Elicitor Concentration Research Articles

Effect of elicitors on physiomorphological and biochemical parameters of Indian mustard (Brassica juncea) and rapeseed (B. napus)

The present investigation was aimed to assess the effect of different concentrations of elicitors viz., salicylic acid (SA) and benzothiadiazole (BTH) on physiomorphological and biochemical parameters of Brassica juncea and B. napus cultivars. The field experiment was conducted during 2012-13 at Punjab Agricultural University, Ludhiana, India. Four different treatments of elicitors and a fungicide treatment were given to 10 week old plants up to four consecutive weeks. Content of total soluble protein, free amino acids, total sugars and reducing sugars ofleaves of both B. juncea and B. napus were evaluated after each spray. The content of total soluble protein increased, whereas the contents of free amino acids, total sugars and reducing sugars decreased in all the treatments after each spray. Among all the treatments, the combinations of elicitors, i.e., BTH (3 ppm) + SA (33 ppm) and BTH (7 ppm) + SA (17 ppm) exhibited maximum contents of total soluble protein, free amino acids, total sugars and reducing sugars than control in both the cultivars. These treatments were more pronounced in increasing plant height, internodal distance and number of pods per plant in both the cultivars. Our results suggest that the combinations of elicitors act synergistically to promote growth and metabolic activities in B. juncea and B. napus cultivars leading to the induction and regulation of disease resistance.

Effects of Streptomyces pactum Act12 on Salvia miltiorrhiza hairy root growth and tanshinone synthesis and its mechanisms.

The fermentation broth and mycelium pellet of Streptomyces pactum Act12 (Act12) may promote the accumulation of soluble sugar when added to Salvia miltiorrhiza hairy roots, increasing the accumulation level by as much as 23.20% compared with the control; it may also inhibit the accumulation of soluble protein in the hairy roots, decreasing it by as much as 17.96% compared with the control. The ACT12 also has a certain promotional effect on the growth of hairy root at an appropriate concentration of elicitors and upregulates the expression of genes 3-hydroxy-3-methyglutary1-CoA reductase (HMGR), 1-deoxy-D-xylulose 5-phosphate synthase (DXS), 1-deoxy-D-xylulose-5-phosphate reductoisomerase (DXR), and geranylgeranyl diphosphate synthase (GGPPS). Among these effects, that of the HMGR gene expression is as high as 33.66 times that of the control, indicating that the test Streptomyces pactum may efficiently adjust the secondary metabolism of S. miltiorrhiza at the level of gene transcription, thereby greatly increasing the accumulation level of tanshinone in the hairy roots; among which, the cryptotanshinone levels increased most significantly, as much as 33.63 times that of the control, and the total tanshinone levels were 12.61 times that of the control.

Effect of plant growth regulators and elicitors on rhinacanthin accumulation in hairy root cultures of Rhinacanthus nasutus (L.) Kurz

Several biologically active secondary metabolites like anthraquinones, sterols, triterpenes, flavonoids and naphthoquinones are present in Rhinacanthus nasutus. Naphthoquinones are important group of compounds generally known as rhinacanthin (RC) consists of 15 derivatives named RC A–D and G–Q of which RC-C, RC-D and RC-N have various medicinal properties. The individual role of two auxins i.e. indole-3-butyric acid (IBA) and α naphthalene acetic acid (NAA) and two elicitors i.e. methyl jasmonate (MJ) and salicylic acid (SA) in Murashige and Skoog medium on hairy root growth and RC (RC-C, RC-D and RC-N) accumulation was investigated in the present study. Time course study revealed that IBA and NAA at 2.5 μM showed maximum fresh weight (FW) and dry weight (DW) 4 weeks after culture. However, RC production was maximum after 6 weeks of culture on both media. A concentration-dependent response was observed when various concentrations of MJ (2.0, 5.0, 10 and 15 μM) and SA (10, 50, 100 and 150 μM) were supplemented in the medium. On MJ and SA media the FW and DW decreased as the concentration of elicitors increased. However, this decrease was more severe in MJ treated cultures. All the MJ and SA treated cultures showed significantly higher amount of RC-C, RC-D and RC-N in hairy roots harvested 7 days after elicitation as compared to control. Of the two elicitors, MJ was more efficient in inducing RC accumulation than SA. The highest RC content (6.3 mg/g DW RC-C; 1.1 mg/g DW RC-D and 0.61 mg/g DW RC-N) was observed after treatment with 10 μM MJ which was about 1.7-, 2.5- and 3.5-fold higher RC-C, RC-D and RC-N respectively than the control.

