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The aim of our long-term research program is to improve the quality and quantity of amaranth production through mutation breeding using γ-radiation. In this paper, we present the characterization of the new variety ‘Zobor’ of A. hypochondriacus × A. hybridus developed by radiation-induced mutagenesis of hybrid K-433. Multiyear phenotypic characterization of an important yield parameter (1000-seed weight) showed that the studied mutant variety ‘Zobor’ has an advantage in seed weight over the nonirradiated control seeds of K-433 with predictable performance of this yield trait. ‘Zobor’ exhibited changes in seed morphometric parameters, starch particle size, and pasting properties with no change in amylose content and swelling power. Moreover, the seeds of ‘Zobor’ showed the significantly highest folate content among selected amaranth varieties. The mutant variety could, therefore, be interesting for the development of functional foods and as a low-management crop, attractive for cultivation in Europe.

Amaranth species represent a diverse group of plants. Many of them are a rich source of secondary metabolites with many positive biological effects. Total phenolic, total flavonoid and rutin content, antioxidant activity against superoxide and hydroxyl radicals, FRAP (Ferric-reducing ability of plasma) assay and DPPH (2,2-Diphenyl-1-picrylhydrazyl) radical scavenging assay were determined in ethanol extracts of dried leaves of the new Slovak amaranth varieties ‘Pribina’ and ‘Zobor’. The amount of total phenolic substances (‘Pribina’ GAE 38.3 mg.g−1 DM and ‘Zobor’ GAE 26.1 mg.g−1 DM), content of total flavonoids (‘Pribina’ QE 26.5 mg.g−1 DM and ‘Zobor’ QE 20.3 mg.g−1 DM) and rutin (‘Pribina’ 50.8 mg.g−1 DM and ‘Zobor’ 15.2 mg.g−1 DM) were higher in the variety ‘Pribina’, compared to the variety ‘Zobor’. A statistically higher antioxidant activity against superoxide radical (1.63%·mg−1g−1 DM), hydroxyl radical (3.20%.mg−1g−1 DM), FRAP assay (292.80 µmol.L−1·mg−1.g−1 DM) and DPPH (54.2 ± 1.78 µg.mL−1 DM) were detected in the ‘Pribina’ variety. Antiradical and antioxidant activities of both extracts showed high positive correlations in relation to the content of total phenolic substances, total flavonoids and rutin. Amaranth is an undemanding crop on specific environmental conditions and is resistant to abiotic and biotic stress.

The aim of this work was to assess the regeneration capacity of Amelanchier alnifolia var. cusickii and Lonicera kamtschatica cv. ‘Jugana’ from different types of explants under various hormonal treatments. The whole leaves, petioles, and internodal segments of in vitro plants were examined as explants. Several plant growth regulators (cytokinins and auxins) were evaluated for their ability to induce adventitious regeneration. Direct and indirect organogenesis was achieved under certain culture conditions in both species. The frequency of shoot regeneration was strongly dependent on concentrations of plant growth regulators in the induction media (L.kamtschatica ‘Jugana’) or concentrations of plant growth regulators in the induction media and type of explant (A. alnifolia var. cusickii). Results showed that leaves were not suitable explants for A. alnifolia var. cusickii. Both species were able to regenerate shoots from internodal segments and petioles. The highest induction of shoots was obtained on Murashige and Skoog (MS) medium enriched with 2 mg/L thidiazuron (TDZ) and 0.5 mg/L indole-3-butyric acid (IBA) for Amelanchier alnifolia and with 1 mg/L TDZ and 0.2 mg/L indole-3-acetic acid (IAA) for L. kamtschatica ‘Jugana’. Obtained adventitious shoots were further proliferated in order to investigate their multiplication capacity. The multiplication of shoots was successful in all cultivars, with the best results reported in A. alnifolia var. cusickii (7.07 shoots/explant on average).

Berry fruit species are, in many countries, considered biologically and economically valuable and important species of small fruits. The aim of this work was to examine the influence of either decreased or increased mesos concentrations (MgSO4, CaCl2, and KH2PO4) on shoot multiplication of five cultivars of three small fruit species (Amelanchier alnifolia var. cusickii, Rubus fruticosus ‘Black Satin’ and ‘Loch Ness’, and Vaccinium corymbosum ‘Brigitta Blue’ and ‘Toro’). Mesos nutrients were manipulated from half to four times their base concentration. The results indicate that mesos manipulation significantly influences the number and length of shoots in most of the studied cultivars. The greatest multiplication rate for A. alnifolia was achieved with tripled mesos, whereas ‘Black Satin’ and ‘Loch Ness’ reacted positively to a lower (1–2x) concentration of mesos. Decreasing the concentration of mesos to half led to worse quality in both blackberry and Saskatoon shoots. ‘Brigitta Blue’ was more sensitive to greater mesos concentrations compared to ‘Toro’. Optimizing the mineral nutrition of plants cultivated in vitro enhances their multiplication rate and contributes to a higher production of good quality plantlets.

