I. A. Shilov1, N. S. Velishaeva1, Yu. V. Aniskina1, O. S. Kolobova1, T. V. Shalaeva1, O. M. Borisenko2, Ya. N. Demurin2, S. S. Frolov2
1All-Russian Research Institute of Agricultural Biotechnology, ul. Timiryazevskaya, 42, Moskva, 127550, Russian Federation
2Federal Scientific Center of the Pustovoit All-Russian Research Institute of Oil Crops, ul. Filatova, 17, Krasnodar, 350038, Russian Federation
Abstract. The multiplex system for genetic identification of lines and hybrids of sunflower Helianthus annuus L. has been developed based on 7 microsatellite loci (ORS815, ORS394, HA140, HA432, ORS546, ORS1144, ORS78). The proposed technique includes one-tube PCR of all loci followed by electrophoresis analysis of DNA fragments using the genetic analyzer. Based on experimental studying of nucleotide structure of each locus the primers designed so that amplified regions contained only short tandem repeat regions. The advantage of the proposed approach is the ability to standardize and automate the procedure using 96-well plates, which allows the possibility for high-throughput analyses. The nine hybrids as well as corresponding parent lines of sunflower selected by All-Russia Research Institute of Oilseed Crops named by V.S. Pustovoit (VNIIMK): Ameli, Aris, Arneb, Clip, Surus, Taizar, Typhoon, Fakel, Factor were analyzed using above multiplex system. The DNA fragment lengths obtained for all loci appeared to be the same for each plant within corresponding line that confirmed the uniformity of all parent lines, while the total allele combinations for each line allowed to reliably distinguish among themselves. All analyzed sunflower hybrids appeared to be uniform. The DNA-profile of each hybrid contained the corresponding maternal and paternal alleles that confirmed the origin of hybrids from declared parent lines. The suggested multiplex system can be used for efficient identification of lines and hybrids of sunflower as well as it seems to be a promising tool for monitoring of seed production.
Keywords: sunflower (Helianthus annuus L.), DNA-profile, microsatellites.
Author Details: I. A. Shilov, D. Sc. (Biol.), head of laboratory (е-mail: Этот адрес электронной почты защищён от спам-ботов. У вас должен быть включен JavaScript для просмотра.); N. S. Velishaeva, Cand. Sc. (Biol.), senior research fellow; Yu. V. Aniskina, Cand. Sc. (Biol.), senior research fellow; O. S. Kolobova, senior research fellow; T. V. Shalaeva, research fellow; O. M. Borisenko, Cand. Sc. (Biol.), head of laboratory; Ya. N. Demurin, D. Sc. (Biol.), head of division; S. S. Frolov, Cand. Sc. (Agr.), deputy director.
For citation: Shilov IA, Velishaeva NS, Aniskina YuV, et al. [Genetic identification of Helianthus annuus L. sunflower lines and hybrids based on multiplex microsatellite analysis]. Dostizheniya nauki i tekhniki APK. 2023;37(1):10-5. Russian. doi: 10.53859/02352451_2023_37_1_10.