Analytical techniques to recognize inclusion complexes formation involving monoterpenes and cyclodextrins: A study case with (-) borneol, a food ingredient.

Monoterpenes are non-polar secondary metabolites widely used by industry due to their excellent therapeutic, food-ingredient and cosmetic properties. However, their low solubility in water limits their use. In this sense, cyclodextrins (CDs) have been widely used to solve these technological challenges. Thus, this study aims to use (-)-borneol as a monoterpene model to prepare inclusion complexes between ß-CD and hydroxypropyl-ß-CD (HP-ß-CD) through different ways and characterize them in order to choose the best inclusion method to improve physicochemical properties of monoterpenes. To achieve this goal, the samples were prepared by physical mixture (PM), paste complex (PA) and freeze-drying complex (FD) and then, extensively characterized by thermal analysis, Fourier-transform infrared spectroscopy, scanning electron microscopy, size particle, X-ray diffraction and nuclear magnetic resonance. The physicochemical results showed that freeze-drying was more effective to form inclusion complexes between (-)-borneol with both CDs. This research highlights the importance of recognizing the best method to prepare inclusion complexes, including food additives as (-)-borneol, to achieve better results in food preparations.

Control of resin production in Araucaria angustifolia, an ancient South American conifer.

Araucaria angustifolia is an ancient slow-growing conifer that characterises parts of the Southern Atlantic Forest biome, currently listed as a critically endangered species. The species also produces bark resin, although the factors controlling its resinosis are largely unknown. To better understand this defence-related process, we examined the resin exudation response of A. angustifolia upon treatment with well-known chemical stimulators used in fast-growing conifers producing both bark and wood resin, such as Pinus elliottii. The initial hypothesis was that A. angustifolia would display significant differences in the regulation of resinosis. The effect of Ethrel(®) (ET - ethylene precursor), salicylic acid (SA), jasmonic acid (JA), sulphuric acid (SuA) and sodium nitroprusside (SNP - nitric oxide donor) on resin yield and composition in young plants of A. angustifolia was examined. In at least one of the concentrations tested, and frequently in more than one, an aqueous glycerol solution applied on fresh wound sites of the stem with one or more of the adjuvants examined promoted an increase in resin yield, as well as monoterpene concentration (α-pinene, ß-pinene, camphene and limonene). Higher yields and longer exudation periods were observed with JA and ET, another feature shared with Pinus resinosis. The results suggest that resinosis control is similar in Araucaria and Pinus. In addition, A. angustifolia resin may be a relevant source of valuable terpene chemicals, whose production may be increased by using stimulating pastes containing the identified adjuvants.

Distinct modes of adventitious rooting in Arabidopsis thaliana.

The literature describes different rooting protocols for Arabidopsis thaliana as models to study adventitious rooting, and results are generally perceived as comparable. However, there is a lack of investigations focusing on the distinct features, advantages and limitations of each method in the study of adventitious rooting with both wild-type (WT) ecotypes and their respective mutants. This investigation was undertaken to evaluate the adventitious rooting process in three different experimental systems, all using A. thaliana, analysing the same rooting parameters after transient exposure to auxin (indole-3-acetic acid) and control conditions: excised leaves, de-rooted plants and etiolated seedlings. The founding tissues and sites of origin of roots differed depending on the system used, whereas all rooting patterns were of the direct type (i.e., without callus formation). None of the systems had an absolute requirement for exogenous auxin, although rooting was enhanced by this phytohormone, with the exception of de-rooted plants, which had adventitious rooting strongly inhibited by exogenous auxin. Root elongation was much favoured in isolated leaves. Auxin-overproducing mutants could not be used in the detached leaf system due to precocious senescence; in the de-rooted plant system, these mutants had a WT-like rooting response, whereas the expression of the 'rooty' phenotype was only evident in the etiolated seedling system. Adventitious rooting of etiolated WT seedlings in the presence of exogenous auxin was inhibited by exogenous flavonoids, which act as auxin transport inhibitors; surprisingly, the flavonoid-deficient mutant chs had a lower rooting response compared to WT. Although Arabidopsis is an excellent model system to study adventitious rooting, physiological and developmental responses differed significantly, underlining the importance of avoiding data generalisation on rooting responses derived from different experimental systems with this species.

Immunoadjuvant and anti-inflammatory plant saponins: characteristics and biotechnological approaches towards sustainable production.