Modulation of artemisinin biosynthesis by elicitors, inhibitor, and precursor in hairy root cultures of Artemisia annua L.

Artemisinin is frequently used in the artemisinin-based combination therapy to cure drug-resistant malaria in Asian subcontinent and large swath of Africa. The hairy root system, using the Agrobacterium rhizogenes LBA 9402 strain to enhance the production of artemisinin in Artemisia annua L., is developed in our laboratory. The transgenic nature of hairy root lines and the copy number of transgene (rol B) were confirmed using polymerase chain reaction and Southern Blot analyses, respectively. The effect of different concentrations of methyl jasmonate (MeJA), fungal elicitors (Alternaria alternate, Curvularia limata, Fusarium solani, and Piriformospora indica), farnesyl pyrophosphate, and miconazole on artemisinin production in hairy root cultures were evaluated. Among all the factors used individually for their effect on artemisinin production in hairy root culture system, the maximum enhancement was achieved with P. indica (1.97 times). Increment of 2.44 times in artemisinin concentration by this system was, however, obtained by combined addition of MeJA and cell homogenate of P. indica in the culture medium. The effects of these factors on artemisinin production were positively correlated with regulatory genes of MVA, MEP, and artemisinin biosynthetic pathways, viz. hmgr, ads, cyp71av1, aldh1, dxs, dxr, and dbr2 in hairy root cultures of A. annua L.

Induced systemic resistance: an innovative control method to manage branched broomrape (Orobanche ramosa L.) in tomato

The biochemical responses of tomato ( Lycopersicon esculentum Mill.) roots to different concentrations of the hormonal elicitors, salicylic acid (SA) and indole acetic acid (IAA), as presoaking treatments, on the healthy and Orobanche ramosa -infected plants were studied in pot experiment under control conditions. Application of SA and IAA activate the production of proline and soluble proteins in tomato roots, where their levels were more pronounced in the infected than the healthy plants. Moreover, both elicitors increased the levels of phenolics and flavonoids accompanied by stimulations in the activities of phenylalanine ammonia lyase (PAL), peroxidase (POX) and polyphenol oxidase (PPO) in tomato roots, where the magnitude of induction were more obvious in infected plants than healthy one. Also, microscopic examination revealed the anatomical differences observed in both untreated healthy and infected plants as well as in plants treated with SA and IAA. The present results indicate the effectiveness of SA and IAA in enhancing resistance of tomato to retard the incidence of infestation by Orobanche. These biochemical and histological changes involved in plant defense mechanisms may be one of the basic ways participate in the action of SA and IAA for increasing resistance in tomato against Orobanche ramosa . Keywords : Orobanche ramosa ; tomato, Induced resistance, Phenolics, PR-proteins . *Corresponding author : Shahnaz Al-Wakeel (e-mail: shahnazalwakeel@yahoo.com) (Received: 12.04.2012 Accepted: 17.07.2012)

Effect of abiotic elicitation on main health-promoting compounds, antioxidant activity and commercial quality of butter lettuce (Lactuca sativa L.)

The study presents changes in the phytochemical levels, antiradical activity and quality of lettuce caused by different chemical elicitors: arachidonic acid (AA), jasmonic acid (JA), and abscisic acid (ABA). The application of 1μM and 100μM JA induced an increase in the concentration of phenolic compounds, including flavonoids and phenolic acids. Flavonoid levels were also increased after treatment with 100μM AA and ABA. Some of the elicitor concentrations used also caused an increase in the levels of other phytochemicals, such as chlorophyll a (1μM and 100μM AA, 50μM ABA); chlorophyll b (100μM AA); carotenoids (100μM AA, 1μM JA and 100μM ABA) and vitamin C (100μM AA, 100μM JA). The highest antiradical activity was noted after treatment with 100μM AA, 100μM JA. 1,1-Diphenyl-2-picrylhydrazyl (DPPH) scavenging ability was positively and significantly correlated with flavonoid, chlorophyll and carotenoid levels. These results may suggest that the antiradical activity of lettuce was determined not only by phenolics, but also by other bioactive compounds. Elicitation did not change the sensory quality of lettuce. Therefore, treatment with elicitors could be a useful tool for improving the health-promoting qualities of lettuce without the loss of sensory quality.

