Evidence that Daminozide, but not Two Other Growth Retardants, Modifies the Fate of Applied Gibberellin A9 in Chrysanthemum morifolium Ramat

Interactions between three growth retardants and gibberellinA₉ (GA₉) in Chrysanthemum morifolium Ramat. cv. Bright GoldenAnne grown in short days have revealed that the particular retardantemployed modifies the response to this gibberellin. When plants were given a foliar spray of daminozide (a substitutedsuccinamic acid), the effects of a subsequent application ofGA₉ to each lateral shoot, on both stem length and the rateof inflorescence development, were relatively small. However,GA₉ was highly active on plants previously sprayed with piproctanylbromide (a quaternary ammonium, piperidinium compound) or onthose given a compost drench of chlorphonium chloride (a quaternaryphosphonium retardant). In the absence of GA₉ all three retardantsreduced stem length and delayed flowering. Irrespective of the extent of the responses to various dosesof GA₉ (1–50 µg per shoot), a close relationshipwas maintained between internode extension and earliness offlowering, as in previous studies with retardants and othergibberellins. On plants which had received chlorphonium chlorideGA₃ and GA₁₃ were highly active and poorly active, respectively. The data suggest that daminozide could act as a retardant notonly by blocking the biosynthesis of gibberellins but also,wholly or partly, by restricting the metabolism or action ofendogenous gibberellins.     

Interactions of the Growth Retardants Daminozide and Piproctanyl Bromide, and Gibberellins A1, A3, A4+7, A5 and A13 in Stem Extension and Inflorescence Development in Chrysanthemum morifolium Ramat

Interactions between the growth retardants daminozide (a substitutedsuccinamic acid) or piproctanyl bromide (a quaternary ammonium,piperidinium compound), and a subsequent application of a singledose (40 µg) of either gibberellin A₁, A₃, A₄₊₇ or A₁₂,showed that, in Chrysanthemum morifolium Ramat. cv. Bright GoldenAnne, a strong interdependence exists between elongation ofthe lateral shoot and the rate of development of its terminalinflorescence. A₁, A₃, and A₄₊₇ were highly active in overcoming the restrictionson both internode extension and the rate of flower-bud developmentimposed by either retardant, suggesting that these two retardanteffects are caused by a deficiency of active gibberellins (GN).In the absence of retardant, A₁, A₃, and A₄₊₇ markedly increasedstem elongation, and flowering occurred earlier than in plantsreceiving neither retardant nor GN. A₁₃ the only 20-carbon GNtested, was much less active, while A₅ had a relatively greatereffect on the time of flowering than on shoot elongation. Thus,it is not necessarily the rate of stem extension which determinesthe rate of inflorescence development. The response to different amounts of A₁, A₃ or A₁₃ (1, 5, 10,20, or 50 µg per shoot) neither suggest that different‘threshold’ levels of a particular GN are requiredto induce increases in either stem elongation or in the rateat which inflorescences develop, nor did a change in the dosegiven lead to any consistent differential effect on these twoprocesses. Chrysanthemum morifolium Ramat., stem extension, inflorescence development, growth retardants, gibberellins     

Size of the Chrysanthemum Shoot Apex in Relation to Inflorescence Initiation and Development

The size of the apical dome of Chrysanthemum morifolium Ramat.at the transition to inflorescence initiation in continuouslight (long days) was not systematically influenced by eitherthe temperature or the irradiance under which the plants weregrown. It was generally 0.26 mm in diameter and c. 3.6 x 10^–3mm³ in volume when the first bract was initiated. The dimensionsof the apical dome of plants in short days were only slightlysmaller at this stage. Similarly, each step in the further developmentof the chrysanthemum inflorescence was associated with a narrowrange of apex sizes, indicating that inflorescence initiationand development are closely related to apex size. Chrysanthemum morifolium Ramat, shoot apex, inflorescence initiation     

Flow Cytometric Determination of Relative Nuclear DNA Contents in Bicellulate and Tricellulate Pollen

