Pilot-Scale Culture of Hypericum Perforatum L. Adventitious Roots in Airlift Bioreactors for the Production of Bioactive Compounds

Hypericum perforatum L. (St. John’s Wort) is an important medicinal plant which is widely used in the treatment for depression and irritable bowel syndrome. It is also used as a dietary supplement. Major bioactive phytochemicals of H. perforatum are phenolics and flavonoids. Quality of these phytochemicals is dramatically influenced by environmental and biological factors in the field grown plants. As an alternative, we have developed adventitious root cultures in large-scale bioreactors for the production of useful phytochemicals. Adventitious roots of H. perforatum were cultured in 500 l pilot-scale airlift bioreactors using half-strength Murashige and Skoog medium with an ammonium and nitrate ratio of 5:25 mM and supplemented with 1.0 mg l^?1 indole butyric acid, 0.1 mg l^?1 kinetin, and 3 % sucrose for the production of bioactive phenolics and flavonoids. Then 4.6 and 6.3 kg dry biomass were realized in the 500 l each of drum-type and balloon-type bioreactors, respectively. Accumulation of 66.9 mg g^?1 DW of total phenolics, 48.6 mg g^?1 DW of total flavonoids, 1.3 mg g^?1 DW of chlorogenic acid, 0.01 mg g^?1 DW of hyperin, 0.04 mg g^?1 DW of hypericin, and 0.01 mg g^?1 DW of quercetin could be achieved with adventitious roots cultured in 500 l balloon-type airlift bioreactors. Our findings demonstrate the possibilities of using H. perforatum adventitious root cultures for the production of useful phytochemicals to meet the demand of pharmaceutical and food industry.     

Production of a new mucilage compound in Lepidium sativum callus by optimizing in vitro growth conditions

The mucilage in Lepidium sativum L. is considered a biologically active compound with diverse medicinal properties. Different explants (hypocotyls and leaf) were transferred to Murashige and Skoog (MS) medium supplemented with twelve different plant growth regulator combinations under two different incubations (light and dark). The best mucilage production from callus (36.76% g g^?1 dry weight) was obtained in the MS medium supplemented with 1 mg L^?1 of 2, 4-D and 2 mg L^?1 of BAP under the light condition. The mucilage produced by callus culture was nearly three times more than the mucilage yield of the seeds. The glucose, arabinose + mannose and galactose were 43.4 (mg g^?1 DW), 195.3 (mg g^?1 DW) and 86.2 (mg g^?1 DW) in the mucilage originated from seed, callus leaf and callus hypocotyl, respectively. The present study proposes an efficient method for producing large scales of mucilage with a favorable sugar aimed at food or pharmaceutical industries.     

Optimization of Elicitation Conditions with Methyl Jasmonate and Salicylic Acid to Improve the Productivity of Withanolides in the Adventitious Root Culture of Withania somnifera (L.) Dunal

Adventitious root cultures derived from leaf derived callus of Withania somnifera (L.) Dunal were treated with methyl jasmonate and salicylic acid independently. Biomass accumulation, culture age, elicitation period, and culture duration were optimized for higher withanolides production in the two best-responding varieties collected from Kolli hills (Eastern Ghats) and Cumbum (Western Ghats) of Tamil Nadu, India. Between the two elicitors, salicylic acid (SA) improved the production of major withanolides (withanolide A, withanolide B, withaferin A, and withanone) as well as minor constituents (12-deoxy withastramonolide, withanoside V, and withanoside IV) in the Kolli hills variety. Treatment of root biomass (11.70?g FW) on 30-day-old adventitious root cultures with 150???M SA for 4?h elicitor exposure period resulted in the production of 64.65?mg?g^?l dry weight (DW) withanolide A (48-fold), 33.74?mg?g^?l DW withanolide B (29-fold), 17.47?mg?g^?l DW withaferin A (20-fold), 42.88?mg?g^?l DW withanone (37-fold), 5.34?mg?g^?l DW 12-deoxy withastramonolide (nine fold), 7.23?mg?g^?l DW withanoside V (seven fold), and 9.45?mg?g^?l DW withanoside IV (nine fold) after 10?days of elicitation (40th day of culture) when compared to untreated cultures. This is the first report on the use of elicitation strategy on the significant improvement in withanolides production in the adventitious root cultures of W. somnifera.     

