Root volatiles in plant–plant interactions II: Root volatiles alter root chemistry and plant–herbivore interactions of neighbouring plants

Volatile organic compounds (VOCs) emitted by plant roots can influence the germination and growth of neighbouring plants. However, little is known about the effects of root VOCs on plant–herbivore interactions of neighbouring plants. The spotted knapweed (Centaurea stoebe) constitutively releases high amounts of sesquiterpenes into the rhizosphere. Here, we examine the impact of C. stoebe root VOCs on the primary and secondary metabolites of sympatric Taraxacum officinale plants and the resulting plant‐mediated effects on a generalist root herbivore, the white grub Melolontha melolontha. We show that exposure of T. officinale to C.stoebe root VOCs does not affect the accumulation of defensive secondary metabolites but modulates carbohydrate and total protein levels in T. officinale roots. Furthermore, VOC exposure increases M. melolontha growth on T. officinale plants. Exposure of T. officinale to a major C. stoebe root VOC, the sesquiterpene (E)‐β‐caryophyllene, partially mimics the effect of the full root VOC blend on M. melolontha growth. Thus, releasing root VOCs can modify plant–herbivore interactions of neighbouring plants. The release of VOCs to increase the susceptibility of other plants may be a form of plant offense.     

基于LC-MS代谢组学技术的丽豆-大豆差异代谢物比较研究

丽豆(Calophaca sinica)是我国华北地区特有的一种珍稀野生植物。为探明丽豆的营养价值，该文以大豆(Glycine max)为参照组，利用液相色谱-质谱联用(LC-MS)技术对其种子进行了比较代谢组学研究。结果表明：(1)丽豆和大豆中共检测到1 857种代谢产物，二者成分相同且含量相似的代谢物有1 698种(>90%),差异代谢物有159种(<10%)。(2)在差异代谢物中，成分差异的有9种，其中有5种为丽豆特有，剩余150种均为含量差异，其中48种(约30%)在丽豆中的含量高于大豆。(3) KEGG注释到8条差异代谢物显著富集(P<0.1)的通路，主要包括初生代谢物的各类氨基酸生物合成途径和次生代谢物的罗汉松脂素、花生四烯酸以及二萜类等生物合成途径。(4)丽豆比大豆含量低的化学组分主要是初生代谢产物，比大豆含量高的化学组分主要是次生代谢物，而这些次生代谢物在调节血糖、骨坏损修复、增强免疫以及消炎抗癌等生理过程中有着积极的作用。综上所述，该研究认为丽豆与大豆具有相近的营养价值，并且对改善人类亚健康状况有积极的影响；此外，该文使我们对丽豆的营养价值和代谢组成...     

Growth,secondary metabolite production and antioxidant enzyme response of <Emphasis Type="Italic">Morinda citrifolia</Emphasis> adventitious root as affected by auxin and cytokinin

Morinda citrifolia adventitious roots were cultured in shake flasks using Murashige and Skoog medium with different types and concentrations of auxin and cytokinin. Root (fresh weight and dry weight) accumulation was enhanced at 5 mg l⁻¹ indole butyric acid (IBA) and at 7 and 9 mg l⁻¹ naphthalene acetic acid (NAA). On the other hand, 9 mg l⁻¹ NAA decreased the anthraquinone, phenolic and flavonoid contents more severely than 9 mg l⁻¹ IBA. When adventitious roots were treated with kinetin (0.1, 0.3 and 0.5 mg l⁻¹) and thidiazuron (TDZ; 0.1, 0.3 and 0.5 mg l⁻¹) in combination with 5 mg l⁻¹ IBA, fresh weight and dry weight decreased but secondary metabolite content increased. The secondary metabolite content (including 1,1-diphenyl-2-picrylhydrazyl activity) increased more in TDZ-treated than in kinetin-treated roots. Antioxidative enzymes such as catalase (CAT) and guaiacol peroxidase (G-POD), which play important roles in plant defense, also increased. A strong decrease in ascorbate peroxidase activity resulted in a high accumulation of hydrogen peroxide. This indicates that adventitious roots can grow under stress conditions with induced CAT and G-POD activities and higher accumulations of secondary metabolites. These results suggest that 5 mg l⁻¹ IBA supplementation is useful for growth and secondary metabolite production in adventitious roots of M. citrifolia.     

