Effect of genotype,explant and medium onin vitro regeneration of red pepper

In vitro plant regeneration was achieved inCapsicum praetermissum, C. baccatum andC. annuum cvs. G4, Bhiwapuri Sweet pepper, Cayenne pepper and Hybrid pepper. Shoots were induced from hypocotyl, cotyledon and leaf explants on Murashige and Skoog medium supplemented with 5.7 M indoleacetic acid (IAA)+13.3 M benzyladenine (BA); 22 M BA; and 44 M BA. Analysis of variance revealed that the most significant effect on shoot regeneration was due to the explant and it accounted for 56.3% of total variation observed. The genotype x explant effect on regeneration was minor relative to all other 2- and 3-way interactions because leaf explants consistently regenerated more shoots than hypocotyls or cotyledons in all the genotypes and thereby reduced the variation among the genotypes. Explant x medium interaction revealed that 22 M BA was the best growth regulator supplement in regeneration medium for optimal shoot regeneration from leaf explants. Rooting of regenerated shoots was achieved on 5.7 M IAA-containing medium, and the rooting response was better from shoots induced on medium fortified with 5.7 M IAA plus 13.3 M BA. Complete plantlets with diploid chromosome number (2n=2x=24) were transferred to soil and 60–70% of these plantlets survived and grew well.     

Swords into plowshares? Nicotiana sylvestris does not use nicotine as a nitrogen source under nitrogen-limited growth

Whether or not a plant can recover its investment of resources in a chemical defense is central to the mobile-immobile metabolite dichotomy of the resource availability theory. Biochemical measures of metabolite turnover have been used to estimate this trait, but they do not address the ecological question of resource recovery. Numerous studies have found that many Nicotiana species, which normally produce the nitrogen-intensive defense metabolite, nicotine, can rapidly take up and metabolize exogenously administered nicotine from hydroponic solutions. However, Baldwin et al. (1994) found no evidence for turnover of endogenously produced nicotine in pulse-chase experiments using ¹⁵NO₃ as the biosynthetic precursor in N. sylvestris. Given that the capacity to metabolize nicotine exists, we asked (1) whether N. sylvestris could metabolize exogenously fed nicotine and sustain growth under nitrogen-limited conditions and (2) whether leaf damage alters the plants' ability to use nicotine as a nitrogen source. We fed plants with sufficient nicotine in hydroponic culture to increase their nitrogen pools by 70% at the time of nicotine feeding; in 6–10 consecutive harvests over 28–35 days, we measured the biomass of roots, leaves and stems, and the total nitrogen pools of these plant parts as well as the pools of nicotine, nornicotine and myosmine of these plant parts in undamaged nicotinefed and control plants and finally, in a separate experiment, in nicotine-fed damaged and undamaged plants. Nicotine feeding increased nicotine pools by 1.2 times, which was not sufficient to significantly increase total nitrogen pools at the end of the experiment. Nicotine-fed plants rapidly demethylated their acquired nicotine pools to nornicotine, but did not process the alkaloid pool further than myosmine over the duration of the experiment. Leaf damage significantly increased the nicotine pool, but did not significantly alter the processing of the exogenously acquired nicotine. We conclude that N. sylvestris does not recover the nitrogen invested in nicotine even under nitrogen-limited growth, that the rapid metabolism of exogenously introduced nicotine is likely a detoxification pathway, and that these plants are homeostatic with regard to their nicotine pools.     

Fast track to the trichome: induction of N-acyl nornicotines precedes nicotine induction in Nicotiana repanda

 Nicotiana repanda Wildenow ex Lehmann acylates nornicotine in its trichomes to produce N-acyl-nornicotine (NacNN) alkaloids which are dramatically more toxic than nicotine is to the nicotine-adapted herbivore, Manduca sexta. These NacNNs, like nicotine, were induced by methyl jasmonate (MeJA) and wounding, but the 2-fold increase in NacNN pools was much faster (within 6 h) than the MeJA-induced increase in nornicotine pools (24 h to 4 d), its parent substrate. When ¹⁵NO₃ ⁻ pulse-chase experiments with intact and induced plants were used to follow the incorporation of ¹⁵N into alkaloids in different plant parts over the plant's lifetime, it was found that the root nicotine pool was most rapidly labeled, followed by the shoot nornicotine and NacNN pools. After 3 d, 3.12% of ¹⁵N acquired was in nicotine (0.93%), nornicotine (0.32%) and NacNNs (1.73%) while only 0.14% was in anabasine. Once NacNNs are externalized to the leaf surface, they are not readily re-distributed within the plant and are lost with senescing leaves. The wound- and MeJA-induced N-acylation of nornicotine is independent of induced changes in nornicotine pools and the rapidity of the response suggests its importance in defense against herbivores. Received: 3 July 1999 / Accepted: 17 September 1999     

