Naslov
An improved method for high-quality DNA extraction from olive embryos - plant tissue rich in polyphenols and polysaccharides

Autori
Vuletin Selak, Gabriela ; Baruca Arbeiter, Alenka ; Perica, Slavko ; Knap, Tea ; Klepo, Tatjana ; Bandelj, Dunja

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
Olivebioteq 2018 ; 6th International Conference on the Olive Tree and Olive Products / - Sevilla, 2018, 81-81

Skup
6th International Conference on the Olive Tree and Olive Products (OLIVEBIOTEQ 2018)

Mjesto i datum
Sevilla, Španjolska, 15.10.2018. - 19.10.2018

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
Olea europaea, microsatellites, DNA extraction, paternity analysis, self-compatibility, cross-compatibility

Sažetak
Different methods have been used to evaluate the success and inter-compatibility in pollination assays. Microsatellites are appropriate markers for paternity analysis of olive embryos to measure the success of the crosses, especially when potential pollen donors are multiple and not applied by hand. In our study, the paternity analysis based on microsatellites was used for genotyping and identification of the potential pollen donors of ʻOblicaʼ, the leading olive variety in Croatia and for assessing the percentage of self-fertilization in multivarietal olive orchard. Embryos accumulate high contents of polysaccharides, polyphenols, or other compounds that bind and/or co-precipitate with DNA. This condition increased the sample viscosity and significantly decreased DNA quality, interfering with the PCR performance. To improve the PCR output, we have modified the extraction method: with repeated extraction procedure based on CetylTrimethylAmmoniumBromide-PolyVinylPyrrolidone protocol, and using the previously extracted DNA samples and dissolved in TE buffer as an initial material, the amplification success was significantly improved. Seven microsatellite loci were used for genotyping olive embryos from ʻOblicaʼ trees and for all potential pollen donor trees grown in the orchard. Seeds have been considered as a product of self-fertilization if only maternal alleles were found at all analysed microsatellite loci. When the presence of paternal alleles for each primer was recorded together with maternal, the progeny has been considered a product of cross-fertilization. The knowledge of self-compatibility and cross-compatibility relationships of olive cultivars contribute to defining the new guidelines for farmers regarding the proper management and provide valuable information for breeders.

Izvorni jezik
Engleski

Znanstvena područja
Biologija, Poljoprivreda (agronomija)

POVEZANOST RADA