Ute Kästner

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.

Regeneration of protoplasts after somatic hybridisation of Hydrangea

Somatic hybridisation of Hydrangea is a promising tool to obtain new basic material for breeding. Viable mesophyll protoplasts were isolated from 21 cultivars and accessions of H. macrophylla, H. paniculata, H. arborescens, H. quercifolia and H. febrifuga. Induction of cell divisions was observed after electromanipulation and fusion by polyethylene glycol. Multi-cellular structures developed in liquid media. Plated microcalli grew on different solid regeneration media. Several phytohormones, their concentration and combination influenced the development of callus and roots. The regular appearance of endophytes challenged successful plant regeneration. As a result of endophytes, the development of microcolonies stopped and they died in liquid protoplast media. Plated microcalli or growing calli turned brown on regeneration medium. Antibiotic Timentin® inhibited expansion of endophytes for a short time. The addition of ascorbic and citric acid to the regeneration media had inhibiting effect on endophyte growth. Calli showed more vitality and grew faster. The supplement of the regeneration media with karrikinolide, a recently discovered new plant growth regulator, brought contradictory results. After addition of karrikinolide microcolonies looked healthier by their shining green colour in liquid media followed by a severe browning of callus soon afterwards. In the further course, karrikinolide promoted the development of endophytes. Shoot induction and plant regeneration succeeded only once from callus that was a result from H. macrophylla ‘Schneeball’ and H. macrophylla ‘Nachtigall’ fusion.

Comparison of in vitro haploid induction in balm (Melissa officinalis)

The generation of haploids in lemon balm (Melissa officinalis L.) is the prerequisite for accelerated development of homozygous lines. Wide experiments to induce sporophytic development were arranged. Donor plants of lemon balm were established in a greenhouse, climate chambers and in vitro. For haploid induction, different variations of media and culture conditions e.g. pretreatment with cold and heat were tested. Sporophytic development of pollen was induced in anther and microspore culture. Callus developed sporadically in anther culture. Multi-cellular structures and microcallus generated from microspores. In vitro cultivated ovules became dark brown and somatic callus generated at the cut surface. Cultivation of ovaries maintained swelling on medium, but undeveloped embryos died.