GA4+7 plus benzyladenine reduce foliar chlorosis of Lilium longiflorum

The effectiveness of two commercial formulations of gibberellin (GA) and benzyladenine (BA) for reducing foliar chlorosis on Easter lily (Lilium longiflorum Thunb.) was compared. On a per liter basis, plants were sprayed with 0, 100, 200, or 400 mg (BA equivalent) of Accel (GA₄₊₇:BA of 1:10) or Promalin (GA₄₊₇:BA of 1:1) when the crop leaf area index (LAI)=3. One group of plants was sprayed with 100 mg of Accel or Promalin (BA equivalent) per liter twice: once at LAI=3 and again 3 weeks later. Plants were harvested when the largest flower bud on each plant measured 13 cm in length, stored for 0 or 3 weeks at 2.5°C in the dark, and then moved into a post-harvest evaluation room at 21°C, where foliar chlorosis was monitored for 3 weeks. Senescence of some lower leaves on plants in every treatment was evident at harvest, and incidence of senescence increased during the 21 days of post-harvest evaluation. Cold storage increased the number of leaves senescing during the subsequent evaluation period. Application of Promalin or Accel significantly reduced leaf senescence compared to that of untreated plants. At harvest, 21% of the leaves on untreated plants were senescent, while plants treated with Promalin or Accel averaged 3 or 9% senescent leaves, respectively. Following 7 days of post-harvest evaluation, Promalin was more effective in preventing chlorosis than Accel at the 400 mg l⁻¹ (BA equivalent) level. Following 14 or 21 days of post-harvest evaluation, Promalin was more effective than Accel for the 100 mg l⁻¹ 2× and 400 mg l⁻¹ (BA equivalent) treatments.Plants in all Promalin and Accel treatments were taller than untreated plants 1 week after sprays were applied. At harvest, plants sprayed with Promalin were between 6 and 14 cm taller than untreated plants, but those treated with Accel were the same height as untreated plants.Neither Promalin nor Accel influenced the occurrence of malformed or aborted flowers in this study. However, cold storage significantly increased the number of plants with aborted buds and malformed flowers. Unstored plants averaged 0.16 aborted buds and 0.02 malformed flowers each, while those stored 3 weeks averaged 0.51 aborted buds and 0.18 malformed flowers each.     

Effect of exogenous growth regulators on flowering and cytokinin levels in azaleas

‘Alaska’ and ‘Redwing’ azaleas having dormant flower buds were sprayed with gibberellins (GA₃ or GA_{4 + 7}) alone and in combination with thiourea, N⁶ benzyl adenine (BA) or kinetin weekly for 3 or 4 weeks to test the efficacy of these materials in breaking bud dormancy. Additional plants received 6 weeks of cold storage at 4.5°C or glasshouse day temperatures of 21°C and above. The 2000 and 3000 mg l^?1 GA₃ and Ga_{4 + 7} sprays were better than 1000 mg l^?1 in promoting flowering, with ‘Redwing’ responding better than ‘Alaska’. GA-treated plants flowered in fewer days than those receiving cold storage. Flower diameter and pedicel length increased with higher levels of GA, and flower uniformity was comparable to cold-stored plants on most GA-treated ‘Redwing’-plants. Thiourea, BA and kinetin applied alone had no effect and considerable cytokinin activity was highest in GA-treated buds 14–21 days after treatment application. No increase in activity occurred on plants not receiving GA.     

The Effects of Some Growth Regulators on Peach Flower Bud Abscission

Applications of growth regulators to detached peach laterals in winter had variable effects on flower bud abscission. GA₃ and GA₄₊₇ increased bud abscission while IAA, BA and ABA inhibited abscission. Ethephon had a variable effect. Interactions were obtained between the compounds generally with IAA or GA playing the major role. GA had a peak of activity during a critical period in late winter indicating a variation in responsiveness. Variations in sensitivity to GA were obtained for different cultivars. Field applications of GA₃, GA₄₊₇, and ethephon to Elberta peach trees in winter, alone and in various combinations, promoted bud abscission.     