Saponins can be classified as triterpenoid (C30) or steroidal (C27), based on their carbon nucleus (aglycone). Sugar residues are linked to the aglycone, conferring an amphiphilic nature on these molecules, which is relevant for their biological activities. Saponins include a large variety of molecules that find several applications in pharmacology. Saponins have been shown to display immunoadjuvant, anti-inflammatory, antiplatelet, hypocholesterolemic, antitumoral, anti-HIV, antibacterial, insecticide, fungicide and anti-leishmanial activities. Anti-inflammatory medicines are increasingly demanded to treat various forms of arthritis in aging and obese populations and to help reduce the doses and duration of conventional corticotherapy with less side effects and without immunosuppression. The vaccine market for both human and veterinary uses is close to US$ 15 billion, progressively inflated by the recurrent threat of global pandemics.This paper provides an overview of recent advances (main focus on the last five years) on plant saponins that show anti-inflammatory and/or immunoadjuvant activities: source plants, isolation procedures, mechanism of action and biotechnological approaches towards sustainable production of bioactive saponins. Special attention is given to ginseng and Quillaja saponins. Strategies based on plant cultivation, cell and tissue culture, elicitation, and metabolic engineering for improved production of saponins are described. Future directions for research in the field and strategies to overcome bottlenecks are also discussed.

Parâmetros de fermentação de silagens de cana-de-açúcar submetidas a diferentes tratamentos / Fermentation parameters of sugar cane silages submitted to different treatments

Os valores de materia seca (MS), pH, teores de N-NH3/NT, ácidos orgânicos e etanol, de silagens de cana-de-açúcar foram determinados, em delineamento experimental em blocos ao acaso. Os tratamentos aplicados à silagens foram: sem aditivo, testemunha (T); 0,5 por cento de uréia (U); 0,5 por cento de zeólita (Z); 0,5 por cento de uréia mais 0,5 por cento de zeólita (U+Z); inoculante bacteriano/enzimático Bio Max® cana (IBB) e inoculante bacteriano Silobac® (IBS). As silagens foram analisadas aos 56 dias após ensilagem. Os tratamentos U e U+Z apresentaram concentrações médias de ácido lático mais altas que o T, respectivamente 8,9 e 4,7g por cento. Os teores de etanol das silagens aditivadas foram semelhantes aos do tratamento T, respectivamente 12,4 e 12,9g por cento, sugerindo que não houve redução da fermentação alcoólica no material tratado. Com exceção da uréia, nenhum dos outros aditivos utilizados neste experimento promoveu melhoria de parâmetros qualitativos das silagens avaliadas. O efeito benéfico da uréia sobre a qualidade da silagem de cana parece ter sido limitado, não suprimindo a ocorrência do processo de fermentação alcoólica no material.

The values of dry matter (DM), pH, amoniacal nitrogen/total nitrogen (N-NH3/NT), organic acids, and ethanol of sugar cane silages were determined using a randomized blocks design. The treatment were: no addtitive (control); 0.5 percent urea (U); 0.5 percent zeolite (Z); 0.5 percent urea plus 0.5 percent zeolite (U+Z); Bio Max® (IBB), and Silobac® (IBS). Silages were analyzed on the 56th day after ensiling. The U and U+Z treatments showed high latic acid concentrations in relation to control, respectively 8.9 and 4.7g percent. The average ethanol contents of the treated silages were similar to control, respectively 12.4 and 12.9g percent, suggesting no reduction of the alcoholic fermentation process in treated materials. Additives used in this experiment did not enhance the qualitative parameters of the silages except for urea treated silages. It was concluded that the beneficial effect of urea on the quality of the sugar cane silages in the conditions of this experiment is limited, not suppressing the occurrence of the alcoholic fermentation process in the material.

Psychollatine, a glucosidic monoterpene indole alkaloid from Psychotria umbellata.

A monoterpene indole alkaloid, psychollatine ( 1), was isolated from Psychotria umbellata leaves. Its structure was characterized by interpretation of spectroscopic data and by comparison of its NMR data with those of croceaine A ( 2) from Palicourea crocea. The configuration of psychollatine ( 1) was established by NOE difference and circular dichroism (CD) techniques, while its conformation was evaluated through molecular modeling studies and NMR coupling constants.