The chemistry of defense and apparency in the corollas ofNicotiana attenuata

The morphological and chemical characteristics of flowers which attract pollinators present a dilemma for plants; advertising may increase the apparency of plants to their predators and some pollinators are also predators. We explore how a self-compatible disturbance species,Nicotiana attenuata, copes with this potential dilemma by examining the changes in emission of chemicals from flowers in response to pollination and herbivory. We propose that chemical changes induced by herbivory and pollination reflect the function of the chemicals in the plant. The emission of a single compound, benzyl acetone (BA, 4-phenyl-2-butanone), by flowers increases dramatically (50x) in the evening, peaking just after dark —a pattern of emission characteristic of moth-pollinated flowers. Pools of BA were found only in the outer lip of the corolla where pollinators come in contact with the flower, and diurnal changes in the size of the corolla pool closely paralleled the amount emitted by intact flowers throughout the day, as determined by headspace sampling. Pollination dramatically decreases both the pools of BA in the corolla and its emission from flowers. Similarly, nicotine, a broadly biocidal defense metabolite and an induced defense in vegetative and reproductive tissues, is also found in the headspace of flowers and is principally localized in the basal parts of the corolla below the attachment of the filaments and the nectar reward. Moreover, the dynamics of the corolla pools of BA and nicotine throughout the day are consistent with their roles in advertisement and defense, respectively. The corolla pools of nicotine are stable throughout the day except during the period of peak BA production and emission when nicotine pools decrease significantly. The coordinated increase in BA emission and decline in nicotine pools are not inexorably linked, because herbivory or mechanical damage to corolla tissue rapidly increases corolla nicotine pools without affecting the increase in BA pools. Similarly, leaf damage results in a slower, systemic increase in corolla nicotine pools during reproductive growth but again does not affect BA pools. Excised flowers emitted BA in a manner similar to that of intact flowers, and excision of a majority of flowers from a plant did not alter the BA emission patterns of the remaining flowers. We conclude that althoughN. attenuata's defensive and advertisement chemistries respond synchronously to some environmental stimuli, the flowers' chemical responses to pollinators and herbivoresare distinct and the differences reflect their ecological roles. We propose that the cost-benefit framework of the optimal defense and apparency theories can be fruitfully applied to the allocation of defense metabolites and floral volatiles that function in pollinator attraction, and that this framework can be readily tested by manipulating the patterns of the emissions of plants in the field.     

Seasonal changes in net photosynthesis rates and photosynthetic capacity in leaves of Cistus salvifolius,a European mediterranean semi-deciduous shrub

Summary During five different periods between Nov. 1982 and Aug. 1983, the diurnal patterns exhibited in photosynthetic CO₂ uptake and stomatal conductance were observed under natural conditions on twigs of Cistus salvifolius, a Mediterranean semi-deciduous shrub which retains a significant proportion of its leaves through the summer drought. During the same periods, net photosynthesis at saturating CO₂ partial pressure was measured on the same twigs as a function of irradiance at different temperatures. From these data, photosynthetic capacity, defined here as the CO₂- and light-saturated net photosynthesis rate, was obtained as a function of leaf temperature. C. salvifolius is a winter growing species, shoot growth being initiated in Nov. and continuing through May. Photosynthetic capacity was quite high in Nov., March and June, exceeding 40 mol m^-2 s^-1 at optimum temperature. In Dec., photosynthetic capacity was somewhat reduced, perhaps due to low night-time temperatures (<5°C) during the measurement period. In Aug., capacity in oversummering shoots at optimum temperature fell to less than 8 mol m^-2 s^-1, due to water trees and perhaps leaf aging. Seasonal changes in maximal photosynthetic rates under ambient conditions were similar, and like those found in co-occurring evergreen sclerophylls. Like the evergreens, Cistus demonstrated considerable stomatal control of transpirational water loss, particularly in oversummering leaves. During each measurement period except Aug. when capacity was quite low, the maximum rates of net photosynthesis measured under ambient conditions were less than half the measured photosynthetic capacities at comparable temperatures, suggesting an apparent excess nitrogen investment in the photosynthetic apparatus.     