Endogenous gibberellins and their effects on flowering and stem elongation in cabbage (Brassica oleracea var. capitata)

Summary

Endogenous gibberellins were extracted from cabbage shoots and were analysed using gas chromatography and mass spectrometry. Nine gibberellins (GA₁, GA₁₉, GA₂₀, GA₄₄, GA₁₂, GA₄, GA_15, GA₂₄ and GA₂₅) were identified. Two gibberellin biosynthesis pathways were suggested, an early-13-hydroxlyated pathway and a non-13-hydroxylated pathway, to operate in cabbage shoots. GA₁, GA₄ and prohexadione calcium, a gibberellin biosynthesis inhibitor, were applied to the shoot tip of cabbage ‘Sousyu’ and ‘Kinkei No.201’ with or without cold treatment. Without cold treatment, stem elongation was increased by gibberellins and was suppressed by prohexadione calcium in both cultivars. But prohexadione calcium treatment, followed by gibberellin, promoted stem elongation more than gibberellin alone. Flowering was not induced by gibberellin or prohexadione calcium without cold treatment. When gibberellin and prohexadione calcium were applied during a cold treatment, stem elongation after the cold treatment was increased by gibberellins and was suppressed by prohexadione calcium in both cultivars. Flower bud appearance was promoted by GA₁ and GA₄ in ‘Sousyu’, but in ‘Kinkei No. 201’ only GA₄ was markedly effective. Inhibition of stem elongation and delay of flower bud appearance by prohexadione calcium were overcome by applying GA₁ or GA₄. Neither gibberellin nor prohexadione calcium treatment changed the number of leaf nodes at anthesis. These results indicated that stem elongation and flower bud development are regulated by gibberellins, but gibberellins might have little effect on flower induction.     

The response of partly cooled tulips to vacuum infiltration with gibberellins

Partly cooled (5°C) tulip ‘Apeldoorn’ were treated with gibberellins GA₃ and GA_{4 + 7} by vacuùm infiltration, with a view to defining conditions suitable for exploiting the effects of GA on forced tulips (faster flowering, control of stem extension, reduction of floral bud blasting).The first experiment showed that GA₃ and GA_{4 + 7} were equally effective in reducing the glasshouse period following 6 or more weeks cold storage; with less than 6 weeks cold storage, effects were less marked. Stem length at flowering was reduced by GA treatments, particularly by GA₃ and following more than 6 weeks cold storage. However, the vacuum infiltration method used (30 min at 10 torr) resulted in serious flower losses.Next, the effect of GA₃ concentration (up to 1500 mg 1^?1) was studied using vacuum infiltration treatments for 1–15 min at 20–510 torr, which resulted in fewer flower losses. Following 4 weeks cold storage, reducing pressure or increasing GA₃ concentration reduced both glasshouse period and stem length, with no effect of duration of treatment; GA₃ concentration was the only factor affecting flower length, which was increased. Following 8 weeks cold storage, increasing GA₃ concentration, vacuum or duration reduced glasshouse period. With all 3 factors at their maximum levels, 16 days earliness was obtained compared with controls. With maximum earliness, stem length was reduced to about 23 cm, compared to about 26 cm for treatments giving about 1 week's earliness, and 32 cm for untreated controls. Increasing vacuum appeared the most economical way of obtaining earliness, 20 torr giving 7 days earliness even at only 250 mg GA₃ 1^?1. Treatments giving earlier flowering also gave larger flowers. For comparison, there was little effect of soaking bulbs at atmospheric pressure even at 500 mg GA₃ 1^?1 for up to 20 h.Further experiments, conducted with vacuum infiltration at 260 torr for 15 min, confirmed these GA effects using formulated GA₃ (as “Berelex”) and GA_{4 + 7} (as “Regulex”). Effects of GA on stem length at flowering had disappeared by the time stems reached their final length. Comparisons with bulb injection of GA showed that this method required less GA than vacuum infiltration for similar effects, and that the greater effectiveness of GA_{4 + 7} compared with GA₃ was less marked using vacuum infiltration.     

Cold treatments enhance responsiveness to gibberellin in stock (Matthiola incana (L.) R. Br.)

Summary

Endogenous GAs have been suggested as regulators of stem elongation and flowering of cold-requiring plants. Here, the relationship between temperature conditions and responsiveness to GA₄ on stem elongation and flowering of stock (Matthiola incana) was investigated. The optimum temperature for induction of flower bud initiation was 10°C, and the minimum duration was 20 d in the late flowering cv. Banrei; the type of cold treatment effect on flowering was classified as a “direct effect”. Stem elongation was markedly promoted by cold treatment regardless of flower bud initiation. The cold treatment amplified the stem elongation response to GA₄. The GA₄ level necessary for flower bud initiation was lower in the 10°C treatment than in the 15°C treatment, and it became lower at longer durations of cold treatment. These results indicate that the cold treatments enhance responsiveness to GA₄ not only in the stem elongation process but also in the flower bud initiation process and that the development of responsiveness to GA₄ may correlate with the temperature and duration of cold treatment.     

