Shoot multiplication in cultures of mature <Emphasis Type="Italic">Alnus glutinosa</Emphasis>

Shoot cultures were initiated from mature trees of Alnus glutinosa. On medium containing 1–5 μM 6-benzylamino purine (BAP), the shoots elongated without branching, formed heavy callus at the base of the stems and readily formed roots. The possibility that these characteristics could be attributed to the strong influence of endogenous auxin was tested on media that contained two auxin transport inhibitors, 1-N- naphthylphthalamic acid (NPA) and 2,3,5-triiodobenzoic acid (TIBA), at concentrations of 0.1–3 μM, in combination with 2 μM BAP. On these media, shoots produced numerous branches, less callus and no roots. After 30 weeks (five subcultures) on this medium, leaves were smaller and showed signs of vitrification. These problems were resolved without detriment to shoot proliferation, by reverting to medium without NPA or TIBA. Shoots rooted readily after transfer to medium without growth regulators and were successfully acclimatised after transfer to soil.     

Distribution of assimilate during the flush cycle of growth in Theobroma cacao L.

The growth of the shoot and roots of seedling plants of cocoa (Theobroma cacao L.) under constant glasshouse conditions showed a rhythmic cycle, with the maximum growth stages of each alternating in a regular sequence. When the growth cycle of the shoot was upset by removing all new leaves immediately after unfolding, the roots showed a high constant growth rate during this period, suggesting that normally the rapidly expanding leaves exert an inhibitory influence on the roots. Conversely removal of portions of the root delayed the production of new leaves in the shoot. The level of soluble and starch carbohydrate in the mature leaves, roots and stem declined during the period of expansion of the flush leaves, but accumulated again at the end of the leaf expansion stage. It is likely that this reserve carbohydrate was remobilised and translocated to the flush leaves during their period of expansion. A large proportion of newly formed photoassimilate, as shown by the distribution of ¹⁴C radioactivity from different source leaves, was also translocated to the young leaves during expansion. The large sink created by these leaves may cause photoassimilate and reserve carbohydrate to be diverted from the roots, thereby inhibiting root growth during the stage of leaf expansion. It is suggested that the rhythmic leaf production at the apex may control the growth cycle of the roots.     

Plant regeneration from callus cultures of Psoralea corylifolia Linn

Plantlet regeneration via organogenesis was achieved in callus cultures derived form mature leaves, stems and leaves, petioles and roots of young seedling of Psoralea corylifolia on Murashige and Skoog medium supplemented with 2.5–3.0 mg L^-1 BA, 1.0 mg L^-1 NAA and 3% (w/v) sucrose. The rate of shoot bud regeneration was positively correlated with the concentration of hormones in the nutrient media. Shoot buds regenerated more readily from juvenile explants (seedling source) as compared to the mature explants. Addition of adenine sulphate (5 mg L^-1) to the culture medium increased the growth of shoot buds. Optimum responses were obtained in hypocotyl and leaf explants using NAA in combination with BA, the highest rate of shoot bud regeneration being in hypocotyl explants. Rooting was readily achieved on the differentiated shoots on MS basal media without growth regulators. Regenerated plantlets were successfully established in the greenhouse.     

In vitro propagation of two species of commelinaceae from bud cultures and repeated subcultures on a growth regulator-free medium

Summary In our wide-ranging research on in vitro propagation of some monocotyledonous plants, two Commelinaceae species were studied: Tradescantia fluminensis var. foliis variegatis and Tradescantia pallida. Initial cultures were established successfully using nodal and apical meristems that produced single shoots, many roots, and no callus, by utilizing growth regulator-free MS medium. Addition of growth regulators did not increase the activity of explants that produced single or multiple shoots, atypical roots, and no callus. Consecutive cultures were possible using the apical and nodal meristems of the previous generation. The behavior of the different generations in culture did not change and was similar to the initial cultures. Their growth capacity was maintained over many months, also on a growth regulator-free medium. In both species, the chromosome number in the root tips of the mother plant and all morphologically stable in vitro plantlets confirmed a constant ploidy level, in T. fluminensis 2n=72, and in T. pallida 2n=24.     

