Articles
Somaclonal variation in regenerated plants derived from callus of peach rootstocks
Article number
1187_23
Pages
233 – 242
Language
English
Abstract
This research aimed to investigate a reliable protocol for callus induction and regeneration of peach (Prunus persica L.) rootstocks; ‘Montclair’, ‘Nemaguard’ and ‘Okinawa’, using mature seeds as in vitro establishment material.
Seeds recorded the highest germination percentage when disinfested using 20% Clorox for 30 min prior to culture on Woody Plant Medium (WPM). Each seed was cultured separately and tagged with a serial code.
The combination of 1.5 mg L-1 benzyladenine (BA) + 0.2 mg L-1 gibberellic acid (GA3) produced sufficient number of shoots with suitable medium height for all tested rootstocks during multiplication stage.
After three consecutive subcultures, all the three tested rootstocks succeeded in callus formation with highest mass by adding 1.0 mg L-1 indolebutyric acid (IBA) + 1.0 mg L-1 naphthaleneacetic acid (NAA) to half-strength Murashige and Skoog (MS) culture medium in the presence of 1.0 g L-1 polyvinyl alcohol (PVA). Callus clusters of ‘Montclair’ rootstock, which recorded the highest callus induction frequency on different auxin treatments, were differentiated when transferred to MS medium free of PVA and growth regulators.
The highest concentration of NAA (2.0 mg L-1) was more effective regarding root formation, especially when combined with the highest IBA concentration (2.0 mg L-1). Inter Simple Sequence Repeat-Polymerase Chain Reaction (ISSR-PCR) results confirmed the role of fragments polymorphism of ‘Montclair’ rootstock regenerants identification and discrimination as compared to directly regenerated one from the original individual.
Seeds recorded the highest germination percentage when disinfested using 20% Clorox for 30 min prior to culture on Woody Plant Medium (WPM). Each seed was cultured separately and tagged with a serial code.
The combination of 1.5 mg L-1 benzyladenine (BA) + 0.2 mg L-1 gibberellic acid (GA3) produced sufficient number of shoots with suitable medium height for all tested rootstocks during multiplication stage.
After three consecutive subcultures, all the three tested rootstocks succeeded in callus formation with highest mass by adding 1.0 mg L-1 indolebutyric acid (IBA) + 1.0 mg L-1 naphthaleneacetic acid (NAA) to half-strength Murashige and Skoog (MS) culture medium in the presence of 1.0 g L-1 polyvinyl alcohol (PVA). Callus clusters of ‘Montclair’ rootstock, which recorded the highest callus induction frequency on different auxin treatments, were differentiated when transferred to MS medium free of PVA and growth regulators.
The highest concentration of NAA (2.0 mg L-1) was more effective regarding root formation, especially when combined with the highest IBA concentration (2.0 mg L-1). Inter Simple Sequence Repeat-Polymerase Chain Reaction (ISSR-PCR) results confirmed the role of fragments polymorphism of ‘Montclair’ rootstock regenerants identification and discrimination as compared to directly regenerated one from the original individual.
Authors
R.A. Mahmoud, E.I. Bakr, S. El-Kosary, E.K. Fayed
Keywords
Prunus persica, in vitro, callus induction, regeneration, ISSR
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