The aim of our long-term research program is to improve the quality and quantity of amaranth production through mutation breeding using γ-radiation. In this paper, we present the characterization of the new variety ‘Zobor’ of A. hypochondriacus × A. hybridus developed by radiation-induced mutagenesis of hybrid K-433. Multiyear phenotypic characterization of an important yield parameter (1000-seed weight) showed that the studied mutant variety ‘Zobor’ has an advantage in seed weight over the nonirradiated control seeds of K-433 with predictable performance of this yield trait. ‘Zobor’ exhibited changes in seed morphometric parameters, starch particle size, and pasting properties with no change in amylose content and swelling power. Moreover, the seeds of ‘Zobor’ showed the significantly highest folate content among selected amaranth varieties. The mutant variety could, therefore, be interesting for the development of functional foods and as a low-management crop, attractive for cultivation in Europe.

The aim of this work was to evaluate shoot multiplication of two blackberry (Rubus fruticosus L.) cultivars ?Black Satin? and ?Loch Ness? on two culture media: Murashige & Skoog (MS) and its modification Murashige & Skoog Van der Salm medium (MS VDS) which differ only in iron source (FeNaEDTA, FeEDDHA, respectively). Statistical analyses showed no significant difference in shoot multiplication between different iron sources for both tested cultivars. For ?Black Satin? it was shown that double concentration of chelates FeNaEDTA and FeEDDHA in culture media negatively affected on shoot growth and multiplication. These results can be very useful for further optimization of micropropagation process for different Rubus cultivars.

Berry fruit species are, in many countries, considered biologically and economically valuable and important species of small fruits. The aim of this work was to examine the influence of either decreased or increased mesos concentrations (MgSO4, CaCl2, and KH2PO4) on shoot multiplication of five cultivars of three small fruit species (Amelanchier alnifolia var. cusickii, Rubus fruticosus &lsquo, Black Satin&rsquo, and &lsquo, Loch Ness&rsquo, and Vaccinium corymbosum &lsquo, Brigitta Blue&rsquo, Toro&rsquo, ). Mesos nutrients were manipulated from half to four times their base concentration. The results indicate that mesos manipulation significantly influences the number and length of shoots in most of the studied cultivars. The greatest multiplication rate for A. alnifolia was achieved with tripled mesos, whereas &lsquo, reacted positively to a lower (1&ndash, 2x) concentration of mesos. Decreasing the concentration of mesos to half led to worse quality in both blackberry and Saskatoon shoots. &lsquo, was more sensitive to greater mesos concentrations compared to &lsquo, Optimizing the mineral nutrition of plants cultivated in vitro enhances their multiplication rate and contributes to a higher production of good quality plantlets.

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.

The introduction of foreign DNA into the plant genome by Agrobacterium tumefaciens is a promising technique of targeted gene transfer which depends on good working regeneration system. The aim of the work was to elaborate the system for efficient adventitious organogenesis and transgenic plant regeneration in Rubus fruticosus L. using explants from mature plants. Regeneration of putative transgenic shoots took place from flag explants cultivated vertically on MS medium with 1 mg l-1 TDZ and 0.02 mg l-1 IBA followed by transfer on MS medium with 1 mg l-1 BAP, 0.02 mg l-1 IBA and 0.1 mg l-1 GA3 supplemented with 10-15 mg l-1 hygromycin after transformation by A. tumefaciens strain LBA 4404 carrying plasmid pCambia 1304. Four putative transgenic plants of cv. 'Čačanska Bestrna' were rooted and acclimatized. [Projekat Ministarstva nauke Republike Srbije, br. TR-31064]

The aim of work was to elaborate the system for Agrobacterium tumefaciens-mediated transformation and transgenic plant regeneration in Rubus fruticosus L. using explants from mature plants. Regeneration of transgenic shoots was achieved from cut ends of petioles completely immersed in the medium using flag explants cultivated vertically on MS medium with 1 mg l−1 TDZ and 0.02 mg l−1 IBA followed by transfer on medium with 1 mg l−1 BAP, 0.02 mg l−1 IBA and 0.1 mg l−1 GA3, both supplemented with 10 and 15 mg l−1 hygromycin after transformation by A. tumefaciens strain LBA 4404/pCambia 1304. Four putative transgenic plants of cv. ‘Cacanska Bestrna’ were rooted, acclimatized and analysed. Transgenic character of the tissue from analysed plants was confirmed by nested PCR and subsequent sequencing.

Methodological Approaches to Simple Enzyme Polymorphism Analyses of Amaranth Species (AmaranthusSP.)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 inAmaranthusspecies led us to test some methodological approaches to enzyme polymorphism analysis. For these experiments genotypes ofAmaranthus cruentusL. (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).

