Effect of temperature on bulb growth capacity and sensitivity to summer sprouting in Lilium longiflorum Thunb.

The production of Lilium longiflorum bulbs in The Netherlands, with its cool climate, has been a problem because of unsatisfactory bulb growth and the risk of premature sprouting of the daughter bulbs (summer sprouting). To investigate the possibilities of breeding a type of L. longiflorum that can be grown under cool climatic conditions, a collection of 27 L. longiflorum cultivars from Japan, The Netherlands and the United States was tested, together with two Asiatic hybrids and a L. speciosum cultivar, under low phytotron temperatures (10, 14, 17°C) and in field experiments.‘Mount Everest’, ‘Saeki’, ‘Indian Summer’ and ‘White American’ were among the best L. longiflorum cultivars, with less than 20% summer sprouting and good bulb production. This was in contrast to ‘Hinomoto’, ‘Ace’ and some American introductions, which showed more than 60% summer sprouting and low bulb production. The lower the phytotron temperature, the more summer sprouting occurred. The differences observed between the cultivars in the field experiments were in agreement with those observed in the phytotron. The genetic variation proved large enough to start a breeding program for L. longiflorum adapted to cool climate conditions.     

Effects of pre- and post-bloom temperature regimes on development of Lilium longiflorum Thunb.

Glasshouse grown ‘Ace’ and ‘Nellie White’ Easter lily plants were subjected to different temperature regimes to determine temperature requirements during pre- and post-bloom development. Rate of leaf- and flower-bud development and stem elongation on the primary (mother) axis were directly proportional to the range of temperatures used (6–24°C), and were equally effective in predicting crop development. Scale initiation on the secondary (daughter) axis during pre-bloom phases was proportional to growing temperature, reaching maximum activity at 18°C in ‘Ace’ and at 12°C in ‘Nellie White’. The shift from scale to leaf initiation and development following anthesis was favored by 12 rather than 18°C with significant reductions in leaf initiation in both cultivars at 24°C. No difference in secondary meristem diameter occurred with temperature during pre-bloom, but large dome size was associated with 12°C or lower during the post-bloom phase. Primary scale weight increase (filling), reached a maximum 50 days following anthesis, and was greatest at 18°C. Secondary scale filling reached a maximum 80 days after anthesis at both 18 and 24°C. The secondary axis became increasingly responsive to sprout-inducing temperatures with increasing age and development. Fifty days after anthesis, 12 and 18°C were equally effective in sprouting ‘Ace’ bulbs, while 12°C was more effective with ‘Nellie White’. Early leaf senescence, associated with high (24°C) temperature, did not favor increased bulb size, daughter leaf primordia count and meristem diameter, or sprouting.     

Plant regeneration from in vitro cultured leaves of Lanzhou lily (Lilium davidii var. unicolor)

Lanzhou lily (Lilium davidii var. unicolor) is one of the best lilies which are edible in China but the efficient shoot regeneration system has not been developed. The purpose of the present study is to establish an efficient and reproducible protocol for induction of shoots in vitro from L. davidii var. unicolor leaves. Shoot regeneration from in vitro cultured leaves of L. davidii var. unicolor was tested on the 26 media based on NN [Nitsch, J.P., Nitsch, C., 1969. Haploid plants from pollen grains. Science 163, 85–87] basal medium, containing different concentrations of thidiazuron (TDZ) in combination with different concentrations of α-naphthaleneacetic acid (NAA). Shoot organogenesis occurred directly from the leaves without forming callus. Shoot regeneration mainly occurred from the cuts across the midvein and the base of the leaf explants. The highest frequency of regeneration (93.3%) and the largest number of shoots per leaf (3.83) were obtained on NN basal medium supplemented with 0.5 mg l⁻¹ TDZ and 1.0 mg l⁻¹ NAA. All the regenerated shoots formed complete plantlets on half-strength MS [Murashige, T., Skoog, F., 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant 15, 473–497] basal medium containing 0.1–0.5 mg l⁻¹ indole-3-butyric acid (IBA) with in 30 days, and 92% of the regenerated plantlets survived in the soil. This study will be useful for Agrobacterium-mediated transformation and exploitation of somaclonal variation of Lanzhou lily.     

