Is “Esterhazy II”, an Old Walnut Variety in the Hungarian Gene Bank, the Original Genotype?

The old walnut (Juglans regia L.) genotype called “Esterhazy II” was well-known in the Austro-Hungarian Monarchy before World War II, and it can still be found in the Austrian, German and Swiss backyard gardens today. Unfortunately, nowadays, vegetatively propagated progenies of the original “Esterhazy II” are not available anymore around the world because walnut grafting started later than this genotype had become well-known. Although various accessions with “Esterhazy II”-“blood“ are available, it is difficult to determine which one can be considered true or the most similar to the original one. In this paper, phenological and nut morphological characteristics of an “Esterhazy II” specimen planted in a Hungarian gene bank were compared to the varieties “Milotai 10” and “Chandler”. Examined characteristics were: budbreak, blossom time, type of dichogamy, ripening time, nut and kernel features. An additional SSR fingerprinting was used to identify identical genotypes and to demonstrate the relatedness of the analyzed “Esterhazy II” genotype to the other Hungarian walnut cultivars. It can be concluded that under the name “Esterhazy II”, several different genotypes can be observed. All the checked characteristics except budbreak fitted well with the previous descriptions. Our results confirmed that the examined “Esterhazy II” genotype shows high similarity to the “original“ “Esterhazy II” described in the literature.

Validating Blackberry Seedling Pedigrees and Developing an Improved Multiplexed Microsatellite Fingerprinting Set

Confirming parentage and clonal identity is an important aspect of breeding and managing germplasm collections of clonally propagated, outcrossing crops, like blackberry (Rubus subgenus Rubus). DNA fingerprinting sets are used to identify off-cross progeny and confirm clonal identity. Previously, a six-simple sequence repeat (6-SSR) fingerprinting set was developed for blackberry using a small number of samples. The usefulness of the 6-SSR fingerprinting set for pedigree confirmation had not been evaluated. Therefore, it was used in this study to validate parentage for 6 and 12 biparental populations from the University of Arkansas (UA) and US Department of Agriculture Agricultural Research Service (USDA-ARS), Horticultural Crops Research Unit (HCRU) breeding programs, respectively. Twenty-seven of the 489 individuals in these breeding populations were identified as off-cross. The 6-SSR fingerprinting set was sufficient for parentage confirmation; however, a total of 61 plants distributed across 28 sets of genotypes could not be distinguished from each other. An 8-SSR fingerprinting set with improved resolution was subsequently developed and used to evaluate 177 Rubus accessions from the USDA-ARS National Clonal Germplasm Repository, UA, and USDA-ARS HCRU programs. The 8-SSR fingerprinting set distinguished all samples expected to have unique genotypes and identified differing DNA fingerprints for two sets of accessions suspected to have identical fingerprints. Cluster analysis grouped the accessions from the eastern and western US breeding programs based on geography and descent. Future work will focus on establishing a database of DNA fingerprints for germplasm identification and for determining pedigree relationships between blackberry accessions.

Genetic variability assessment in ‘Muscat’ grapevines including ‘Muscat of Alexandria’ clones from selection programs

Genetic variability is needed to face environmental changes and pathogen constrains. In addition, the search for intravarietal variability contributes to the avoidance of genetic erosion, preserving clones that are adapted to particular conditions. Variability is also important to diversify grapevine-derived products. In this work, we have analyzed the genetic variability of ‘Muscat germplasm’ including samples from neglected vineyards from Alicante and Valencia provinces, accessions of the germplasm collections of ‘Colección de Vides de El Encín’ (Alcalá de Henares, Madrid) and ‘La Casa de las Vides’ (Agullent, Valencia), accessions supplied by nurseries of Valencia province, and ‘Muscat of Alexandria’ clones selected using differential ampelographic characteristics in selection programs (La Marina, Alicante). Fifteen microsatellites (SSRs) were used to study intervarietal variability. The SSR fingerprinting allowed the identification of some accessions, variants, and synonymies. Amplified Fragment Length Polymorphisms (AFLPs) markers and Microsatellite-AFLPs were used to determine the variability attended in ‘Muscat of Alexandria’ accessions. A CAPs (Cleaved Amplified Polymorphic Sequences) marker, recently developed for the discrimination of ‘Muscat’ flavor genotypes using the SNP1822 G>T, was assessed and showed that all the analyzed accessions were ‘Muscat’ flavored. The variation found among the analyzed germplasm is very interesting because variants within ‘Muscat of Alexandria’, ‘Muscat Italia’, and ‘Muscat d’Istambul’ have been identified. In addition, intravarietal genetic variation was found among the analyzed accessions in ‘Muscat of Alexandria’ from selection programs.

Genetic identification of 43 elite clonal accessions of Populus deltoides by SSR fingerprinting

In this study, 43 elite clones of Populus deltoides were fingerprinted with ABI 3730 capillary electrophoresis by using six SSR primer pairs. Based on the fingerprinting profiles, 62 polymorphic bands were generated with a mean number of 10 alleles per locus, and allele numbers amplified by each primer in these clones ranged from 5 (NJFUP-poly10) to 16 (NJFUP-poly07). The power of discrimination values for these primer pairs ranged from 0.80 to 0.94, with an average value of 0.89. Among the six primer pairs, the most efficient primer pair for genetic discrimination of these elite clones was NJFUP-poly02, which could identify 22 of the 43 elite clones directly. In conclusion, all the 43 elite clones of P. deltoides could be discriminated unambiguously based on their genotypes at the six SSR loci. The combination of all six loci gave considerable reliability and accuracy for genetic identification of these clonal accessions. Subsequently, genetic relationship of these clones was plotted by the UPGMA clustering and principle component analysis. Results of both analyses indicated that clone “C7-1” had the largest genetic distance from the other clones, followed by clone “C51-1” and a subgroup comprising clone “C100-3” and “C62-3”. These clones are proposed to be possibly more affected by the inter-specific gene introgression.