In vitro response of pistachio nodal explants to silver nitrate

The effect of silver nitrate on shoot differentiation and shoot growth was examined in order to improve the regeneration efficiency of pistachio (Pistacia vera L. cv. Kirmizi) in vitro. Nodal explants of in vitro-grown seedlings were used to test various concentrations and combinations of 6-benzyladenine (BA), kinetin (KIN), gibberellic acid (GA₃) and silver nitrate (AgNO₃). Addition of AgNO₃ up to 48.0 μM to the culture medium improved the regeneration frequency and shoot growth, and reduced basal callus formation in all regenerated explants. The highest regeneration frequency (100%) was recorded on Murashige and Skoog (MS) medium containing 9.0 μM BA, 0.2 μM GA₃ and 24.0 or 48.0 μM AgNO₃ in combination. The best proliferation response in terms of both shoot formation and low callus production was obtained in the medium containing a combination of 9.0 μM BA, 0.2 μM GA₃ and 12.0 μM AgNO₃. Regenerated shoots, coming from three cycles of subculturing in proliferation media, were rooted in half-strength MS medium containing 12.0 μM indole-3-butyric acid (IBA). Well rooted plantlets were acclimatized and eventually established in peat and perlite. The development and optimization of an effective micropropagation protocol that is presented in this paper can give an important contribution to improve the quality of pistachio plants and, as a consequence, of orchard production in Middle East countries.     

Effects of bud scales and gibberellins on dormancy of in vitro cultured Japanese pear leaf buds

Leaf buds of Japanese pear were collected in early June and early November and regarded as summer and winter dormant buds, respectively. Bud explants with and without scales were prepared from each of them, and cultured in vitro for 75 days at 25°C with 14 h photoperiod, on a medium either without growth regulators, or supplied with BA and GAs (GA₃ and GA₄₊₇), singly or in combination.When either BA or GA₄₊₇ was contained in the medium, bud expansion occurred. Thereafter, summer dormant buds grew into shoots in the presence of BA, while winter dormant buds, although they swelled profusely, remained in a rosette. In the presence of BA, GA₄₊₇ markedly stimulated shoot elongation of summer dormant buds, but GA₃ did not. In winter dormant buds, GA₄₊₇ not only failed to stimulate shoot elongation, but also interfered with the BA-induced swelling described above.The presence of bud scales delayed expansion of summer dormant buds, while it had little effect on winter dormant buds. The delaying effect of scales on expansion of summer buds was effectively removed by application of GA₄₊₇ to the medium.     

Effects of gibberellins and benzylandenine on dormancy and flowering of Lilium speciosum

GA_{4 + 7} (1000 mg/l), alone or in combination with BA (100 mg/l), was found to induce shoot emergence and flowering in dormant bulbs of L. speciosum, while GA₃, alone or in combination with BA, had no effect. BA had a significant influence on increasing flower numbers, particularly when combined with GA_{4 + 7}.     

Day and night temperatures,daily light integral,and CO2 enrichment affect growth and flower development of Campanula carpatica ‘Blue Clips’

Campanula carpatica Jacq. ‘Blue Clips’ plants were grown in a greenhouse under nine combinations of day and night temperatures created by moving plants every 12 h among three day/night temperatures (15, 20, and 25°C). At each temperature, there were three daily light integrals (DLI; 4.2, 10.8, and 15.8 mol m⁻² per day, averaged over the experimental period) created with varying supplemental light, and ambient (≈400 μmol mol⁻¹) and enriched (≈600 μmol mol⁻¹) CO₂ concentrations. Time to flower was closely related to average daily temperature (ADT), and was not significantly affected by the day or night temperatures delivered to achieve a specific ADT. Time to flower was not largely affected by DLI or CO₂ enrichment. As plant ADT increased between 15 and 25°C, flower diameter decreased about 1 mm per degree and was not related to the difference between day and night temperatures (DIF). Flower diameter was smallest and least sensitive to changes in temperature at lower DLI and at ambient CO₂ levels. There were 10 less flower buds and 0.3 g less dry mass per plant at first flower for every 1° increase in plant ADT at high and medium DLIs. Flower bud number and dry mass were relatively low and less sensitive to changes in ADT at low DLI, and increased slightly with CO₂ enrichment at medium and high but not at low DLI. Plant height was not related to ADT, but increased linearly as DIF increased from −6 to 12°C at all DLIs, but the response was stronger under low DLI than high and medium DLIs. Flower bud number and dry mass were correlated closely with the ratio of DLI to daily thermal time (base temperature of 0°C). Flower bud number and dry mass were highest when C. carpatica plants were grown at 15°C with a DLI of 10–15 mol m⁻² per day.     

