Gibberellin-cytokinin interaction in the correlation between the cotyledon and its axillary bud in pea (Pisum sativum L.)

If one cotyledon is removed from decapitated pea seedlings and the remaining one is treated with the cytokinin BA, then a complete correlation reversal occurs in numerous cases: instead of the bud belonging to the removed cotyledon, the bud belonging to the remaining cotyledon starts to grow. However, if GA is applied to the remaining cotyledon together with BA, then the number of these correlation reversals sharply drops. This in respect to cytokinin morphogenetically (correlatively) contradictory effect could play a significant role in apical dominance in plants.     

Estimation of substances with the cytokinin activity in studies on the correlation between the cotyledon and its axillary bud in pea (Pisum sativum L.)

The content of substances showing the cytokinin activity was estimated on decapitated and one-cotyledon-deprived pea plants during that period when the promoting effect of the cotyledon excision had not yet been manifested. Results of the bioassay showed that after the excision an increase in the level of substances with cytokinin activity occurred only in cotylars growing in axillas of these excised cotyledons. These results coincide with earlier data about the content of gibberellins and auxins in the same object.     

High frequency plant regeneration from cotyledon and hypocotyl explants of ornamental kale

A high frequency shoot regeneration system for ornamental kale [Brassica oleracea L. var. acephala (D.C.) Alef.] was firstly established from seedling cotyledon and hypocotyl explants. The ability of cotyledon and hypocotyl to produce adventitious shoots varied depending upon genotype, seedling age and culture medium. The maximum shoot regeneration frequency was obtained when the explants from cv. Nagoya 4-d-old seedlings were cultured on Murashige and Skoog (MS) medium supplemented with 3 mg dm⁻³ 6-benzylaminopurine (BA) and 0.1 mg dm⁻³ naphthaleneacetic acid (NAA). The frequency of shoot regeneration was 65.0 % for cotyledons, 76.1 % for hypocotyls; and the number of shoots per explant was 4.3 for cotyledons, 8.2 for hypocotyls. Hypocotyl explants were found to be more responsive for regeneration when compared with cotyledons. Among the 4 cultivars tested, Nagoya showed the best shoot regeneration response. The addition of 3.0 mg dm⁻³ AgNO₃ was beneficial to shoot regeneration. Roots were formed on the base of the shoots when cultured on half-strength MS medium.     

In vitro plant regeneration from cotyledon and hypocotyl segments in two bell pepper cultivars

In vitro plant regeneration has been obtained from Capsicum annuum cvs. Pico and Piquillo. Shootbuds were induced from hypocotyl and cotyledon segments after 15–20 days of culture on MS basal medium supplemented with IAA and BAP or Zeatin. Shoot-buds grew into rosettes that rooted in MS plus NAA (0.1 mg/l) and IBA (0.05 mg/l) after 15 days. The small plantlets were successfully transferred to pots with a mixture of peat and perlite and maintained under greenhouse conditions. Elongation took place when the plantlets were growing in the greenhouse.Abbreviations BAP 6-benzylaminopurine - 2,4-D 2,4-dichlorophenoxyacetic acid - IAA indole-3-acetic acid - IBA indole-3-butyric acid - Kn Kinetin - MS Murashige and Skoog (1962) - NAA alpha-naphthalenacetic acid - Z Zeatin     

On the role of exogenous gibberellin and cytokinin in the correlation between the leaf and its axillary bud in hortensia (Hydrangea opuloides C. KOCH)

The role of exogenously supplied gibberellin (GA₃) and cytokinin (benzyladenin -BA) in the correlation between the mature leaf and its axillary bud was investigated in one-node segments ofHydrangea. When both leaves were left on the segments, then both GA and BA were able to determine the dominance between axillary buds, that means that the bud treated with the corresponding growth regulator grew more vigorously. When one of the leaves was removed, the bud belonging to the removed leaf grew more vigorously, but GA applied onto the axillary bud belonging to the remaining leaf caused a complete correlation reversal: the bud belonging to the remaining leaf grew more vigorously. On the contrary, the application of BA onto the bud of the remaining leaf resulted in only insignificantly stimulated growth of the bud belonging to this leaf.     

