammerschlag, Freddi
item BERTOZZI, TERRY - ADELAIDE UNIVERSITY
item COLLINS, GRAHAM - ADELAIDE UNIVERSITY
item SEDGLEY, MARGARET - ADELAIDE UNIVERSITY

Submitted to: Plant Cell Tissue and Organ Culture
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 3/6/2001
Publication Date: 11/20/2001
Citation: Ainsley, P.J., Hammerschlag, F.A., Bertozzi, T., Collins, G.G., Sedgley, M. 2001. Regeneration of almond (prunus dulcis mill.) from immature seed cotyledons. Plant Cell Tissue And Organ Culture. 67:221-226

Interpretive Summary: Improvement of almond (Prunus dulcis Mill.)is a long term process through conventional breeding methods. Modern biotechnology techniques have the potential to hasten the production of improved cultivars and broaden the gene pool available for crop improvement in woody fruit and nut species, however, a limiting step to using these techniques is the inability to generate almond plants from cell cultures. In other Prunus species, the efficiency generating shoots from cell cultures has been improved by using juvenile tissues including immature embryos. In this study, we investigated the influence of juvenile tissue (cotyledons), cultivar, plant growth regulator type and concentration, and a dark treatment on the efficiency of shoot formation from cell cultures. A protocol for high efficiency in vitro shoot formation from juvenile almond tissues was developed for all cultivars. This information will be of use to scientists interested in improving almond using either tissue culture or gene transfer techniques.

Technical Abstract: Reported is the regeneration of adventitious shoots from immature cotyledons of four almond cultivars (Ne Plus Ultra, Nonpareil, Carmel and Parkinson). Open-pollinated fruit was collected from orchard-grown trees 100-115 days after full bloom. The proximal ends of the cotyledons were excised and the embryonic axes discarded. These explants were subjected to a range of tissue culture conditions designed to investigate the effects of different concentrations of thidiazuron (TDZ) and indole-3-butyric acid (IBA) and the presence or absence of light for the first 7 days of culture. Shoot regeneration rates were highest for cotyledons cultured for 8 weeks on Murashige and Skoog (MS) basal medium containing TDZ (10.0 uM), followed by 4 weeks on medium without plant growth regulators. Regeneration levels were further improved if cotyledons were maintained in darkness for the first 7 days. The inclusion of IBA (0.5 uM) in media significantly reduced the development of adventitious shoots. The frequency of cotyledons that developed adventitious shoots under the optimum tested conditions for Ne Plus Ultra, Nonpareil, Carmel, and Parkinson were 80.0%, 73.3%, 100.0%and 86.7%, respectively.