In vitro Clonal Propagation of Neem (Azadirachta indica)

The micropropagation of neem (Azadirachta indica) was accomplished by culture of buds from crown branches of a mature tree, basal-sprouts of another mature tree and a single juvenile plant. Cultures derived from these three different sources showed significant variation in in vitro response. In case of the crown and basal-sprout explants, addition of 12.5 μM PVP-40 in the establishment medium controlled leaching of phenol growth inhibitors. Phenolic leaching was not observed in juvenile explants. DKW medium (with 0.22 μM BA) was significantly better than MS for shoot proliferation. Shoot cultures of crown branch origin did not elongate and eventually died after the third subculture. In the presence of 4.9 μM IBA in half-strength DKW, 90% of the shoots and 100% of basal-sprout and seedling explant origin, formed roots. Plantlets from both explant types showed 90% survival after acclimatization.     

Interactive effects of elevated atmospheric CO2, mycorrhization and drought on long-distance transport of reduced sulphur in young pedunculate oak trees (Quercus robur L.)

Pedunculate oak (Quercus robur L.) was germinated and grown under nutrient non-limiting conditions for a total of 10–15 weeks at ambient CO₂ concentration and 1100 μmol mol^–1 CO₂ either in the presence or the absence of the mycorrhizal fungus Laccaria laccata. Half of the oak trees of these treatments were exposed to drought during final growth by suspending the water supply for 21 d. Mycorrhization and elevated atmospheric CO₂ each enhanced total plant biomass per tree. Whereas additional biomass accumulation of trees grown under elevated CO₂ was mainly attributed to increased growth of lateral roots, mycorrhization promoted shoot growth. Water deficiency reduced biomass accumulation without affecting relative water content, but this effect was more pronounced in mycorrhizal as compared to non-mycorrhizal trees. Elevated CO₂ partially prevented the development of drought stress, as indicated by leaf water potential, but did not counteract the negative effects of water deficiency on growth during the time studied. Enhanced biomass accumulation requires adaption in protein synthesis and, as a consequence, enhanced allocation of reduced sulphur produced in the leaves to growing tissues. Therefore, allocation of reduced sulphur from oak leaves was studied by flap-feeding radiolabelled GSH, the main long-distance transport form of reduced sulphur, to mature oak leaves. Export of radiolabel proceeded almost exclusively in basipetal direction to the roots. The rate of export of radioactivity out of the fed leaves was significantly enhanced under elevated CO₂, irrespective of mycorrhization. A higher proportion of the exported GSH was transported to the roots than to basipetal stem sections under elevated CO₂ as compared to ambient CO₂. Mycorrhization did not affect ³⁵S export out of the fed leaves, but the distribution of radiolabel between stem and roots was altered in preference of the stem. Trees exposed to drought did not show appreciable export of the ³⁵S radioactivity fed to the leaves when grown under ambient CO₂. Apparently, drought inhibited basipetal transport of reduced sulphur at the level of phloem loading and/or phloem transport. Elevated CO₂ seemed to counteract this effect of drought stress to some extent, since higher leaf water potentials and improved ³⁵S export out of the fed leaves was observed in oak trees exposed to drought and elevated CO₂ as compared to trees exposed to drought and ambient CO₂.     

Agrobacterium rhizogens mediated transformation of Rauvolfia serpentina: Regeneration and alkaloid synthesis

Shoot cultures of Rauvolfia serpentina infected with Agrobacterium rhizogenes 15834 produced tumourous tissue at the site of injection, which eventually developed callus with hairy roots. Sporadic shoot formation occurred from the hairy roots. The shoots were grown to maturity in the green house. The mature transformed plants (RST) showed distinct variations in their physiological characteristics. The flowers of the transformed plants were more delicate and less pigmented when compared to the flowers of the mature normal plants. The roots of the transformed plants were hairy with a number of lateral branches, whereas the roots of the normal plant had very few lateral branches. The biomass of the transformed plant was 86.39 g/plant (fresh wt), significantly higher than the normal plant which was 77.3 g/plant (fresh wt). The total alkaloid content in the mature transformed plant (0.073 g per plant) was similar to the normal plant (0.078 g per plant), although the hairy roots contained little alkaloid.Abbreviations MS Murashige & Skoog's basal medium - MLS modified Linsmaier & Skoog's basal medium - BA benzyladenine - NAA naphthalene acetic acid - TLC thin layer chromatography - HPLC high performance liquid chromatography     

Survival and growth responses of Myricaria laxiflora seedlings to summer flooding