The role of gibberellins and cytokinins in the growth correlative effect of cotyledons in flax and pea

It is wellknown that following the amputation, or darkening of one cotyledon in decapitated flax seedlings, the opposite remaining, or illuminated, cotyledon exerts a stimulatory effect on the growth of its axillary bud. For the induction of this stimulating effect a 21–72 h continuous darkening of the cotyledon is sufficient. Endogenous gibberellins take part in the stimulation effect of the illuminated cotyledon, since their level in the illuminated cotyledon increases as early as 12–48 h following the darkening of the opposite cotyledon. The apical part of the cotyledon has a higher growth stimulatory effect on the growth of the cotyledonary axillary bud than the basal half. This again is associated with endogenous gibberellins the level of which is higher in the apical half of the cotyledon than in the basal one. Upon removal of the root and hypocotyl base in decapitated flax seedlings deprived of one cotyledon, the remaining cotyledon loses its stimulatory influence, so that the bud of the amputated cotyledon grows more vigorously (Dostál 1955). In this growth correlative phenomenon the root may be substituted by cytokinin BA applied in the form of a 0.1–1.0 per cent paste onto the remaining cotyledon, for again in this case the bud of the preserved cotyledon grows more vigorously. Following the decapitation of the axillary of the amputated cotyledon in decapitated pea seedlings with an intact root and deprived of one cotyledon, the axillary of the remaining cotyledon grows more intensively than the serial of the removed one. If the plants operated on in the same way are deprived of the root, the serial of the removed cotyledon gains a correlative growth predominance. If the plants deprived of root are cultivated at the same time in a solution of BA (10–20 mg 1⁻¹), the correlative predominance is acquired by the axillary of the remaining cotyledon. In growth correlations between cotyledons and their axillary buds in pea seedlings the root may thus be substituted by exogenous cytokinin, as well.     

In vitro plantlet regeneration from cotyledon, hypocotyl and root explants of hybrid seed geranium

In vitro plantlet regeneration systems for the seed geranium (Pelargonium x hortorum Bailey) using cotyledon, hypocotyl and root explants were optimized by studying the influence of seedling age, growth regulators and excision orientation on organogenesis. Indole-3-acetic acid combined with zeatin yielded the highest rate of shoot production on cotyledon explants (0.2–2 shoots per explant). More shoots were produced on explants cut from the most basal region of cotyledons from 2 to 4-day-old seedlings than from older seedlings or more distal cut sites. Hypocotyl explants produced the highest number of shoots, up to 40 shoots per explant, on indole-3-acetic acid (2.8–5.6 mM) + zeatin (4.6 mM) or thidiazuron (4.5 mM). Maximum shoot formation (0.3–1.4 shoots per explant) on root explants occurred when they were cultured on medium containing zeatin. Regenerated shoots rooted best on a basal medium containing no growth regulators. There were substantial differences among cultivars in shoot formation from each of the explant systems.Abbreviations BA 6-benzylaminopurine - 2,4-d 2,4-dichlorophenoxyacetic acid - IAA indole-3-acetic acid - NAA naphthaleneacetic acid - TDZ thidiazuron     

Postharvest changes in endogenous cytokinins and cytokinin efficacy in potato tubers in relation to bud endodormancy