Autotoxicity and chemical defense: nicotine accumulation and carbon gain in solanaceous plants

We quantified the accumulation of and tolerance to exogenously-fed nicotine by monitoring photosynthetic capacity and growth in two nicotine producing species of Solanaceous plants (Nicotiana sylvestris andN. glauca) as well as two Solanaceous species (Datura stramonium andLycopersicon esculentum) that do not produce nicotine to examine the relationship between tolerence and the ability to produce nicotine in defensive quantities. SinceN. sylvestris uses nicotine as an inducible defense, we examined whether nicotine tolerance is induced by damage to examine further the relationship between nicotine tolerence and synthesis. All species were grown in a 1 mM nicotine-containing hydroponic solution. Reductions in the photosynthetic capacity of nicotine-fed plants were found in all species tested. Nicotine-producing species showed no greater tolerance as measured by photosynthetic capacity than the two non-producing species. Leaf damage marginally increased the tolerence ofN. sylvestris to exogeneouslyfed nicotine suggesting that photosynthetic tolerance is coordinated with nicotine production in this nicotine-producing species.N. glauca plants regained photosynthetic capacity after their accumulated nicotine was demethylated to form nornicotine. Leaf nicotine pools in the other three species did not decrease, suggesting that for these species alkaloid metabolism does not play a major role in tolerance. Tolerance, as measured by biomass gained, was higher in the two non-producing species than in the nicotine-producing species suggesting that nicotine may also be functioning as a growth regulator. These results do not support the hypothesis that tolerance is as important as biosynthetic ability in determining which species accumulate defensively significant quantities of nicotine.     

Effects of deer,mice and dwarf bamboo on the emergence,survival and growth of <Emphasis Type="Italic">Abies homolepis</Emphasis> (Piceaceae) seedlings

In the subalpine mixed forest of Mt. Ôdaigahara, mid-western Japan, the understory is dominated by dwarf bamboo (Sasa nipponica), which is the major forage of overly populous sika deer (Cervus nippon). In the present study, we monitored the survival and growth of Abies homolepis seedlings over 5years to determine how they responded to the experimental removal of dwarf bamboo and to the exclusion of sika deer and mice (Apodemus argenteus and A.speciosus). Deer and dwarf bamboo reduced the survival of seedlings but had different effects on growth. The stems of seedlings were shorter in the presence of deer, indicating that taller seedlings were apt to be browsed by deer, whereas the diameters of seedlings were smaller in the presence of dwarf bamboo, mainly owing to its shading effect. The presence of mice decreased the number of seedlings germinating in a particular site, but had no effect on seedling survival after germination. There was no significant indirect effect whereby the survival of seedlings was predicted to be facilitated by the decreased biomass of bamboo because of grazing by deer. We supposed that this might be because the direct negative effect of deer was so large as to conceal the positive indirect effect.     

Adventitious shoot regeneration from vegetative shoot apices in pear and putative role of cytokinin accumulation in the morphogenetic process

Adventitious shoot regeneration was obtained from callus produced from main vegetative apices of pear of in vitrogrown shoots of Italian cultivars Spadona and Precoce di Fiorano and wild pear genotypes ISF54 and ISF61. The highest morphogenetic response was obtained on a medium containing 8.8 M 6-benzyladenine, 1.0 M -naphthaleneacetic acid and 250 mg l^–1cefotaxime. The explants were maintained for 30 days in darkness and then transferred to an auxin-free medium and to the light. Histological studies revealed that the new vegetative buds originated from callus that completely altered the morphology of the explant tissues by the 30th day of culture. The in situ localisation of cytokinins, performed using antibodies with marked specificity against zeatin (Z) and isopentenyladenine, revealed an accumulation of Z in the cambiform cells of the leaf primordia and in the shell zone of the new forming buds showing a primary role of this cytokinin in cell differentiation of in vitro pear organogenesis.     