Characterisation of two phenotypes of <Emphasis Type="Italic">Centella asiatica</Emphasis> in Southern Africa through the composition of four triterpenoids in callus,cell suspensions and leaves

Two morphologically distinct phenotypes of Centella asiatica (Type-1 and Type-2) in South Africa were compared in relation to the levels of triterpenoid saponins with the aim of assessing their potential for biotechnological manipulation of triterpenoid synthesis. The metabolites investigated included madecassoside and asiaticoside and their sapogenins madecassic—and asiatic acid; produced in cultured undifferentiated cells (cell suspensions and calli) and leaves. Weight determination in plant cell suspensions and the accumulation of secondary metabolites after 16 days for Type-1 and 20 days for Type-2 were investigated since these secondary metabolites accumulate during the period that follows the active growth phase. The four triterpenoids of interest were analysed and quantified by HPLC in crude ethanolic extracts. A difference in bioactive triterpenoids was exhibited that was tissue specific and varied between the two phenotypes. The triterpenoids from leaf tissue were more easily quantifiable in each phenotype than in the case of the undifferentiated cells (callus and cell suspensions), which had lower, but still quantifiable, levels of these targeted secondary metabolites. Leaves contained the highest triterpenoid levels (ranging from 1.8 to 5% dry weight for the triterpenoid acids and their glycosides, respectively), with the free acids occurring in a ratio of approximately 1:2.5 in relation to the glycoside content.     

UPLC‐HRMS‐based untargeted metabolic profiling reveals changes in chickpea (Cicer arietinum) metabolome following long‐term drought stress

Genetic improvement for drought tolerance in chickpea requires a solid understanding of biochemical processes involved with different physiological mechanisms. The objective of this study is to demonstrate genetic variations in altered metabolic levels in chickpea varieties (tolerant and sensitive) grown under contrasting water regimes through ultrahigh‐performance liquid chromatography/high‐resolution mass spectrometry‐based untargeted metabolomic profiling. Chickpea plants were exposed to drought stress at the 3‐leaf stage for 25 days, and the leaves were harvested at 14 and 25 days after the imposition of drought stress. Stress produced significant reduction in chlorophyll content, F_v/F_m, relative water content, and shoot and root dry weight. Twenty known metabolites were identified as most important by 2 different methods including significant analysis of metabolites and partial least squares discriminant analysis. The most pronounced increase in accumulation due to drought stress was demonstrated for allantoin, l ‐proline, l ‐arginine, l ‐histidine, l ‐isoleucine, and tryptophan. Metabolites that showed a decreased level of accumulation under drought conditions were choline, phenylalanine, gamma‐aminobutyric acid, alanine, phenylalanine, tyrosine, glucosamine, guanine, and aspartic acid. Aminoacyl‐tRNA and plant secondary metabolite biosynthesis and amino acid metabolism or synthesis pathways were involved in producing genetic variation under drought conditions. Metabolic changes in light of drought conditions highlighted pools of metabolites that affect the metabolic and physiological adjustment in chickpea that reduced drought impacts.     

Cardiotonic glycosides from biomass of <Emphasis Type="Italic">Digitalis purpurea</Emphasis> L. cultured in temporary immersion systems

Cardiotonic glycosides are extracted mostly from leaves of Digitalis plants. Commercial production of bioactive secondary metabolites by traditional agriculture is an inefficient process and can be affected by climatic and soil conditions. Strategies, based on in vitro culture methods, have been extensively studied to improve the production of specific plant derived chemicals. The aim of the present research was to obtain biomass of D. purpurea using the temporary immersion system (TIS) and to determine the content of cardiotonic glycosides (digitoxin, digoxin and lanatoside C) as secondary metabolites of commercial value for the pharmaceutical industry. Shoots were cultured in 1,000 ml TIS during 28 days. The effect of four immersion frequencies (once every 2, 4, 6, and 12 h) was studied. Biomass accumulation was influenced by immersion frequency. The maximum biomass accumulation (values in respect of fresh and dry weight) was obtained with immersions every 4 h (six immersions per day). HPLC analysis revealed the presence of digoxin and digitoxin for all immersion frequencies. No lanatoside C was detected in the biomass cultured in TIS. Digoxin concentrations varied depending on the frequencies tested. In contrast, the digitoxin content showed no dependency on the immersion frequency. Net production of digoxin and digitoxin per TIS were found to be higher with immersions every 4 h. The best net production of digitoxin and digoxin per TIS were 167.6 and 119.9 μg, respectively. The development of organ culture based on temporary immersion system can be a reliable method for the steady production of biomass for cardiotonic glycosides production, which is reported for the first time for Digitalis genus in this communication.     