Using an indirect enzyme‐linked immunosorbent assay (ELISA), the effects of postharvest storage duration and temperature on endogenous cytokinins in potato ( Solanum tuberosum L. cv. Russet Burbank) tuber apical bud tissues in relation to endodormancy status were determined. Following fractionation by HPLC, a total of eight cytokinins were detected and these were: zeatin riboside‐5'‐monophosphate (ZRMP), zeatin‐ O ‐glucoside (ZOG), zeatin (Z), zeatin riboside (ZR), isopentenyl adenosine‐5'‐monophosphate (IPMP), isopentenyl adenine‐9‐glucoside (IP‐9‐G), isopentenyl adenine (IP) and isopentenyl adenosine (IPA). Regardless of postharvest storage temperature or endodormancy status, IP‐9‐G was the most abundant cytokinin detected while ZRMP and ZOG were the least abundant ones. In tubers preincubated at a growth‐permissive temperature (20°C) prior to extraction, the loss of endodormancy was preceded by significant increases in the endogenous levels of Z, ZR, IPMP and IP‐9‐G. When stored continuously at a growth‐inhibiting temperature (3°C), significant increases in ZR, IP‐9‐G and IP + IPA were observed. The total content of cytokinins increased by over 7‐fold during postharvest storage and this increase was a result of de novo biosynthesis. Dose‐response studies using IPA and ZR demonstrated a time‐dependent increase in apparent cytokinin sensitivity during postharvest storage. With the exception of IP‐9‐G, injection of any of these endogenous cytokinins resulted in the rapid and complete termination of tuber endodormancy. The significance of these results with respect to endodormancy regulation and the possible mechanisms controlling cytokinin levels in potato tubers are discussed.     

Plantlet regeneration from cotyledon,cotyledon petiole,and hypocotyl explants via somatic embryogenesis pathway in roquette (Eruca sativa Mill)

Abstract

A high-efficiency plant regeneration protocol based on somatic embryo formation for Huining Roquette, an interesting ecotype of Eruca sativa Mill, was established for future transgenic applications. On Murashige and Skoog (MS) medium containing 2,4-dichlorophenoxyacetic acid (2,4-D), alone or in combination with 6-benzylaminopurine (BA) or kinetin (KT), the cotyledon explants, cotyledon petioles, and hypocotyls all produced embryogenic callus (ECs) or somatic embryos (SEs) to different extents. After transferring onto hormone-free MS medium, the ECs or SEs from the different explants and media, all of them developed shoots with a frequency of 6–48%, and then produced roots with a frequency of 2–29%. As regards the probability of shoot differentiation, cotyledon explants appeared similar to hypocotyls, but superior to cotyledon petioles; 2,4-D + KT worked more effectively than 2,4-D alone and 2,4-D + BA for callus induction and shoot differentiation. The optimal hormone combinations for plant regeneration of cotyledon, cotyledon petiole, and hypocotyl explants were 1.0 mg/l 2,4-D + 0.1 mg/l KT, 0.8 mg/l 2,4-D + 0.3 mg/l BA, and 1.0 mg/l 2,4-D + 0.3 mg/l KT, respectively. MS medium with 60–80 g/l sucrose was the most effective for improving SE maturation and germination.     

Callus induction and high-frequency plant regeneration from hypocotyl and cotyledon explants of Arctium lappa L.

Summary This study describes a protocol for plant regeneration from cultured seedling explants of Arctium lappa. Hypocotyls and cotyledons of A. lappa were induced to form callus by culturing on Murashige and Skoog (MS) medium supplemented with 2.0mg l^−1 2,4-dichlorophenoxyacetic acid and 0.5–2.0 mg l^−1 benzyladenine (BA). Formation of adventitious buds could be induced from calluses or explants directly by culturing on MS medium containing 1.0–2.0 mg l^−1 α-naphthaleneacetic acid (NAA) and 0.5–2.0 mg l^−1 BA. These regenerated shoots were rooted on MS medium with 1.0 mg l^−1 indole-3-butyric acid or indole-3-acetic acid in combination with 1.0 mgl^−1 NAA. The regenerated plants acclimatized in soil were normal morphologically and in growth characters. They flowered and set seeds in the following year after acclimatization.     

