Articles
ADVENTITIOUS SHOOT REGENERATION IN PRUNUS DULCIS – A MOLECULAR APPROACH TO THE REGENERATION PROCESS
Article number
738_87
Pages
663 – 668
Language
English
Abstract
The understanding of de novo regeneration process in almond may provide clues regarding the critical steps in adventitious organogenesis that require optimisation.
Transgenic almond shoots have already been recovered, however the rate of success has been very low (Miguel and Oliveira, 1999). The main bottleneck of this system, apparently, is the fact that the host cells targeted to introduce the transgene are often different from those acquiring competence to re-enter cell cycle and therefore undergo organogenesis.
It is difficult to understand which cells do actually become competent and by which mechanism.
We have studied indirect shoot induction in almond using genomic tools.
Two main strategies were used: a candidate gene approach for the almond Knotted-1 and CDKA:1 (putative markers for organogenesis events) and a transcriptomic approach with micro-array technology, using two suppression subtractive hybridisation libraries constructed from two defined time frames of organogenesis, to screen and discover novel markers.
Several candidate genes were identified and studied by Real-Time PCR expression analyses.
The collected data may help to answer questions long ago standing by, and to which fundamental knowledge obtained from model plants has been clearly insufficient.
Finding genes directly involved in the competence for shoot neo-formation in woody species may help to study regulatory mechanisms underlying organogenesis.
Transgenic almond shoots have already been recovered, however the rate of success has been very low (Miguel and Oliveira, 1999). The main bottleneck of this system, apparently, is the fact that the host cells targeted to introduce the transgene are often different from those acquiring competence to re-enter cell cycle and therefore undergo organogenesis.
It is difficult to understand which cells do actually become competent and by which mechanism.
We have studied indirect shoot induction in almond using genomic tools.
Two main strategies were used: a candidate gene approach for the almond Knotted-1 and CDKA:1 (putative markers for organogenesis events) and a transcriptomic approach with micro-array technology, using two suppression subtractive hybridisation libraries constructed from two defined time frames of organogenesis, to screen and discover novel markers.
Several candidate genes were identified and studied by Real-Time PCR expression analyses.
The collected data may help to answer questions long ago standing by, and to which fundamental knowledge obtained from model plants has been clearly insufficient.
Finding genes directly involved in the competence for shoot neo-formation in woody species may help to study regulatory mechanisms underlying organogenesis.
Authors
A.M. Santos, M.J. Oliver, A.M. Sanchez, P. Payton, N. Saibo, M.M. Oliveira
Keywords
adventitious organogenesis, PdKn1, PdCdc2a, micro-arrays, suppression subtractive hybridisation, Real-Time PCR
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