PAL-mediated SA biosynthesis pathway contributes to nematode resistance in wheat

The pathogen cereal cyst nematode (CCN) is deleterious to Triticeae crops and is a threat to the global crop yield. Accession no. 1 of Aegilops variabilis, a relative of Triticum aestivum (bread wheat), is highly resistant to CCN. Our previous study demonstrated that the expression of the phenylalanine ammonia lyase (PAL) gene AevPAL1 in Ae. variabilis is strongly induced by CCN. PAL, the first enzyme of phenylpropanoid metabolism, is involved in abiotic and biotic stress responses. However, its role in plant–CCN interaction remains unknown. In the present study, we proved that AevPAL1 helps to confer CCN resistance through affecting the synthesis of salicylic acid (SA) and downstream secondary metabolites. The silencing of AevPAL1 increased the incidence of CCN infection in roots and decreased the accumulation of SA and phenylalanine (Phe)-derived specialized metabolites. The exogenous pre-application of SA also improved CCN resistance. Additionally, the functions of PAL in phenylpropanoid metabolism correlated with tryptophan decarboxylase (TDC) functioning in tryptophan metabolism pathways. The silencing of either AevPAL1 or AevTDC1 exhibited a concomitant reduction in the expression of both genes and the contents of metabolites downstream of PAL and TDC. These results suggested that AevPAL1, possibly in coordination with AevTDC1, positively contributes to CCN resistance by altering the downstream secondary metabolites and SA content in Ae. variabilis. Moreover, AevPAL1 overexpression significantly enhanced CCN resistance in bread wheat and did not exhibit significant negative effects on yield-related traits, suggesting that AevPAL1 is valuable for the genetic improvement of CCN resistance in bread wheat.     

Simultaneous determination and quantification of three pentacyclic triterpenoids—betulinic acid,oleanolic acid,and ursolic acid—in cell cultures of <Emphasis Type="BoldItalic">Lantana camara</Emphasis> L.

A simple procedure has been described for simultaneous determination and improved yield of three pentacyclic triterpenoids—betulinic, oleanolic, and ursolic acids—from callus cultures of Lantana camara. Cell biomass was obtained from leaf disk explants cultured on Murashige and Skoog (Physiol Plant 15:473–497, 1962) medium supplemented with 5 μM 6-benzylaminopurine, 1 μM 2,4-dichlorophenoxyacetic acid, and 1 μM α-naphthaleneacetic acid. Optimum separation of the three compounds was achieved by reverse-phase high-pressure liquid chromatography on a C₁₈ column with 80:20 (v/v) acetonitrile/water as mobile phase. With this route, a yield of 3.1% betulinic acid, 1.88% oleanolic acid, and 4.12% ursolic acid per gram dry weight was obtained from cultures. Leaves from the parent plant, used as control, showed total absence of betulinic acid, and the quantities of oleanolic and ursolic acids present in them were only marginally higher than that found in in vitro-raised cultures. Presence of the three compounds was further confirmed by electrospray ionization mass spectrometry.     

Nitric Oxide Elicitation Induces the Accumulation of Secondary Metabolites and Antioxidant Defense in Adventitious Roots of<Emphasis Type="Italic">Echinacea purpurea</Emphasis>

Nitric oxide and reactive oxygen species are important signal molecules that play key roles in plant defense responses. We investigated the involvement of nitric oxide elicitation in the synthesis of secondary metabolites within the adventitious roots ofEchinacea purpurea. When roots were treated with 100 μM sodium nitroprusside (SNP), an exogenous nitric oxide producer, the accumulation of phenolics, flavonoids, and caffeic acid derivatives was enhanced. This level of SNP also induced an antioxidant defense, as indicated by increases in Superoxide dismutase, ascorbate peroxidase, and ascorbic acid, along with decreases in hydrogen peroxide, lipid peroxidation, and dehydroascorbate/ascorbic acid. However, a higher concentration (250 μM SNP) acted as a pro-oxidant, thereby raising the levels of hydrogen peroxide, lipid peroxidation, and dehydroascorbate/ascorbic acid while diminishing ascorbic acid, ascorbate peroxidase, and the accumulation of secondary metabolites compared with our observations at 100 μM SNP. Therefore, we conclude that elicitingE. purpurea adventitious roots with a concentration of 100 μM SNP is beneficial to their accumulation of secondary metabolites.     

