Effect of different sucrose and nitrogen salt levels in the medium and temperature on in vitro propagation of Helleborus niger L.

Helleborus niger L. is a rhizomatous, herbaceous perennial with overwintering, divided, basal leaves. The objective of the study was to investigate the influence of different levels of sucrose (10, 20, 30, 40, 50, 60, 70, and 80 g l−1) and nitrogen salts (25%, 50%, and 100% according to MS medium) as well as temperature (15°C, 20°C) on in vitro multiplication and rooting and ex vitro acclimatization of H. niger. The growth and multiplication of axillary shoots were performed on modified MS medium supplemented with various growth regulators (2iP, BAP and kinetin – each at a concentration of 1.0 mg l−1, GA3 2.5 mg l−1). For the induction of roots, the medium was supplemented with IBA 1 mg l−1 and NAA 0.1 mg l−1. Rooted plants were transplanted in a peat–perlite substrate (4:1) in a heated greenhouse for ex vitro acclimatization. The multiplication rate of H. niger shoots, in vitro rooting, and ex vitro acclimatization were strongly dependent on the sucrose/nitrogen salt relationship in the medium. The highest multiplication rate of axillary shoots (3.7) was found at a temperature of 15°C or 20°C, on the medium with cytokinins and GA3 supplemented with sucrose 20–30 g l−1 and nitrogen salts at 50%. Sucrose at a concentration of 50 g l−1 strongly stimulated the number of roots per microplant (5.8–6.0) on the media with a reduced level of nitrogen salts (25% and 50%) when the temperatures were 20°C and 15°C, respectively. The plants rooted on the media with a high sucrose/nitrogen salt ratio showed acclimatization rates which ranged from 82% to 100%. Morphological observation of plantlets revealed obvious differences in leaf shape and size and the architecture of the root system as well as differences in the developmental stages of shoots grown on media with different sucrose and nitrogen salt concentrations

Effect of different sucrose and nitrogen salt levels in the medium and temperature on in vitro propagation of Helleborus niger L.

Helleborus niger L. is a rhizomatous, herbaceous perennial with overwintering, divided, basal leaves. The objective of the study was to investigate the influence of different levels of sucrose (10, 20, 30, 40, 50, 60, 70, and 80 g l−1) and nitrogen salts (25%, 50%, and 100% according to MS medium) as well as temperature (15°C, 20°C) on in vitro multiplication and rooting and ex vitro acclimatization of H. niger. The growth and multiplication of axillary shoots were performed on modified MS medium supplemented with various growth regulators (2iP, BAP and kinetin – each at a concentration of 1.0 mg l−1, GA3 2.5 mg l−1). For the induction of roots, the medium was supplemented with IBA 1 mg l−1 and NAA 0.1 mg l−1. Rooted plants were transplanted in a peat–perlite substrate (4:1) in a heated greenhouse for ex vitro acclimatization. The multiplication rate of H. niger shoots, in vitro rooting, and ex vitro acclimatization were strongly dependent on the sucrose/nitrogen salt relationship in the medium. The highest multiplication rate of axillary shoots (3.7) was found at a temperature of 15°C or 20°C, on the medium with cytokinins and GA3 supplemented with sucrose 20–30 g l−1 and nitrogen salts at 50%. Sucrose at a concentration of 50 g l−1 strongly stimulated the number of roots per microplant (5.8–6.0) on the media with a reduced level of nitrogen salts (25% and 50%) when the temperatures were 20°C and 15°C, respectively. The plants rooted on the media with a high sucrose/nitrogen salt ratio showed acclimatization rates which ranged from 82% to 100%. Morphological observation of plantlets revealed obvious differences in leaf shape and size and the architecture of the root system as well as differences in the developmental stages of shoots grown on media with different sucrose and nitrogen salt concentrations

Differential effects of N-1-naphthylphthalamic acid (NPA) and 2,3,5-triiodobenzoic acid (TIBA) on auxin control of swelling of the shoots of Bryophyllum calycinum Salisb.