Effect of photoperiod on flower bud initiation and development in myoga (Zingiber mioga Roscoe)

Selection of suitable production locations in Australia and New Zealand for production of myoga (Zingiber mioga Roscoe) has been limited by lack of information on climatic influences on flowering. This study focused on photoperiod as potential production sites within Australia differ considerably in daylength due to the geographical range. The two cultivars available in Australia (Inferior and Superior) were examined in this trial due to previously observed differences in vegetative and reproductive development.Plants grown under long-day conditions (16 h) and short-day conditions (8 h) with a night break produced flower buds, while those under short-day conditions (8 h) did not. The failure of plants under short-day conditions to produce flower buds was due to abortion of developing floral primordia rather than a failure to initiate inflorescences. It was concluded that for flower development in myoga a qualitative long-day requirement must be satisfied, but that flower initiation was day-neutral.Short-day conditions resulted in abortion of flower primordia, premature senescence of foliage and reduced foliage dry weight in both cultivars. Early senescence and low flower bud yield of the Inferior cultivar, but not the Superior cultivar have been observed in crop evaluation trials in Southern Australia and New Zealand. Differences in critical photoperiod between the two cultivars may explain this observation and therefore photoperiodic requirements may be an important consideration in site and planting date selection for different cultivars.     

Modification of episodic growth in Skimmia japonica ‘Rubella’ by gibberellic acid treatment

Applications of GA₃ in May/June, when the first flush of growth was maturing, induced an extra flush of vegetative growth and initiated lateral bud break. Flower initiation was delayed and flower size reduced by the GA₃. These responses were only slightly modified by additional sprays with BAP and TIBA. GA₄₊₇ was ineffective in promoting either extra extension growth or lateral bud break, but it did reduce flower size.     

Induction of somatic embryogenesis in lotus (Nelumbo nucifera Geartn.)

Callus induction and somatic embryogenesis of lotus (Nelumbo nucifera Gaertn.) cv. Satabankacha were studied. Callus was initiated by culturing bud, cotyledon, and young leaf explants on Murashige and Skoog (MS) (1962) medium containing a combination of 0, 4, 8 and 10 μM 2,4-dichlorophenoxy acetic acid (2,4-D) and 0, 1, 2 and 3 μM 6-furfuryl amino purine (kinetin) or substituting 0, 0.5 and 1 μM benzyladenine (BA) for kinetin. Bud explants cultured on medium containing 4 μM 2,4-D and 1 μM BA gave the best callus growth. For somatic embryogenesis, the calli initiated on MS medium containing a combination of 4, 6, 8 and 10 μM 2,4-D and 1 μM BA and subsequently transferred to media containing 2–4 μM 2,4-D and 0 or 0.5 μM BA produced the most somatic embryos. When cultures were 12-week-old, callus produced on medium with 6 μM 2,4-D and 1 μM BA showed the best growth for somatic embryo regeneration. When transferred to a medium with 2 μM 2,4-D and 0.5 μM BA somatic embryos were produced from 33% of the calli. Embryos developed to the stage proembryo within 4 weeks and formed globular, heart, torpedo and mature embryos within 16 weeks.     

Cultivation of cut flower and bulb species with saline water

The long-term effect of saline water irrigation on flower yield and quality was investigated in three herbaceous cut flower crops of commercial importance, the Emily cultivars of Japanese limonium, Trachelium caeruleum and Eustoma grandiflorum (lisianthus), and in two bulb species, Hippeastrum hybridum and Ornithogalum arabicum. Among the tested crops, limonium showed the highest resistance to salinity. Irrigation water with an electrical conductivity of up to 11.5 dS m⁻¹ had little or no effect on stem yield and length of limonium flowering stems. In Trachelium, salinity had no effect on the yield of flowering stems or the size of the inflorescence, but it markedly reduced stem weight and length. The concomitant reduction in the number of nodes to flowering was reflected in earlier flower initiation. Since delayed flower differentiation and over-elongation of Trachelium stems is a serious problem during the winter months, application of mildly saline irrigation for winter production could be used to induce earlier flower initiation and to control stem height. In lisianthus subjected to salinity from bud appearance onwards, a salinity level of 6.0 dS m⁻¹ increased stem weight and the number of flowers per stem without affecting other quality parameters. The work carried out with Trachelium and lisianthus, although limited, indicates that salinity may be used for improving the quality of some cut flowers. In contrast to its beneficial effect on the herbaceous species, salinity led to a significant reduction of bulb, leaf, and root weight of the two bulbous species, H. hybridum and O. arabicum.     