Distribution and expression characteristics of triterpenoids and OSC genes in white birch (<Emphasis Type="Italic">Betula platyphylla</Emphasis> suk.)

Betulin and oleanolic acids (pentacyclic triterpenoid secondary metabolites) have broad pharmacological activities and can be potentially used for the development of anti-cancer and anti-AIDS drugs. In this study, we detected the accumulation and the distribution characteristics of betulin and oleanolic acid in various organs of white birch at different ages. We also determined the expression of 4 OSC genes (LUS, β-AS, CAS1 and CAS2) involved in the triterpenoid synthesis pathways by real time RT-PCR. The result showed that the 1-year old birch can synthesize betulin and oleanolic acid. In addition, betulin and oleanolic acids were mainly distributed in the bark, while the content in the root skin and leaf was very low. The content of betulin and oleanolic acid in birch varied in different seasons. The content of betulin and oleanolic acid and their corresponding LUS and β-AS gene expression were very low in 1-year old birch. With increasing age of birch, betulin content was increased, while oleanolic acid was decreased. Similar changes were also observed for their corresponding synthesis genes LUS and β-AS. In the leaf of 1-year old plant, the highest expression of CAS1 and CAS2 occurred at end of September, while expression of LUS and the β-AS was low from June to October. In the stem skin,high expression of β-AS and the LUS genes occurred from the end of July to September. In the root, high expression of the β-AS gene was observed at the end of October. These results indicated that triterpenoid gene expression was similar to the triterpene accumulation. Expression of LUS gene and β-AS gene in birch with different ages were corresponding to the betulinic and oleanolic acid accumulation. Expression of CAS1 and CAS2 genes were elevated with increasing age of birch. This study provides molecular mechanisms of triterpenes synthesis in birch plants.     

Effect of long-term cultivation on resveratrol accumulation in a high-producing cell culture of Vitis amurensis

Resveratrol is a polyphenol, present in grapes, peanuts, and other plant sources, with a wide range of valuable biological activities. We established a Vitis amurensis cell culture accumulating high levels of resveratrol by introducing the rolB gene of Agrobacterium rhizogenes in the V. amurensis genome, and studied the stability of resveratrol accumulation during 27 months of continuous subculturing. This study demonstrates a decline in the high level of resveratrol production by the rolB transgenic cell line during its long-term cultivation. Elicitation of the rolB transgenic calli with methyl jasmonate and salicylic acid, which are known to stimulate the production of plant secondary metabolites, resulted in a recovery of resveratrol accumulation in the rolB transgenic cell culture, while the empty vector-transformed culture with trace starting content of resveratrol exhibited low inducibility to the treatment.     

Metabolic shift from secondary metabolite production to induction of anti-oxidative enzymes during NaCl stress in <Emphasis Type="Italic">Swertia chirata</Emphasis> Buch.-Ham.

Swertia chirata Buch.-Ham. is an endangered medicinal plant having high medicinal value. Present study was envisaged to understand in vitro induction of marker secondary metabolites using NaCl elicitation. 50 and 100 mM of NaCl concentrations were applied on 1-month-old static culture shoots. Plants were assessed for cellular damage, anti-oxidative enzymatic system and production of secondary metabolites. There was significant (p ≤ 0.05) increase in secondary metabolites at 50 mM NaCl without any cellular damage or induction of anti-oxidative enzymes. Initial increase in metabolic content of secondary metabolites was observed during 100 mM NaCl treatment, which falls back to normal levels at the seventh day. There was concurrent induction of scavenging enzymes during this period. Results suggest channelling of different defence strategies in response to differential NaCl treatment. Biochemical relationship between induction of anti-oxidative enzymes and production of secondary metabolites has further been discussed in light of physiological requirements.     

Effect of low light intensity on growth and accumulation of secondary metabolites in roots of Glycyrrhiza uralensis Fisch

Effects of low light intensity on growth and accumulation of secondary metabolites of a medicinal plant Glycyrrhiza uralensis Fisch. were investigated. Hydroponic-cultivated one year-old rhizome seedlings were grown under three low irradiances, 200, 100, and 50 μmol m⁻² s⁻¹ for 135 days. Control plants were cultured under natural light conditions. Low light intensity stress decreased leaf thickness, photosynthesis and biomass, but increased leaf area and chlorophyll concentrations. Low light intensity also significantly increased accumulation of glycyrrhizic acid and liquiritin in the root, while the maximum values of both secondary metabolites were obtained under an irradiance of 100 μmol m⁻² s⁻¹. Concentrations of both secondary metabolites were negatively correlated with root biomass. The results suggested that G. uralensis could endure an environment with low light intensity and suitable light control might increase the secondary metabolite contents within agroforestry systems.     