Shoot growth dynamics and photosynthetic response to increased nitrogen availability in the alpine willow Salix glauca

Plants subjected to increases in the supply of resource(s) limiting growth may allocate more of those resources to existing leaves, increasing photosynthetic capacity, and/or to production of more leaves, increasing whole-plant photosynthesis. The responses of three populations of the alpine willow, Salix glauca, growing along an alpine topographic sequence representing a gradient in soil moisture and organic matter, and thus potential N supply, to N amendments, were measured over two growing seasons, to elucidate patterns of leaf versus shoot photosynthetic responses. Leaf-(foliar N, photosynthesis rates, photosynthetic N-use efficiency) and shoot-(leaf area per shoot, number of leaves per shoot, stem weight, N resorption efficiency) level measurements were made to examine the spatial and temporal variation in these potential responses to increased N availability. The predominant response of the willows to N fertilization was at the shoot-level, by production of greater leaf area per shoot. Greater leaf area occurred due to production of larger leaves in both years of the experiment and to production of more leaves during the second year of fertilization treatment. Significant leaflevel photosynthetic response occurred only during the first year of treatment, and only in the dry meadow population. Variation in photosynthesis rates was related more to variation in stomatal conductance than to foliar N concentration. Stomatal conductance in turn was significantly related to N fertilization. Differences among the populations in photosynthesis, foliar N, leaf production, and responses to N fertilization indicate N availability may be lowest in the dry meadow population, and highest in the ridge population. This result is contrary to the hypothesis that a gradient of plant available N corresponds with a snowpack/topographic gradient.     

Effect of CO2 enrichment and nitrogen availability on resource acquisition and resource allocation in a grass,Bromus mollis

Summary The effects of CO₂ enrichment on the growth, biomass partitioning, photosynthetic rates, and leaf nitrogen concentration of a grass, Bromus mollis (C₃), were investigated at a favorable and a low level of nitrogen availability. Despite increases in root: shoot ratios, leaf nitrogen concentrations were decreased under CO₂ enrichment at both nitrogen levels. For the low-nitrogen treatment, this resulted in lower photosynthetic rates measured at 650 l/l for the CO₂-enriched plants, compared to photosynthetic rates measured at 350 l/l for the non-enriched plants. At higher nitrogen availability, photosynthetic rates of plants grown and measured at 650 l/l were greater than photosynthetic rates of the non-enriched plants measured at 350 l/l. Water use efficiency, however, was increased in enriched plants at both nitrogen levels. CO₂ enrichment stimulated vegetative growth at both high and low nitrogen during most of the vegetative growth phase but, at the end of the experiment, total biomass of the high and low CO₂ treatments did not differ for plants grown at low nitrogen availability. While not statistically significant, CO₂ tended to stimulate seed production at high nitrogen and to decrease it at low nitrogen.     

Sediment addition enhances transpiration and growth of Spartina alterniflora in deteriorating Louisiana Gulf Coast salt marshes

Transpiration, leaf conductance, net photosynthesis, leaf growth, above-ground biomass and regeneration of new culms were studied in a rapidly subsiding Spartina alterniflora Lois. salt marsh following the addition at 47 and 94 Kg m^–2 of new sediment. Plant growth was enhanced in response to sediment addition as was evident by a significant increase in leaf area, above-ground biomass production and regeneration of new culms (p 0.05). Leaf conductance and transpiration rates were significantly greater in sediment treated plants than in control plants (p 0.05). Enhanced production of culms per unit area of marsh resulted in increased leaf area which allowed a greater capacity for net photosynthesis and contributed to increases in above-ground biomass of sediment treated plots.     

Lateral bud culture of papaya (Carica papaya L.) for clonal propagation

Lateral buds may be preferred to shoot tips for in vitro propagation of papaya because of its unbranched nature. Proliferating shoot cultures from lateral buds appeared extremely compact with shortened internodes and leaf lamina of the cytokinin level (BAP 2 M) reported for multiple shoot production from shoot tips. ZEA (4 M) and 2iP (8 M) although reduced the proliferation rate, resulted in better growth of the shoot from lateral bud. Rooting was observed with IBA 20 M but plantlets so produced remained stunted.     

