Usefulness of RAPD, RFLP and SCAR molecular markers and AGPaseB gene methylation level in the screening of resistance to the golden cyst nematode (Globodera rostochiensis) pathotype Ro1 in different Polish potato genotypes

Abstract The aim of this study was to assess the application of various DNA markers as well as the ADP-glucose pyrophosphorylase small subunit (AGPaseB) gene methylation for the screening of potato cultivars and breeding lines with different resistance to Globodera rostochiensis. Tetraploid genotypes were included. The 2 kb and 0.7 kb random amplified polymorphic (RAPD) fragments as well as a PCR-amplified 1.6 kb fragment of AGPaseB gene positioned close to the Gro1 locus were tested. Two novel sequence characterised amplified region (SCAR) fragments were generated from 2 kb RAPD product of susceptible and resistant genotypes. Significant correlation (r=0.32) was detected only between the presence of SCARI and SCARII fragments markers in all the investigated genotypes. Three different allelic forms of the AGPaseB gene were detected in tetraploid potato genotypes. One of these allelic forms may be closely linked with Gro1. A strong methylation occurred at promoter region and the 5' part of AGPaseB gene which was generally absent at its 3' UTR region. The methylation pattern of AGPaseB investigated by restriction with MspI and HpaII varied between some G. rostochiensis resistant and sensitive genotypes. We concluded that methylation profiling of AGPaseB gene should be promising for the development of practical assays for resistance to nematodes in potato

Preliminary Genetic Map of a New Recombinant Inbred Line Population for Narrow-leafed Lupin (Lupinus angustifolius L.)

Genetic maps are an essential tool for investigating molecular markers’ linkage with traits of agronomic importance. Breeders put a lot of emphasis on this type of markers, which are used in breeding programs implementation and speed up the process of a new variety development. In this paper, we construct a new, high-density linkage genetic map for Polish material on narrow-leafed lupin. The mapping population originated from crossing the Polish variety ‘Emir’ and the Belarusian breeding line ‘LAE-1’. A new map was constructed based on DArTseq markers—a new type of marker generated with the next-generation sequencing (NGS) technique. The map was built with 4602 markers, which are divided into 20 linkage groups, corresponding with the number of gametic chromosomes in narrow-leafed lupin. On the new map there are 1174 unique loci. The total length of all linkage group is 3042 cM. This map was compared to the reference genome of narrow-leafed lupin and the CDS sequence for model legume species: emphMedicago truncatula, emphLotus japonicus and Glycine max. Analysis revealed the presence of the DArTseq marker common for all investigated species. We were able to map 38 new, unplaced scaffolds on the new genetic map of narrow-leafed lupin. The high-density genetic map we received can be used for quantitative trait locus (QTL) mapping, genome-wide association study analysis and assembly of the reference genome for the whole genome sequencing (WGS) metho

The linear regression between weight of 1000 seeds and seed coat for analysed genotypes <i>Lupinus angustifolius</i>.

<p>The linear regression between weight of 1000 seeds and seed coat for analysed genotypes <i>Lupinus angustifolius</i>.</p

Mean squares from variance analysis for 1000-seed weight and seed coat percentage.

<p>*significance at the level of α = 0.05.</p><p>**significance at the level of α = 0.01.</p

Nucleotide sequence and annealing temperatures of primers generating polymorphic ISSR products in the analysed genotypes.

<p>*R purine, Y pyrimidine; N any nucleotide; B indicates C, G or T; D as A, G, or T; H as A, C, or T; and V as A, C, or G.</p

Weight of 1000 seed and seed coat percentage in analysed genotypes of <i>Lupinus angustifolius</i> L.

<p>Means for each year based of three replications ± standard errors.</p><p>A,B,C …-uniform groups based on the Duncan test.</p><p>AU-originated from Australia.</p><p>PL-originated from Poland.</p><p>BY - originated from Belarus.</p><p>SEM – standard error of the means.</p