Ethylene treatment improves diosgenin accumulation in in vitro cultures of Dioscorea zingiberensis via up-regulation of CAS and HMGR gene expression

Background: The perennial medicinal herb Dioscorea zingiberensis is a very important plant used for steroid drug manufacturing for its high level of diosgenin in rhizome. Although the stimulation of diosgenin accumulation by ethylene has been reported in a few of plant species, its regulation is not yet characterized at the molecular level, the underlying molecular mechanism remains elusive. Results: In this study, the effects of ethylene on diosgenin biosynthesis in in vitro cultures of D. zingiberensis were described. The results showed that, in samples treated with ethylene at concentration E3 (10 4 dilution of 40% ethephon), the diosgenin biosynthesis was significantly promoted in comparison with the control samples. Treatment with high concentrations of ethylene had inhibitory effect, whereas with low concentration of the gas elicitor brought about no detectable deleterious effect on the growth rate and diosgenin content of the cultures. The considerable increase of diosgenin level in in vitro cultured Dioscorea zingiberensis by ethylene application is accompanied by the concomitant increase of soluble proteins and chlorophyll content. The gene expressions of cycloartenol synthase and 3-hydroxy-3-methylglutaryl-CoA reductase but not of squalene synthase or farnesyl pyrophosphate synthase were up-regulated by applied ethylene. Conclusions: Our results suggest that ethylene treatment enhanced diosgenin accumulation via up-regulation of the gene expressions of cycloartenol synthase and 3-hydroxy-3-methylglutaryl-CoA reductase.

Elicitation of pharmacologically active phenolic compounds from Rauvolfia serpentina Benth. Ex. Kurtz.

Quality of medicinal plants used for the production of galenics or pharmacologically useful compounds is usually assessed by the content of biologically active compounds. Most of these plants are grown in fields, this study focused to evaluate the effect of three natural elicitors salicylic acid, methyl salicylate and acetyl salicylic acid (SA, MSA, ASA) on phenolic compound content of Rauvolfia serpentina, and to compare the efficiency of several solvent system in phenolic compound extraction from this plant. A tremendous increase of phenolics and stimulation of the biomass yield were achieved. Tuning of organ specificity by modification of the concentration of elicitor was also observed. This methodology represents a convenient alternative to cell suspension or hydroponic cultures being applicable in wide agricultural practice and to improve phenolic concentration during extraction.

Study of Various Factors for Enhancement of Artemisinin in Artemisia Annua Hairy Roots

Artemisinin is an alternate anti-malarial drug which is widely used in the cure of multi-drug resistant Plasmodium falciparum malaria. In this study, enhancement of artemisinin content by using several elicitors and precursors was attempted initially by using one at a time (OVAT) approach. The most potent elicitor(s) & precursor were thereafter identified by the detailed analysis of the responses with respect to biomass and bioactive compound with respect to the concentrations of different effectors. Central Composite Design (CCD) was thereafter used to identify the antagonistic or synergistic effects of high and low concentrations of different precursors and elicitors. The maximum artemisinin content obtained was 3.45 mg/g on 15d by using the elicitor methyl jasmonate (40 µg/l) and the precursors casein acid hydrolysate (50 µg/l), sodium acetate (500 µg/l). Effect of addition time of elicitors and precursors on overall optimum biomass growth and artemisinin yield and productivity was also established.

English

The effects of an endophytic fungal elicitor from Cunninghamella sp. (AL4) on the volatile oil production and defence responses of host plant Atractylodes lancea were investigated. The aim was to improve the capability of endophytic fungi resources in promoting medicinal plant secondary metabolites. The results showed that the endophytic fungal elicitor significantly stimulated volatile oil accumulation. The best inducing concentration of the elicitor for volatile oil production was 30 mgL-1. The highest amount of volatile oil was harvested 9 days (d) after elicitor treatment. The activity of 3-hydroxy-3-methylglutaryl CoA reductase increased by 2.79-folds compared with the control of 7 d after treatment. The elicitor also enhanced the activities of phenylalanine ammonia-lyase, peroxidase superoxide dismutase, polyphenol oxidase, and catalase. These enhanced activities led to improvedplant defence-related secondary metabolite synthesis ability. In addition, the elicitor-treated cells were consistently healthy, and had slightly increased biomass. The results demonstrated for the first time the optimal induction time and concentration of the endophytic fungal elicitor, as well as the changes in physiological indices after endophyte-host interaction. The information provided in the present study may be applied in large-scale medicinal plant suspension cultures.   Key words: Endophytic fungus, fungal elicitor, Atractylodes lancea, volatile oil,Cunninghamella sp.