Nuclear DNA content in mature pollen was measured with a flowcytometer Pollen of Lilium longiflorum, Dendranthema grandiflora(syn Chrysanthemum monfolium) and Zea mays was chopped and stainedwith the DNA fluorochrome DAPI DNA levels, expressed as arbitraryC values, were compared with those of nuclei isolated from leafor root material of the same plants In mature tricellulate pollen the generative cell is dividedafter second pollen mitosis into two sperm cells Tricellulatepollen from maize and chrysanthemum gave rise to one large 1Cpeak and, only in the case of chrysanthemum, a much smallerone at the 2C level These results suggest that the haploid nucleiof the vegetative as well as both sperm cells in tricellulatepollen are arrested in the G₁ stage of nuclear division Thesmall 2C peak in the case of chrysanthemum probably arose froma fraction of pollen with the sporophytic chromosome number(2n pollen) In contrast to this, mature bicellulate lily pollengave rise to two identical peaks at the 1C and the 2C levelFrom this result it was concluded that in bicellulate pollen,the 1C peak is caused by the signal of the haploid vegetativenucleus arrested in the G₁ stage of nuclear division, whereasthe 2C peak originates from the haploid generative nucleus whichhas already undergone DNA synthesis and is arrested in G₂ Lilium longiflorumThunb, lily, Dendranthema grandiflora Tzelev (syn Chrysanthemum morifolium Ramat ), chrysanthemum, Zea maysL, maize, male gametophytic cells, vegetative cells, generative cells, sperm cells, unreduced pollen, sporophytic cells, relative nuclear DNA contents, replication stage     

Effect of Potato-2 Medium on Anther Culture of Interspecific Rice Hybrids

The effect of two culture media, potato-2 and N₆ supplementedwith kinetin and either 2,4-dichlorophenoxyacetic acid (2,4-D)or -naphthalene acetic acid (NAA) on anther culture responseof two interspecific rice hybrids was studied. While calluscould be successfully induced and plants regenerated from theF₁ of O. saliva x O. rufipogon, the other hybrid, O. salivax O. longistaminala did not respond to the anther culture. Nevertheless,some success in callus induction was achieved when anthers froma few selected F₂ plants were cultured from the latter cross.No interaction effects between the media (potato-2, N₆ and growthhormones (2,4-D and NAA) for anther response to callusing wereobserved. Potato-2 medium proved to be superior to N₆ in termsof increased anther response, early callus induction, multiplecalli formation and also overall green plant regeneration Oryza saliva L., O. rufipogon Griff., O. longistaminata A. Chev. et Roehr, interspecific hybrid, anther culture, potato-2 medium, N₆ medium     

Inflorescence Development in Chrysanthemum morifolium as a Function of Light on the Inflorescence

Covered, developing flower buds of Chrysanthemum morifoliumcv. Bright Golden Anne did not atrophy, although their dry weightwas lower than that of uncovered buds at 21, 28, 35 and 42 dafter the start of short days. This reduced dry weight was primarilydue to a reduction in the dry weight of the bracts, which arephotosynthetically active. The reduction in dry weight was notdue to a decrease in the number of bracts or florets or to alag in development of the covered buds. At 49 d the weight ofboth covered and uncovered buds was not significantly different,although the weight of the covered bracts was still reducedcompared with uncovered bracts. At 28 d uncovered buds fixedabout 40 times more ¹⁴CO₂ than covered buds. Both covered anduncovered buds had the same sink intensity and relative specificactivity, but the first bract had a greater sink intensity andrelative specific activity when covered than when uncovered,owing to photosynthesis by the bract itself. Chrysanthemum morifolium, flower development, assimilate partitioning, light, bract photosynthesis     

