First report about the identification and preliminary analysis of a partial sequence of dihydropyrimidine dehydrogenase (NADP plus) in Thermopsis turcica during floral development using degenerate primers

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Official URL: http://dx.doi.org/10.1007/s11738-017-2458-x

Thermopsis turcica Kit Tan, Vural & Kucuko-duk is an herbaceous perennial, endemic and listed as endangered species by IUCN in Turkey. This plant is noted for its unusual floral structure characterized by a 2-4 carpellary ovary. In this study, a DPD (NADP+)-like gene was partially sequenced for the first time in T. turcica. Since there is no previous molecular genetic information available for T. turcica, RT-PCR was performed using degenerate primers targeted to conserved sequences of WUS protein homologues from related legume species. Amplified cDNAs of the expected size were sequenced and analyzed using bioinformatics tools. The analysis strongly suggested that a 283 bp PCR product was part of a dihydropyrimidine dehydrogenase (NADP+)-like coding sequence with a Flavin mononucleotide binding domain. The putative gene was named Tt-DPD and the partial CDS submitted to the NCBI database (accession number KT182937). This gene has not been identified previously in T. turcica. The DPD (NADP+) enzyme is the rate-limiting step in pyrimidine degradation, which is essential for the biosynthesis of beta-alanine and pantothenates in plants; it has also been shown to be required for normal seed development in Arabidopsis. Expression of Tt-DPD was monitored by both endpoint and real-time RT-PCR. High expression of the identified gene was observed in the mid-developmental stage of the pistil of T. turcica. The findings presented here provide a starting point for understanding the roles of this gene in pyrimidine catabolism in T. turcica.