Otto Schrader

Ryd4 Hb : a novel resistance gene introgressed from Hordeum bulbosum into barley and conferring complete and dominant resistance to the barley yellow dwarf virus

Barley yellow dwarf virus (BYDV) causes high yield losses in most of the major cereal crops worldwide. A source of very effective resistance was detected within the tetraploid wild species of Hordeum bulbosum. Interspecific crosses between a resistant H. bulbosum accession and H. vulgare cv. ‘Igri’ were performed to transfer this resistance into cultivated barley. Backcrosses to H. vulgare resulted in offspring which carried a single subterminal introgression of H. bulbosum chromatin on barley chromosome 3HL and proved to be fully resistant to BYDV-PAV, as inferred by ELISA values of zero or close to zero and lack of BYDV symptoms. Genetic analysis indicated a dominant inheritance of the BYDV-PAV resistance factor, which we propose to denote Ryd4Hb. The identity and effect of Ryd4Hb are discussed in relation to other known genes for BYDV resistance or tolerance, as well as the relevance of this gene for resistance breeding in barley.

Interspecific hybrids between onion (Allium cepa L.) with S-cytoplasm and leek (Allium ampeloprasum L.)

Abstract Interspecific hybrids between Allium cepa and A. ampeloprasum have been generated as a first step for the introduction of S-cytoplasm from onion into leek. Pre-zygotic barriers of crossability were observed after the arrival of pollen tubes at the end of the style when entering the cavity. Nevertheless, micropyle penetration of pollen tubes and the formation of hybrid embryos were also observed. After accomplishing in vitro culture of ovaries and ovules successively, triploid hybrid plants with 24 chromosomes were obtained. Their hybrid nature was confirmed by RAPD analysis, genomic in situ hybridization, and morphological analysis. Southern hybridization with a cytoplasmic probe indicated the transfer of unaltered S-cytoplasm into the hybrid plants.

Characterization of centromeric histone H3 (CENH3) variants in cultivated and wild carrots (Daucus sp.)

In eukaryotes, centromeres are the assembly sites for the kinetochore, a multi-protein complex to which spindle microtubules are attached at mitosis and meiosis, thereby ensuring segregation of chromosomes during cell division. They are specified by incorporation of CENH3, a centromere specific histone H3 variant which replaces canonical histone H3 in the nucleosomes of functional centromeres. To lay a first foundation of a putative alternative haploidization strategy based on centromere-mediated genome elimination in cultivated carrots, in the presented research we aimed at the identification and cloning of functional CENH3 genes in Daucus carota and three distantly related wild species of genus Daucus varying in basic chromosome numbers. Based on mining the carrot transcriptome followed by a subsequent PCR-based cloning, homologous coding sequences for CENH3s of the four Daucus species were identified. The ORFs of the CENH3 variants were very similar, and an amino acid sequence length of 146 aa was found in three out of the four species. Comparison of Daucus CENH3 amino acid sequences with those of other plant CENH3s as well as their phylogenetic arrangement among other dicot CENH3s suggest that the identified genes are authentic CENH3 homologs. To verify the location of the CENH3 protein in the kinetochore regions of the Daucus chromosomes, a polyclonal antibody based on a peptide corresponding to the N-terminus of DcCENH3 was developed and used for anti-CENH3 immunostaining of mitotic root cells. The chromosomal location of CENH3 proteins in the centromere regions of the chromosomes could be confirmed. For genetic localization of the CENH3 gene in the carrot genome, a previously constructed linkage map for carrot was used for mapping a CENH3-specific simple sequence repeat (SSR) marker, and the CENH3 locus was mapped on the carrot chromosome 9.

Construction of a chromosome-assigned, sequence-tagged linkage map for the radish, Raphanus sativus L. and QTL analysis of morphological traits

The radish displays great morphological variation but the genetic factors underlying this variability are mostly unknown. To identify quantitative trait loci (QTLs) controlling radish morphological traits, we cultivated 94 F4 and F5 recombinant inbred lines derived from a cross between the rat-tail radish and the Japanese radish cultivar ‘Harufuku’ inbred lines. Eight morphological traits (ovule and seed numbers per silique, plant shape, pubescence and root formation) were measured for investigation. We constructed a map composed of 322 markers with a total length of 673.6 cM. The linkage groups were assigned to the radish chromosomes using disomic rape-radish chromosome-addition lines. On the map, eight and 10 QTLs were identified in 2008 and 2009, respectively. The chromosome-linkage group correspondence, the sequence-specific markers and the QTLs detected here will provide useful information for further genetic studies and for selection during radish breeding programs.