Uptake and Distribution of Copper in Chrysanthemum morifolium

The effects of various levels of copper on the uptake and distributionof copper in Chrysanthemum morifolium grown in solution cultureand peat-sand have been examined. Whole plants growing in shortdays were sampled at regular intervals, divided into roots,stem, leaves and lateral shoots, and analysed for copper. Thepartitioning of copper between these tissues showed that a relativelylarge proportion (30–40 per cent) of the total plant copperwas accumulated in the roots of normal plants during the harvestingperiod, compared with approximately 10 per cent in the rootsof copper deficient plants. Whilst the copper content (ug g^–1) of leaves and stemfrom normal plants was negatively correlated with the amountof dry matter produced (P < 0·001), the correspondingcopper deficient tissues showed little variation in copper contentwith increases in tissue dry weight. A more detailed investigationof the copper content of leaves from normal plants showed thatgradients existed within the plant with respect to both leafposition and time of harvest which could be described by a singlecubic surface equation (P < 0·001).     

Physiological activity of a viridicatin derivative, 3-(4-phenylcarbostyriloxy) acetic acid

A carboxymethylene derivative (V-OCH₂COOH) of viridicatin (V-OH)promoted the root growth of rice and sesame seedlings. V-OCH₂COOHhad no known hormonal activities, per se, but did have an inhibitoryeffect on IAA and 2,4-D-induced growth of Avena coleoptile sectionsand of carrot root callus. However, inhibition by VOCH2COOHof 2,4-D-induced growth in carrot root callus was to some extentreversed by increasing the concentration of 2,4-D. V-OCH₂C0OHseemed to competitively inhibit IAA-induced elongation of Avenacoleoptile sections. (Received September 14, 1970; )     

Nitrogen Utilization in N-limited Barley During Vegetative and Generative Growth: III. POST-ANTHESIS KINETICS OF NET NITRATE UPTAKE AND THE ROLE OF THE RELATIVE ROOT SIZE IN DETERMINING THE CAPACITY FOR NITRATE ACQUISITION

Barley (Hordeum vulgare L., cvs Golf, Mette, and Laevigatum)was grown under nitrogen limitation in solution culture untilnear maturity. Three different nitrogen addition regimes wereused: in the ‘HN’ culture the relative rate of nitrate-Naddition (RA) was 0·08 d^–1 until day 48 and thendecreased stepwise to, finally, 0·005 d^–1 duringgrain-filling; the ‘LN’ culture received 45% ofthe nitrogen added in HN; the ‘CN’ culture was maintainedat RA 0·0375 d^–1 throughout. Kinetics of net nitrateuptake were measured during ontogeny at 30 to 150 mmol m^–3external nitrate. V_max (which is argued to reflect the maximuminflux rate in these plants) declined with age in both HN andLN cultures. A pronounced transient drop was observed just beforeanthesis, which correlated in time with a peak in root nitrateconcentration. Similar, but less pronounced, trends were observedin CN. The relative V_max (unit nitrogen taken up per unit nitrogenin plants and day) in all three cultures declined from 1·3–2·3d^–1 during vegetative growth to 0·1–0·7d^–1 during generative growth. These values are in HN andLN cultures 15- to more than 100-fold in excess of the demandset by growth rates throughout ontogeny. Predicted balancingnitrate concentrations (defined as the nitrate concentrationrequired to support the observed rate of growth) were below6·0 mmol m^–3 in HN and LN cultures before anthesisand then decreased during ontogeny. In CN cultures the balancingnitrate concentration increased during grain-filling. Apartfrom the transient decline during anthesis, most of the effectof ageing on relative V_max can be explained in terms of reducedcontribution of roots to total biomass (R:T). The loss in uptakeper unit root weight is largely compensated for by the declinewith time in average tissue nitrogen concentrations. The quantitativerelationships between relative V_max and R:T in ageing plantsare similar to those observed for vegetative plants culturedat different RAs. The data support the contention that the capacity for nitrateacquisition in N-limited plants is under general growth control,rather than controlled by specific regulation of the biochemicalpathway of nitrate assimilation. Key words: Barley, nitrogen concentration, root: total plant biomass ratio, V_max     

Establishment and physiological analyses of photoautotrophic callus cultures of the fern Platycerium coronarium (Koenig) Desv. under CO2 enrichmen