Myricaria laxiflora, an endangered shrub species distributed along the banks of the Yangtze River in the Three Gorges area, is completely submerged from June to October every year. It is generally assumed that summer flooding has a strong impact on the survival and growth of seedlings. We designed an outdoor randomized block experiment on the responses of seedling survival and growth to different flooding depth and flooding duration treatments during the flood season in the Three Gorges area. Seedling survival rate, aboveground biomass, belowground biomass, total biomass, root depth, length of primary branch and the number of primary and secondary branches were examined.M. laxiflora was found to acclimate to summer flooding by becoming dormant and losing biomass. Seedlings of M. laxiflora ceased growing during the summer flooding season, regardless of the flooding depth and flooding duration they were subjected to. The number of primary and secondary branches, aboveground biomass and total biomass of seedlings was reduced with prolonged flooding. The length of primary branches and aboveground biomass were more sensitive to flooding than other measured parameters and differed significantly between the onset and the end of flooding.In each flooding treatment most seedlings of M. laxiflora survived a flooding period of 2 months and recovered rapidly after the flooding was terminated in September. After 3 months of recovery, aboveground biomass, total biomass and the number of the primary branches increased significantly. Furthermore, seedling survival and growth in the flooding treatments were not significantly different from the controls both during the summer flooding stage and in the recovery stage. All of these results suggest that summer flooding does not affect seedling survival and growth in this species. On the contrary, flooding released seedlings from the stress of drought during summer and facilitated seedling establishment. M. laxiflora appears to cope adaptively with the flooding cycle by going into a state of dormancy during the flood season.     

Disturbance of forest by trampling: Effects on mycorrhizal roots of seedlings and mature trees of Fagus sylvatica

The effects of disturbance by recreational activities (trampling) on changes in soil organic matter (SOM) and on mycorrhizal roots of seedlings and mature trees were studied in four stands of a beech (Fagus sylvatica L.) forest near Basel, Switzerland. At each site, comparable disturbed and undisturbed plots were selected. Disturbance reduced ground cover vegetation and leaf litter. Beech seedlings had lower biomass after disturbance. Ergosterol concentration in seedling roots, an indicator of mycorrhizal fungi, was lower in two of the four disturbed plots compared to undisturbed plots; these two disturbed sites had especially low litter levels. Based on ergosterol measurements, mycorrhizas of mature trees did not appear to be negatively affected by trampling. Total fine roots and SOM were higher in the disturbed than in the undisturbed plots at three sites. At the fourth site, fine roots and SOM in the disturbed areas were lower than in the undisturbed areas most probably due to nutrient input following picnic activities. Principal component analysis revealed a close correlation between SOM and fine roots of mature trees as well as litter and seedling biomass. Trampling due to recreational activities caused considerable damage to the vegetation layer and in particular to the beech seedlings and their mycorrhizal fine roots, whereas, roots of mature trees were apparently resilient to trampling.     

Evidence for an interaction between basal medium and plant growth regulatiors during adventitious or axillary shoot formation of cauliflower

Summary Leafy shoots of cauliflower were produced from hypocotyl segments, curd branches (debarassed of existing meristems) and intact curd pieces submitted to various basal media. Adventitious shoot production and axillary bud break exhibited very different nutritive requirements. In each case, an interaction between the plant growth regulators and the composition of the basal medium was discovered. Propagation from curd pieces was dependent on the interaction between plant growth regulators and high sucrose-to-macronutrients ratio in the medium. A medium containing 168 mM sucrose, supplemented with 2.5 μM N⁶-benzyladenine and 5 μM naphthaleneacetic acid allowed the production of numerous shoots without any hyperhydric symptoms.     

Precociously germinating rapeseed embryos retain characteristics of embryogeny

We compared the germination of Brassica napus L. embryos at three stages of development-mid-cotyledon, maturation and mature dry-to determine at which stage they acquired the capacity for normal germination and seedling development. Embryos were removed from the seed and cultured on hormone-free medium, allowing them to germinate. The transition from embryogeny to germination was monitored both morphologically and biochemically, using synthesis of 12 S storage protein as a marker of embryogeny. The mature embryos (dry seeds) set the standard for normal seedling development: radicle emergence, hypocotyl extension and cotyledon expansion occurred within 2 d and true leaves were formed within a week of germination. Rocket immunoelectrophoresis indicated that the storage proteins in seedlings from mature dry embryos were completely degraded within a week. In contrast, the midcotyledon-stage embryos appeared to germinate abnormally, retaining many embryonic characteristics. Although the roots emerged, the hypocotyls did not elongate and secondary cotyledons instead of leaves were formed at the shoot apex. Also, the seedlings continued to synthesize and accumulate storage proteins. The maturation-stage embryos did develop into normal-looking seedlings, but complete degradation of storage proteins required several weeks, presumably reflecting continued synthesis and turnover. We conclude that embryogenic and germination-specific processes can occur concurrently and that the capacity to develop as normal seedlings is acquired gradually during the maturation process.Abbreviations dpa days post anthesis - EDTA ethylenedi-aminetetraacetic acid - FW fresh weight     