Immobilization mediated enhancement of phyllanthin and hypophyllanthin from Phyllanthus amarus

ABSTRACT Phyllanthus amarus plant is used in the traditional system of medicine as a hepatoprotective drug for which the major lignans phyllanthin and hypophyllanthin are responsible. So far, no significant work has been done on the culture of this plant. Realizing the hepatoprotective potential, the present investigation was undertaken. A cost effective process was developed for enhancing phyllanthin and hypophyllanthin utilizing the immobilization technique. HPTLC was used to compare the phyllanthin and hypophyllanthin contents in calcium alginate immobilized cells obtained from fresh grown plants and MS medium was supplemented with different abiotic elicitors, under aseptic conditions for the treatment with chitosan, copper sulphate, phenylalanine and silver nitrate solution to make the whole process commercially viable. It was revealed that silver nitrate and phenylalanine at low concentration enhances phyllanthin and hypophyllanthin yield as compared to control immobilized cell culture. The study revealed that an increase in the content of phyllanthin and hypophyllanthin was elicitor concentration dependent and silver nitrate treatment gave a maximum yield of hepatoprotective bioactives as compared to the other abiotic elicitors used.

Effect of Elicitor Spray at Different Reproductive Stages on Saponin Content of Soybean

The beneficial health effects of soybeans may be enhanced by increasing bioactive compounds including soyasaponins (ssp). The objective of this study is to elucidate the effect of elicitors sprayed on Ozark variety soybeans, on ssp content. Different concentrations of elicitors, ethyl acetate (EA) and methyl jasmonate (MJ), were sprayed at 4 different growth stages (1-bloom, 2-pod development, 3-seed development, and 4-seed maturity). Seeds were ground, defatted, ssp was extracted and identified and quantified with HPLC. Elicitor and growth stage had an effect on βg and βa contents of soybeans compared with control (P < 0.05). Elicitor had an effect on total ssp content (P < 0.001) and αg and γg content of soybeans compared with control (P < 0.05). Total ssp content of EA 0.05 M, MJ 0.001 M, and 0.005 M sprayed soybeans were higher than EA 0.001 M, which is higher than control (P < 0.05; 3.62, 3.56, 3.56, 3.29, and 2.98 μmol/g soybean, respectively). The overall effect of elicitor on total ssp content was not dependent on growth stage, however, elicitors sprayed at growth stages 1, 2, and 3 showed differences among elicitor applied soybeans. Elicitors applied at growth stage 4 did not have any effect on total ssp content compared to control. Elicitors EA 0.05 M, MJ 0.001, and 0.005 M can be applied on any growth stage to increase total saponin content of soybean variety Ozark. Higher saponin content may improve the beneficial health effects of soybean consumption.

Effects of elicitors on the enhancement of asiaticoside biosynthesis in cell cultures of centella (Centella asiatica L. Urban)

AbstractIn this work, the effects of elicitor concentration and elicitation day on the growth and asiaticoside production of centella cells were investigated. The results showed that 2-hydroxybenzoic acid from 50–200 μM and yeast extract from 2–5 g L−1 had different eliciting influences. The addition of 2-hydroxybenzoic acid and yeast extract to the cultures strongly enhanced asiaticoside production in centella cells. The increase in asiaticoside content induced by the addition of 100 μM of 2-hydroxybenzoic acid and 4 g L−1 of yeast extract at day 10 of inoculation was about 5- and 3.5-fold, respectively, as compared with that of the reference cells. In general, 2-hydroxybenzoic acid (abiotic elicitor) was more effective in enhancing asiaticoside biosynthesis than yeast extract (biotic elicitor).

Proteomic Insight Into the Molecular Principles of Grapevine Habituation

Proteomic Insight Into the Molecular Principles of Grapevine Habituation Two-dimensional gel electrophoresis coupled to protein microarray analysis was used to examine, for the first time, the molecular mechanisms of grapevine (Vitis vinifera L., cv. Limberger) habituation. The examination of 2-D maps derived from control and habituated cell culture revealed the presence of 55 protein spots displaying a differential expression pattern. These facts have provide a molecular evidence suggesting that the habituated cells can be used as a model for study of cell differentiation and plant defense mechanisms. Cell death, extra-cellular alkalinization and expression of genes responsible for the formation of the defense-related proteins were analyzed in suspension cultures with hormonal autonomy (habituation). Results obtained using habituated grapevine cells compared with non-habituated cells were different and strongly depended on the concentration of elicitor applied.