Gametophyte-derived callus cultures of Platycerium coronariumcould be maintained under photoautotrophic conditions on Murashigeand Skoog medium supplemented with 2µM 2,4-dichlorophenoxyaceticacid (2,4-D) and with CO₂ enrichment. Progressive reductionof sucrose from the medium resulted in a reduction in growth,but an increase in total chlorophyll content. When subculturingwas delayed beyond 2 weeks, callus cells differentiated intogametophytes on the medium with 0.2 sucrose and no CO₂ enrichment.Enriching the photoautotrophic cultures on 2µM 2, 4-Dwith 1% CO₂ resulted in about 1.7-fold increase in fresh weightwithin 42 d. Total chlorophyll content was generally higherwith 1% CO₂ enrichment than with 10%. F_v/F_m ratio was higherfor callus on low levels of sucrose (>0.5%) than that onsucrose 1.0%. An increase in autofluorescence of chloroplasts,but not the size, was observed with decreasing sucrose levelsin the medium. Autofluorescence decreased with increase in CO₂from 0.03%. Our data are in agreement with the view that long-termexposure to high levels of decrease in photosynthetic capacity. Key words: Platycerium coronarium, stag's horn fern, autofluorescence of chloroplasts, confocal laser scanning microscope, F_v/F_m ratio, photoautotrophic callus     

Mass Propagation of Eucalyptus camaldulensis in a Scaled-up Vessel Under In Vitro Photoautotrophic Condition

A scaled-up culture vessel was designed for the large-scalephotoautotrophic micropropagation of chlorophyllous plants.The culture vessel (volume 20 l) contained a plug cell traywith 448 plantlets, and had a forced ventilation system to supplyCO₂-enriched air. A nutrient-reservoir was connected to theculture vessel from which nutrient solution was circulated tothe culture vessel every 24 h. Nodal leafy cuttings of Eucalyptuscamaldulensis L. were cultured photoautotrophically in thissystem without sugar in the nutrient medium, but with an enrichedCO₂concentration and a high photosynthetic photon flux. Thegrowth and the net photosynthetic rate of the in vitro grownplantlets and the survival percentage of the plantlets aftertransplanting to ex vitro conditions were compared with thoseof plantlets grown photoautotrophically under natural ventilationin conventional small culture vessels (Magenta-type vessels;volume 0.4 l). Fresh and dry masses and net photosynthetic ratewere significantly higher in plantlets grown in the scaled-upvessel compared to plantlets grown in the conventional smallvessels (control). The environmental conditions created in thisscaled-up vessel (with forced ventilation) also facilitatedacclimatizationin vitro . Importantly, after transplanting tothe ex vitro condition, plantlets grew well without any specializedexvitro acclimatization treatment. Copyright 2000 Annals of BotanyCompany CO₂enrichment, Eucalyptus camaldulensis L., ex vitro, forced ventilation, natural ventilation, photoautotrophic, scaled-up vessel, survival percentage     

Growth and Biomass Allocation, with Varying Nitrogen Availability, of Near-isogenic Pea Lines with Differing Foliage Structure

The single-gene mutation afila in pea (Pisum sativum L.) resultsin the replacement of proximal leaflets with branched tendrils,thereby reducing leaf area. This study investigated whethertheafila line could adjust biomass partitioning when exposedto varying nutrient regimes, to compensate for reduced leafarea, compared with wild-type plants. Wild-type and afila near-isogeniclines were grown in solution culture with nitrate-N added toinitially N-starved seedlings at relative addition rates (R_N)of 0.06, 0.12, 0.15 and 0.50 d^-1. The relative growth rate (R_W)of the whole plants closely matched R_Nat 0.06 and 0.12 d^-1,but higher R_Nresulted in a slightly higher growth rate. At agiven R_N, the wild-type line had lower plant nitrogen statusthan the afila line. R_Wof the roots of the afila line was lessthan R_Wof the roots of the wild-type at the three higher ratesof N supply despite a greater accumulation of N in the rootsof the afila plants. Consequently, plant nitrogen productivity(growth rate per unit nitrogen) was lower for afila. Dry matterallocation was strongly influenced by nitrogen status, but nodifferences in shoot–root dry matter allocation were foundbetween wild-type and afila with the same plant N status. Theseresults imply that decreased leaf area as a result of the single-genemutation afila affects dry matter allocation, but only accordingto its effect on the nitrogen status. Copyright 2000 Annalsof Botany Company Pisum sativum, pea, nitrogen limitation, growth, shoot–root allocation, relative growth rate, nitrogen productivity, isolines     