Optimised isolation of polysaccharides from Lentinula edodes strain NCBI JX915793 using response surface methodology and their antibacterial activities

Mycelial growth in a defined medium by submerged fermentation is a rapid and alternative method for obtaining fungal biomass of consistent quality. Biomass, exopolysaccharides (EPS) and intracellular polysaccharides (IPS) production were optimised by response surface methodology in Lentinula edodes strain LeS (NCBI JX915793). The optimised conditions were pH 5.0, temperature 26°C, incubation period of 25 days and agitation rate of 52 r/min for L. edodes strain LeS. Under the calculated optimal culture conditions, biomass production (5.88 mg mL^? 1), EPS production (0.40 mg mL^? 1) and IPS production (12.45 mg g^? 1) were in agreement with the predicted values for biomass (5.93 mg mL^? 1), EPS (0.55 mg mL^? 1) and IPS production (12.64 mg g^? 1). Crude lentinan exhibited highest antibacterial effects followed by alcoholic, crude and aqueous extracts. The results obtained may be useful for highly effective yield of biomass and bioactive metabolites.     

Influence of Culture Medium Composition and Light Conditions on the Accumulation of Bioactive Compounds in Shoot Cultures of <Emphasis Type="Italic">Scutellaria lateriflora</Emphasis> L. (American Skullcap) Grown In Vitro

Methanolic extracts from in vitro grown Scutellaria lateriflora shoots cultured on five Murashige and Skoog (MS) medium variants supplemented with different combinations of 6-benzylaminopurine (BAP) and α-naphthaleneacetic acid (NAA) under different light conditions (monochromatic light, white light and no light) were analysed by HPLC for three groups of metabolites: flavonoids (26 compounds), phenolic acids and their precursors (19+2) and phenylethanoid glycosides (2). The analyses revealed the presence of baicalein, baicalin, wogonin, wogonoside, 3,4-dihydroxyphenylacetic acid and verbascoside. There was clear evidence of the influence of plant growth regulators and light conditions on the accumulation of the analysed groups of secondary metabolites. The amounts of the compounds changed within a wide range—for the total flavonoid content, 30.2-fold (max. 1204.3 mg·100 g^?1 dry weight (DW)); for 3,4-dihydroxyphenylacetic acid, 5.5-fold (max. 33.56 mg·100 g^?1 DW); and for verbascoside, 1.5-fold (169.15 max. mg·100 g^?1 DW). The best medium for the production of most of the compounds was the Murashige and Skoog variant with 1 mg l^?1 BAP and 1 mg l^?1 NAA. For verbascoside, the best ‘productive’ medium was the MS variant supplemented with 0.5 mg l^?1 BAP and 2 mg l^?1 NAA. The accumulation of the metabolites was stimulated to the greatest extent by blue light, under which the extracts were found to contain the highest total amount of flavonoids and the highest amounts of flavonoid glucuronides, baicalin and wogonoside, as well as of verbascoside. Their amounts were, respectively, 1.54-, 1.49-, 2.05- and 1.86-fold higher than under the control white light.     

Effect of Elicitors and Precursors on Azadirachtin Production in Hairy Root Culture of Azadirachta indica

The present study involved strategies for enhancement in in vitro azadirachtin (commercially used biopesticide) production by hairy root cultivation of Azadirachta indica. Improvement in the azadirachtin production via triggering its biosynthetic pathway in plant cells was carried out by the exogenous addition of precursors and elicitors in the growth medium. Among the different abiotic stress inducers (Ag⁺, Hg⁺², Co⁺², Cu⁺²) and signal molecules (methyl jasmonate and salicylic acid) tested, salicylic acid at 15 mg l^?1 of concentration was found to enhance the azadirachtin yield in the hairy roots to the maximum (up to 4.95 mg g^?1). Similarly, among the different biotic elicitors tested (filter-sterilized fungal culture filtrates of Phoma herbarium, Alternaria alternata, Myrothecium sp., Fusarium solani, Curvularia lunata, and Sclerotium rolfsii; yeast extract; and yeast extract carbohydrate fraction), addition of filter-sterilized fungal culture filtrate of C. lunata (1 %?v/v) resulted in maximum azadirachtin yield enhancement in hairy root biomass (up to 7.1 mg g^?1) with respect to the control (3.3 mg g^?1). Among all the biosynthetic precursors studied (sodium acetate, cholesterol, squalene, isopentynyl pyrophosphate, mavalonic acid lactone, and geranyl pyrophosphate), the overall azadirachtin production (70.42 mg l^?1 in 25 days) was found to be the highest with cholesterol (50 mg l^?1) addition as an indirect precursor in the medium.     

