Differences in genomic DNA extracted from bark and from wood of different zones in Robinia trees using RAPD-PCR

Determination of genetic differences and levels of gene expression in mature and old tissues (e.g. wood) is often difficult based on morphological and anatomical characteristics, levels of metabolites or enzymatic activity. The use of molecular markers allows assessment of polymorphic (genetic) vari...

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Bibliographic Details
Published in:Trees (Berlin, West) West), 1998-04, Vol.12 (6), p.377-384
Main Authors: DE FILIPPIS, L, MAGEL, E
Format: Article
Language:English
Subjects:
Agronomy. Soil science and plant productions
Bark
Biological and medical sciences
Deoxyribonucleic acid
DNA
Enzymatic activity
Fundamental and applied biological sciences. Psychology
Gel electrophoresis
Gene expression
Generalities. Genetics. Plant material
Genetic diversity
Genetic resources, diversity
Genetics and breeding of economic plants
genomics
Heart
Metabolites
Oligonucleotides
Plant material
Polymerase chain reaction
Post-transcription
Primers
Random amplified polymorphic DNA
Robinia
Robinia pseudoacacia
Structure-function relationships
Trees
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Summary:Determination of genetic differences and levels of gene expression in mature and old tissues (e.g. wood) is often difficult based on morphological and anatomical characteristics, levels of metabolites or enzymatic activity. The use of molecular markers allows assessment of polymorphic (genetic) variation amongst individuals and between closely related species directly at the DNA level, but such techniques have not been generally applied to the bark and wood of mature trees. In this study we have applied the technique of random amplification of polymorphic DNA (RAPD) by the polymerase chain reaction (PCR) to analyse the relationship between the bark and variously aged wood zones of Robinia pseudoacacia. The use of micro polyacrylamide gel electrophoresis coupled to silver staining for DNA provided a quick, reliable and sensitive method of detecting polymorphisms. It was necessary to test a small number of ten-base synthetic oligonucleotide primers before arriving at a set of five which clearly identified post-transcriptional differences between bark, sapwood, transition zone and heartwood even in the one individual tree. The variability of the technique, and in particular the origin and quality of the DNA extracted was analysed. We demonstrated that the procedures and protocols developed are applicable to all tissue types tested from bark to the inner heartwood zones. Our results show that RAPD-PCR technology is a versatile and sensitive method of detecting genomic changes in trees.[PUBLICATION ABSTRACT]
ISSN:0931-1890
1432-2285
DOI:10.1007/PL00009723