To study the expression of pathogenicity-related genes inErwinia amylovora, seven candidate reference genes (ffh,glyA,gyrA,proC,pykA,recA,rpoB) were selected and validated with the following five different mathematic algorithms: geNorm, NormFinder, BestKeeper, the delta CT method and the RefFinder web-based tool. An overall comprehensive ranking output from each of the selected software programs revealed thatproCandrecA, followed byffhandpykA, were the most stably expressed genes and can be recommended for the normalization of RT-qPCR data. A combination of the three reference genes,proC,recAandffh, allowed for the accurate expression analysis ofamsBandhrpNgenes and the calculation of their fold change inE. amylovoraafter its infection of susceptible and resistant apple cultivars. To the best of our knowledge, this is the first study presenting a list of the most suitable reference genes for use in the relative quantification of target gene expression inE. amylovora in planta, selected on the basis of a multi-algorithm analysis.

The purpose of the study was to support the selection process of the most valuable currant and gooseberry accessions cultivated in Northern Europe, in order to establish a decentralized core collection and, following the selection, to ensure sufficient genetic diversity in the selected collection. Molecular analyses of the material from nine project partners were run at seven different laboratories. The results were first analysed for each partner separately, and then combined to ensure sufficient genetic diversity in the core collection.

Genetic variation between five apple cultivars (‘Golden Delicious’, ‘Gala’, ‘Jonagold’, ‘Šampion’, and ‘Idared’) and ten of their sports (‘Golden Delicious Reinders’, ‘Goldrosio’, ‘Gala Must’, ‘Gala Schniga Schnitzer’, ‘Jonagored’, ‘Jonagold Excel’, ‘Szampion Arno’, ‘Szampion Reno Malinowy’, ‘Idaredest’, and ‘Red Idared’) was investigated using five types of DNA markers: Inter-Simple Sequence Repeats (ISSR), Simple Sequence Repeats (SSR), Amplified Fragment Length Polymorphism (AFLP), Sequence-Specific Amplified Polymorphism (S-SAP), and Inter-Primer Binding Site (iPBS) amplification. In total, 941 polymorphic amplified fragments were obtained using 12 ISSR, 12 SSR, ten AFLP, 19 iPBS, and 15 S-SAP primers or primer pairs. Four of the above-described techniques (except for SSRs with the primer pairs used in this study) were able to distinguish between the sports and their parental cultivar. The most effective technique to distinguish between the genotypes analysed was S-SAP, which detects variations in DNA regions flanking retrotransposon insertion sites.The combined use of ISSR,AFLP, iPBS, and S-SAP markers identified and distinguished all of the sports tested.

Obtaining a sufficient quantity of high-quality, intact RNA is the first crucial step in its study by RNA sequencing on next-generation sequencing platforms or quantitative PCR. Different RNA extraction methods or commercial kits vary in yield and in the quality and integrity of the RNA obtained, which may affect the results of downstream applications. Often, these factors depend on the organism under study and nature of the sample. Therefore, the selection of an appropriate RNA isolation method is critical. In this study, we present the results of an evaluation of three different commercial kits for the isolation of total RNA from Erwinia amylovora in apple tissue as well as the usefulness of different kinds of Deoxyribonuclease I for DNA removal and kits for rRNA depletion. To our knowledge, this is the first report on the method of isolation of high-quality E.amylovora mRNA for RNA-seq.

Correlations between the sowing value of primed tomato seeds (mean germination time – MGT, germination uniformity – T_75-25) and selected physiological events (total activity of dehydrogenases – TAD, activity of catalase – AC, activity of the cell cycle – ACC, and leakage of electrolytes – LE) were analyzed in order to find useful physiological markers of seed quality. To accomplish this purpose, we analyzed the effects of different environment-friendly priming methods, the levels of seed moisture content (MC), and incubation times during priming on germination properties (MGT, T_75-25) in relation to TAD, AC, ACC, and LE. In addition, we evaluated the effects of used seed conditioning methods on the growth and development of plants obtained from these seeds. The results showed positive effects of seed conditioning on the growth and development of plants obtained from these seeds, and that MGT and T_75-25 were significantly correlated with TAD, AC, ACC, and LE from the seeds. The correlations between MGT and the physiological parameters were the strongest (r² = 77.0 - 97.2%). Somewhat lower values of the determination coefficient (r² = 62.8 - 92.7%) were noted between T_75-25 and the physiological events. Therefore, the investigated physiological parameters of seed quality can be used as markers of germinability and sowing value reached by primed tomato seeds.