Sources of variation in rapidly inducible responses to leaf damage in the mountain birch-insect herbivore system

Summary Studies on rapidly inducible resistance in trees against insect herbivores show substantial variation in the strength of responses. Here we report the results of a study which examined causes of this variation. We bioassayed the quality of leaves of two developmental phases (young vs. mature) of the mountain birch Betula pubescens ssp. tortuosa by measuring the growth of two instars of Epirrita autumnata larvae. We used only short shoot leaves from trees of a natural stand, uniform in size and age. Damage was caused by larvae and artificial tearing of leaf lamina, varying the scale and time. We separated seasonal changes in plants from instar-dependent effects of the animals by testing experimental larvae in two subsequent growth trials. We found that only larval-made damage induced responses in leaves that made the leaves significantly poorer quality for the test larvae. Artificial damage induced only weak responses, and artificial canopy-wide damage even caused slight improvement of leaf quality. Cumulative leaf damage did not strengthen birch responses. Leaves that were in the expansion phase responded to damage while fully-expanded, mature leaves showed no response. The pattern of responses indicated that there might be physiological constraints: small-scale damage induced resistance against the larvae but largescale damage did not. Prevalent weather conditions might have modified these responses. Larvae of two instars and sexes, of low- and high-density populations responded to leaf damage similarly. However, prior experience of larvae with the host plant may have affected subsequent larval performance. Variation in rapidly inducible responses in birches was caused by plant characters rather than by test animals.     

Mycorrhizal networks and distance from mature trees alter patterns of competition and facilitation in dry Douglas-fir forests

The distribution of dry Douglas-fir forests in western North America is expected to shift northward with climate change and disappear from the grassland interface in the southern interior of British Columbia. This shift may be accentuated by clearcutting, a common harvesting practice that aims to reduce the competitive effects of residual mature trees on new regeneration, but in so doing, ignores their facilitative effects. In this study, we investigated the net effects of competition from and facilitation by mature trees retained on harvested sites on seedling establishment in the dry interface Douglas-fir forests. We demonstrate that access to a mycorrhizal network (MN) and proximity to trees have important influences on seedling performance. On six sites, we established trenched plots around 24 mature Pseudotsuga menziesii var. glauca (Douglas-fir) trees, then planted Douglas-fir seedlings into four mesh treatments that served to restrict MN access (i.e., planted into mesh bags with 0.5-, 35-, or 250-μm pores, or without mesh) or into impermeable bags (grown in isolation) at four distances (0.5, 1.0, 2.5, or 5.0 m). Seedling survival tended to be greater and water stress lower where seedlings had full access to the MN. Seedling height, shoot biomass, needle biomass, and nutrient uptake peaked at 2.5–5.0 m from mature trees. Seedlings 0.5 m from mature trees had lower CO₂ assimilation rates and wood δ¹³C compared to seedlings 5.0 m away. Competition for soil resources was highest near mature trees but facilitation was relatively greater at further distances, resulting in a zone of net benefit for seedlings. These results show that intraspecific tree-seedling interactions are both competitive and facilitative in dry Douglas-fir forests, and that they are spatially dependent. After disturbance, maintaining residual mature trees may be important for their beneficial regeneration zones. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Factors influencing production of inflorescence-derived somatic seedlings of sweetgum

Somatic embryos derived from staminate inflorescence tissues of three mature sweetgum (Liquidambar styraciflua) trees were tested for germination and conversion frequencies and early growth variables following pregermination cold treatments. Individual mature embryos were selected from repetitively embryogenic cultures maintained on Woody Plant Medium (WPM) with 1 mg l^–1 naphthaleneacetic acid and cultured on basal WPM at 10 8C for 0, 4 or 8 weeks prior to being transferred to WPM germination medium in the light. After 4 weeks, germinated embryos were planted in potting mix in an acclimatization chamber, grown for an additional 8 weeks and evaluated for conversion and growth. Conversion frequency, which ranged up to 80%, was affected by ortet and clonal line, while the number of first-order lateral roots was affected by ortet, clonal line and cold treatment, with 8 weeks of cold promoting the highest number.     