Evaluation of antioxidant activities of the edible and medicinal Acacia albida organs related to phenolic compounds

This study compared phenolic contents and antioxidant activity in different organs of Acacia albida (leaves and bark) and focuses on identification of phenolic compounds of leaves by HPLC-DAD. The analysed organs exhibited differences in total polyphenol contents (100 and 59.5 mg GAE g^? 1 DW). Phenolic contents of leaves were two times higher than those in bark. Ethanolic extracts exhibited good antioxidant activities with IC₅₀ = 26 μg mL^? 1 for DPPH and EC₅₀ = 50 μg mL^? 1 for FRAP. Identification by HPLC-DAD revealed the presence of nine phenolic compounds known for their high antioxidant activity. The results suggested that this species can be used as source of natural antioxidants.     

Optimization of Adventitious Root Culture for Production of Biomass and Secondary Metabolites in Prunella vulgaris L.

Adventitious root cultures of Prunella vulgaris L. were established in shaking flask system for the production of biomass and secondary metabolites. Adventitious root cultures were induced from callus cultures obtained from leaf explants on solid Murashige and Skoog (MS) medium containing combination of 6-benzyladenine (BA; 1.0 mg l^?1) and naphthalene acetic acid (NAA; 1.5 mg l^?1). Thereafter, 0.49 g inoculum was transferred to liquid MS medium supplemented with different concentrations of NAA (0.5–2.0 mg l^?1). Growth kinetics of adventitious roots was recorded with an interval of 7 days for 49 days period. Highest biomass accumulation (2.13 g/l) was observed in liquid medium containing 1.0 mg l^?1 NAA after 21 days of inoculation. However, other concentrations of NAA also showed similar accumulation pattern but the biomass gradually decreases after 49 days of inoculation. Adventitious roots were collected and dried for investigation of total phenolics (TP), total flavonoids (TF), and antioxidant activities. Higher TPC (0.995 GAE mg/g-DRB) and TFC (6.615 RE mg/g-DRB) were observed in 0.5 mg l^?1 NAA treated cultures. In contrast, higher antioxidant activity (83.53 %) was observed 1.5 mg l^?1 NAA treated cultures. These results are helpful in up scaling of root cultures into bioreactor for secondary metabolites production.     

Mineral Supplementation Increases Erythrose Reductase Activity in Erythritol Biosynthesis from Glycerol by Yarrowia lipolytica

The aim of this study was to examine the impact of divalent copper, iron, manganese, and zinc ions on the production of erythritol from glycerol by Yarrowia lipolytica and their effect on the activity of erythrose reductase. No inhibitory effect of the examined minerals on yeast growth was observed in the study. Supplementation with MnSO₄·7H₂O (25 mg l^?1) increased erythritol production by Y. lipolytica by 14.5 %. In the bioreactor culture with manganese ion addition, 47.1 g l^?1 of erythritol was produced from 100.0 g l^?1 of glycerol, which corresponded to volumetric productivity of 0.87 g l^?1 h^?1. The addition of Mn²⁺ enhanced the intracellular activity of erythrose reductase up to 24.9 U g^?1 of dry weight of biomass (DW), hence, about 1.3 times more than in the control.     

A Rapid Method for the Analysis of Ten Compounds in Psoralea corylifolia L. by UPLC

The objective of this study was to develop a simple, efficient and reliable method for routine quantitative analysis for Psoralea corylifolia L. An ultra performance liquid chromatography with DAD detector system was employed for simultaneous quantification of ten compounds. The chromatographic analysis was performed by UPLC with C₁₈ column and gradient elution of 0.05% formic acid aqueous solution and acetonitrile in 16 min. All calibration curves were linear (R ² ≥ 0.9990) over the tested ranges. The LOD and LOQ were lower to 13.07 and 39.22 ng mL^?1 with 2 μL of injection volume, respectively. The intra- and inter-day precisions as determined from sample solutions were below 4.1 and 4.2%. The average recoveries were ranged from 94.2 to 108.8% with RSDs ≤ 4.6%. This validated method was applied for the analysis of ten analytes in P. corylifolia L. from different origins. The variation of the content of ten compounds was remarkable among the tested samples: psoralenoside increased from 7.42 to 17.04 mg g^?1, isopsoralenoside from 6.05 to 14.34 mg g^?1, psoralen from 2.37 to 3.90 mg g^?1, isopsoralen from 2.53 to 3.65 mg g^?1, neobavaisoflavone from 1.59 to 2.96 mg g^?1, bavachin from 1.02 to 2.35 mg g^?1, psoralidin from 0.45 to 1.91 mg g^?1, isobavachalcone from 1.33 to 4.71 mg g^?1, corylifol A from 1.02 to 2.40 mg g^?1 and bakuchiol from 28.10 to 63.89 mg g^?1.     

