Design of degenerate primers for studying the expression of alcohol dehydrogenase in the psamophyte Alyssum desertorum

DOI: https://doi.org/10.7124/FEEO.v38.1820
Keywords: Alyssum desertorum, waterlogging, alcohol dehydrogenase, degenerate primers, RT-PCR

Abstract

Aim. To address the question of how psammophytes (plants of sandy soils) can tolerate waterlogging, we chose a representative of the flora of Ukraine, Alyssum desertorum Stapf. An important component of plant resistance to excess water is rapid activation of anaerobic metabolism, the key enzyme of which is alcohol dehydrogenase (ADH). The genetic sequence of ADH gene in this species is unknown. The aim of the research was to design and evaluate efficiency of degenerate primers for studying ADH expression by RT-qPCR. Methods. Primers were selected based on an analysis of the literature and bioinformatic databases. Results. The position of A. desertorum was analysed taking into account molecular systematics, and the structure of ADH in related species was elucidated. A consensus sequence of the conserved region of the gene and primers have been created. A separate task was to design degenerate primers for the reference genes ACT2 and APRT. Testing of the primers by RT-PCR using two Brassicaceae species: A. desertorum and Arabidopsis thaliana (L.) Heynh., showed their effectiveness. Conclusions. It was concluded that the designed degenerate primers can be used for RT-qPCR analysis of ADH expression in A. desertorum under waterlogging.

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