Marion Nachtigall

The recipient potato cultivar influences the genetic makeup of the somatic hybrids between five potato cultivars and one cloned accession of sexually incompatible species Solanum bulbocastanum Dun.

Somatic hybridization is a biotechnological tool, which allows the transfer of multiple resistance genes from sexually incompatible Solanum species into cultivated potato. Here we report the effect of the recipient commercial tetraploid potato cultivar on the genetic make-up of the somatic hybrids (SHs) with an accession of the incongruent diploid species Solanum bulbocastanum Dun. The SHs were produced by mesophyll protoplast electrofusion. The analysis of ploidy by flow cytometry was first used to select hexaploid putative SH shoots but SSR and AFLP markers, DAPI staining and later flow cytometry evaluation of ploidy reveals symmetric and asymmetric SH plants regeneration in proportions that depend on the potato cultivar. The growth and fertility of the SHs support the effect of recipient cultivar. Out of five different fusion combinations, the highest number of SHs and tuberosum morphology was recorded for the combinations: blb41 (+) ‘Delikat’ 235 SHs (104 symmetric and 131 asymmetric) and blb41 (+) ‘Rasant’ 33 SH plants (22 symmetric, 11 asymmetric). There were fertile SHs of these combinations and BC1 and BC2 progenies were obtained. Less successful were the combinations: blb41 (+) ‘Quarta’ 64 SHs (57 symmetric and 7 asymmetric), blb41 (+) ‘Baltica’ 25 SHs (16 symmetric and 9 asymmetric), which were infertile and blb41 (+) ‘Agave’ with only one highly asymmetric non-viable SH plant. The production of a large number of SHs with diverse commercial cultivars is a prerequisite for further selection of useful pre-breeding material. The causes of nuclear constitution asymmetry and somatic incompatibility of the two species are also discussed.

Genetic Diversity Analysis of Date Palm (Phoenix dactylifera L.) Cultivars from Morocco Using SSR Markers

Abstract In Morocco, date palm is the most important arboricultural crop and little is known about its germplasm. Thus, this work aimed at analyzing genetic diversity among 200 date palms sampled from three oases (Figuig, Tata and Zagora) of Morocco using microsatellite markers. Among these palms, 191 were females, belonging to 26 cultivars, and 9 were males. Eighteen primers were used for the analysis of their genetic diversity. Only 15 primers amplified successfully all the samples. The total number of alleles was 116 and the percentage of polymorphic loci was high and ranged between 60 and 100% with an average of 93.33%. The genetic similarity values ranged from 0.146 to 0.745. The molecular variance analysis showed 64% of variability among cultivars. The obtained dendrogram showed three groups and generally, a good structuring of cultivars. However, we noticed one case of homonymy among cultivars. In fact “Tadmant” cultivar of Figuig was different from “Tadmant” of Tata and Zagora. Males were clustered in two main subgroups.

Development of SSR Markers for the Genus Patellifolia (Chenopodiaceae)

Premise of the study: Microsatellite primers were developed to promote studies on the patterns of genetic diversity within Patellifolia patellaris (Chenopodiaceae) and the relationship between the three species of the genus Patellifolia. Methods and Results: The genomic sequence from P. procumbens was screened for simple sequence repeats (SSRs), and 3648 SSRs were identified. A subset of 53 SSR markers was validated, of which 25 proved to be polymorphic in the three species except for the P. webbiana–specific marker JKIPat16. The number of alleles ranged from 85 in P. patellaris, 187 in P. procumbens, and 202 in P. webbiana. Conclusions: The set of 25 new markers will facilitate studies of the relationships between the three Patellifolia species and of the spatial and temporal distribution of genetic diversity within the species.

Genetic diversity and differentiation in Patellifolia (Amaranthaceae) in the Macaronesian archipelagos and the Iberian Peninsula and implications for genetic conservation programmes

This is the first comprehensive investigation of the patterns of genetic diversity of Patellifolia species. The main objective of our research work is to determine Most Appropriate crop Wild relative Populations (MAWP) suited to conserve in situ wild relatives of the sugar beet. Individual plant samples of P. patellaris were collected at 26 and of P. procumbens/P. webbiana at seven sites and analysed with 24 and 22 microsatellite markers, respectively. On average 15 alleles per locus were found within the set of 581 P. patellaris and an average of 12 alleles per locus in the set of 172 P. procumbens/P. webbiana individuals. The factorial analysis showed diversity patterns which agree well with the geographic origin of the samples. The genetic data suggest that P. patellaris reproduces mainly by self-fertilisation while P. procumbens/P. webbiana have the signature of out-breeders. The measure Δ was used to calculate the genetic distance of each occurrence to the pooled remaining occurrences, the complement. Occurrences with either the lowest or the highest genetic distance to the complement are particularly suited to conserve the genetic diversity of the species. Eight occurrences of P. patellaris, two of P. procumbens and one for P. webbiana were determined according to this scheme, proposed as MAWP and recommended for the establishment of genetic reserves.