A comparison between methods used to control nutrient supply

Experimental methods to supply nutrients to culture solutionsin order quantitatively to control plant nutrition are compared.In experiments with tomato and birch plants, for which the dataare available in databases (Ingestad et al., 1994a, b), thenutrients were supplied at constant relative addition rates(R_A over sufficiently long periods of time to achieve acclimatedplants and reliable measurements of plant responses. The plantswere maintained under steady-state conditions, i.e. the internalnutrient concentrations (c₁) remained constant, as a resultof a numerical equality between the relative uptake rate (R_U)and the relative growth rate (R_G). These results are comparedto experiments with pea plants (Macduff et al., 1993). In oneseries (a), R_A was applied, but without strict control of internalsteady-state, and in the other series (b), the external concentration(c_e) was maintained constant. With limiting nitrogen, in bothseries, there was a substantial deviation from equality betweenR_U and R_G. In (a), c_I changed during the experimental periodand the purpose of the R_A approach was lost. In (b), a constantc_e had little effect on nitrogen uptake and plant growth. Atthe three highest concentrations, steady-states were obtainedat non-limiting uptake rates. At the lowest concentration, theuptake rate of nitrogen was about the same, but there was adecrease of R_a, which apparently was not caused by reduced uptake.Clear-cut relationships can not therefore be established betweentreatment variables and plant responses and the conclusionsreached by Macduff et al. (1993) have little support in theirexperimental results. This indicates an urgent need to updateboth theories and experimental methods together: in particular,it is important to identify the system under investigation andto distinguish between control of the medium and control ofthe plant. Key words: Experimental control, external nutrient concentration, non-limiting and limiting nutrient supply, relative addition rate, relative uptake rate, relative growth rate, steady-state     

Effect of Ethylene and Culture Environment on Rice Callus Proliferation

Modifications to the gaseous envelope by callus during culturein Petri dishes were shown to reduce growth and promote necrosisof several rice (Oryza sativa L.) cultivars. Incubatingcallusunder a continuous flow of gas mixtures of known compositionsuggested that the inhibition of growth was caused by the accumulationof ethylene, the depletion of oxygen and, to a lesser extent,the accumulation of carbon dioxide. In order to evaluate theimportance of ethylene accumulation aminoethoxyvinylglycine(AVG), 1-aminocyclopropane-l-carboxylic acid (ACC and silvernitrate (AgNO₃), were added to the nutrient medium and ethylenemeasurements performed during callus culture. Ethylene restrictedcallus growth particularly under high (35 °C) as comparedto moderate (25 °C) temperatures and under illuminated ascompared to darkened incubation. Under illuminated incubationat 25 °C AVG (5 mmol m^–3) and AgNO°(50 mmol m^–3)significantly improvedcallus growth (100 and 60% respectively)while ACC (200 mmol m^–3) significantly decreased growth(40%). AVG and AgNO₃ were less effective under dark incubationat 25 °C where ethylene production was lower. Furthermore,callus growth was significantly better in large as comparedto small culture vessels since the ethylene concentration wasdiluted and more oxygen was available for respiration. Bettercontrol of ethylene and increased oxygen availability couldbe a way ofproducing healthy callus for the formation of embryogenictissues of otherwise recalcitrant cultivars of rice (e.g. IndicaIR42) and may be a way of improving manipulation of other cerealspecies. Key words: 1-Aminocyclopropane-1-carboxylic acid, aminoethoxyvinylglycine, callus, ethylene, Oryza sativa, silver nitrate     