Características de fermentação da silagem de cana-de-açúcar tratada com uréia, zeólita, inoculante bacteriano e inoculante bacteriano/enzimático / Fermentation of sugarcane silage treated with urea, zeolita, bacteria inoculant and bacteria/enzymatic inoculant

Estudaram-se as características de fermentação da cana-de-açúcar (RB72454) submetida aos tratamentos: controle; 0,5 por cento uréia; 0,5 por cento zeólita; 0,5 por cento uréia e 0,5 por cento zeólita; inoculante bacteriano comercial; inoculante bacteriano/enzimático comercial. O material ensilado foi aberto com um, três, cinco, sete, 14, 28 e 56 dias de fermentação e analisado quanto aos teores de MS, carboidratos solúveis, PB, NNH3/NT, pH, FDN, FDA, celulose, hemicelulose, lignina e DIVMS. Observou-se redução do conteúdo de MS em todas as silagens, com média de 21,1 por cento, em relação ao material original (28,7 por cento). A concentração média de carboidratos solúveis no material original foi de 19,7 por cento e, após 56 dias de fermentação, foi de 0,92 por cento. A concentração de PB das silagens testemunha e tratadas com zeólita, inoculante comercial bacteriano e inoculante comercial bacteriano/enzimático variou entre 2,1 por cento e 3,1 por cento e naquelas que receberam uréia e uréia+zeólita foi de 8,4 por cento. Os teores de NNH3/NT foram inferiores a 10 por cento nas silagens testemunha e tratadas com zeólita, inoculante bacteriano comercial e inoculante bacteriano/enzimático comercial, entretanto foi de 30,4 por cento e 31,1 por cento nas silagens com uréia e uréia+zeólita, respectivamente. No primeiro dia de fermentação, o pH apresentou média de 3,75. Após 56 dias de fermentação, as concentrações de FDN, FDA, celulose e hemicelulose aumentaram, apresentando média entre os tratamentos de 68,6 por cento, 39,6 por cento, 34,5 por cento e 29,1 por cento, respectivamente. O coeficiente de DIVMS reduziu-se com a ensilagem, em todos os tratamentos avaliados, sendo de 57,6 por cento no material original e média de 47,6 por cento nas silagens.

The characteristics of fermentation of sugarcane (RB72454) submitted to the treatments control; 0.5 percent urea; 0.5 percent zeolita; 0.5 percent urea and 0.5 percent zeolita; commercial bacterial inoculant and commercial bacterial/enzymatic inoculant were studied. The material stored in silos was opened on 1, 3, 5, 7, 14, 28 and 56 days of fermentation and was analysed for grades of MS, soluble carbohydrates, PB, NNH3/NT, pH, FDN, FDA, cellulosis, hemicellulosis, lignin and DIVMS. The reduction of the MS content of the silos was observed, with average of 21.1 percent compared to the original material (28.7 percent). The concentration of carbohydrates soluble on the original material was 19.7 percent and, after 56 days of fermentation, it was 0.92 percent. The concentration of PB in the control silage and treated with zeolita, commercial bacterial inoculant and commercial bacterial/enzymatic inoculant varied between 2.1 percent and 3.1 percent and in thoses that received urea and urea+zeolita was 8.4 percent. The grades of NNH3/NT were lower than 10 percent in the control silage and treated with zeolita, commercial bacterial inoculant and commercial bacterial/enzymatic inoculant. However, they were 30.4 percent and 31.1 percent in the silage with urea and urea+zeolita, respectively. On the first day of fermentation, the pH presented average of 3.75. After 56 days of fermentation, the concentration of FDN, FDA, cellulosis and hemicellulosis increased, presenting averages considering the treatments of 68.6 percent, 39.6 percent, 34.5 percent and 29.1 percent, respectively. The coefficient of DIVMS reduced with the fermentation, in all the evaluated treatments, being of 57.6 percent in the original material and averaging 47.6 percent in the silages.

Chemical and mechanical changes during leaf expansion of four woody species of dry Restinga woodland.

Young leaves are preferential targets for herbivores, and plants have developed different strategies to protect them. This study aimed to evaluate different leaf attributes of presumed relevance in protection against herbivory in four woody species (Erythroxylum argentinum, Lithrea brasiliensis, Myrciaria cuspidata, and Myrsine umbellata), growing in a dry restinga woodland in southern Brazil. Evaluation of leaf parameters was made through single-point sampling of leaves (leaf mass per area and leaf contents of nitrogen, carbon, and pigments) at three developmental stages and through time-course sampling of expanding leaves (area and strength). Leaves of M. umbellata showed the highest leaf mass per area (LMA), the largest area, and the longest expansion period. On the other extreme, Myrc. cuspidata had the smallest LMA and leaf size, and the shortest expansion period. Similarly to L. brasiliensis, it displayed red young leaves. None of the species showed delayed-greening, which might be related to the high-irradiance growth conditions. Nitrogen contents reduced with leaf maturity and reached the highest values in the young leaves of E. argentinum and Myrc. cuspidata and the lowest in M. umbellata. Each species seems to present a different set of protective attributes during leaf expansion. Myrciaria cuspidata appears to rely mostly on chemical defences to protect its soft leaves, and anthocyanins might play this role at leaf youth, while M. umbellata seems to invest more on mechanical defences, even at early stages of leaf growth, as well as on a low allocation of nitrogen to the leaves. The other species display intermediate characteristics.