Pyrazinecarboxamides as Potential Elicitors of Flavonolignan and Flavonoid Production in Silybum marianum and Ononis arvensis Cultures In Vitro

The effect of new synthetic pyrazinecarboxamide derivatives as potential elicitors of flavonolignan and flavonoid production in Silybum marianum and Ononis arvensis cultures in vitro was investigated. Both tested elicitors increased the production of flavonolignans in S. marianum callus and suspension cultures and flavonoids in O. arvensis callus and suspension cultures. Compound I, 5-(2-hydroxybenzoyl)-pyrazine-2-carboxamide, has shown to be an effective elicitor of flavonolignans and taxifoline production in Silybum marianum culture in vitro. The maximum content of silydianin (0.11%) in S. marianum suspension culture was induced by 24 h elicitor application in concentration of 1.159 × 10−3 mol/L. The maximum content of silymarin complex (0.08%) in callus culture of S. marianum was induced by 168 h elicitor application of a concentration 1.159 × 10−4 mol/L, which represents contents of silydianin (0.03%), silychristin (0.01%) and isosilybin A (0.04%) compared with control. All three tested concentrations of compound II, N-(2-bromo-3-methylphenyl)-5-tert-butylpyrazin-2-carboxamide increased the flavonoid production in callus culture of O. arvensis in a statistically significant way. The best elicitation effect of all elicitor concentrations had the weakest c3 concentration (8.36 × 10−6 mol/L) after 168 h time of duration. The maximum content of flavonoids (about 5,900%) in suspension culture of O. arvensis was induced by 48 h application of c3 concentration (8.36 × 10−6 mol/L).

Enhanced extracellular production of trans-resveratrol in Vitis vinifera suspension cultured cells by using cyclodextrins and methyljasmonate

In this work, the effect of different inducing factors on trans-resveratrol extracellular production in Monastrell grapevine suspension cultured cells is evaluated. A detailed analysis provides the optimal concentrations of cyclodextrins, methyljasmonate and UV irradiation dosage, optimal cell density, elicitation time and sucrose content in the culture media. The results indicate that trans-resveratrol production decreases as the initial cell density increases for a constant elicitor concentration in Monastrell suspension cultured cells treated with cyclodextrins individually or in combination with methyljasmonate; the decrease observed in cell cultures elicited with cyclodextrins alone is far more drastic than those observed in the combined treatment. trans-Resveratrol extracellular production observed by the joint use of cyclodextrins and methyljasmonate (1,447.8 ± 60.4 μmol trans-resveratrol g(-1) dry weight) is lower when these chemical compounds are combined with UV light short exposure (669.9 ± 45.2 μmol trans-resveratrol g(-1) dry weight). Likewise, trans-resveratrol production is dependent on levels of sucrose in the elicitation medium with the maximal levels observed with 20 g l(-1) sucrose and the joint action of cyclodextrins and 100 μM methyljasmonate. The sucrose concentration did not seem to limit the process although it affects significantly the specific productivity since the lowest sucrose concentration is 10 g l(-1), the highest productivity is reached (100.7 ± 5.8 μmol trans-resveratrol g(-1) dry weight g(-1) sucrose) using cyclodextrins and 25 μM methyljasmonate.

Chitosan physico–chemical properties modulate defense responses and resistance in tobacco plants against the oomycete Phytophthora nicotianae

Enzymatic defense responses and protection against Phytophthora nicotianae were studied in tobacco plants treated with chitosan of different molecular weights (MW) and degrees of acetylation (DA). The concentration and mode of chitosan application affected enzymatic induction in tobacco leaves. β-1,3-Glucanase (EC 3.2.1.6) activity required 10 times the polymer concentration relative to the oligochitosan mixture to induce the highest activity above control when treated by foliar spray, indicating the influence of molecular weight in this response. PAL (EC 4.3.1.5) and POD (EC 1.11.1.6) activities increased above control as a result of the influence of polymer degree of acetylation when treatments were applied by foliar spray. A higher DA favored PAL activity, whereas a lower DA induced higher POD activity. Using an in vitro bioassay, it was found that the three chitosan compounds caused a reduction of the infection index of P. nicotianae in tobacco plants that was dependent on elicitor concentrations. There was a significant relationship between the reduction of the infection index and an increase in PAL activity when chitosan was applied by foliar spray and by substrate drench to the plant rhizosphere. These results demonstrate the influence of chitosan physico–chemical properties in plant-induced resistance and the relevance of particular responses in plant protection against pathogens.