The effects of explant and cytokinin type on regeneration of Prunus microcarpa

We investigated in vitro regeneration ability of Prunus microcarpa subsp. tortusa using various explants (root, cotyledon and hypocotyl pieces) and cytokinins [benzyladenine (BA), meta-Topolin (mT) and thidiazuron (TDZ)]. Sectioned cotyledon, root and hypocotyl pieces of in vitro grown seedlings were cultured on Nas and Read Medium (NRM) containing BA (7.5, 10, 12.5, 15 or 17.5 μM), mT (2.5, 5.0, 7.5, 10 or 12.5 μM) or TDZ (2.5, 5.0, 7.5, 10 or 12.5 μM). As a measurement of morphogenetic reaction, the ratios of regenerating explants and the numbers of primary adventitious shoots per regenerating explant were analyzed. Cotyledon explants exhibited higher regeneration ratios than hypocotyl explants, and the root explants were inappropriate for regeneration. Both BA and mT were effective on shoot regeneration but higher regenerating explant ratios were obtained when BA was used. In comparison with BA and mT, the effect of TDZ on enhancing explant regeneration ability was insignificant. Mean number of adventitious shoot per regenerating explant was between 1 and 4, and regenerating explant ratios were between 0% and 77%. The practical appliacations of the results are discussed.     

Growth regulator requirements for adventitious regeneration from Lilium bulb-scale tissue in vitro,in relation to duration of bulb storage and cultivar

Addition of NAA (1-naphthylacetic acid) to the nutrient medium strongly influenced the adventitious regeneration of plantlets from bulb-scale tissue of lilies in vitro. The percentage of regenerating explants, the number of plantlets per explant, and also the bulblet growth of the resulting plantlets were affected by the auxin, low concentrations being stimulatory and high concentrations inhibitory.The cytokinins BA (6-benzyladenine) and 2iP (N6-[Δ2-isopentenyl]-adenine) did not influence the adventitious regeneration, but caused aberrant growth of the regenerated plantlets.Prolonged cold storage of the mother bulbs decreased the demand for exogenous auxin for maximum adventitious regeneration. The optimum for bulblet growth was not affected.Results of experiments with cultivars ‘Pirate’, ‘Enchantment’ and ‘Connecticut King’ suggest an inverse relationship between the in vivo propagation rate by scaling and the NAA demand for optimal adventitious regeneration in vitro.     

Effect of bulb storage temperature on leaf emergence and plant development during scale propagation of Lilium longiflorum ‘White American’

Effects of temperature and duration of bulb storage on leaf emergence of scale bulblets and on the type of plant development were investigated in Lilium longiflorum ‘White American’. After the parent bulbs had been stored for 0, 5, 10 or 15 weeks at 30, 20, 10, or 0°C, scales collected from different parts of a bulb were scale-propagated at 25 or 14°C, or under the temperature regimes normally used in The Netherlands. Independently of the scale position, a higher storage temperature promoted leaf emergence from scale bulblets, whereas a lower temperature delayed it. Higher storage temperatures produced more epigeous type plants, especially from outer and middle scales. Sprouting of the parent bulb had no effect on leaf emergence from the scale bulblets nor on the type of plant development.     

In vitro bulblet formation from leaf segments of lilies,especially Lilium rubellum Baker

Bulblets were induced from 5-mm lily leaf segments, cultured on Murashige and Skoog's mineral salts medium (1962) together with α-naphthalene acetic acid (NAA) 1 mg/l, 6-benzylaminopurine (BA) 0.1 mg/l and several organic elements.Bulblet formation was affected by several factors, especially the time of excision and the origin of the explant; bulblets developed well in leaf explants excised before flowering, and mainly in those excised within 15 mm of the leaf base. However, bulblets were seldom induced in explants excised after flowering, regardless of their origin.NAA was almost indispensable for bulblet formation, but BA had a slight stimulatory effect only when NAA was also present. Sucrose concentration and light also affected bulblet regeneration.     

The effects of growth substances and photoperiod on the development of shoot apices of Vitis cultured in vitro

‘Rougeon’ grapevines were recovered from shoot apices using in vitro culture. Shoot development from apices 0.5–1 mm in length, which contained 2–4 leaf primordia was obtained with 5 × 10^?6M benzylaminopurine and 5 × 10^?7M naphthaleneacetic acid (NAA) under a 10-hour photoperiod, but not under continuous light, in a 15-hour photoperiod, or in darkness. At least one vigorous shoot was produced by 100% of the cultures.Rooting of these shoots was induced under the same photoperiod with the same basal culture media containing only 10^?7 M NAA. Roots were produced by 91% of the cultures. Zero-, 16- or 24-h photoperiods were ineffective in promoting root development.Rooted plantlets were transferred into soil with 80% of them surviving. The grapevines were allowed to reach an average length of 2 m in a growth chamber before growth conditions were modified to induce dormancy. Vines were planted in a vineyard; after 2 growing-seasons we were unable to distinguish the micropropagated vines from ‘Rougeon’ vines propagated in the standard manner, on the basis of fruit or vegetative morphology.     