The effects of N-1-naphthylphthalamic acid (NPA) and 2,3,5-triiodobenzoic acid (TIBA) on the swelling of the stem in intact and decapitated plants of Bryophyllum calycinum in relation to the interaction with auxin, indole-3-acetic acid (IAA), are described. NPA induced conspicuous local internode swelling only in the area of its application in intact plants and in the decapitated internode in the case of simultaneous application of IAA on the top of the internode. By contrast, TIBA applied to an internode of intact plants induced swelling along the entire internode above the treatment area, and similar results were obtained in the decapitated internode when TIBA was applied in the middle of the internode and IAA was applied onto the top of the internode. The differential effect of NPA and TIBA on stem swelling in B. calycinum is discussed in relation to their differential mode of action on auxin transport

Plant regeneration of Alstroemeria in vitro

The regenerative ability of explants from various organs of Alstroemeria plants was investigated. Rhizome apical and axillary tips cultured on the Murashige and Skoog medium with BA - 2. mgl-1 and NAA - 0,5 mgl-1 were the best among the tissue tested as initial explants. Five weeks after isolation the rhizome with 1-4 upright growing shoots were obtained. The types of rhizome explants influenced development and growth of lateral rhizomes and upright growing shoots. There were no significant differences in number of roots formed on various kind of rhizome explants. Rooting was strongly influenced by NAA. Subapical segments of vegetative stem, segments of flower pedicels and parts of ovary did not regenerate rhizome or roots but occasionally callus was formed on the medium with kinetin - 2 mgl-1 and NAA - 2 mgl-1. Segments excised from vegetative stem sporadically developed roots on the medium with NAA or IBA in concentrations 3 and 9 mgl-1

Effect of growth retardants, cytokinins and auxins on the multiplication and rooting in vitro of Alstroemeria × hybrida "Juanita"

Rhizome cultures of “Jiianita” Polish cultivar of Alstroemeria × hybrida were used to enhance an effectiveness of micropropagation method of new cultivars and selections. The effect of cytokinins (BAP, kinetin and 2iP), auxins (IAA, IBA and NAA), growth retardants (paclobutrazol and flurprimidol alone or in combination were studied in relation to rhizome branching. aerial shoot production and rooting of rhizome. The greatest number of aerial shoots as well as the shortest shoots were observed at the highest BAP concentration (6 mg l-1). However, the rhizonies had the poorest rooting ability. BAP at low concentrations combined with kinetin or 2iP also strongly stimulated aerial shoot formation and rhizome branching. Unfortunately. those shoots were of poor qualily. Application of BAP at low concentration with paclobutrazol (0,1–0,5 mg l-1) or flurprimidol (0,01–1 mg l-1) in presence of 1 mg l-1 NAA resulted in high number of aerial shoots (5–6), reduction of their length and higher rooting ability of the rhizomes. Growth retardants applied with NAA strongly stimulated formation of the roots but suppressed their elongation

Hormonal regulation of the growth of leaves and inflorescence stalk in Muscari armeniacum Leichtl.

It is known that chilling of Muscari bulbs is necessary for the growth of the inflorescence stalk and flowering, but not for the growth of leaves. Gibberellic acid (GA) accelerated stem growth and flowering in chilled Muscari bulbs. In the present experiment it was shown that in unchilled derooted Muscari bulbs the growth of leaves, but not the growth of the inflorescence stalk, was observed when bulbs were stored in water, GA at a concentration of 50 and 100 mg/L, benzyladenine (BA) at a concentration of 25 and 50 mg/L, or a mixture of GA+BA (50+25 mg/L), but abscisic acid (ABA) at a concentration of 10 mg/L greatly inhibited the growth of leaves. In chilled derooted Muscari bulbs the growth of leaves and inflorescence stalk was observed when bulbs were stored in water or GA, but BA and GA+BA treatments totally inhibited the growth of the inflorescence stalk without an effect on the growth of leaves. These results clearly showed that the growth of leaves and inflorescence stalk in Muscari bulbs are controlled by plant growth regulators in different ways. ABA totally inhibited the growth of leaves and inflorescence stalk in chilled derooted Muscari bulbs. It was shown that after the excision of the inflorescence bud in cultivated chilled Muscari bulbs, the inflorescence stalk died, but application of indole-3-acetic acid (IAA) 0.5% in the place of the removed inflorescence bud induced the growth of the inflorescence stalk. IAA applied under the inflorescence bud inhibited the development of flowers (flower-bud blasting) and induced the growth of the inflorescence stalk below the treatment site. These results are discussed with reference to hormonal regulation of stem (stalk) growth in tulip, narcissus, hyacinth, and Hippeastrum