Transfer of a male-sterility-inducing cytoplasm from onion to leek (Allium ampeloprasum)

Abstract.Two interspecific triploid (AAC) hybrids (84/1-94 and 99/1-94) from crosses between onion [Allium cepa (2n=2x=16, CC)] and leek [A. ampeloprasum (2n=4x=32, AAAA)] were backcrossed to leek in order to transfer a male-sterility-inducing cytoplasm from onion that would enable the production of hybrid leek. GISH evaluations of meiosis in the interspecific hybrids revealed irregularities due to univalent onion chromosomes producing micronuclei from onion chromatin, whereas the pairing of the two sets of leek chromosomes was nearly normal. Attempts to use colchicine to double the chromosome number of the hybrids failed. Backcrosses of 84/1-94 to leek as the pollen parent were not successful. The first backcross of 99/1-94 to tetraploid leek produced 11 BC1 plants with chromosome numbers between 38 and 41. Identification of parental chromosomes by GISH showed that all eight onion chromosomes and 30–33 leek chromosomes were transmitted to the backcross progenies due to unreduced egg cells. Onion chromosomes were eliminated during the second backcross. Southern hybridization confirmed the transfer of the T-cytoplasm like source of CMS from onion to the BC2 progenies. After the third backcross to leek, 158 plants were obtained with varying numbers of onion chromosomes and some intergenomic recombinant chromosomes. Alloplasmic leek plants without onion chromatin were selected for further characterization of male sterility and quality traits.

Chromosome number and ploidy level of balm (Melissa officinalis)

BackgroundLemon balm (Melissa officinalis L.) is of increasing importance resulting in rising growth area. Improved knowledge on the genome structure, number of chromosomes in connection with the taxonomical structure of balm is indispensable for improved new varieties.ResultsA collection of 40 balm accessions (M. officinalis) was characterized by flow cytometry and FISH (18/25S and 5S rDNA) to determine the chromosome number and ploidy level. Three different types were found: diploid genotypes with 2n = 2× = 32 chromosomes; tetraploid 2n = 4× = 64 chromosomes and triploid 2n = 3× = 48 chromosomes. A haploid base number of × = 16 chromosomes is likely. First time described triploid accessions are sterile but cytologically and morphologically stable for many years. Triploids express better winter hardiness and regeneration after harvesting cuts as well as bigger leaves and internodes.ConclusionsA basic chromosome number of x = 16 is reported for the first time for the species M. officinalis.AbstractDie wachsende Bedeutung von Zitronenmelisse (Melissa officinalis L.) führt zur Ausdehnung des hierfür erforderlichen Anbauumfanges. Ein verbesserter Kenntnisstand der Genomstruktur, der Chromosomenzahl und der hiermit in Zusammenhang stehenden taxonomischen Struktur der Melisse sind unerlässliche Voraussetzungen für verbesserte, neue Sorten.Eine Kollektion von 40 Melisseherkünften (M. officinalis) wurde durchflusszytometrisch und durch FISH (18/25S and 5S rDNA) untersucht, um den Ploidiegrad und die Chromosomenzahl zu ermitteln. Drei unterschiedliche Typen wurden konnten bestimmt werden: diploide Genotypen mit 2n = 2× = 32 Chromosomen; tetraploide mit 2n = 4× = 64 Chromosomen und triploide mit 2n = 3× = 48 Chromosomen. Die haploide Chromosomenzahl ist mit x = 16 anzunehmen. Die erstmalig beschriebenen triploiden Herkünfte sind steril aber zytologisch und morphologisch über viele Jahre stabil. Sie zeigen eine bessere Winterhärte und einen schnelleren Wiederaufwuchs nach Ernteschnitten, wie auch größere Blätter und Internodien.Die Basischromosomenzahl von x = 16 wird erstmalig für die Art M. officinalis beschrieben.