Поліморфізм довжини інтронів генів β-тубуліну у Deschampsia antarctica Desv. з морської Антарктики
Aim. The use of molecular markers for DNA-Fingerprinting provides an efficient way to carry out precise and rapid species identification. Tubulin-Based-Polymorphism (TBP) was originally introduced as a novel method for assaying genetic diversity in plants. Antarctic hair grass (Deschampsia antarctica E Desv.) is the only representative of the grass family (Poaceae) distributed in Antarctic. Antarctic species range is extended in the longitudinal direction and includes a number of islands with different areas. The aim of this study was to develop primers for amplification of the second intron of D. antarctica β-tubulin genes and to investigate molecular genetic differentiation D. antarctica populations from two distant regions of maritime Antarctic using TBP-analysis and its variations. Methods. D. antarctica plants from the Argentine islands and King George Island (South Shetland Islands) were studied. To analyze genetic variation, different methods of tubulin polymorphism analysis were used, such as TBP, с-ТВР, and h-TBP. Amplified fragments were fractionated by electrophoresis on non-denaturing polyacrylamide gel and DNA bands were detected using silver staining. Results. The size of amplified fragments of β-tubulin genes were ranged from 370 bp to 1300 bp for TBP method, from 1100 bp to 1700 bp for h-TBP, from 320 bp to 1750 bp for c-TBP with degenerate primers, and from 345 bp to 1180 bp for c-TBP with specific primers. No variation was detected in profiles of PCR-products generated with different primers. Conclusions. D. antarctica from two distant regions of maritime Antarctic demonstrate genetic ho-mogeneity by β-tubulin genes intron length.
Keywords: Deschampsia antarctica E Desv., molecular markers, β-tubulin, tubulin based polymorphism.