Mean labor time of a leaf

The mean labor time of a leaf (hour/day^–1) is defined as the ratio of mean daily photosynthetic rate of a leaf (D_a; molm^–2day^–1) to the mean value of potential hourly photosynthetic rate (6060A_max mol m^–2h^–1) of the leaf. A model was proposed to estimate mean labor time of leaves. Mean labor time was obtained as the product of 24 (hours/day^–1) and the four effects, each of which reduces production of a leaf: diel change in light (Diel Effect), reduction in light during cloudy and rainy days (Cloudy Effect), shading on the focal leaves (Shading Effect), and midday and afternoon depression in photosynthesis (Depression Effect). These four effects were estimated for open grown saplings of alder (Alnus sieboldiana), by measuring instantaneous photosynthetic rate and photon flux density above each leaf. The potential daily photosynthetic rate calculated from diel light condition in a clear day was 46.5% of hypothetical daily photosynthetic rate where maximum instantaneous photosynthetic rate was assumed to last throughout the life of the leaf (Diel Effect). The average of the daily photosynthetic rate considering clear, cloudy and rainy days was 79.7% of the clear day (Cloudy Effect). The photosynthetic rate estimated from light condition on the leaf was 85.6% of that in the open site (Shading Effect). Midday depression reduced the daily photosynthetic rate to 72.1% of the potential daily photosynthetic rate (Depression Effect). The product of the four effects multiplied by 24h gave the estimate of mean labor time of leaves to be approximately 5.5 (h/day^–1).     

Comparative physiology and demography of three Neotropical forest shrubs: alternative shade-adaptive character syndromes

A suite of functionally-related characters and demography of three species of Neotropical shadeadapted understory shrubs (Psychotria, Rubiaceae) were studied in the field over five years. Plants were growing in large-scale irrigated and control treatments in gaps and shade in old-growth moist forest at Barro Colorado Island, Panama. Irrigation demonstrated that dry-season drought limited stomatal conductance, light saturated photosynthesis, and leaf longevity in all three species. Drought increased mortality of P. furcata. In contrast, irrigation did not affect measures of photosynthetic capacity determined with an oxygen electrode or from photosynthesis-CO₂ response curves in the field. Drought stress limited field photosynthesis and leaf and plant survivorship without affecting photosynthetic capacity during late dry season. Leaves grown in high light in naturally occurring treefall gaps had higher photosynthetic capacity, dark respiration and mass per unit area than leaves grown in the shaded understory. P. furcata had the lowest acclimation to high light for all of these characters, and plant mortality was greater in gaps than in shaded understory for this species. The higher photosynthetic capacity of gap-grown leaves was also apparent when photosynthetic capacity was calculated on a leaf mass basis. Acclimation to high light involved repackaging (higher mass per unit leaf area) as well as higher photosynthetic capacity per unit leaf mass in these species. The three species showed two distinct syndromes of functionally-related adaptations to low light. P. limonensis and P. marginata had high leaf longevity (3 years), high plant survivorship, low leaf nitrogen content, and high leaf mass per unit area. In contrast, P. furcata had low leaf survivorship (1 year), high plant mortality (77–96% in 39 months), low leaf mass per unit area, high leaf nitrogen content, and the highest leaf area to total plant mass; the lowest levels of shelf shading, dark respiration and light compensation; and the highest stem diameter growth rates. This suite of characters may permit higher whole-plant carbon gain and high leaf and population turnover in P. furcata. Growth in deep shade can be accomplished through alternative character syndromes, and leaf longevity may not be correlated with photosynthetic capacity in shade adapted plants.     

Thidiazuron-induced in vitro shoot formation from roots of intact seedlings of Albizzia julibrissin

Seedlings of silktree (Albizzia julibrissin Durrazz.) were grown in vitro on MS-media containing B5 vitamins, 3% sucrose, 0.25% phytagel and various concentrations (0.1–10 M) of thidiazuron (TDZ). Addition of TDZ to the culture medium greatly reduced shoot and root elongation but did not influence shoot production from the cotyledonary node or apex. Within 8–10 days the seedling roots split open, formed large masses of callus, and developed green patches which eventually grew into normal shoots while still within the culture medium containing TDZ at 0.1–1.0 M. Such callus and shoot formation did not occur in control cultures lacking TDZ. At higher TDZ concentrations (2.5–10 M), the green patches formed in the callus did not further develop into shoots. Addition of other cytokinins (kinetin, benzylaminopurine, zeatin) to the culture medium also induced some shoot formation from the roots, but higher concentrations than TDZ were required to induce regeneration. Isopentenyladenine failed to induced shoot formation. Following excision and transfer to MS media with or without 4.9 M IBA, the shoots induced by kinetin or benzylaminopurine rooted 4–7 days earlier than those induced by TDZ, but all excised shoots developed into normal rooted plantlets within 3 weeks.Abbreviations TDZ thidiazuron     