Screening of bioconstituents and in vitro cytotoxicity of Clematis gouriana leaves

Clematis gouriana (Ranunculaceae), a perennial herb, is used by the local inhabitants of the western Himalayan region for its medicinal properties. Major bioconstituents of C. gouriana leaves using different solvent extracts were obtained and analysed. The results revealed promising contents of phenolics (from 18.19 ± 0.10 to 22.17 ± 0.10 mg g^? 1) as gallic acid and flavonoids (from 2.83 ± 0.01 to 6.52 ± 0.08 mg g^? 1) as quercetin equivalent in different extracts. Aqueous acetone extract showed higher antioxidant activity with IC₅₀ value of 129.11 and 25.35 μg mL^? 1 against DPPH and ABTS free radicals, respectively. Antioxidant yield ranged from 16.87 ± 0.27 to 24.48 ± 0.13 mg g^? 1 of Trolox equivalent in different extracts as measured by the FRAP assay. Furthermore, ethylacetate extract exhibited strong in vitro cytotoxicity against Chinese hamster ovary and glioma cell lines. Proximate composition (proteins, fats, ash and minerals) of C. gouriana leaves was also assessed. Results demonstrated the potential of C. gouriana bioconstituents as nutraceuticals.     

In vitro antifungal activity of extracts obtained from Hypericum perforatum adventitious roots cultured in a mist bioreactor against planktonic cells and biofilm of Malassezia furfur

Xanthone-rich extracts from Hypericum perforatum root cultures grown in a Mist Bioreactor as antifungal agents against Malassezia furfur.

Extracts of Hypericum perforatum roots grown in a bioreactor showed activity against planktonic cells and biofilm of Malassezia furfur. Dried biomass, obtained from roots grown under controlled conditions in a ROOTec mist bioreactor, has been extracted with solvents of increasing polarity (i.e. chloroform, ethyl acetate and methanol). The methanolic fraction was the richest in xanthones (2.86 ± 0.43 mg g^? 1 DW) as revealed by HPLC. The minimal inhibitory concentration of the methanol extract against M. furfur planktonic cells was 16 μg mL^? 1. The inhibition percentage of biofilm formation, at a concentration of 16 μg mL^? 1, ranged from 14% to 39%. The results show that H. perforatum root extracts could be used as new antifungal agents in the treatment of Malassezia infections.     

Preparation of activated carbon from organic fraction of municipal solid wastes by ZnCl2 activation method and use it for elimination of chromium(VI) from aqueous solutions

In this study, the use of the organic fraction of municipal solid waste as an abundant and low-cost raw material for producing activated carbon was investigated. For this purpose, ZnCl₂ was used as a chemical activation agent and the carbonization process took place at 800 °C in N₂ atmosphere. Seven sorbents were prepared by chemical activation (pyrolysis under N₂ atmosphere at temperature of 800 °C after impregnation with ZnCl₂) with different ratios of ZnCl₂. The optimum ratio of organic fraction of municipal solid waste to ZnCl₂ was inspected via methylene blue number and iodine number (ASTM Designation: D4607–94). The results showed that the adsorbent with 60 % ZnCl₂/raw material was the most appropriate one with a satisfactory adsorption capacity, 112.4 mg g^?1 for methylene blue and 134.0 mg g^?1 for iodine. In addition, the structural analysis of this sorbent was performed using FT-IR, BET surface area, SEM–EDX and thermal analysis. Application of this sorbent to remove Cr(VI) from wastewater was studied to find an adsorption capacity of 66.7 mg g^?1. The experimental adsorption equilibrium data were fitted to Langmuir adsorption model with an acceptable adsorption capacity of 66.7 mg g^?1.     