The effect of day length,source of light and growth regulators on growth and flowering of Clerodendrum thomsonae Balf

The formation of flower buds in Clerodendrum seems not to be affected by day length, but the development of the buds is delayed in long days. When long days were established by means of low-intensity illumination with incandescent lamps, few flowers developed and the stems elongated considerably even at a day length of 16 hours. When fluorescent lamps were used for day-length extension, short shoots with many flowers were obtained even in 24-hour days. Flower development was also delayed by gibberellic acid (GA₃), but promoted by chlormequat both in short and long days. Shoot elongation was retarded by chlormequat and promoted by GA₃.Plants obtained from commercial greenhouses varied considerably with respect to growth and flowering. By selection a clone was obtained which flowered richly on short shoots. Shoot elongation stopped when flowering began. A ‘negative’ selection gave rise to a clone which flowered sparsely and in which shoot elongation was not influenced by flowering.     

The effect of temperature and light on flower development in Pelargonium × domesticum

Factors influencing the number of flowers produced in Pelargonium × domesticum cultivar ‘Lavender Grand Slam’, and their rate of development after the plants had been given a period of low-temperature flower induction, were studied.Under short days progressive abortion reduced flower numbers as the temperature regime during the forcing-period was increased. Under long days this effect was less marked and the high-temperature regime advanced flowering by over a month. When long days and high temperatures were used for forcing, it was necessary to maintain a high light intensity, 335 J/cm²/day giving the best results in terms of earliness of flowering and the number of flowers produced. At lower irradiance there was some risk of flower abortion, particularly in the first inflorescence.     

Effect of light,temperature and CO2 on the growth of Campanula isophylla stock plants and on the subsequent growth and development of their cuttings

Stock plants of Campanula isophylla Moretti were subjected to different temperature and light conditions and to various CO₂ regimes. The number and the fresh and dry weight of the cuttings produced were recorded. The after-effect of stock-plant treatment on root formation and growth of cuttings was studied.Increasing light intensity and CO₂ supply strongly promoted cutting production and increased both fresh and dry weight of the cuttings. These factors also markedly influenced root formation and root growth of the cuttings. Stock plant conditions also strongly influenced the growth and lateral shoot formation of the rooted cuttings. It is concluded that cuttings from stock plants grown under favourable light (10 Klx) and CO₂ conditions (900 v.p.m.) contain factors beneficial for root formation, growth and shoot formation. The results are discussed in relation to the carbohydrate content of the cuttings.     

Establishment of efficient regeneration protocol from leaf explants of Iris pumila shoots cultured in vitro

An efficient plant regeneration protocol via somatic embyogenesis by leaf base culture of in vitro grown Iris pumila shoots was developed. Induction of embryogenic calli was achieved on MS media supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D), kinetin (4.5 μM, each) and some additives (L-proline, casein hydrolysate, adenine sulphate and tyrosine). Further differentiation of embryogenic calli was achieved on MS hormone-free media, and on media supplemented with either BAP (4.5 μM) or BAP + zeatin (4.5 and 0.2 μM, respectively), which allowed somatic embryos, as well as shoot-like structures to form. Fully developed somatic embryos germinated on an MS hormone-free medium. An anatomical study confirmed that shoot-like structures represented early germinating stages of somatic embryos. Acclimatization of plants derived from somatic embryos was 64% after 1 year and no morphological variation was observed.     

Factors affecting the growth of in vitro cultured lateral buds from Phalaenopsis flower stalks

Phalaenopsis flower-stalk buds cultured in vitro show 3 modes of growth: dormant, vegetative and reproductive. The effects of bud position on the stalk, temperature, and benzyladenine (BA) on the mode of bud growth were examined.Flower stalks were cut into 1-node sections, each with 1 bud, and inserted into solid culture medium, top side up. Buds on the upper sections had a tendency to remain dormant regardless of temperature. Sprouting buds placed at 20°C or 25°C showed reproductive growth except for some on the basal sections which grew vegetatively. At 28°C, all buds developed vegetatively independent of their original position on the stalk. The buds on the sections taken from both stalks which had elongated at min 18°C and min 28°C were subcultured; the mode of growth of the cultured buds was not affected by the temperature during flower-stalk elongation but was affected by the subsequent culture temperature.Cultured buds which had remained dormant were stimulated to sprout by addition of BA to the medium. At 5 p.p.m. and above, all buds began to develop, but malformations developed on the leaves of the shoots. All factors considered, BA at 2.5 p.p.m. was found to be the optimum level. The further development of the buds released from dormancy by BA was also affected by the position on the flower stalk and by cultural temperature.     