Phase Change in Hedera helix L: II. THE POSSIBLE BOLE OF ROOTS AS A SOURCE OF SHOOT GIBBERELLIN-LIKE SUBSTANCES

When synthesis was estimated by the agar diffusion techniqueboth basal and lateral adventitious roots of Hedera helix L.in the juvenile growth phase were shown to synthesize gibberellin-likesubstances. Seedlings and cuttings from juvenile ivy could begrown in water culture for several weeks. When roots were excisedfrom these plants shoot growth was reduced in comparison withthat of intact plants. The stems, apices, and leaves of derootedseedlings and cuttings contained lower levels of extractablegibberellin-like substances than comparable organs of intactplants. The major zone of gibberellin-like activity in intactplants co-chromatographed with gibberellins A₁ and A₃. Whenthese gibberellins were applied to plants grown in culture theywere found to promote growth of intact but not derooted plants.These findings are discussed in relation to the role of rootfactors and particularly gibberellins in phase change.     

Cold-Tolerant Crop Species Have Greater Temperature Homeostasis of Leaf Respiration and Photosynthesis Than Cold-Sensitive Species

Some plant species show constant rates of respiration and photosynthesismeasured at their respective growth temperatures (temperaturehomeostasis), whereas others do not. However, it is unclearwhat species show such temperature homeostasis and what factorsaffect the temperature homeostasis. To analyze the inherentability of plants to acclimate respiration and photosynthesisto different growth temperatures, we examined 11 herbace-ouscrops with different cold tolerance. Leaf respiration (R_area)and photosynthetic rate (P_area) under high light at 360 µll^–1 CO₂ concentrations were measured in plants grown at15 and 30°C. Cold-tolerant species showed a greater extentof temperature homeostasis of both R_area and P_area than cold-sensitivespecies. The underlying mechanisms which caused differencesin the extent of temperature homeostasis were examined. Theextent of temperature homeostasis of P_area was not determinedby differences in leaf mass and nitrogen content per leaf area,but by differences in photosynthetic nitrogen use efficiency(PNUE). Moreover, differences in PNUE were due to differencesin the maximum catalytic rate of Rubisco, Rubisco contents andamounts of nitrogen invested in Rubisco. These findings indicatedthat the temperature homeostasis of photosynthesis was regulatedby various parameters. On the other hand, the extent of temperaturehomeostasis of R_area was unrelated to the maximum activity ofthe respiratory enzyme (NAD-malic enzyme). The R_area/P_area ratiowas maintained irrespective of the growth temperatures in allthe species, suggesting that the extent of temperature homeostasisof R_area interacted with the photosynthetic rate and/or thehomeostasis of photosynthesis.     

Dynamics of Carbon Photosynthetically Assimilated in Nodulated Soya Bean Plants under Steady-state Conditions 1. Development and Application of 13CO2 Assimilation System at a Constant 13C Abundance

A long-term, steady-state ¹³CO₂ assimilation system at a constantCO₂ concentration with a constant ¹³C abundance was designedand applied to quantitative investigations on the allocationof photoassimilated carbon in nodulated soya bean (Glycine maxL.) plants. The CO₂ concentration in the assimilation chamberand its ¹³C abundance were maintained constant with relativevariances of less than ±0.5 per cent during an 8-h assimilationperiod. At the termination of 8-h ¹³CO₂ assimilation by plantsat early flowering stage, the currently assimilated carbon relativeto total tissue carbon (measured by the degree of isotopic saturation)were for young leaves (including flower buds), 13.9 per cent;mature leaves, 15.7 per cent; stems+petioles, 5.9 per cent;roots, 5.4 per cent and nodules, 6.9 per cent, 48 h after theend of the ¹³CO₂ assimilation period, they were 12.3, 7.5, 7.4,6.8 and 6.1 per cent, respectively. The treatment with a highconcentration of nitrate in the nutrient media significantlydecreased the allocation of ¹³C into nodules. Experiments on¹³CO₂ assimilation by plants at the pod-filling stage were alsoconducted. Labelling by ¹³C was weaker than at the early floweringstage, but an intense accumulation of ¹³C into reproductiveorgans was observed. Glycine max L., nodulated soya bean plants, ¹³CO₂ assimilation, carbon dynamics     

