The results of studies of maize lines by SSR and morphological markers are presented. The polymorphism of the studied maize lines was determined by 8 SSR markers: phi064, umc1448, umc1792, bnlg1782, bnlg1129, phi084, phi015, phi083. According to the PCR results obtained alleles were used to calculate their frequencies and the locus polymorphic information content (PIC). It was identified from 2 to 9 alleles in the studied lines in accordance with the used markers. The frequencies of the identified alleles were 0.01—0.72. It was determined that the umc1448 marker (PIC 0.83) turned out to be the most polymorphic, the phi015 (PIC 0.46) the least polymorphic. A high PIC value was noted for the phi064 and bnlg1782 markers. The presence of unique alleles in the studied lines was determined for the umc1448 markers — the 132 bp allele, for the bnlg1782 marker — alleles with sizes of 240 and 242 bp. As the results of cluster analysis, based on the degree of note of morphological traits, the differences between 91 lines were determined. The pairs of lines with a different type of cytoplasm (fertile line and its sterile analogue) turned out to be identical in 36 traits. It was determined that genetic distances calculated according to the presence/absence of identified alleles revealed 88 distinct lines. It was established that one pair of lines, the fertile line and its sterile analogue, have no differences in the two marker systems studied, except for the type of cytoplasm, which is determined in the field by the presence of pollen. This indicates the need to attract at least one more SSR marker to determine the differences in this sample of lines. It was determined that lines that turned out to be identical in terms of SSR markers are morphologically distinct, which shows the effectiveness of an integrated approach to determining the difference of lines in the process of qualification examination for distinctness, uniformity and stability (DUS).
Keywords: i> Zea mays L., SSR markers, genetic distances, line difference
Full text and suplimented materialsFree full text: PDF
1. State register of plant varieties suitable for distribution in Ukraine. URL: https:// sops.gov.ua/reestr-sortiv-roslin (01.10.2019)
2. Drozdov, V.I. (2010). Manual for using the Statistica 6.0. Kursk: Izdatelstvo YuZGU [in Russian].
3. Ermantraut, E.R., Prysiazhniuk, O.I. & Shevchenko, I.L. (2007). Statistical analysis of agronomic study data in the software suite Statistica 6.0. Kyiv: PolihrafKonsaltynh [in Ukrainian].
4. Tkachik, S.O. (Ed.) (2015). Methods of examination of plant varieties of cereals, cereals and legumes for suitability for distribution in Ukraine Vinnytsia: FOP Korzun D. Yu. [in Ukrainian].
5. Syvolap, Yu.M., Kalendar, R.N. & Verbytskaia, T.H. (1998). The use of PCR analysis in genetic selection studies. Kiev: Ahrarna nauka [in Russian].
6. Babu, R., Nair, S.K., Kumar, A., Venkatesh, S., Sekhar, J.C., Singh, N.N. & Gupta, H.S. (2005). Two-generation marker-aided backcrossing for rapid conversion of normal maize lines to quality protein maize (QPM). Theoretical and Applied Genetics, No. 5, pp. 888-897. https://doi.org/10.1007/s00122-005-0011-6
7. Everitt, B.S., Landau, S., Leese, M. & Stahl, D. (2011). Cluster analysis (5th ed.). Chichester: Wiley. https://doi.org/10.1002/9780470977811
8. Greveniotis, V.A., Giourieva, V.S., Bouloumpasi, E.C., Sioki, E.J. & Mitlianga, P.G. (2018). Morpho-physiological Characteristics and Molecular Markers of Maize Crosses Under Multi-location Evaluation. Journal of Agricultural Science, No. 11. http:// doi.org/10.5539/jas.v10n11p79 https://doi.org/10.5539/jas.v10n11p79
9. Guidelines for the conduct of tests for distinctness, uniformity and stability. Mays. URL: https://www.upov.int/edocs/tgdocs/en/tg002.pdf (01.10.2019)
10. Gunjaca, J., Buhinicek, I., Jukic, M., Sarcevic, H., Vragolovic, A., Kozic, Z. & Pejic, I. (2008). Discriminating maize inbred lines using molecular and DUS data. Euphytica, No. 1-2, pp. 165-172. https://doi.org/10.1007/s10681-007-9518-z
11. Gurung, D., George, M. & Dela Cruz, Q. (2010). Analysis of genetic diversity within Nepalese maize populations using SSR markers. Nepal J. Sci. Technol., No. 11, pp. 1-8. https://doi.org/10.3126/njst.v11i0.4082
12. Hossain, F., Muthusamy, V., Pandey, N., Vishwakarma, A.K., Baveja, A., Zunja- re, R.U. & Gupta, H.S. (2018). Marker-assisted introgression of opaque2 allele for rapid conversion of elite hybrids into quality protein maize. Journal of Genetics, No. 1, pp. 287-298. https://doi.org/10.1007/s12041-018-0914-z
13. International Rules for Seed Testing 2019. URL: https://www.seedtest.org/en/international-rules-for-seed-testing-_content--1-1083.html (01.10.2019) https://doi.org/10.15258/istarules.2019.10
14. Liu, X., Zhang, Y., Zheng, Z., Li, Z., He, C., Liu, D. & Tan, Z. (2010). QTL Mapping for Controlling Days to Pollen Shed under Different Nitrogen Regimes in Maize. Proceeding of 2010 4th International Conference on Bioinformatics and Biomedical Engineering. https://doi.org/10.1109/ICBBE.2010.5518247
15. Liu, Z., Peter, S.O., Long, M., Weingartner, U., Stamp, P. & Kaeser, O. (2002). A PCR assay for rapid discrimination of sterile cytoplasm types in maize. Crop science, No. 2, pp. 566-569. https://doi.org/10.2135/cropsci2002.0566
16. Lu, H. & Bernardo, R. (2001). Molecular marker diversity among current and historical maize inbreds Theoretical and Applied Genetics, No. 4, pp. 613-617. https://doi.org/10.1007/PL00002917
17. Molecular biomarker analysis - SSR analysis of maize (E) : ISO/TR 17623:2015. Geneva, 2015.
18. Nagy, E., Timar, I., Hegyi, Z., Spitko, T. & Marton, L. C. (2009). SSR markers as tools in maize breeding for high starch content. Maydica, No. 2, p. 253. http:// doi.org/10.3389/fpls.2016.00223
19. Namorato, H., Miranda, G.V., de Souza, L.V., Oliveira, L.R., DeLima, R.O. & Eder E. (2009). Comparing Biplot Multivariate Analyses with Eberhartand Russell' method for genotype x environmentinteraction Crop. Breeding and Applied Biotechnology, No. 9, pp. 299-307. https://doi.org/10.12702/1984-7033.v09n04a03
20. TGP/15 Guidance on the Use of Biochemical and Molecular Markers in the Examination of Distinctness, Uniformity and Stability (DUS). URL: https:// www.upov.int/edocs/tgpdocs/en/tgp_15.pdf (01.10.2019)
21. Yadav, V. K. & Singh, I. S. (2010). Comparative evaluation of maize inbred lines (Zea mays L.) according to DUS testing using morphological, physiological and molecular markers. Agricultural Sciences, No. 3, p. 131. https://doi.org/10.4236/as.2010.13016