Gabriela Libiaková

Micropropagation in vitro of highbush blueberry (Vaccinium corymbosum L.)

Murashige and Skoog medium (MS) and modified Anderson’s Rhododendron medium (mAN) were compared for in vitro shoot multiplication of three highbush blueberries ‘Berkeley’, ‘Bluecrop’ and ‘Goldtraube’. All media contained 0.5 mg l−1 zeatin applied either alone or combined with 0.1, 1 and 5 mg l−1 IBA. In vitro rooting was induced using mAN medium supplemented with 0.8 mg l−1 IBA and 4 g l−1 activated charcoal. The results obtained showed that mAN medium is more suitable for in vitro multiplication of the selected highbush blueberry cultivars than MS medium. Low concentration of IBA (≤1 mg l−1) added in zeatinsupplemented mAN medium increases shoot multiplication efficiency of highbush blueberries in vitro and can be recommended for large-scale propagation of high-quality plants. MS medium induced partial or full necrosis of stems and leaves, which was more pronounced on media containing zeatin combined with increasing concentration of IBA. Rooting capacity of shoots varied widely among the tested blueberry cultivars. The highest rooting and acclimatization rates were achieved in ‘Goldtraube’ (82.8% and 91.8% respectively), and the lowest (10% and 66.7% respectively) were in ‘Berkeley’.

Methodological Approaches to Simple Enzyme Polymorphism Analyses of Amaranth Species (Amaranthus SP.)

Methodological Approaches to Simple Enzyme Polymorphism Analyses of Amaranth Species (Amaranthus SP.) Amaranths have drawn a great interest in last years as an agronomic crop, alternative energy source and ornamental plants around the world. Several of them are cultivated as the leaf vegetables, cereals, or as the colourful, quick growing garden plants. Increasing demand for the breeding and proteomic study in Amaranthus species led us to test some methodological approaches to enzyme polymorphism analysis. For these experiments genotypes of Amaranthus cruentus L. (genotype Ficha) and mutant line of hybrid K-433 were selected, characterized by a good seed quality and quantity, suitable for food production. The attention was devoted to test of published methodology, which were slightly modified for enzyme (ACP, ADH, CAT, DIA, GOT, IDH, MDH, PGD, PGI and PGM) multiplicity analysis for amaranths and testing of feasibility of chosen analysed organ weights, dimensions of Whatman No. 2 wicks and different volumes of extract buffer. Results from enzyme multiplicity analysis are presented by means of photographed fingerprints and phenotypes are expressed in diagrams where positions of zymograme bands are marked by using factors of relative mobility (Rm).

Shoot proliferation ability of selected cultivars of Rubus spp. as influenced by genotype and cytokinin concentration

The aim of this work was to develop protocols for micropropagation of selected Rubus cultivars – ‘Tulameen’ and ‘Black Jewel’ (Rubus idaeus L.), ‘Black Satin’ (Rubus fruticosus L.) and ‘Tayberry’ (Rubus fruticosus x Rubus idaeus). Nodal segments carrying dormant buds were used for in vitro culture establishment. For shoot initiation, MS medium containing 8 g*L-1 phytoagar, 30 g*L-1 sucrose, 1 mg*L-1 6-benzylaminopurine (BAP), 0.2 mg*L-1 indole-3-butyric acid (IBA) and 150 mg*L-1 cefotaxime was used. The shoot proliferation ability of each cultivar was evaluated by multiplication coefficient (number of shoots formed per explant) during 4 subcultures. Statistical analyses confirmed the significantly higher shoot multiplication ability in cvs. ‘Black Satin’ and ‘Tulameen’ in comparison with cvs. ‘Tayberry’ and ‘Black Jewel’, as well as significant differences in shoot proliferation between subcultures (1st and 2nd versus 3rd and 4th). Testing of influence of different BAP concentrations on shoot formation showed that BAP 1-2 mg*L-1 is the best for shoot proliferation with the higher multiplication rate obtained in cv. ‘Black Satin’ in comparison with ‘Black Jewel’. Statistical evaluation showed that genotype and BAP concentration significantly influenced the ability of shoot proliferation in Rubus cultivars.

Revealing the seed proteome of the health benefitting grain amaranth (Amaranthus cruentus L.)

Amaranth, a staple food source in ancient Aztec, Maya and Inca cultures, has been recognised as a 21st century crop. This superfood, known as Inca wheat, attracts the worldwide attention of researchers and farmers for its superior agronomical and technological properties but especially because of its exceptional nutritive value. A combination of two-dimensional electrophoresis (2-DE) with MS facilitating the effective differentiation of 13 classes of Amaranthus cruentus L. “Ficha” mature seed proteins was used in the current study. The best resolution results in the 2-DE were obtained using immobilised pH gradients strips with a pH range of 5–8. Out of the 461 spots detected, 249 were successfully identified by LC-MS/MS analysis, making this the most inclusive protein profile of mature amaranth seed. Unknown proteins represented the most abundant class of proteins (59), the second most abundant category was related to energy (46) and then to seed storage proteins (43). These results expand the knowledge of the amaranth seed proteome and it is expected that the data presented here will contribute to further in-depth characterisation of Amaranthus seeds.