Rapid Determination of Coumarins in Plants by Capillary Electrophoresis

The goal of this study was to optimize the parameters for capillary electrophoresis to separate and determine aesculin, aesculetin, umbelliferone, and dihydrocoumarin in plant materials. A simple and rapid capillary zone electrophoresis method is reported for this separation that required less than nine minutes. A bare fused silica capillary was employed using 20 mM borax in 5% methanol at pH 10.1 as the background electrolyte with an applied voltage of 30 kV at 27°C. Good linearity and reproducibility were obtained with high correlation coefficients. The analytes were determined by molecular absorption at 194 and 206 nm. Coumarins were determined in Aesculus hippocastanum L., Cichorium intybus L., Melilotus officinalis L., and Juniperus communis L. “Pendula.” The concentrations of aesculin and aesculetin were 3.07 and 6.31 mg g^?1 in Aesculus hippocastanum. In Cichorium intybus, the aesculetin concentration was 2.42 mg g^?1. The dihydrocoumarin concentration was 0.54 mg g^?1 in Melilotus officinalis, and the concentration of umbelliferone was 0.58 mg g^?1 in Juniperus communis “Pendula.”     

Characterisation of phenolic compounds of the ethyl acetate fraction from Tabernaemontana catharinensis and its potential antidepressant-like effect

This study evaluates the antidepressant-like effect and analysed the qualitative and quantitative 74 phenolic standards of ethyl acetate fraction from Tabernaemontana catharinensis leaves. Acute administration of fraction in mice reduced the immobility time in forced swimming and tail suspension tests confirming its antidepressant-like activity. The anti-immobility effect elicited by this fraction was prevented by the pretreatment of mice with PCPA (100 mg kg^?1), ketanserin (5 mg kg^?1), SCH 23,390 (0.05 mg kg^?1) or yohimbine (1 mg kg^?1). A sub effective dose of the fraction produced a synergistic effect with fluoxetine (5 mg kg^?1). Chromatographic analysis identified 4-hydroxybenzoic and p-coumaric acids in the ethyl acetate fraction from T. catharinensis. Capillary electrophoresis presented 7.34 ± 0.02 mg g^?1 of p-coumaric acid concentration in the fraction. Therefore, it is possible that antidepressant-like effect elicited by ethyl acetate fraction from T. catharinensis be dependent on the p-coumaric acid.     

Octahedral copper(II) carboxylate complex: synthesis,structural description,DNA-binding and anti-bacterial studies

Self-assembly of CuSO₄, para-methyl-2-phenyl acetate and 1,10-phenanthroline afforded good-quality crystalline complex in quantitative yield. The complex was characterized by FTIR and UV-visible spectroscopy, electrochemistry, and powder and single-crystal XRD studies. Its structure was found to possess axially elongated octahedral symmetry with CuO₄N₂ chromophore. Its purity was assessed by powder XRD spectrum. Absorption study yielded a broad band corresponding to ²E_g→²T_2 g transition. Electrochemical solution study indicated diffusion-controlled irreversible electron transfer process corresponding to Cu(II)/Cu(I) redox couple with diffusion coefficient = 7.89(±0.1)×10^?9 cm²s^?1. Results of spectroscopic techniques support each other. Complex exhibited excellent DNA-binding ability through UV-visible spectroscopy and cyclic voltammetry yielding K_b values 1.399 × 10⁴ M^?1 and 5.81 × 10³ M^?1, respectively. The complex exhibited significant activity against bacterial strains Escherichia coli, Micrococcus luteus and Staphylococcus aureus and good activity against Bacillus subtilis. These preliminary studies impart good biological relevance on the synthesized complex.     

Water-insoluble β-cyclodextrin polymer crosslinked by citric acid: synthesis and adsorption properties toward phenol and methylene blue

Water-insoluble β-cyclodextrin polymer (β-CDP) crosslinked by citric acid was obtained with a yield of 65% through an environment friendly synthesis procedure. FT-IR spectra disclosed that the hydroxyl groups of β-CD had reacted and condensated with the carboxyl groups of citric acid, and at the same time the structural characteristics of β-CD were essentially maintained in β-CDP. The β-CDP exhibited notable adsorption capability toward phenol (q _max = 13.8 mg g^?1) and especially large adsorption capability toward methylene blue (q _max = 105 mg g^?1). The concentration of methylene blue in water could be reduced to 0.11 mg L^?1 by the β-CDP, indicating the excellent adsorption sensitivity of β-CDP toward methylene blue. The adsorption results disclosed that the interior cavity and inclusion property of β-CD were maintained in the synthesized β-CDP.     