Analgesic and anti-inflammatory activity of a crude root extract of Pfaffia glomerata (Spreng) Pedersen.

The anti-inflammatory and antinociceptive effects of the crude hydroalcoholic extract (PE) of Pfaffia glomerata roots was assessed in the carrageenan-induced rat paw edema at the doses of 100, 200 and 300 mg/kg, using different animal models. An anti-inflammatory dose effect response correlation of r=0.997 and Y=11.67x+0.02 was found. At the same doses, the extract-inhibited acetic acid-induced writhing in mice, but no dose response correlation was found. Oral administration of 100 mg/kg of PE and 0.5 mg/kg of dexamethazone inhibited by 29 and 61%, the granulomatous tissue formation (p>0.05), respectively. These results indicate the potential of this plant extract to treat chronic inflammation. At the assayed doses no significant activity was found in the hot plate test, as well as in the cell migration-induced by carrageenan.

Control of development and valepotriate production by auxins in micropropagated Valeriana glechomifolia.

Valeriana glechomifolia is a plant species endemic to southern Brazil that accumulates valepotriates, which are terpene derivatives, in all of its organs. Valepotriates are the presumed sedative generic components of the pharmaceutically used species of Valeriana. The influence of various concentrations of the auxins indole-3-acetic acid, indole-3-butyric acid and alpha-naphthaleneacetic acid on the growth of micropropagated V. glechomifolia was investigated under conditions of transient and continuous exposure. Changes in the development of roots and shoots as well as the production of the valepotriates acevaltrate, valtrate and didrovaltrate (analyzed by high-performance liquid chromatography) were evaluated. The best performance in valepotriate production, growth and survival under ex vitro conditions following plant acclimatization was achieved in the continuous presence of 5.71 microM IAA. When cultured in medium containing IAA plants produced stable levels of valepotriates throughout the entire cultivation period.

Evaluation of the trypanocidal and leishmanicidal in vitro activity of the crude hydroalcoholic extract of Pfaffia glomerata (Amarathanceae) roots.

Three different concentrations (1, 10 and 50 microg/ml) of lyophilized hydroalcoholic crude extract of Pfaffia glomerata roots were assayed in vitro against strains of Trypanosoma cruzi (Y) and Leishmania braziliensis. It was observed that P. glomerata hydroalcoholic extract was relatively active within the tested concentrations for L. (V) braziliensis, but inactive against T. cruzi. Despite the fact that both protozoans belong to the Trypanosomatidae family, we suggest that the difference observed for activity should be related to the biological differences between the two parasite species.

Sinonasal tract seromucous adenocarcinomas: a report of 12 cases.

Sinonasal seromucous adenocarcinomas may originate from the surface epithelium or from the submucosal glands. We reviewed the clinicopathologic material from 12 patients with sinonasal tract seromucous adenocarcinomas at the University of Texas M. D. Anderson Cancer Center (Houston, TX). There were nine men and three women age 30 to 87 years (mean age, 56.3 years). The clinical presentation included nasal obstruction, nasal mass, and epistaxis. Eight tumors were located in the nasal cavity, three in the ethmoidal sinuses, and one involved the nasal cavity and ethmoid. Histologically, in nine cases the neoplastic glands were lined by a single cell type, arranged back to back without intervening stroma and often inducing desmoplastic reaction. The remaining three tumors also had a cribriform and papillary pattern. All patients were treated by surgical resection. Three patients had recurrences, which occurred at 36, 36, and 48 months after initial therapy. Their treatment involved surgery and irradiation. Eleven patients are alive and free of disease at 36 to 108 months after diagnosis. One patient died 48 months after diagnosis of another cause. Sinonasal tract seromucous adenocarcinomas arise purely from submucosal seromucous glands. The diagnosis is facilitated by their anatomic location, the absence of tumor within the mucosal surface epithelium, and the striking similarity to terminal tubules of the seromucous glands.

