PCR-RFLP analysis of rbcL chloroplast gene

Olga Sheikina,N.A Shyurova,O.V Tkachenko

doi:10.1051/bioconf/20224303007

Olga Sheikina, N.A Shyurova + Show 1 more

Open Access

https://doi.org/10.1051/bioconf/20224303007

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Abstract

To identify intergeneric, interspecific, and intraspecific polymorphisms, PCR-RFLP studies of the chloroplast rbcL gene profiles were carried out using five restriction endonucleases (AluI, HpaII, BsuRI, BstHHI, BstMBI) in 14 species belonging to 6 different genera and 7 Thuja occidentalis cultivars. For all the genera studied (Pinus sp., Abies sp., Picea sp., Microbiota sp., Syringa sp., Thuja sp.), A genus-specific restriction profile characterized by a unique combination of restriction DNA fragments was revealed. The combination of enzymes used in the study did not reveal the interspecies and intraspecific variability of the rbcL gene. Species belonging to the same genus and all varieties of western thuja were characterized by identical PCR-RFLP profiles. It is assumed that further research, with the inclusion of a larger number of species and enzymes, will reveal polymorphism at the interspecies and intraspecific levels.

Highlights

PCR-RFLP analysis (Polymerase Chain Reaction - Restriction Fragment Length Polymorphism) of chloroplast DNA is widely used for phylogenetic studies, especially at the interspecies level and above [1-3]
Species belonging to the same genus in all cases had an identical restriction profile
An intergeneric polymorphism of the rbcL gene nucleotide sequence was revealed based on the analysis of the frequency of restriction sites for five restriction endonucleases (AluI, HpaII, BsuRI, BstHHI, BstMBI)

Summary

Introduction

PCR-RFLP analysis (Polymerase Chain Reaction - Restriction Fragment Length Polymorphism) of chloroplast DNA (chlDNA) is widely used for phylogenetic studies, especially at the interspecies level and above [1-3]. Studies of interspecies polymorphism have been carried out using restriction analysis of amplified chloroplast DNA from different species of the Pinaceae family [1] and different species of the genus Abies [2]. In a number of studies, it was possible to detect intraspecific variability of different parts of the chloroplast genome [2,5]. Not all restriction endonucleases make it possible to establish differences in the nucleotide sequences of chlDNA [6]. Research is needed to search for restriction enzymes that allow detecting polymorphism at various levels - intraspecific, interspecific and intergeneric

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