A Model of Flowering in Chrysanthemum

A mathematical model of flowering in Chrysanthemum morifoliumRamat. is described which may be used to predict quantitiessuch as the number of primordia initiated by the apex, plastochronduration and apical dome mass before, during and after the transformationof the apical meristem from vegetative to reproductive development.The model assumes that primordial initiation is regulated byan inhibitor present in the apical dome. Within each plastochronthe apical dome grows exponentially, and the inhibitor concentrationdeclines through chemical decay and dilution. When the inhibitorconcentration falls to a critical level a new primordium isinitiated. There is instantaneous production of inhibitor, anda decrease in dome mass corresponding to the mass of the newprimordium. The process continues until the apical dome attainsa particular mass when the first bract primordium is produced.Subsequent primordia compete with the apical dome for substrates,and the specific growth rate of the dome declines with successiveplastochrons. Eventually, the net mass of the dome starts todecline until it is entirely consumed in the production of floralprimordia. Chrysanthemum morifoliumRamat, flowering, primordial initiation     

Regulation of Tracheary Element Differentiation by Exogenous L-Methionine in Callus of Soya Bean Cultivars

The relationship between the induction of tracheary elementdifferentiation and exogenous L-methionine was examined in agar-growncultures of soya bean callus initiated from Glycine max L. ‘Wayne’and ‘Clark 63’. Although Wayne is a normal cultivarsoya bean, seedlings of Clark 63 exhibit abnormal growth at25 °C due to exessive ethylene biosynthesis at this temperature.Wayne callus showed increased xylogenesis in the presence ofexogenous L-methionine (3.7 µg 1^–1) in comparisonto IAA–KN controls at both 20 and 25 °C. Clark 63callus produced greater numbers of tracheary elements in responseto exogenous L-methionine only at 25 °C. The induction ofxylem differentiation was independent of the maintenance temperatureof the stock cultures of both cultivars. Xylogenesis initiatedbyan IAA–KN medium was inhibited by the addition of AgNO₃(20 mg 1^–1) to the extent of 76.5 per cent in cv. Wayneand 6 per cent in cv. Clark 63. The inhibitory effect was partiallyreversed by the addition of L-methionine (3.7 µg 1^–1)to the IAA–KN–AgNO₂ medium. These data support thehypothesis that xylogenesis in vitro involves auxin, cytokininand ethylene. differentiation, xylogenesis, L-methionine, ethylene, Glycine max L., soya bean, callus culture, auxin, kinetin     

Biosynthesis of furano-terpenes by sweet potato cell culture

Callus was induced from sweet potato root tissue on an agarmedium containing Heller's minerals, vitamins, 2,4-D, yeastextract and sucrose. Furano-terpenes were scarcely detectedin the callus. However, when the callus was transferred to aliquid culture medium and incubated with reciprocal shaking,furano-terpenes were rapidly produced mainly in the culturemedium. Furano-terpene production by the cell culture was suppressedby addition of Ceratocystis fimbriata spores or HgCl₂ to theculture medium. Yeast extract and sucrose in the culture mediumwere important for furano-terpene production. 3-Hydroxy-3-methylglutarylcoenzyme A (HMG-CoA) reductase activity increased in the cells,followed by the production of furano-terpenes. The TLC patternof furano-terpenes produced by the cell culture was essentiallythe same as that produced by sweet potato root tissue infectedby C. fimbriata or treated with HgCl₂, but the quantitativeproportion of the individual furano-terpenes in the former differedmarkedly from that in the latter. (Received January 11, 1979; )