In vitro regeneration of sesame (Sesamum indicum L.) from seedling cotyledon and hypocotyl explants

The goal of this study was to develop an efficient regeneration protocol to be used for genetic transformation of sesame. Published regeneration methods using benzyladenine (BA) and 1-naphthalene acetic acid (NAA) were unsuccessful for the cultivars used herein. Experiments were carried out using cotyledon and hypocotyl explants from the cultivar Mtwara-2. Later the optimised culture conditions were used to investigate the regeneration response of different genotypes. There was significant interaction between hormone treatments and macronutrients for shoot and root regeneration. Results also showed that shoot regeneration was significantly influenced by explant type. Shoots were only obtained from cotyledons whereas both cotyledons and hypocotyls could produce roots. Modified Murashige and Skoog (MS) medium with N6 macronutrients resulted in twice the shoot regeneration frequency obtained with ½MS macronutrients in the presence of thidiazuron (TDZ). The shoot regeneration frequency was significantly reduced when BA was used in place of TDZ. On shoot regeneration medium containing BA and NAA, only roots were formed. Replacing NAA with indole-3-acetic acid (IAA) greatly improved the regeneration of shoots. The optimum growth regulator combination for shoot regeneration was 20 μM TDZ together with 2.5 μM IAA, which gave a frequency of 63% and 4.4 shoots per regenerating explant for the best cultivar Ex-El. Genotypic differences were significant both for the number of explants regenerating shoots and the number of shoots produced per regenerating explant.     

Shoot regeneration from cotyledon explants of fibre flax (Linum usitatissimum) and their subsequent rooting

In this study, the morphogenic ability of cotyledon explants of fibre flax (Linum usitatissimum L.) was investigated in relation to their physiological age and genotype and the effect of growth regulators in the medium. Among four investigated genotypes, the explants of cvs. A-29 and Alexim manifested the highest and the lowest morphogenic ability, respectively. The optimum ratio of growth regulators for shoot regeneration was shown to depend on the age of donor plantlets from which the explants were obtained. For explants excised from younger plantlets, the lower concentration of benzyladenine (BA) and the absence of auxin were preferable. For explants from older plantlets, the higher BA concentration and NAA presence in the medium were required for shoot regeneration. The addition of ABA to the regeneration medium usually reduced shoot regeneration frequency; however, the inhibitory effect of ABA depended on other growth regulators.     

Protoplasts from cotyledon and hypocotyl of sunflower (Helianthus annuus L.): shoot regeneration and seed production

Summary Cotyledon and hypocotyl protoplasts of Helianthus annuus inbred line 47 302 bcd were embedded in alginate and plated on L4 medium (Lenée and Chupeau 1986). After one month, the calli were transferred on MSSH regeneration medium (Murashige and Skoog 1962; Schenk and Hildebrandt 1972) where they regenerated shoots (overall efficiency 10^–2%). The shoots were elongated on B5 (Gamborg et al. 1968) medium first without hormones, then supplemented with GA3 and BAP (both 0.05 mg/l). In order to overcome the difficulty to induce rooting by classical methods, the elongated shoots were grafted on a sunflower rootstock. The grafted shoots produced flowers and seeds. Different factors have been shown to have an important influence on the capacity to regenerate shoots: the genotype, the physical culture conditions at the callus regeneration step (e.g. protoplasts embedded in alginate), and the media composition.Abbreviations BAP 6-benzylaminopurine - GA3 gibberellic acid - IBA indole-3-butanoic acid - IAA indole acetic acid - MES 2-N-morpholinoethane sulfonic acid - NAA 1-naphthalene acetic acid - 2,4D 2,4 dichlorophenoxyacetic acid     

Role of endogenous cytokinins and ethylene in adventitious shoot regeneration in lemon (Citrus limon)

In Vitro Cellular & Developmental Biology - Plant - In this study, adventitious organogenesis from mature tissues of ‘Verna 51’ and ‘Fino 49’ lemon cultivars has been...     