Seasonal changes in CO2 assimilation in leaves of five major Greek olive cultivars

Leaf CO₂ assimilation rate, stomatal conductance (g_s), internal CO₂ concentration (C_i), chlorophyll (a + b) content, specific leaf weight (SLW) and stomatal density were measured during the season, under field conditions, for five major Greek olive cultivars, ‘Koroneiki’, ‘Megaritiki’, ‘Konservolia’, ‘Lianolia Kerkiras’, and ‘Kalamon’, with different morphological and agronomic characteristics and diverse genetic background. Measurements were taken from current-season and 1-year-old leaves, and from fruiting and vegetative shoots, throughout the season, from March to November in years 2001 and 2002. CO₂ assimilation rates showed a substantial seasonal variation, similar in all cultivars, with higher values during spring and autumn and lower values during summer and late autumn. Stomatal conductance (g_s) followed similar trends to leaf CO₂ assimilation rates, increasing from March to July, following by a decrease during August and increasing again in autumn. ‘Koroneiki’ had the highest leaf CO₂ assimilation rate and g_s values (21 μmol m⁻² s⁻¹ and 0.45 mol m⁻² s⁻¹, respectively) while ‘Lianolia Kerkiras’ and ‘Kalamon’ showed the lowest leaf CO₂ assimilation rate and g_s values (13–14 μmol m⁻² s⁻¹ and 0.22 mol m⁻² s⁻¹, respectively). One-year-old leaves had significantly higher leaf CO₂ assimilation rate than current-season leaves from April to June, for all cultivars. From August and then, leaf CO₂ assimilation rate in current-season leaves was higher than in 1-year-old leaves. There were no significant differences in leaf CO₂ assimilation rate between fruiting and vegetative shoots. Total chlorophyll (a + b) content increased with leaf age in current-season leaves. In 1-year-old leaves chlorophyll content increased in spring, then started to decrease and increased slightly again late in the season. Chlorophyll content was higher in 1-year-old leaves than in current-season leaves throughout the season. Total specific leaf weight (SLW) increased throughout the season for all cultivars. Stomatal density in lower leaf surface was lowest for ‘Koroneiki’ (399 mm⁻²) and highest for ‘Megaritiki’ (550 mm⁻²). Our results showed differences in leaf CO₂ assimilation rate among the five different olive cultivars, with a diverse genetic background, ranging from 12 to 21 μmol m⁻² s⁻¹. From the five cultivars examined, ‘Koroneiki’, a drought resistant cultivar, performed better and was able to maintain higher leaf CO₂ assimilation rate, even under high air vapor pressure deficit. All cultivars had a pronounced seasonal variation in leaf CO₂ assimilation rate, affected by date of the year, depending on ambient conditions. The high temperatures and high air vapor pressure deficit occurring during summer caused a reduction in leaf CO₂ assimilation rate in all cultivars. Leaf CO₂ assimilation rate was also affected by leaf age for all cultivars, with old leaves having significantly higher leaf CO₂ assimilation rate than young leaves early in the season.     

Factors controlling high efficiency adventitious bud formation and plant regeneration from in vitro leaf explants of roses (Rosa hybrida L.)

Studies were conducted to improve adventitious bud regeneration in roses (Rosa hybrida L.), specifically to extend the protocol to different genotypes and to initiate production of multiple shoots per explant. The best results were obtained by using a two-stage procedure where excised leaflets were incubated on Murashige and Skoog (MS) (1962) induction medium with 6.8 μM TDZ plus 0.49 μM IBA in the dark for 7 days and subsequently transferred to an MS-based regeneration medium with 2.22 μM BA plus 0.049 μM IBA exposed to a PPFD of 15 μmol m⁻² s⁻¹ PAR. Bud formation capacity was also significantly affected by the genotype and the environment, such as the use of bottom cooling creating a lower RH in the vessel. The addition of silver nitrate to the induction medium also significantly improved the percentage of regeneration in three genotypes tested. Regenerated shoots failed to elongate when transferred to MS proliferation medium containing 0.5 mg l⁻¹ BA, however maximum bud development and elongation were achieved when kinetin in the range 1–2 mg l⁻¹ was used. Elongated shoots were excised and rooted best on zero growth regulator half-strength MS modified medium. Rooted plantlets were acclimatized under greenhouse conditions for evaluation of somaclonal variation.     

