Keywords
Rosa alberta
Rosa spinosissima
CTAB-based method
DNA
How to Cite
Abstract
The Rosa genus includes several species of ecological and conservation importance. However, obtaining high-quality genomic DNA from Rosa leaf extracts can be difficult because secondary metabolites can interfere with DNA extraction and subsequent molecular analysis. This study evaluated the performance and versatility of the CTAB-based DNA extraction protocol for leaves of Rosa acicularis Lindl., Rosa alberta Regel and Rosa spinosissima L. from the Altai and Tarbagatai regions of Kazakhstan. Genomic DNA was extracted using a CTAB-based method and compared with the DNA obtained using the primary extraction method. DNA concentrations were measured spectrophotometrically. Differences between extraction methods were analyzed using Welch’s t-test.
The CTAB protocol yielded the highest DNA amounts in the analysed samples. Average DNA concentration ranged from 188.95 to 385.24 ng/µl, depending on species and location. The highest average concentration was observed in R. acicularis from the Altai Mountains (385.24 ± 142.22 ng/µl), and the lowest in R. spinosissima (188.95 ± 89.38 ng/µl). Significant differences were found between individual samples, especially in R. alberta from Tarbagatai and R. spinosissima from the Altai Mountains. Samples processed with the primary extraction method generally showed lower and more variable DNA content; differences between extraction methods were statistically significant (p < 0.05).
The CTAB-based protocol provided sufficient DNA extraction for molecular analysis of the studied Rosa species and proved applicable to plant material from different regions of Kazakhstan. However, the large variability between samples suggests that DNA concentration alone does not fully reflect DNA quality. Further verification of DNA purity and PCR amplification efficiency is recommended for subsequent molecular biological applications.