<Emphasis Type="Italic">In vitro</Emphasis> regeneration and bulblet growth from lily bulbscale explants as affected by retardants,sucrose and irradiance

Bulbscales of oriental lily hybrid Star Gazer were used as the explants. Bulblets were formed on the basal portion of the excised bulbscales on MS medium supplemented with growth retardants, different sucrose concentrations and exposed to continuous light or dark. ^{Alar, Cycocel} and Paclobutrazol in concentration 1 mg dm⁻³ produced higher number of bulblets as compared to the control. The number of bulblets, however, decreased with the increase in concentration of the growth retardants. The number of bulblets was higher at 90 than at 60 g dm⁻³ sucrose and when the bulbscales were exposed to continuous light than to darkness. The growth retardants, higher sucrose concentration and continuous dark stimulated fresh mass of bulblets. The number of bulblets having roots and leaves decreased in medium with Alar, Cycocel and Paclobutrazol as compared to the control. A few bulblets produced roots and leaves in medium with 90 g dm⁻³ sucrose and none of the regenerated bulblets produced leaves under continuous dark.     

Developmental changes induced by gamma irradiation in Ipomoea batatas L. Lam (sweet potato).

Storage roots of Ipomoea batatas L. Lam were exposed to gamma irradiation at 90, 180, and 240 Gy. The two highest doses caused a delay in the initiation of rooting and especially of shooting; they also caused formation of rosette-like, stemless shoots without apical meristems. The storage roots irradiated with 90 Gy did form stems, but 50% of them had no apical meristem. The irradiated storage roots produced absorbent roots which developed shoots after 70 days of growth. Nonirradiated storage roots did not differentiate shoots from the absorbent roots. When grown in vitro, phytomers, stem segments with leaves and an axillary meristem, separated from the shoots of irradiated storage roots and exhibited growth aberrations, very intense rooting, and a delay in shooting. Phytomers from nonirradiated normal plants were irradiated with 10, 20, 30, 40, and 90 Gy and grown in a hormone-free medium. The 40- and 90-Gy doses delayed shooting as well as rooting. Only phytomers exposed to 40 and 90 Gy differentiated shoots from the absorbent roots. A stimulation of growth revealed in the accumulation of dry mass was found in the shoots of phytomers irradiated with 10 to 30 Gy. The long after-effects of irradiation as well as possible causes of growth stimulation are discussed.     

Production and distribution of dry matter in trees of Coffea arabica L. in Kenya as affected by seasonal climatic differences and the presence of fruits

Standard growth analysis procedures were used to study the production and distribution of dry matter in 3 1/2 to 5-year-old coffee trees through their first and into their second commercial fruiting year. The trees were growing in the field and were treated according to normal commercial practice. Up to fifteen fruiting and deblossomed trees were harvested on each of seven occasions, at intervals of 63–90 days. The dry weights of four aerial fractions, five root fractions and all fallen, picked and pruned material were recorded. The net assimilation rate (E) of deblossomed trees was as large as that recorded in East Africa for coffee seedlings (0·13 g dm^-2wk^-1). Fruiting trees increased in dry weight faster than deblossomed trees, even when their leaf area was 30% smaller. Their E was up to 0·19 g dm^-2wk^-1when expressed on a leaf area basis and up to 0·16 g dm^-2wk^-1even when the total green fruit surface area was included in the calculation. In the hot, dry season, January–February 1968, all parts of the trees increased in dry weight relatively slowly, except the thin roots (< 3 mm diameter) which, in this season, took about 10% of the dry weight increment. The trees had been pruned in December and E of deblossomed trees was only 0·09 g dm^-2wk^-1. Flushes of shoot growth occurred at the beginning of the Long Rains 1967 and 1968 (February–March). During the 1967 shoot growth flush, 61% of the dry weight increment was used in the production of new, large leaves with a mean specific leaf area of over 140 cm²g^-1. The thin roots took about 10 % of the increment, but the thicker roots increased in weight very slowly. The branches and trunk extended rapidly, but their radial growth was relatively slow. Following the shoot growth flushes, the leaf area ratio of the trees was large (0·38 dm²g^-1in 1967) and during the Long Rains (April–June 1967, 1968), when conditions were favourable for photosynthesis (E 0·13 g dm^-2wk^-1), all parts increased in dry weight relatively rapidly, although in 1968 the thin roots took as little as 3 % of the total increment. Leaf area ratio decreased during the Long Rains owing to a large decrease in specific leaf area (in 1967: 118 to 95 cm²g^-1). Pruning was carried out in June and new leaf production during the cool, dry season, July to mid-September, was very slow. Consequently, the total dry weight increase of the trees during this season was relatively small, although the E of deblossomed trees was 0·13 g dm^-2wk^-1. The leaves, which took 33 % of the increment, decreased further in specific leaf area (to 83 cm²g^-1) and most root fractions increased in dry weight rapidly, the thin ones taking 17 % of the increment. Light fruiting in 1967 did not affect the seasonal periodicity in growth described. Both light and heavy fruiting tended, eventually, to lessen the dry weight increase of leaves and thin roots proportionately more than that of the trunk and thick roots. In 1968 fruiting trees retained over 8000 fruits per tree, which took over 70 % of the dry weight increment during the 1968 Long Rains, and became 36 % of the trees' dry weight. Some rootlets on these trees decreased in weight. Aspects of the productivity, growth periodicity and fruiting of coffee are discussed, and some management implications are noted.     

