Abstract
A general description of molecular genetic markers is provided in the present article. The classification of DNA markers used for identification of raw materials of plant origin is presented. The most appropriate method for identifying raw materials derived from fruit and berries is chosen using study reports. The use of PCR for determining the quantitative and qualitative composition of the raw materials is considered. DNA regions used for PCR diagnostics of raw material derived from fruit and berries are characterized. The significance of amplification for a PCR test is outlined. The optimal PCR conditions have been selected and the advantages of this method have been revealed in the present study. Amplification profiles of DNA from the samples have been analyzed using different primers. Experiments with different primers allowing for identification of the raw material were carried out. The possibility of using a complex including a common gene and a variable gene for the identification of raw materials derived from fruit and berries has been considered. The sequence of DNA regions has been analyzed. The possibility of using a ribosomal RNA gene for generic and interspecific differentiation of DNA samples has been demonstrated. The significance of oligonucleotide primers and PCR product length for the reliability of the whole genotyping system has been elucidated. A scheme of PCR-based DNA profiling has been developed. Two types of the procedure were compared and the most appropriate type was chosen according to cost efficiency.
Highlights
Identification of plant species from which raw material was derived is among the main areas of application of molecular genetic markers
The aim of the present study was to investigate the possibility of using polymerase chain reaction (PCR) for DNA diagnostics of raw materials derived from fruit and berries and to analyze target DNA regions, profiles generated by amplification, and PCR products
The method is based on repeated copying of specific DNA fragments; the specificity is determined by complementarity of DNA sequences and the copying is mediated by the enzyme DNA polymerase
Summary
Identification of plant species from which raw material was derived is among the main areas of application of molecular genetic markers. DNA fragments associated with a specific nucleotide sequence constitute a new class of molecular markers. The number of such markers is severalfold higher than that of the markers characterized previously (isozymes, storage proteins, and morphological features). Expression of DNA markers is neither dependent on the phenotype nor tissue-specific and can be detected at any stage of plant development. The possibility of detecting identical multiband DNA profiles for two randomly selected individuals is close to 2∙10-9 for avocado, 1.5∙10-9 for papaya varieties, 4.2∙10-5 for apple varieties, and 2.4∙10-3 for varieties of raspberries and blackberries [12, 13]
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