Effect of light on cardenolide production by Digitalis lanata tissue cultures

The influence of light on the accumulation of cardenolides by Digitalis lanata S-1 was studied. Optimal radiant flux for chlorophyll accumulation w     

Cardenolide glycoside production in Digitalis lanata

Leaves of Digitalis lanata metabolize progesterone more rapidly than pregnenolone when applied to the surface of the leaves. Their transformation into digitoxigenin, gitoxigenin, digoxigenin, digitoxin, gitoxin, digoxin and the corresponding cardenolides was demonstrated.     

Embryogenesis and cardenolide formation in tissue cultures of Digitalis lanata

Digitalis lanata strain VII from filament callus grew in small cell colonies in nutrient media with high auxin activity. Upon increasing the cytoki     

Biosynthesis of cardenolides in Digitalis lanata

[8-³H]-Cholesterol was synthesized. A doubly labelled sample of [8-³H, 4-¹⁴C]-cholesterol was administered to Digitalis lanata plants and the cardenolides were isolated. Biosynthesized digitoxigenin and digoxigenin retained all the tritium. Barring the migration of the tritium in biosynthesis the results are interpreted as indicative that neither intermediates with δ7, δ⁷ or δ⁸⁸⁽¹⁴⁾ are participating in the elaboration of cardenolides.     

Structure of cyclohexanone derivatives from Digitalis purpurea

Two cyclohexanone derivatives isolated from the growing tips of Digitalis purpurea were probably 8-oxabicyclo(3.3.1)- 5-hydroxy-4-oxo-non-2-ene (A) and 2-hydroxy-2-(2-hydroxyethyl)-cyclohexanone (B).     

Carbohydrates in Digitalis purpurea at various stages of development

The concentrations of carbohydrates in Digitalis purpurea have been determined at various stages of development. An increase of the carbohydrate content was observed during growth. Glucose was the major carbohydrate component in the leaves and stem, whereas fructose predominated in the inflorescence. The growing tips and other actively growing parts of the plant contained relatively high concentrations of two new compounds.     

Hormonal effects on diosgenin biosynthesis and growth in Dioscorea deltoidea tissue cultures

Analysis of Dioscorea deltoidea tissue cultures grown in the presence of 2,4-D, indole-3-butyric acid, isopentenyladenine, benzyladenine and GA singly and in combination showed that the medium with 2,4-D most consistently favored diosgenin production. GA and high benzyladenine concentrations were toxic.     

Efficient and simple plant regeneration via organogenesis from leaf segment cultures of persimmon (Diospyros kaki thunb.)

Summary An efficient and simple plant regeneration system via organogenesis from leaf segments of persimmon (Diospyros kaki Thunb.) cultivars ‘Fuyu’ and ‘Nishimurawase’ has been developed. The regeneration capacity was influenced by the culture vessels, gelling agents, plant growth regulators, and light conditions. Leaf explants taken from in vitro shoots were cultured on a modified Murashige and Skoog medium (MS1/2N), for 16 wk without transfer to fresh medium. Adventious shoots appeared after 4 and 8 wk in culture of ‘Nishimurawase’ and ‘Fuyu’ tissues, respectively. The culture of leaf explants in Erlenmeyer flasks with medium containing 4 g l⁻¹ agar enhanced shoot formation in comparison to media with increased agar concentrations. Optimal shoot regeneration was obtained with 5 mg l⁻¹ (22.8 μM) zeatin and 0.1 mg l⁻¹ (0.05 μM) indole-3-butyric acid (IBA) for ‘Nishimurawase’, and 10 mg l⁻¹ (45.6 μM) zeatin and 0.1 mg l⁻¹ (0.05 μM) IBA for ‘Fuyn’. Shoot regeneration frequencies in both cultivars were 100%, and shoot numbers per explant reached up to 9.2 for ‘Nishimurawase’ and 2.2 for ‘Fuyu’. Dark incubation during the first 4–5 wk was the most effective condition to successfully influence shoot regeneration in both cultivars. While dark incubation was essential for adventitious shoot formation by ‘Fuyu’, it was only slightly beneficial to ‘Nishimurawase’. More than 80% of the regenerated shoots rooted within 4 wk on hormone-free MS1/2N demium after having been dipped for 30 s in 250 mg l⁻¹ (1.1. mM) IBA solution.     