Woodland strawberry axillary bud fate is dictated by a crosstalk of environmental and endogenous factors

Plant architecture is defined by fates and positions of meristematic tissues and has direct consequences on yield potential and environmental adaptation of the plant. In strawberries (Fragaria vesca L. and F. × ananassa Duch.), shoot apical meristems can remain vegetative or differentiate into a terminal inflorescence meristem. Strawberry axillary buds (AXBs) are located in leaf axils and can either remain dormant or follow one of the two possible developmental fates. AXBs can either develop into stolons needed for clonal reproduction or into branch crowns (BCs) that can bear their own terminal inflorescences under favorable conditions. Although AXB fate has direct consequences on yield potential and vegetative propagation of strawberries, the regulation of AXB fate has so far remained obscure. We subjected a number of woodland strawberry (F. vesca L.) natural accessions and transgenic genotypes to different environmental conditions and growth regulator treatments to demonstrate that strawberry AXB fate is regulated either by environmental or endogenous factors, depending on the AXB position on the plant. We confirm that the F. vesca GIBBERELLIN20-oxidase4 (FvGA20ox4) gene is indispensable for stolon development and under tight environmental regulation. Moreover, our data show that apical dominance inhibits the outgrowth of the youngest AXB as BCs, although the effect of apical dominance can be overrun by the activity of FvGA20ox4. Finally, we demonstrate that the FvGA20ox4 is photoperiodically regulated via FvSOC1 (F. vesca SUPPRESSOR OF OVEREXPRESSION OF CONSTANS1) at 18°C, but at higher temperature of 22°C an unidentified FvSOC1-independent pathway promotes stolon development.

Environmental conditions and apical dominance dictate woodland strawberry plant architecture by regulating axillary bud fate.     

The synthesis, transport and metabolism of endogenous cytokinins

Abstract Present evidence indicates that only the root systems of plants have been shown conclusively to synthesize cytokinins. Although most of these compounds are apparently exported to the shoot via the xylem, there are indications that more attention should be given to the possibility of translocation through the phloem. Within mature leaves the cytokinins derived from the roots are converted to inactive or storage forms by means of glucosylation. While it would appear that glucosylation could occur in all living plant cells whenever the cytokinins are no longer required for active growth, and could provide the plant with a potential reservoir of cytokinins, very little is known with regard to the transport and reutilisation of these compounds.     

The hormonal regulation of axillary bud growth in Arabidopsis

Apically derived auxin has long been known to inhibit lateral bud growth, but since it appears not to enter the bud, it has been proposed that its inhibitory effect is mediated by a second messenger. Candidates include the plant hormones ethylene, cytokinin and abscisic acid. We have developed a new assay to study this phenomenon using the model plant Arabidopsis. The assay allows study of the effects of both apical and basal hormone applications on the growth of buds on excised nodal sections. We have shown that apical auxin can inhibit the growth of small buds, but larger buds were found to have lost competence to respond. We have used the assay with nodes from wild-type and hormone-signalling mutants to test the role of ethylene, cytokinin and abscisic acid in bud inhibition by apical auxin. Our data eliminate ethylene as a second messenger for auxin-mediated bud inhibition. Similarly, abscisic acid signalling is not to be required for auxin action, although basally applied abscisic can enhance inhibition by apical auxin and apically applied abscisic acid can reduce it. By contrast, basally applied cytokinin was found to release lateral buds from inhibition by apical auxin, while apically applied cytokinin dramatically increased the duration of inhibition. These results are consistent with cytokinin acting independently to regulate bud growth, rather than as a second messenger for auxin. However, in the absence of cytokinin-signalling mutants, a role for cytokinin as a second messenger for auxin cannot be ruled out.     

Different endogenous cytokinins between male and female Mercurialis annua L.

The endogenous pool of cytokinin metabolites during sexual differentiation of Mercurialis annua L. was studied with a computerized gas-chromatography-mass spectrometry system. Certain metabolites were common to both sexes: ribosides (isopentenyl-adenosine, ribosylzeatin) and the nucleotide of I₆-Ade. Zeatin could be detected only in females while its nucleotide was present in males. The results were obtained with differentiating apices and whole plants. The high Z concentration and the low level of its nucleotide are related to the absence of two dominant complementary genes, determining maleness. Study of the regulation of cytokinin metabolism now seems possible.Abbreviations IPA isopentenyl adenosine - I₆-Ade isopentenyl adenine - Z zeatin - RZ ribosylzeatin