The ability of Bipolaris sorokiniana to modify geraniol and (-)-alpha-bisabolol as exogenous substrates.

The biocatalytic potential of Bipolaris sorokiniana was investigated in its ability to modify the monoterpene geraniol and the sesquiterpene alpha-bisabolol as exogenous substrates, using phosphate buffer as reaction medium. The cultures showed a promising oxidative profile, with conversion of geraniol to 6-methyl-5-hepten-2-one (74.9% yield) in a 5-day incubation and alpha-bisabolol to bisabolol oxide B (84.2% yield), in a 7-day incubation.

Qualidade das silagens de seis genótipos de sorgo (Sorghum bicolor (L. ) Moench) em diferentes estádios de maturaçäo / Quality of silages of six sorghum genotipes in different stages of maturation

Com o objetivo de avaliar a qualidade de silagens de sorgo, realizou-se o plantio, a colheita e a ensilagem de seis genótipos dessa forrageira utilizando-se delineamento inteiramente ao acaso com 18 tratamentos, sendo seis híbridos e três épocas de corte (leitoso, corte 1, pastoso, corte 2 e farináceo, corte 3) e quatro repetições. O material original foi analisado quanto ao teor de matéria seca (MS). Na abertura dos silos determinaram-se o pH e o N-NH3/NT no suco das silagens. As silagens foram analisadas quanto aos níveis de proteína bruta (PB), MS, frações fibrosas e teor de ácidos orgânicos. Os genótipos de sorgo avaliados foram: BR 601, BR 303, BR 304, AG 2006, BR 700 e BRS 701. Todos os híbridos apresentaram diferenças no teor de matéria seca no material original entre alguns cortes (P<0,05). Para essa característica, houve diferenças entre híbridos (P<0,05) para um mesmo corte. Quanto ao teor de MS da silagem, todos os híbridos apresentaram diferenças entre os cortes 1 e 2 e entre os cortes 1 e 3. Apenas os híbridos AG 2006 e BR 700 apresentaram diferenças entre os corte 2 e 3. Houve diferenças entre híbridos (P< 0,05) quanto ao teor de MS em um mesmo corte. O menor valor numérico para essa característica foi obtido no corte 1 do BR 601 (20,8 por cento), e o maior valor foi para o BR 304 no corte 3 (49,1 por cento). Os valores de pH do suco da silagem variaram entre 3,6 e 3,9. Os híbridos BR 303 e BR 304 apresentaram os maiores teores de PB. Conclui-se que os indicadores de padröes de fermentaçäo (pH, N-NH3/NT, e teores de ácidos orgânicos) demonstram que as silagens em todos os tratamentos apresentaram bom padräo de fermentaçäo final. De maneira geral, näo houve alteraçäo das frações fibrosas com o avanço do estádio de maturaçäo. Quanto aos teores de matéria seca, todos os híbridos devem ser ensilados no estádio leitoso, exceto o BR 601, que deve ser ensilado no estádio de pastoso ou farináceo

Digestibilidade in situ das frações fibrosas de silagens de seis genótipos de sorgo (Sorghum bicolor (L. ) Moench) em diferentes estádios de maturaçäo / In situ digestibility of fibrous fractions of six sorghum genotipes in different stages of maturation

Com o objetivo de avaliar a digestibilidade in situ das frações fibrosas de silagens de seis híbridos de sorgo em diferentes estádios de maturaçäo, foram utilizados quatro bovinos machos, fistulados no rúmen, castrados, mestiços Holandês-Zebu, alimentados com feno de gramínea à vontade. Amostras de silagens secas e moídas foram colocadas em sacos de náilon que foram introduzidos nas fístulas dos bovinos. Após períodos de incubaçäo de 6, 12, 24, 48, 72 e 96 horas, os sacos foram removidos e as amostras de um mesmo híbrido, animal e período de incubaçäo foram transformadas em um "pool" homogêneo, moído e armazenado para análises posteriores das frações fibrosas. O delineamento experimental utilizado foi o de parcelas sub-subdivididas no qual as parcelas eram as silagens de híbridos, as sub-parcelas os estádios de maturaçäo e as sub-sub-parcelas os períodos de incubaçäo. No corte 1, exceto nos tempos de incubaçäo de 6, 24 e 96 horas, os híbridos näo diferiram quanto ao desaparecimento médio de FDN. No corte 2 houve diferenças entre híbridos apenas no tempo de 12 e 96 horas e no corte 3, apenas no tempo de 12 horas. Em todos os híbridos registrou-se estabilizaçäo do processo fermentativo a partir de 72 horas, exceto nos híbridos BR 304 nos cortes 1 e 3 e BRS 701 no corte 3. Näo houve diferenças entre híbridos quanto ao desaparecimento médio de FDA em nenhum dos tempos estudados, exceto para o tempo de 96 horas no corte 1 e no tempo de 72 horas no corte 3. As curvas de degradaçäo demonstraram tendência de estabilizaçäo do processo fermentativo a partir de 72 horas, à exceçäo dos híbridos BR 304 no corte 1 e BRS 701 no corte 3. A participaçäo de gräos nas silagens näo influenciou o desaparecimento das frações fibrosas. Entretanto, houve influência do estádio de maturaçäo das plantas sobre sua degradabilidade efetiva, favorecendo os cortes mais precoces