The occurrence of nitrate reductase in apple leaves

Nitrate reductase utilizing NADH or reduced flavin mononucleotide (FMNH₂) as electron donor was extracted from the leaves, stems and petioles, and roots of apple seedlings. Successful extraction was made possible by the use of insoluble polyvinylpyrrolidone (Polyclar AT) which forms insoluble complexes with polyphenols and tannins. The level of nitrate reductase per gram fresh weight was highest in the leaf tissue although the nitrate content of the roots was much higher than that of the leaves. Nitrite reductase activity was detected only in leaf extracts and was 4 times higher than nitrate reductase activity. Nitrate was found in all parts of young apple trees and trace amounts were also detected in mature leaves from mature trees. Nitrate reductase was induced in young leaves of apple seedlings and in mature leaves from 3 fruit-bearing varieties. An inhibitor of polyphenoloxidase, 2-mercaptobenzothiazole was used in both the inducing medium and the extracting medium in concentrations from 10⁻³ to 10⁻⁵m with no effect upon nitrate reductase activity.     

Somatic Mutation Analysis in Salix suchowensis Reveals Early-Segregated Cell Lineages

Long-lived plants face the challenge of ever-increasing mutational burden across their long lifespan. Early sequestration of meristematic stem cells is supposed to efficiently slow down this process, but direct measurement of somatic mutations that accompanies segregated cell lineages in plants is still rare. Here, we tracked somatic mutations in 33 leaves and 22 adventitious roots from 22 stem-cuttings across eight major branches of a shrub willow (Salix suchowensis). We found that most mutations propagated separately in leaves and roots, providing clear evidence for early segregation of underlying cell lineages. By combining lineage tracking with allele frequency analysis, our results revealed a set of mutations shared by distinct branches, but were exclusively present in leaves and not in roots. These mutations were likely propagated by rapidly dividing somatic cell lineages which survive several iterations of branching, distinct from the slowly dividing axillary stem cell lineages. Leaf is thus contributed by both slowly and rapidly dividing cell lineages, leading to varied fixation chances of propagated mutations. By contrast, each root likely arises from a single founder cell within the adventitious stem cell lineages. Our findings give straightforward evidence that early segregation of meristems slows down mutation accumulation in axillary meristems, implying a plant “germline” paralog to the germline of animals through convergent evolution.     

Regeneration of plants from Antirrhinum majus L. callus

Somaclone production in Antirrhinum majus plants by regeneration of plants from callus cultures has been achieved using three types of explant tissue. Regeneration from mature stem internode-derived callus was extremely poor. Callus derived from seedling shoot tips could be induced to form new shoots in six of seven cultivars tested. Regeneration was achieved in all seven cultivars when callus was produced from segments of hypocotyl and was most effective using agar-solidified medium containing 0.25 mgl^-1 naphthoxyacetic acid + 10% coconut milk. In this case, five of the cultivars produced shoots directly, one produced leaves from the petioles of which new shoots emerged, and one regenerated plants chiefly through the production of embryoids.