Carbon fixation in eucalypts in the field

Summary The rate of CO₂ assimilation at light saturation and an intercellular CO₂ concentration of 350 l l^-1 (photosynthetic capacity), measured in leaves of Eucalyptus pauciflora, E. behriana, E. delegatensis and Acacia melanoxylon, declined over the course of cloudless days under naturally varying environmental conditions as well as under constant optimal conditions for high CO₂ uptake. Since the capacity did not recover during the light period, it was different from the midday depression of gas exchange. The change appeared to be caused neither by the diurnal variation of total leaf water potential, by photoinhibition of redox-reaction centres in photosystems nor by changes in the intrinsic properties of Ribulose-bisphosphate carboxylase-oxygenase. The decline was more pronounced in winter than in summer. It was related to the duration of illumination or the cumulative carbon gain. It was reversible in the following dark phase, and it did not occur on changeable days with short peaks of high light.Despite the decline in photosynthetic capacity, the initial slope of the CO₂ response of net photosynthesis, as obtained at low intercellular CO₂ concentrations, remained constant during the day, but declined at night when photosynthetic capacity recovered. In all cases stomatal conductance varied in parallel with photosynthetic capacity. The relevance of changes in photosynthetic capacity for the intercellular CO₂ concentration is discussed.Abbreviations and symbols A CO₂ assimilation - ABA abscisic acid - A_c350 photosynthetic capacity at c_i=350l l^-1 - c_i intercellular CO₂ concentration - g leaf conductance to water vapour - I photon flux density (irradiance) - P air pressure - P_i inorganic phosphate - R_d net CO₂ release at _* - Rubisco Ribulose-bisphosphate carboxylase-oxygenase - RuBP Ribulose-bisphosphate - T leaf temperature - w leaf-to-air water vapour concentration difference - A/c_i carboxylation efficiency at low c_i - _* light-independent CO₂ compensation point - total leaf water potential     

The activity of nitrate reductase and the pool sizes of some amino acids and some sugars in water-stressed maize leaves

The activity of nitrate reductase and the pool sizes of some amino acids and some sugars were measured in relation to the leaf water potential () of maize leaves. The activity of nitrate reductase was severely inhibited in water-stressed maize leaves. This was not due to substrate shortage or the presence of an inhibitor at reduced leaf water potential. While the typical proteinogenic amino acids valine, tyrosine, leucine and isoleucine were almost undetectable in the leaves of the control plants, their concentrations markedly increased with declining , thus indicating protein degradation. The concentrations of serine, glycine and glutamate increased upon water stress, their total amount in severely stressed leaves ranging 5- to 6-fold higher than the total amount of valine, tyrosine, leucine and isoleucine at this stage of water deficit. The pool sizes of glucose, fructose and sucrose decreased in relation to decreasing . The total amount of organic solutes remained almost constant at least up to a of approx.—1.0 MPa and then dropped to about 50% when reached –1.25 MPa.Abbreviations PCR photosynthetic carbon reduction cycle - PCO photosynthetic carbon oxidation cycle - PAR photosynthetically active radiation     

Uptake and metabolism of benzyladenine during shoot organogenesis in Petunia leaf explants