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}.     

Influence of growth regulators and light on in vitro shoot regeneration in M.26 apple roostock

Summary

The effects of different BA and IBA concentrations and dark/light combinations, applied during both the last proliferating subculture (LPS) and the regeneration phase (RP), on shoot regeneration from leaves were evaluated in M.26 apple rootstock. A positive influence on caulogenesis was found with a low cytokinin/auxin ratio in the medium during the LPS. The increase in IBA concentration from 0.49 μM to 4.92 μM in the LPS, along with the absence or the use of a low cytokinin concentration (0.89 μM) in the medium, enhanced the subsequent shoot regeneration from leaves. During the RP, a concentration of 22.2 μM BA gave the best caulogenesis results. During both the LPS and the RP, high IBA concentrations were able to replace the combined effect of a low IBA concentration and dark treatment; this could indicate that dark treatments interact with the auxin metabolism in the leaf caulogenesis response. Auxin application could not reproduce all the effects of dark treatments, suggesting that dark also affects other biochemical and/or physiological aspects, such as gibberellin metabolism. Moreover, cytokinins applied during both the LPS and the RP influenced the size of regenerated shoots.     

The influence of naphthaleneacetic acid on the growth of in vitro cultivated seedlings of Bromeliaceae

The role of NAA during in vitro seed germination and subsequent growth was examined in three Bromeliaceae: Guzmania minor var. ‘Vella’ Mez, Guzmania lingulata var. ‘Splendens’ Mez and Vriesea splendens var. ‘Fire’ Lem. NAA incorporation in the medium resulted in a strong stimulation of root and shoot growth, with an optimum response at 0.5–0.8 mg l^?1. It is hypothesized that the usual slow growth of bromeliad seedlings in soil is primarily due to poor rooting, which is the result of auxin deficiency.     

The effect of nitrogen source and concentration,medium pH and buffering on in vitro shoot growth and rooting in Eucalyptus marginata

This work examined the effect of nitrogen source and medium buffering on the micropropagation of Eucalyptus marginata Donn ex Sm. The number of shoots was increased when media contained 2-(N-morpholino) ethanesulfonic acid (MES) but this increase was minor and only applied to one of the two clones tested. Highest root production was obtained when the medium contained 7.5 mM nitrogen in a ratio of 2NO₃⁻:1NH₄⁺ and was buffered with 10 mM MES. In the rooting medium the pH was influenced most significantly by the nitrogen source, and then whether the medium was buffered. The media pH remained relatively constant when nitrate was the sole nitrogen source and this was assisted by the addition of 10 mM MES. Lower concentrations (<10 mM) of MES were less effective in buffering media over a four-week culture period in both shoot multiplication and rooting medium.     

Direct regeneration of plants derived from in vitro cultured shoot tips and leaves of three Lysimachia species

The regenerability of three ornamental species—Lysimachia christinae, Lysimachia rubinervis and Lysimachia nummularia ‘Aurea’, were investigated using in vitro leaves and shoot tips. 6-Benzylaminopurine (BAP) and α-naphthalene acetic acid (NAA) added to Murashige and Skoog (MS) medium were tested for their effect on organogenesis. On the medium, shoot regeneration occurred directly without callus formation. In these species, L. christinae developed the highest regeneration rate and numbers of shoots/explant from shoot tips (100%, 12.25) and leaf bases (100%, 13.01) on the MS medium containing 3.0 mg l⁻¹ BAP and 0.1 mg l⁻¹ NAA. For L. rubinervis, the highest shoot induction rate and number of shoots/explant were obtained from shoot tip (100%, 16.87–17.20) on the MS medium with 0.1 mg l⁻¹ NAA and 3.0–5.0 mg l⁻¹ BAP. L. nummularia ‘Aurea’, however, showed the highest regeneration rate and number of shoots/explant (100%, 12.73) from leaf bases on MS medium supplemented with 1.0 mg l⁻¹ BAP and 0.1 mg l⁻¹ NAA. All in vitro shoots rooted well on half macronutrient MS medium containing 0.1 mg l⁻¹ NAA. After acclimatization, transplanted plantlets grew normally and flowered in the field.     

Neoformation of bulbils in Allium porrum L. cultured in vitro

A procedure has been developed to obtain bulbils on stem explants of leek (Allium porrum L. cv. ‘Elboeuf’) cultured in vitro. It is also possible to differentiate shoots or roots.