The effect of gibberellic acid on sprouting and flowering of some tulip cultivars

The effect of gibberellic acid, GA₃, on breaking of dormancy, sprouting and flowering of 9 cultivars of tulips was investigated, using 4 cultivars recommended for forcing and 5 not suitable for forcing. GA₃ was applied to the basal plates of the bulbs in a lanolin paste before planting, or was injected into the bulbs prior to rooting or after 38 or 64 days of cold treatment. The application of GA₃ stimulated sprouting, growth of floral stalks and flowering in all investigated cultivars. Injection of GA₃ was more effective than topical application. The effect of GA₃ on sprouting and flowering of plants was especially effective after 38 or 64 days of cold treatment. The possibility of applying GA₃ in commercial horticulture is considered.     

The Influence of Foliar Applications of GA3 GA4,7 and PBA on Breaking Flower Bud Dormancy on Azalea CVS Redwing and Dogwood

Foliar applications of gibberellins (GA₃, GA₄,₇) and 6-benzylamino-9-(2-tetra- hydropyranyl)-9H-purine (PBA) were compared with untreated controls and the conventional low temperature treatment for breaking flower bud dormancy. GA₃ and GA₄, ₇ were compared for their role in promoting flower development. Treatments were initiated when microscopic examination showed the flower buds had reached Stage 6 (style elongated and closed). Plants that did not receive a dormancy-breaking treatment failed to reach anthesis. Application of GA₃ or GA₄, ₇ hastened flowering and increased flower size and pedicel length.     

Effect of fruit load on 9,10-ketol-octadecadienoic acid (KODA), GA and jasmonic acid concentrations in apple buds

The relationship between 9,10-ketol-octadecadienoic acid (KODA), GAs and jasmonic acid (JA) and flower bud formation influenced by fruit load in apples (Malus domestica Borkh.) was investigated. The endogenous KODA and JA concentrations in apical buds in plants subjected to flower thinning treatment (FTT), under which all flowers were removed were higher than those in plants subjected to heavy crop treatment (HCT), under which the number of leaves per fruit was adjusted to 20 from 60 to 120 days after full bloom (DAFB). In contrast, the gibberellic acid concentrations [total of gibberellin A₁ (GA₁) and gibberellin A₄ (GA₄)] in FTT plants were low compared with those in HCT plants. The result suggests that KODA and JA in contrast to GAs may have opposite effects on flower bud formation which is significantly influenced by fruit load in apples and that KODA and JA may also be associated with flower bud formation in apples.     

An investigation on the effect of different plant growth regulating compounds in in vitro shoot tip and node culture of lemon seedlings

The possible application of some less commonly used in vitro growth regulating compounds is outlined. A number of treatments were applied to determine the best way of inducing in vitro shoot proliferation and rooting on a modified Driver–Kuniyuki [HortScience 19 (1984) 507] basal medium of lemon (Citrus limon (L.) Burm, f. cv. Interdonato) seedlings. 6-Benzyladenine (BA) alone (1, 2 and 4 mg l⁻¹) and in combination with either orange juice (10%, v/v), silver nitrate (3 mg l⁻¹), gibberellic acid (GA₃) (0.1 mg l⁻¹ at the establishment stage and 0.5 mg l⁻¹ at all combinations during the proliferation stage) or abscisic acid (ABA) (0.2 mg l⁻¹ only at the establishment stage) were used to stimulate shoot formation during the establishment and the proliferation stage. The combination of BA with ABA gave a high rate of shoot formation, while GA₃ and silver nitrate enhanced shoot elongation. When these shoots were transferred to the rooting stage, the effect of application of two different auxins (indole-3-butyric acid (IBA) and α-napthaleneacetic acid) was examined, as well as different methods of application (auxin added to the basal medium and auxin application by dipping the base of the explant in auxin solution). Dipping the base of the explants in a 50% ethanol solution of IBA at 1000 mg l⁻¹ for 5 s resulted in 80% rooting with subsequent 90% survival of these explants, during acclimatization under mist.