V procese indukcie a multiplikacie výhonkov in vitro zohrava, okrem terminu odberu explantatu, doležitu ulohu zloženie zakladneho kultivacneho media a pritomnosť rastových regulatorov. Specificke naroky roznych druhov drevin na pritomnosť rastových regulatorov v kultivacnom mediu vyžaduju pred etablovanim technologickeho postupu siroke spektrum výskumných prac. Experimentalne prace sa preto zameriavali na vypracovanie optimalneho protokolu pre in vitro multiplikaciu výhonkov Acer davidii ssp. grosseri., dosiaľ komercne nereprodukovaneho javora v podmienkach in vitro. Letorasty sa odoberali v intenzivnej faze rastu v polovici jula. Odber explantatov sa realizoval zo stvorrocných rastlin, ktore boli pestovane v kontajneroch na pokusnej ploche. Ako explantaty sa použili jedno nodalne segmenty, Kultivacia sa realizovala v definovaných podmienkach pri teplote 21 °C a intenzite žiarenia 40 μmol . m-2 . s-1. Pokus sa realizoval v rozsahu patnastich opakovani pre každý variant pri zavedeni explantatu do kultury. V ramci jedneho opakovania (jednej kultivacnej nadoby) sa použili tri explantaty. Ako zakladne kultivacne medium bolo použite MS medium. Použili sa styri varianty kultivacneho media, ktore sa odlisovali roznymi koncentraciami a kombinaciami cytokininov. Reakcia explantatov na kultivacne medium, iniciacia a multiplikacia výhonkov, sa zaznamenala po siestich a dvanastich týždňoch od založenia pokusu. Výsledky sa podrobili matematicko-statistickemu vyhodnoteniu, na ktore sa použila analýza rozptylu a LSD test statistickej významnosti rozdielov v priemerných hodnotach (Statgraphic software). Explantaty Acer davidii ssp. grosseri reagovali odlisne na rozne typy cytokininov. Explantaty kultivovane na MS mediu obohatenom o 2,6 μM BAP vykazovali najnižsiu multiplikacnu schopnosť. Reakcie explantatov na ďalsich kultivacných mediach sa statisticky navzajom neodlisovali. Priemerna multiplikacia výhonkov na mediu MS + 0,5 μM TDZ bola na urovni 1,2 výhonku. Statisticky nevýznamne zvýsenie multiplikacnej schopnosti (1,49 výhonkov na explantat) bolo dosiahnute po aplikacii MS + 2,6 μM BAP + 0,5 μM TDZ. Mutiplikacia výhonkov na mediach so samotným TDZ a na mediach s kombinaciou BAP + TDZ sa statisticky neodlisovala od multiplikacie zaznamenanej na zakladnom MS mediu, kde priemerna multiplikacna schopnosť dosahovala 1,35 výhonku na explantat.Napriek pozitivnemu vplyvu spolocnej aplikacie TDZ a BAP v kultivacnom mediu, zistene rozdiely v pocte výhonkov na explanat ako aj v pocte listov na výhonok neboli v nasich experimentoch statisticky preukazne v porovnani so zakladným MS mediom.

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., Cieľom výskumu bolo vyvinúť protokol pre mikropropagáciu vybraných kultivarov Rubus – ‘Tulameen’ a ‘Black Jewel’ (Rubus idaeus L.), ‘Black Satin’ (Rubus fruticosus L.) a ‘Tayberry’ (Rubus fruticosus x Rubus idaeus). Pre založenie in vitro kultúr boli použité nodálne segmenty nesúce dormantné púčiky. Iniciácia rastu výhonkov prebiehala na MS médiu s obsahom 8 g*L-1 fytoagaru, 30 g*L-1 sacharózy, 1 mg*L-1 6-benzylaminopurín (BAP), 0.2 mg*L-1 kyselina indolyl-3-maslová (IBA) a 150 mg*L-1 cefotaxímu. Schopnosť proliferácie výhonkov jednotlivých kultivarov bola hodnotená pomocou koeficientu multiplikácie (počet výhonkov vytvorených na explantát) počas 4 subkultivácií. Štatistické analýzy potvrdili významne vyššiu schopnosť multiplikácie výhonkov pri kultivaroch ‘Black Satin’ a ‘Tulameen’ v porovnaní s ‘Tayberry’ a ‘Black Jewel’, ako aj významné rozdiely v proliferácii výhonkov medzi subkultiváciami (1. a 2. versus 3. a 4.). Testovanie vplyvu rôznych koncentrácií BAP na tvorbu výhonkov ukázalo, že BAP 1-2 mg*L-1 je najlepší pre proliferáciu výhonkov s vyššou hodnotou multiplikácie pri kultivare ‘Black Satin’ v porovnaní s ‘Black Jewel’. Štatistické hodnotenie ukázalo, že genotyp a koncentrácia BAP štatisticky významne ovplyvnili schopnosť proliferácie výhonkov pri kultivaroch druhov Rubus.

L. cv. Koralle and Red Pearl. Shoot prolifera-tion using in-vitro-derived single-node segments was markedly influenced by the cultivar and medium pH. Shootproliferation was significantly higher in cv. Koralle (6.92 shoots/explant) than in cv. Red Pearl (5.61shoots/explant) at different culture medium pH. The study confirmed the importance of properly adjusted initialculture medium pH for effective shoot proliferation. For cv. Koralle, medium pH 5.5 was the most favorable forshoot proliferation. For cv. Red Pearl, the number of shoots formed was highest at pH 4.