In vitro isolation, elicitation of psoralen in callus cultures of Psoralea corylifolia and cloning of psoralen synthase gene

Psoralen, an important furanocoumarin occurring abundantly in seeds of Psoralea corylifolia is used as an anticancerous compound against leukemia and other cancer cell lines. Evaluation and isolation of psoralen from the calluses derived from different plant parts, viz. cotyledons, nodes, leaves and roots have been done in the present case for the first time. Amongst all, a maximum of 1934.75 μg/g f.w. of psoralen was recorded in callus derived from cotyledons, followed by 1875.50 and 1465.75 μg/g f.w. of psoralen in node and leaf derived calluses, respectively. Amount of psoralen enhanced further when cotyledonary calluses were exposed to different concentrations of organic elicitors (yeast extract, proline, inositol, casein hydrolyzate (CH), glycine, glutamine and sucrose) and precursors of psoralen (umbelliferone, cinnamic acid and NADPH). Isolation of psoralen was done using methanol as solvent through column chromatography and TLC. FT-IR and NMR further characterized and confirmed the structure of psoralen. In addition, the putative gene, psoralen synthase involved in psoralen synthesis pathway has been isolated, cloned and sequenced which comprised 1237 bp length. BLAST analysis of the gene sequence of psoralen synthase revealed that its nucleotide sequence showed 93% homology with psoralen synthase isolated from Ammi majus. This is the first report of isolation, cloning and characterization of psoralen synthase from Psoralea corylifolia.

Response of somatic embryos of Scots pine to fungal cell wall elicitors

SummaryFungal cell wall elicitors have notable effects on germination of pine seeds, defence response and many other aspects of metabolic processes. In this study, we investigated morphological, physiological and molecular response of somatic embryos of Scots pine to fungal cell wall elicitors from ectomycorrhizal fungus Suillus bovinus, pathogenic fungus Heterobasidion annosum or weak Scots pine pathogen (Heterobasidion parviporum). Results from the embryo rooting experiment indicated that the genotypic origin of the conifer tree and the source of the elicitors affected the root development. Elicitors from either S. bovinus or H. parviporum increased the root formation but increased concentration of elicitors from H. annosum decreased the rooting and survival rate of the embryos. At the molecular level, altered transcript changes were observed among genes encoding proteins involved in cell division, cell wall modification, and stress following exposure of embryos to elicitors. Increased levels of actin and peroxidase gene expression were detected at 7 days after exposure to S. bovinus or H. parviporum elicitors. H. annosum elicitors provoked significant induction of the ethylene synthesis gene PsACS‐1. Generally the gene expression patterns at early stages observed in the subset of genes investigated after treatment with H. parviporum elicitor were found to be similar to that documented for S. bovinus. In contrast, somatic embryos exposed to H. annosum elicitor had different patterns of regulation marked by delayed response, and in some cases gradual degradation and death of the embryos were observed.

Marked effect of Cuscuta on puerarin accumulation in cell cultures of Pueraria tuberosa grown in shake flasks and a bioreactor

Isoflavonoid production in cell cultures of Pueraria tuberosa as influenced by an angiospermic parasite, Cuscuta reflexa, was studied. During the time course, maximum isoflavonoid content was recorded when Cuscuta elicitor was added on day 15 of culture. Among various concentrations of elicitor tried, 1 g l−1 of Cuscuta elicitor was found to be the most effective. The optimized elicitation conditions were used in vessels of varying capacity where maximum yield of ~91 mg l−1 of isoflavonoid was recorded in a 2-l bioreactor which was about 19% higher than the control cultures. In this case, puerarin content increased up to 11 mg l−1 which was 580% higher that the value recorded in the control cultures. In the bioreactor, 8 days of elicitation was optimal for the high accumulation of isoflavonoid, giving productivity of ~4 mg l−1 day−1. The study showed persistent high isoflavonoid yield even during scale-up. Use of a preparation of Cuscuta reflexa as an elicitor is reported for the first time. The increase in isoflavonoid content was elicitor dose-dependent and can be explored to trigger high yields of isoflavonoid/secondary metabolites in production.