Поліморфізм довжин інтронів генів актину у різних сортів льону-довгунця української селекції
Анотація
Aim. Intron length polymorphism (ILP) is an effective tool to identify genotypes and a reliable source of information with high interspecies and intraspecies variability. As the flax is one of the most widespread crops in Ukraine, to ex-plore the various members of the genus Linum L. are very useful. The main aim of this paper is to develop of molecular genetic markers based on the variability of flax actin genes to detect intron length polymorphism in different varieties of Ukrainian selection flax. Methods. CTAB-method for the isolation of DNA was used. Intron length polymorphism was identified by PCR amplification with specific primers, electrophoretic analysis under non-denaturing polyacrylamide gel was done. Results. 16 varieties of Ukrainian selection flax have been analyzed. It was shown that polymorphic am-plicons were available in analyzed plants. The lengths of fragments corresponding actin introns have varied from 676 bp to 1530 bp. Conclusions. The analysis of 16 varieties of Ukrainian selection flax demonstrated the ability of this method to detect the difference between genotypes at intraspecies level. From these results we can make a conclusion that method based on actin intron length polymorphism can be a source of useful information about genetic differentia-tion of taxonomic groups of plants.
Keywords: molecular marker, actin, ILP (intron length polymorphism), Linum usitatissimum L.
Посилання
Karg S., Hist V. New research on the cultural history of the useful plant Linum usitatissimum L. (flax), a resource for food and textiles for 8,000 years. Archaeobot. 2011. V. 20. P. 507-508. doi: 10.1007/s00334-011-0326-y
Jhala A.J., Hall L.M. Flax (Linum usitatissimum L.): Current uses and future applications. Austral. J. Basic Appl. Sci. 2010. V. 4 (9). P. 4304-4312.
Lemesh V.A. Molekuliarnye markery v izuchenii geneticheskikh resursov. Mol. i prikl. genetika: Sb. nauch. tr. 2008. V. 8. P. 94-104. [in Russian]
Matveeva T.V., Pavlova O.A., Bogomaz D.I. Molekuliarnye markery dlia vidoidentifikatsii i filogenetiki rasteniy. Ekol. genetika. 2011. V. 9. P. 32-43. [in Russian] doi: 10.17816/ecogen9132-43
Wang X., Zhao X., Zhu J., Wu W. Genome-wide investigation of intron length polymorphisms and their potential as molecular markers in rice (Oryza sativa L.). DNA Res. 2005. V. 12. P. 417-427. doi: 10.1093/dnares/dsi019
Breviario D., Giani S., Ponzoni T., Mastromauro F., Morell L. Plant tubulin intronics. Cell Biol. Int. 2008. V. 32. P. 571-573. doi: 10.1016/j.cellbi.2007.11.013
Braglia L., Manca A., Mastromauro F., Breviario D. cTBP: A successful intron length polymorphism (ILP)-based genotyping method targeted to well defined experimental needs. Diversity. 2010. V. 2. P. 572-585. doi: 10.3390/d2040572
Rabokon' A.N., Pirko Ia.V., Demkovich A.E., Blium Ia.B. Polimorfizm dliny intronov genov beta-tubulina kak effektivnyi instrument genotipirovaniia rasteniy. Mol. i prikl. genetika: Sb. nauch. tr. 2015. V. 19. P. 35-44. . [in Russian]
Bardini M., Lee D., Donini P., Mariani A., Giani S., Toschi M., Lowe C., Breviario D. Tubulin-based polymorphism (TBP): a new tool, based on functionally relevant sequences, to assess genetic diversity in plant species. Genome. 2004. V. 47. P. 281-291. doi: 10.1139/g03-132
Nick P. Signaling to the microtubular cytoskeleton in plants. Int. Rev. Cytol. 1998. V. 184. P. 33-80. doi: 10.1016/S0074-7696(08)62178-6
McKean P.G., Vaughan S., Gull K. The extended tubulin superfamily. J. Cell Sci. 2001. V. 114. P. 2723-2733.
McCundy D.W., Kovar D.R., Staiger C.J. Actin and actin-binding proteins in higher plants. Protoplasma. 2011. V. 215. P. 89-104. doi: 10.1007/BF01280306
Postovoytova A.S., Baier H.Ya., Pydiura M.O., Pastukhova N.L., Pirko Ya.V., Iemets' A.I., Blium Ya.B. Poshuk ta analiz poslidovnostey heniv aktynu v henomi l'onu. Naukovi dopovidi NUBiP. 2015. V. 8 (57). Available from: http://nd.nubip.edu.ua/2015_8/index.html. [in Ukrainian]
Gupta P.K., Rustgi S. Molecular markers from the transcribed/expressed region of the genome in higher plants. Funct. Integr. Genomics. 2004. V. 4. Р. 62-139. doi: 10.1007/s10142-004-0107-0
Schulman A.H. Molecular markers to assess genetic diversity. Euphytica. 2007. V. 158. P. 313-321. doi: 10.1007/s10681-006-9282-5
Postovoytova A.S., Pirko Ya.V., Blium Ya.B. Polimorfizm dovzhyn druhoho intronu heniv aktynu v henomi Linum usitatissimum L. Faktory eksperymental'noi evoliutsii orhanizmiv. K.: Lohos, 2016. V. 19. S. 38-42. [in Ukrainian]
Rabokon' A.N., Demkovich A.E., Pirko Ia.V., Blium Ia.B Issledovanie polimorfizma dliny intronov genov β-tubulina u rasteniy roda Linum L.. Faktori eksperimental'noi evoliutsii organizmiv. K.: Logos, 2016. V. 19. P. 43-46. [in Russian]
Sambrook J., David W.R. Molecular Сloning: A Laboratory Manual. Cold Spring Harbor. 2001. V. 2. 763 p.
Rahman M.H., Jaquish B., Khasa P.D. Optimization of PCR protocol in microsatellite analysis with silver and SYBR stains. Plant Mol. Biol. Rep. 2000. V. 18. P. 339-348. doi: 10.1007/BF02825061