Transgressive segregation of primary and secondary metabolites in F<Subscript>2</Subscript> hybrids between <Emphasis Type="Italic">Jacobaea aquatica</Emphasis> and <Emphasis Type="Italic">J. vulgaris</Emphasis>

Hybridization between plant species can have a number of biological consequences; interspecific hybridization has been tied to speciation events, biological invasions, and diversification at the level of genes, metabolites, and phenotypes. This study aims to provide evidence of transgressive segregation in the expression of primary and secondary metabolites in hybrids between Jacobaea vulgaris and J. aquaticus using an NMR-based metabolomic profiling approach. A number of F₂ hybrid genotypes exhibited metabolomic profiles that were outside the range encompassed by parental species. Expression of a number of primary and secondary metabolites, including jacaronone analogues, chlorogenic acid, sucrose, glucose, malic acid, and two amino acids was extreme in some F₂ hybrid genotypes compared to parental genotypes, and citric acid was expressed in highest concentrations in J. vulgaris. Metabolomic profiling based on NMR is a useful tool for quantifying genetically controlled differences between major primary and secondary metabolites among plant genotypes. Interspecific plant hybrids in general, and specifically hybrids between J. vulgaris and J. aquatica, will be useful for disentangling the ecological role of suites of primary and secondary metabolites in plants, because interspecific hybridization generates extreme metabolomic diversity compared to that normally observed between parental genotypes.     

Establishment of adventitious root co-culture of Ginseng and Echinacea for the production of secondary metabolites

We have successfully established the co-culture of ginseng (Panax ginseng C.A. Meyer) and echinacea [Echiancea purpurea (L.) Moench.] adventitious roots for the production secondary metabolites. Adventitious roots of ginseng and echinacea were cultured in different proportions (5 g L⁻¹; 4:1, 3:2 and 2:1 ginseng and echinacea, respectively) in 5-L capacity airlift bioreactors containing 4 L Murashige and Skoog medium supplemented with 25 μM indole-3-butyric acid and 50 g sucrose L⁻¹ and maintained at 25°C in the dark for 40 days. Results showed the negative effect of echinacea adventitious roots on the growth of ginseng roots, however, by limiting the inoculum density of echinacea, it was possible to establish the co-cultures. To enhance the accumulation of secondary metabolites, co-cultures were treated with 200 μM methyl jasmonate after 30 days of culture initiation. Methyl jasmonate elicitation promoted the accumulation of ginsenosides in the co-cultures. It was possible to produce ginsenosides and caffeic acid derivatives in higher amounts by establishing co-cultures with higher inoculum proportion of ginseng to echinacea (4:1 and 3:2) followed by elicitation treatment. This work demonstrates the effectiveness of interspecies adventitious root co-cultures for the production of plant secondary metabolites.     

Induction,characterization, and NMR-based metabolic profiling of adventitious root cultures from leaf explants of <Emphasis Type="Italic">Gynura procumbens</Emphasis>

Gynura procumbens is a medicinal plant used in South East Asia to treat various ailments such as rash, hemorrhoids, inflammation, and diabetes. In order to develop a large-scale culture system for G. procumbens biomass containing bioactive compounds, adventitious root cultures were initiated from leaf explants. Murashige and Skoog (MS) media containing different compositions of indole-3-butyric acid (IBA), 1-naphthalene-acetic acid (NAA), and combinations of both plant growth regulators (PGRs) were evaluated for root induction. A combination of 3 mg/l NAA + 1 mg/l IBA gave the highest root induction (48%) as compared to other PGRs treatments after 9 weeks of incubation period. Subsequently, the adventitious roots were established in liquid culture containing MS medium and the combination of 3 mg/l NAA + 1 mg/l IBA. A study on the medium strength, sucrose concentration, pH, and light versus dark was conducted to optimize the in vitro culture conditions. The results showed that differences in MS medium strength from half to double strength, and light or dark condition did not significantly affect the biomass production, while the initial medium pH of 5.5 and 2% w/v sucrose concentration were most suitable for the root culture growth. Nuclear magnetic resonance (NMR) spectroscopy was performed to characterize the metabolite content in the root cultures of G. procumbens. Among the elucidated metabolites were some phenylpropanoids identified as caffeic acid, chlorogenic acid, and 3,5-di-O-caffeoylquinic acid which might be the bioactive compounds associated to the folk use of this plant.     