Auxin effectively induces the formation of the secondary abscission zone in Bryophyllum calycinum Salisb. (Crassulaceae)

We have found that auxin, indole-3-acetic acid (IAA) substantially induces the formation of the secondary abscission zone in stem and petiole explants and in decapitated stem and petiole after excision of blade in intact plants of Bryophyllum calycinum when IAA at a concentration of 0.1% as lanolin paste was applied in the middle of these organs. The secondary abscission zone was formed at a few mm above of the treatment with IAA, and senescence of the part above abscission zone was observed. IAA additionally applied on the top of explants or top of the dacapitated stem or the debladed petiole totally prevented the secondary abscission zone formation and senescence induced by IAA applied in the middle of these organs. Possible mechanisms of the formation of the secondary abscission zone are discussed in terms of the interaction of auxin and ethylene

Hormonal regulation of the growth of leaves and inflorescence stalk in Muscari armeniacum Leichtl.

It is known that chilling of Muscari bulbs is necessary for the growth of the inflorescence stalk and flowering, but not for the growth of leaves. Gibberellic acid (GA) accelerated stem growth and flowering in chilled Muscari bulbs. In the present experiment it was shown that in unchilled derooted Muscari bulbs the growth of leaves, but not the growth of the inflorescence stalk, was observed when bulbs were stored in water, GA at a concentration of 50 and 100 mg/L, benzyladenine (BA) at a concentration of 25 and 50 mg/L, or a mixture of GA+BA (50+25 mg/L), but abscisic acid (ABA) at a concentration of 10 mg/L greatly inhibited the growth of leaves. In chilled derooted Muscari bulbs the growth of leaves and inflorescence stalk was observed when bulbs were stored in water or GA, but BA and GA+BA treatments totally inhibited the growth of the inflorescence stalk without an effect on the growth of leaves. These results clearly showed that the growth of leaves and inflorescence stalk in Muscari bulbs are controlled by plant growth regulators in different ways. ABA totally inhibited the growth of leaves and inflorescence stalk in chilled derooted Muscari bulbs. It was shown that after the excision of the inflorescence bud in cultivated chilled Muscari bulbs, the inflorescence stalk died, but application of indole-3-acetic acid (IAA) 0.5% in the place of the removed inflorescence bud induced the growth of the inflorescence stalk. IAA applied under the inflorescence bud inhibited the development of flowers (flower-bud blasting) and induced the growth of the inflorescence stalk below the treatment site. These results are discussed with reference to hormonal regulation of stem (stalk) growth in tulip, narcissus, hyacinth, and Hippeastrum

Effect of benzyladenine (BA) on auxin-induced stem elongation and thickening in tulip (Tulipa gesneriana L.)

It is well known that stem elongation in tulip is induced by the auxin produced in the leaves and gynoecium. Excision of the flower bud and all the leaves in an early stage of tulip growth resulted in almost total inhibition of stem growth, but the inhibition was completely recovered by the exogenous application of auxin to the place from which the flower bud had been removed. Hormonal control of stem thickening in tulip is much less known. Additional application of benzyladenine (BA) to the tulip stem by soaking a cotton wick wrapped around all the internodes only slightly inhibited stem growth induced by IAA at a concentration of 0.1 and 2.0%, but substantially stimulated the thickening of all the internodes. The treatment of the tulip stem with benzyladenine enabled direct contact of the cytokinin with the epidermis, which is an important factor in stem elongation. The experiment conducted in field conditions also showed that BA only slightly inhibited the elongation of the fourth internode induced by IAA, but stimulated the thickening of that internode. IAA applied at a concentration of 2.0% stimulated ethylene production to a much higher extent than IAA at a concentration of 0.1%, and BA affected the auxin-induced ethylene production only to a small extent. Metabolic significance of these findings is discussed