Identifizierung, Aufbau und Ausbau genetischer Schutzgebiete für wildlebende Verwandte unserer Kulturarten (WVK)

Mit Kultur- und Nutzpflanzenarten verwandte Wildarten sind ungeachtet ihres grosen Potenzials als Quellen neuartiger genetischer Variation fur die Pflanzenzuchtung nicht ausreichend geschutzt. Zur Verbesserung ihres langfristigen Schutzes fordern sowohl internationale als auch nationale Biodiversitatsstrategien die Umsetzung einer In-situ-Erhaltungsstrategie, insbesondere fur jene Arten, die fur den Menschen von unmittelbarer Bedeutung sind. Zur Operationalisierung dieser Strategie wurde im Rahmen des vom Julius Kuhn-Institut koordinierten AEGRO-Projektes die Konzeption des genetischen Schutzgebietes erprobt. Fur vier Kulturpflanzengattungen (Avena, Beta/Patellifolia, Brassica, Prunus) wurde das Konzept bis zur Anwendungsreife weiterentwickelt. Nur vier Entscheidungsschritte sind notwendig, um aus einer Vielzahl in Europa verbreiteter Arten jene Vorkommen auszuwahlen, fur die der Ausbau eines genetischen Schutzareals erforderlich ist. Zur Unterstutzung dieser Vier-Schritte-Methode wurden das Informationssystem „Population Level Information System“ (PLIS) und zur Dokumentation der Ergebnisse des Auswahlverfahrens das Informationssystem „Genetic Reserve Information System (GenResIS) entwickelt. In GenResIS sind detaillierte okogeographische Informationen zu 52 genetischen Schutzgebieten und den darin vorkommenden Arten zu finden. Diese Schutzgebiete dienen nicht nur zur Bewahrung oder Wiederherstellung eines guten Erhaltungszustandes von Pflanzenarten in ihrem naturlichen Lebensraum, sondern auch der Erhaltung von Genen fur die Pflanzenzuchtung, wie exemplarisch am Beispiel von Beta vulgaris subsp. maritima als Quelle der Resistenz gegen das Beet Necrotic Yellow Vein Virus (BNYVV) erlautert wird. Stichworter: Genetische Ressourcen, In-situ-Erhaltungsstrategie, genetische Schutzgebiete, Wildarten Identification, establishment and development of genetic reserves for crop wild relatives (CWR) Abstract Despite their great potential as sources of novel genetic variation for plant breeding crop wild relatives are not protected sufficiently. International as well as national biodiversity strategies call therefore the implementation of the in situ conservation strategy, in particular for those species of immediate relevance to mankind. The genetic reserve concept was tested in the context of the AEGRO project, coordinated by the Julius Kuhn-Institut, to operationalise the in situ conservation strategy and to develop the concept for the genera Avena, Beta/Patellifolia, Brassica, Prunus to the point that it can be put into use. Only four decision steps are required to determine those occurrences among the multitude of species distributed in Europe for which the establishment of a genetic reserve is essential. The information system “Population Level Information System” (PLIS) was developed to support the four step methodology; the “Genetic Reserve Information System” (GenResIS) was established to document the results of the selection procedure. GenResIS provides detailed ecogeographic information on 52 genetic reserves as well as on species occurring within these areas. The reserves do not only serve the protection or recovery of species in their natural habitat. They also serve the maintenance of genes for plant breeding as explained using Beta vulgaris subsp. maritima, the source of resistance against the Beet Necrotic Yellow Vein Virus (BNYVV), as an example. Keywords: Genetic resources, in situ conservation strategy, genetic reserve, wild species

Development and application of molecular and bioinformatic tools for the genetic monitoring of wild beets

Crop Wild Relatives (CWR) are an economically important plant genetic resource (PGR) for plant breeding. One strategy to conserve PGR in the natural habitat (in situ) is the genetic reserve technique. This approach protects the existing intraspecific diversity and allows, in contrast to the ex situ approaches, the emergence of new diversity. The genetic reserve conservation technique will be tested in practice with the genus Beta within the framework of the EU project AEGRO. B. patula (Ait.) (BP) and B. vulgaris (L.) maritima (Arcang.) (BVM) are used as models. Both species are important resources for sugarbeet breeding. BP is a rare and endangered species with a very limited habitat. In contrast, BVM is widespread along the shores of Western Europe. This subspecies is expected to migrate in a northward direction as a consequence of the global climate change. The impact of this migration on the genetic diversity will be examined in future analyses and will establish a basis for future research. 3. Nachwuchswissenschaftlerforum Julius-Kühn-Archiv, 430, 2011 49 A total of 557 plants from 14 sampling plots were analysed using 25 SSR markers (Simple Sequence Repeat markers). Additionally, a database was modeled and implemented to support the genetic monitoring by storing and post-processing all laboratory and field data in a well documented and structured way. This makes the data available for the analysis of time series. For the first time the geographic pattern of genetic diversity in the wild beet BP was described and decision criteria for the establishment of a genetic reserve for BP were postulated. The knowledge of geographic patterns of genetic diversity in the wild beet BVM was consolidated as a baseline for further investigations on the potential influence of climate changes and the corresponding geographic range shift on genetic diversity.