Benzyladenine (BAP) uptake and metabolism were characterized during the key stages of shoot organogenesis in leaf explants of Petunia MD1. Using leaf explant transfer experiments, it was shown that exposure to 2.2 M BAP for 6, 8 or 10 days induced shoot formation on 27, 80 and 100% of the explants respectively, with a concomitant increase in the number of shoots per explant. BAP uptake and metabolism were characterized in leaf explants after 1, 3, 6 or 10 days exposure to [³H]BAP or 10 days exposure plus an additional 2 days on basal medium (10+2). BAP and 9--D-ribofuranosyl-BAP ([9R]BAP) were detected at days 1 and 3 only. Therefore, the BAP free base was not detectable during the shoot induction period between days 6 and 10, as defined by leaf transfer experiments. The BAP ribotide pool was largest on day 1 and decreased to day 10+2. It is possible that the BAP ribotide pool provided either the active cytokinin itself or acted as a short-term storage form for the active cytokinin in petunia shoot organogenesis. Other metabolites detected in petunia leaf tissue included 7--D-glucopyranosyl-BAP ([7G]BAP), 9--D-glucopyranosyl-BAP ([9G]BAP) and an unidentified metabolite C.Abbreviations BAP benzyladenine - [7G]BAP 7--D-glucopyranosyl-BAP - [9G]BAP 9--D-glucopyranosyl-BAP - [9R]BAP 9--D-ribofuranosyl-BAP - [9R-5P]BAP 5-monophosphate of [9R]BAP - [9R-5PP]BAP 5-diphosphate of [9R]BAP - [9R-5PPP]BAP 5-triphosphate of [9R]BAP - TEA Triethylamine This research was supported in part by NSF Grant DCB-8917378 to J.D.C. and USDA-CRGO Grant 89-37261-4791 to T.J.C.     

Photosynthetic formation of inorganic pyrophosphate in phototrophic bacteria

In this paper we report studies on photosynthetic formation of inorganic pyrophosphate (PP_i) in three phototrophic bacteria. Formation of PP_i was found in chromatophores from Rhodopseudomonas viridis but not in chromatophores from Rhodopseudomonas blastica and Rhodobacter capsulatus. The maximal rate of PP_i synthesis in Rps. viridis was 0.15 mol PP_i formed/(min*mol Bacteriochlorophyll) at 23°C. The synthesis of PP_i was inhibited by electron transport inhibitors, uncouplers and fluoride, but was insensitive to oligomycin and venturicidin. The steady state rate of PP_i synthesis under continuous illumination was about 15% of the steady-state rate of ATP synthesis. The synthesis of PP_i after short light flashes was also studied. The yield of PP_i after a single 1 ms flash was equivalent to approximately 1 mol PP_i/500 mol Bacteriochlorophyll. In Rps. viridis chromatophores, PP_i was also found to induce a membrane potential, which was sensitive to carbonyl cyanide p-trifluoromethoxyphenylhydrazone and NaF.Abbreviations BChl Bacteriochlorophyll - F₀F₁-ATPase Membrane bound proton translocating ATP synthase - FCCP Carbonyl cyanide p-trifluoromethoxyphenylhydrazone - H⁺-PPase Membrane bound proton translocating PP_i synthase - TPP⁺ Tetraphenyl phosphonium ion - TPB^- Tetraphenyl boron ion - Transmembrane electrical potential difference     

Rapid adventitious shoot regeneration from leaf explants of European birch

The goal of this research was to develop a rapid and efficient system for regenerating shoots from leaf explants of European birch, Betula pendula Roth. Single-node stem explants were established in culture, and microshoots were subcultured every 4 weeks through 12 subcultures. Leaves from glasshouse plants or subcultured shoots were excised from stems, cut into approximately 35-mm² pieces, and placed on Woody Plant Medium (WPM) containing different combinations of naphthaleneacetic acid (NAA) (0, 3, 6 or 9 M) and benzyladenine (BA) (0, 7.5, 15 or 22.5 M) in a 4×4 factorial design. The percentage of leaf pieces forming shoots and the number of shoots regenerated per explant were recorded after 4 weeks. Only media containing BA without NAA stimulated shoot formation on leaf explants. Fifteen micromolar BA induced the most shoots to form on leaf explants compared to 30, 45 or 60 M of this cytokinin. In addition, shoot regeneration was enhanced up to four-fold between the first and eleventh subculture. Over 90% of the leaf explants regenerated shoots with an average of 18 buds formed per explant for the eleventh subculture. Almost twice as many explants formed shoots if their adaxial side was in contact with the medium rather than oriented away from it. The ability to regenerate shoots from leaves varied among plants, regardless of stock plant age. This reliable shoot regeneration system can be used for rapid shoot proliferation and potentially for genetic engineering of European birch.