Distinct effects of auxin and light on adventitious root development in Eucalyptus saligna and Eucalyptus globulus.

Adventitious rooting is essential for vegetative propagation of woody species. We studied the effects of auxin and light on the development of adventitious roots in cuttings obtained from seedlings of Eucalyptus saligna Smith and E. globulus Labill in an attempt to characterize the adventitious rooting process and identify factors controlling rhizogenesis. Root development was scored as rooting percentage, root density (roots per rooted cutting), mean rooting time and root length. In both species, rooting time was reduced in the presence of auxin. Cuttings from 2-month-old E. saligna seedlings were responsive to lower auxin concentrations than comparable cuttings from E. globulus seedlings. Cuttings from 3-month-old E. saligna seedlings rooted promptly and rooting was not significantly affected by light conditions. In contrast, rooting of cuttings from 3-month-old E. globulus seedlings exhibited recalcitrant behavior and no roots were formed if illuminated during the root formation phase. Effective root regeneration of E. globulus cuttings was obtained by a 4-day exposure to 10 mg l(-1) IBA and culture in darkness during the root formation step. Loss of rooting capacity with seedling age was more pronounced in E. globulus than in E. saligna. The possibility of switching adventitious rooting off and on by manipulating light regime and exogenous auxin supply in E. globulus, and the constitutive nature of rooting in E. saligna may provide useful models for examining the rooting process at the biochemical and molecular levels in Eucalyptus.

Brachycerine, a novel monoterpene indole alkaloid from Psychotria brachyceras.

Brachycerine (1), an unusual alkaloid from the leaves of Psychotria brachyceras, was characterized through spectroscopic data interpretation and its stereochemistry established by NOE difference techniques. Brachycerine (1) was found to be restricted to shoots in rooted cuttings of P. brachyceras (0.018 +/- 0.004% dry weight), and accumulation was unaffected by root induction treatment with auxin.

Improved taxol yield by aromatic carboxylic acid and amino acid feeding to cell cultures of taxus cuspidata.

Cell culture of Taxus cuspidata represents an alternative to whole plant extraction as a source of taxol and related taxanes. Feeding phenylalanine to callus cultures was previously shown to result in increased taxol yields, probably due to the involvement of this amino acid as a precursor for the N-benzoylphenylisoserine side chain of taxol. Inthis study, we have examined the effect of various concentrations of phenylalanine, benzoic acid, N-benzoylglycine, serine, glycine, alanine, and 3-amino-3-phenyl-propionic acid on taxol accumulation in 2-year-old cell suspensions of Taxus cuspidata, cell line FCL1F, and in developing callus cultures of T. cuspidata. All compounds tested were included in media at stationary phase (suspensions) or after the period of fastest growth (calli). Alanine and 3-amino-3-phenyl-propionicacid were tested only in callus cultures and did not affect taxol accumulation. Significant increases or trends toward increases in taxol accumulationin callus and suspensions were observed in the presence of phenylalanine, benzoic acid, N-benzoylglycine, serine, and glycine. The greatest increases in taxol accumulation were observed in the presence of various concentrations of phenylalanine (1 mM for callus; 0.05, 0.1, and 0.2 mM for suspensions) and benzoic acid (0.2 and 1 mM for callus and 0.05, 0.1, and 0.2 mM for suspensions). Increases in taxol yields of cell suspensions in the presence of the most effective precursors brought taxol amounts at stationary phase from 2 mug . g(-1) to approximately 10 mug . g(-1) of the extracted dry weight. The results are discussed in termsof possible implications to taxol biosynthesis and in terms of practical applications to large-scale cell culture systems for the production ofthis drug. (c) 1994 John Wiley & Sons, Inc.