Adventitious shoot formation from leaf explants of Rhododendron

A protocol for high frequency adventitious shoot regeneration adventitious shoot regeneration from leaf explants of Rhododendron spp. has been developed. The highest percentage of regeneration and the greatest number of shoots were obtained when leaf explants were cultured on Anderson's medium containing 4.9 M IBA and 73.8 M 2iP. Genotypic variation was observed for adventitious shoot regeneration potential among the seven cultivars tested. Regeneration frequencies ranged from 0 to 96%. Lodestar had the highest rate of regeneration after 3 months of culture with 96% shoot regeneration and an average of 14 shoots per explant. Regenerated shoots were rooted in soil in about 2 months. This protocol should be useful in applying gene transfer techniques to Rhododendron improvement.Abbreviations IAA 1-H-indole-3-acetic acid - NAA 1-naphthaleneacetic acid - IBA 1-H-indole-3-butyric acid - 2,4-d 2,4-dichlorophenoxyacetic acid - BA 6-benzyladenine - 2iP N-(3-methyl-2-butenyl)-1-H-purine-6-amine     

Incorporation of 14CO2, into cardenolide and sapogenin steroids of Digitalis purpurea

The relative rates of biosynthesis of cardenolide and sapogenin steroids of Digitalis purpurea were estimated by their uptake of ¹⁴CO₂. The incorporation of label into both groups, although initially slow, indicated that biosynthesis occurs even at the end of the growing season. The sapogenins were produced more rapidly than the cardenolides at this stage of plant development. Within the group of sapogenins, digitogenin, the trihydroxy compound, was produced at a greater rate than the dihydroxy steroid gitogenin. In the case of the cardenolides, the trihydroxy gitoxigenin was produced at a slower rate than the dihydroxy digitoxigenin.     

In vitro plant regeneration from leaf callus of Grindelia robusta Nutt

Abstract

A regeneration protocol from leaf explants of Grindelia robusta Nutt. was developed. The combination of 0.5 mg l^?1 IBA plus 0.5 mg l^?1 or 1 mg l^?1 BA added to Murashige-Skoog (MS) medium resulted in the best callus induction frequency; the combination of 0.4 or 0.9 mg l^?1 BA plus 1.2 mg l^?1 GA₃ resulted in the best shoot regeneration. Rooting was successful on MS medium supplemented with 0.5 mg l^?1 IBA. Hardening of G. robusta plants was accomplished in 30 days with 85% survival rate.     

Effects of cytokinin on adventitious root formation in callus cultures ofVigna unguiculata (L.) walp

Summary Yellowish compact callus, induced from cowpea hypocotyls on Murashige and Skoog(MS) medium (1962) containing 0.2 mg/l(0.93 μM) kinetin and 0.4 mg/l (1.81 μM) 2,4-dichlorophenoxyacetic acid (2,4-D), was subcultured on MS medium containing cytokinin alone, auxin alone, or auxins plus cytokinins in order to determine the effect of cytokinins on root organogenesis in callus cultures. The callus actively proliferated on the same medium but did not show any organogenic activity macroscopically as well as microscopically. On medium with N⁶-benzyladenine (BA) and 1-naphthaleneacetic acid (NAA), the yellowish compact callus first changed to pale green compact callus and then many green spots appeared on its surface under light culture. But the yellowsih compact callus remained yellowish and white spots appeared on its surface in dark culture. These spots gradually became white nodular structures. Adventitious root formation from the nodular structures occurred not only on the same medium, but also on medium with either auxin or cytokinin but not both. Yellowish compact callus on medium with auxin alone was transformed to yellowish friable callus, which did not develop adventitious roots. The yellowish friable callus could gain rhizogenic activity only after morphological modification to pale green compact callus on medium with auxin plus cytokinin. The modified callus did not form adventitious roots on medium with auxins but only with cytokinins. Therefore, it is suggested that cytokinins have stimulating effects on root formation from callus that previously did not show rhizogenic activity on medium with auxins alone. In addition, the rhizogenic potential of cowpea callus was discriminated from that of leaf explants, which formed adventitious roots directly on medium with auxin alone.     