Cadmium Accumulation in Relation to Organic Acids and Nonprotein Thiols in Leaves of the Recently Found Cd Hyperaccumulator <Emphasis Type="Italic">Rorippa globosa</Emphasis> and the Cd-accumulating Plant <Emphasis Type="Italic">Rorippa islandica</Emphasis>

It has been proposed that organic acid and nonprotein thiol are involved in the hyperaccumulation of metals. In this study, Cd accumulation, organic acid, and nonprotein thiol production and their relationships in the leaves of Cd-hyperaccumulator Rorippa globosa were examined and compared with a closely related species, Rorippa islandica. The results showed that there was no reduction in biomass of R. globosa when treated with 25 μg Cd g⁻¹ (T₂), despite Cd accumulation in the leaves was up to 158.2 μg g⁻¹ DW. On the other hand, the growth of Cd-treated R. islandica was obviously inhibited as it accumulated more than 100 μg g⁻¹ DW in the leaves. Therefore, R. islandica behaved as a Cd-accumulating plant. The Cd treatments could significantly induce the synthesis of acetic acid in both species, suggesting that acetic acid, as the most abundant organic acid, might be related to the Cd accumulation. Significant positive correlations between Cd concentrations and both tartaric and malic acid concentrations in the leaves of R. globosa were observed. There was a significant positive correlation between Cd concentrations and acetic acid concentrations in the leaves of R. islandica. This trend of tartaric and malic acids in the leaves of R. globosa and acetic acid in the leaves of R. islandica might be related to Cd accumulation. In addition, a quadratic relationship was obviously observed for NP-SH contents and total Cd concentrations in the leaves of R. globosa, indicating that NP-SH was significantly related to Cd accumulation and tolerance.     

Pleiotropic physiological consequences of feedback‐insensitive phenylalanine biosynthesis in Arabidopsis thaliana

A large proportion of plant carbon flow passes through the shikimate pathway to phenylalanine, which serves as a precursor for numerous secondary metabolites. To identify new regulatory mechanisms affecting phenylalanine metabolism, we isolated Arabidopsis thaliana mutants that are resistant to the phytotoxic amino acid m‐tyrosine, a structural analog of phenylalanine. Map‐based cloning identified adt2‐1D, a dominant point mutation causing a predicted serine to alanine change in the regulatory domain of ADT2 (arogenate dehydratase 2). Relaxed feedback inhibition and increased expression of the mutant enzyme caused up to 160‐fold higher accumulation of free phenylalanine in rosette leaves, as well as altered accumulation of several other primary and secondary metabolites. In particular, abundance of 2‐phenylethylglucosinolate, which is normally almost undetectable in leaves of the A. thaliana Columbia‐0 accession, is increased more than 30‐fold. Other observed phenotypes of the adt2‐1D mutant include abnormal leaf development, resistance to 5‐methyltryptophan, reduced growth of the generalist lepidopteran herbivore Trichoplusia ni (cabbage looper) and increased salt tolerance.     

Oxytocin influences the production of glycyrrhizin from cell cultures of Abrus precatorius Linn.

Oxytocin, a peptide animal hormone, was used as a growth regulator to test its effect on biomass accumulation and production of secondary plant constituent glycyrrhizin in the cell cultures of Abrus precatorius. Glycyrrhizin is an important phytoconstituent of liquorice which is widely used in the pharmaceutical and food industries. Cell suspension cultures of A. precatorius were developed from leaf explant of in vitro germinated plant in Murashige and Skoog medium supplemented with 30 g/l sucrose, 1 mg/l naphthalene acetic acid and 1 mg/l kinetin. The influence of oxytocin on biomass accumulation as well as on the production of glycyrrhizin was observed in the cell cultures of A. precatorius. Treatment of A. precatorius cell cultures with 100 μg/l oxytocin, improved glycyrrhizin production up to 34.27 mg/l on the dry cell weight basis third day after oxytocin treatment, which is over four times that of the control cultures, simultaneously nearly two fold increase in the biomass 2 days after the oxytocin treatment was recorded over the control cultures.