Identification and Quantification of Heterologous Compounds Parthenin and Organic Acids in Parthenium Hysterophorus L. Using HPLC-PDA-MS-MS

Parthenium hysterophorus L., is an obnoxious weed known for its environmental health hazards and medicinal uses. These characteristics are due to presence of sesquiterpene lactones and organic acids; therefore a rapid and sensitive analytical procedure using HPLC-PDA-MS-MS was developed and optimized for separation, identification, and quantification of parthenin and six organic acids. Separation and characterization of compounds was achieved on a RP-C18 column with 1% acetic acid in water (A) and acetonitrile (B) as a mobile phase at a flow rate of 0.6 mL min^?1 and by matching their UV and mass spectra with reference compounds. Six organic acids (ferulic acid, 0.1 mg g^?1 to coumaric acid, 13.6 mg g^?1) and parthenin (27.4 mg g^?1) were characterized within 26 minutes of chromatographic separation in plant extract. The calibration curves are linear with correlation coefficients from 0.985 to 0.998, limit of detection and quantification ranged between 1.0 µg mL^?1 (anisic acid) to 2.2 µg mL^?1 (parthenin) and 2.5 µg mL^?1 (coumaric acid) to 5.2 µ g mL^?1 (parthenin) and recovery ranged between 90.9% to 97.3%. To the best of our knowledge this is the first report for the simultaneous separation of parthenin and organic acids. The method is applicable for screening of commercial crops, medicinal plants, and their products which might be mixed with P. hysterophorus during harvesting period.     

Purification and Characterisation of a β-1,4-Xylanase from Remersonia thermophila CBS 540.69 and Its Application in Bread Making

This work reports the first investigation of Remersonia thermophila hemicellulosic hydrolytic enzyme production, with subsequent purification of an extracellular endo-β-1,4-xylanase (RtXyl) and its application in bread making. The research describes RtXyl purification from sorghum-induced submerged liquid cultures of this moderately thermophilic, aerobic, ascomycete fungus. The purified enzyme is a single subunit protein with a molecular mass of 42 kDa and exhibits glycosyl hydrolase family-10-like activity over a broad pH and temperature range. Optimal activity was measured at pH 6.0 and 65 °C respectively, which is suitable for bread making applications. Substrate specificity studies revealed that RtXyl is purely xylanolytic with no side-activities against other plant polysaccharides. The RtXyl catalytic efficiency (K _cat/K _m) was highest with oats spelt xylan (810.90 mg mL^?1 s^?1), wheat arabinoxylan (809.52 mg mL^?1 s^?1) and beechwood xylan (417.40 mg mL^?1 s^?1) with less efficiency towards insoluble oats spelt xylan (236.40 mg mL^?1 s^?1). Hydrolysis products analysed by thin layer chromatography yielded a range of xylosaccharides, predominantly xylotriose and xylobiose. RtXyl application in a basic wheat bread recipe at low dosages (0.297 XU/g) showed its suitability to increase loaf volume by 8.0 % compared with the control bread. RtXyl increased loaf softness by 19.6 % while reducing bread staling by 20.4 % up to 4 days of storage.     

A Phenylurea Cytokinin,CPPU, Elevated Reducing Sugar and Correlated to Andrographolide Contents in Leaves of <Emphasis Type="Italic">Andrographis paniculata</Emphasis> (Burm. F.) Wall. Ex Nees

Cytokinins are phytohormones that play multiple roles to control plant growth and development. In this study, leaf biomass and the production of andrographolide compounds in a medicinal plant Andrographis paniculata were significantly increased after exogenously treating with the synthetic cytokinin cytokinin-1-(2-chloro-4-pyridyl)-3-phenylurea (CPPU) at 0 (water), 5, or 10 mg L^?1 and observed the results for 24 h, 48 h, and 7 days of treatment. It was found that CPPU could significantly enhance new axillary bud formation and further promote branching 4.6–5.6-fold higher, resulting in higher fresh weight (FW) and dry weight (DW) than the control. Application of CPPU at 5 mg L^?1 significantly promoted the highest contents of total reducing sugar at 2.5-fold in leaves and at 1.5-fold in roots. Although treatments of CPPU significantly affected the increasing contents of chlorophyll and carotenoid (1.2–1.6-fold), CPPU at 10 mg L^?1 slightly caused leaf stress and chlorophyll reduction. Interestingly, 5 mg L^?1 CPPU could enhance andrographolide content, an active anti-infectious compound in Andrographis paniculata (2.2-fold higher than the control) that reached the highest content at 24 h after treatment. This study suggested that CPPU should be suitable for field application to promote leaf yields and induce the production of useful pharmaceutical compounds in Andrographis paniculata.