Glucosylation of phenolic compounds by Datura innoxia suspension cultures

Datura innoxia grown in suspension cultures can glucosylate simple phenols. Three isomers of dihydroxybenzene (hydroquinone, resorcinol and catechol) were readily converted into their corresponding mono-β-glucosides. Both salicyl alcohol and salicylaldehyde fed to the cells were transformed specifically to isosalicin instead of salicin. Furthermore, the analysis of the cells treated with salicylic acid suggested the formation of its glucose ester in addition to the corresponding monoglucoside. Feeding experiments showed that the cultured cells possess a remarkably high capacity for glucosylation of hydroquinone, which was totally converted into arbutin within 10 hr after administration. The in vitro glucosylation of hydroquinone carried out by the cell-free extract demonstrated that this enzymic reaction requires the presence of UDPG as a high energy donor of glucose.     

Beneficial effects of activated charcoal on plant tissue and organ cultures

Summary Addition of activated charcoal to the medium for plant tissue cultures improves growth by adsorbing toxic metabolites. This research was supported in part by the National Science Council, Republic of China     

Production of ferruginol by cell suspension cultures of Salvia miltiorrhiza

The time-course production of ferruginol, a diterpene, was examined during growth of Salvia miltiorrhiza in cell culture. Ferruginol was produced in the lag and stationary phases of growth and so production of this secondary metabolite was inversely related to active cell division. The effects of auxins and light on ferruginol production were also examined.     

Variation in pigment production in Lithospermum erythrorhizon callus cultures

Different strains of callus cultures of Lithospermum erythrorhizon showed wide variations in the production of shikonin derivatives. From these cultures, two high pigment-producing strains, whose content of shikonin derivatives are stable and similar to that of intact plant root, have been established by repeated selection.     

Characterization of polyphenols in cell walls of cultured Populus trichocarpa tissues

The amount and composition of cell wall-bound polyphenol (lignin) in cultured Populus trichocarpa tissues which formed numerous xylem elements (xylogenic) or no xylem (non-xylogenic) were compared. Polyphenol accounted for ca 15% of the dry wt of the cell wall and did not differ significantly in amount in xylogenic and non-xylogenic tissues. The syringic acid derivatives, 3,4.5-trimethoxybenzoic acid, was identified as one of the oxidation products of methylated cell walls and was recovered in similar amounts irrespective of xylem formation. In contrast, lignin from xylogenic cultures contained more p-coumaryl alcohol derivatives and less coniferyl alcohol derivatives than lignin from non-xylogenic cultures. In this respect the lignin composition of xylogenic tissues closely resembled that from stems.     

Effects of root restriction and growth regulator treatments on the growth of carrot (Daucus carota L.) seedlings

The growth of carrot (Daucus carota L.) seedlings in clear polystyrene vials (7.5 × 2.5 cm diameter) which were either perforated or unperforated and suspended in a large container containing nutrient solution was compared with that of similar seedlings grown directly in the main container. After 28 days plants grown in the perforated, aerated vials had smaller shoots, tap roots (c 20 percent reduction) and fibrous roots (c 60 percent reduction) than unrestricted plants. In the absence of adequate aeration, the effects of root restriction were greater. Foliar applications of ethephon to the leaves of unrestricted plants reduced shoot weights but had little effect on either tap root or fibrous root weights, or even increased tap root weight at lower concentrations. Silver thiosulphate, aminovinylglycine and a highly mobile cytokinin (SD8339) all slightly enhanced shoot growth of restricted plants but had no effect on either tap roots or fibrous root development. It is suggested that although root restriction effects may be partly mediated by endogenous hormones, it seems unlikely that such effects are due solely to stimulation of ethylene production or a reduction in endogenous cytokinin levels.Abbreviations AVG Aminovinylglycine - BA N⁶-Benzyladenine - Ethephon 2-Chloroethylphosphonic Acid - SD8339 6-Benzylamino-9-(Tetrahydropyran-2-yl)-Adenine - STS Silver Thiosulphate