Molekularna identyfikacja ustalonych i mieszańcowych form Capsicum annuum L. z wykorzystaniem markerów RAPD

Celem badań była molekularna analiza odmian C. annuum L.: ‘Sono’, ‘Anchi’, ‘Mino’, androdiploidalnych linii: R1 i R18 oraz mieszańców pokolenia F1: (‘Sono’בAnchi’), (‘Anchi’בSono’), (‘Sono’בMino’), (‘Mino’בSono’), (‘Sono’×R1), (R1בSono’), (‘Sono’×R18), (R18בSono’). Wyboru komponentów rodzicielskich do krzyżowań zwrotnych dokonano na podstawie oceny biometrycznej obejmującej najważniejsze cechy roślin i owoców papryki. Analiza statystyczna otrzymanych wyników wykazała wysoką wartość użytkową ocenianych genotypów oraz ich zróżnicowanie pod względem analizowanych parametrów. Do identyfikacji form rodzicielskich i mieszańców zwrotnych zastosowano 21 starterów RAPD. Uzyskano 174 produkty, z których 16 (9,2%) stanowiły prążki polimorficzne. Ich obecność pozwoliła na rozróżnienie wszystkich odmian i linii C. annuum L. oraz potwierdziła mieszańcowy charakter analizowanych pokoleń F₁. W przypadku mieszańców zwrotnych możliwe było rozróżnienie form: (‘Sono’×R18)F₁, (R18בSono’)F₁ oraz (‘Sono’×R1)F₁, (R1בSono’)F₁.

The most important objective of breeding programmes is increasing the genetic diversity of cultivated plants. This objective can be achieved through intraspecies crossings and through crossings of different species. The source material used in the presented study comprised of C. annuum L. cultivars: ‘Sono’, ‘Anchi’, ‘Mino’ and androdiploid lines: R1 and R18. Their utility for interspecies crossings was assessed on the basis of statistical analysis of the most important agromorphological traits of pepper. ‘Anchi’ variety proved to be the most productive when it comes to yield of fruit and the number of fruit from a plant. The Tukey’s test shown that as long as these parameters are concerned, it was statistically a separate group. The biggest weight of fruit without the stalk, technological weight and pericarp weight were found in ‘Mino’ variety traits. High results of fruit weight, technological weight and pericarp weight, comparable with ‘Anchi’ and ‘Sono’ variety were achieved in case of androgenic lines R1 and R18. Because of these economically crucial traits, the androdiploid R1 and R18 lines were included in the crossings. 21 RAPD markers were used in the molecular analysis of the hybrids: (‘Sono’בAnchi’), (‘Anchi’בSono’), (‘Sono’בMino’), (‘Mino’בSono’), (‘Sono’×R1), (R1בSono’), (‘Sono’×R18), (R18בSono’) and their parental forms. DNA amplification was done with the application of Taq polymerase and the method for C. annuum L. was used for PCR-RAPD reaction. To define the size of the received products GPB 3000bp DNA Ladder Mix molecular mass marker and GelAnalyzer 2010a computer programme was used. Genetic distance between the genotypes from the study was defined using the formula of Nei and Li (1979). Dendrogram was constructed with UPGMA method with the application of TREECON computer programme. The starters generated 174 products and 16 of them (9.2%) were polymorphic bands. The size of the products ranged between 118 and 2,122 bp. Coefficients of the genetic distance were calculated on the basis of the analyses, which ranged from 0 to 0.037 for the assessed genotypes. A dendrogram illustrating the degree of genetic diversity between parental forms and reciprocal hybrids was also constructed. The results allowed to identify all genotypes used for crossings and indicated their close relationship with R1 and R18 lines forming separate groups. Moreover, hybrid nature of the analyzed F₁ generations was confirmed through their distinction from maternal forms that allowed to exclude self-pollination. In case of reciprocal hybrids identification within the following pairs was possible: (‘Sono’×R18)F₁, (R18בSono’)F₁ and (‘Sono’×R1)F₁, (R1בSono’)F₁. In both variants androdiploid lines R1 and R18 were used as they had a bigger genetic distance than the other genotypes.