Skip to main content
SpringerLink
Log in

Prolonged incubation of callus on auxin herbicide 2,4-D displayed significant effect on alkaloid production in callus of the woody medicinal plant Polyalthia bullata

  • Bioactive/Medicinal Compounds
  • Published:
In Vitro Cellular & Developmental Biology - Plant Aims and scope Submit manuscript

Abstract

The production of alkaloids in the intact plant is usually low, and some of them are tissue-specific and growth stage–specific. In this study, the total alkaloid content (TAC) in Polyalthia bullata was determined using different extraction solvents, and the extracts were subjected to LC-MS analysis. The manipulation of alkaloid production in P. bullata callus was further carried out by growing the callus on two basal media (Murashige and Skoog (MS) and Woody Plant Medium (WPM)) containing auxins (2,4-dichlorophenoxyacetic acid (2,4-D), α-naphthaleneacetic acid (NAA), 4-amino-3,5,6-trichloropicolinic acid (picloram), 3,6-dichloro-o-anisic acid (dicamba), indole-3-acetic acid (IAA), and indole-3-butyric acid (IBA)), precursors (L-phenylalanine, L-tyrosine, and L-tryptophan), and elicitors (methyl jasmonate (MeJA), salicylic acid (SA), and chitosan). The methanolic extracts exhibited the highest alkaloid content in leaf, stem, and root tissues (4.61 ± 0.04 to 7.71 ± 0.04 mg/g DW), and nine alkaloids were detected in P. bullata. The callus grown on MS + 30 μM 2,4-D exhibited the highest alkaloid accumulation (31.07 ± 0.05 μg/mg DW) after 6 weeks of culture compared to other media: MS media containing auxins, MS media containing auxins and precursors, and MS media containing auxins and elicitors. Therefore, we conclude that the MS + 30 μM 2,4-D is potent in enhancing alkaloid production in P. bullata callus.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Figure 1.
Figure 2.
Figure 3.
Figure 4.

Similar content being viewed by others

References

  • Adil M, Ren X, Jeong BR (2019) Light elicited growth, antioxidant enzymes activities and production of medicinal compounds in callus culture of Cnidium officinale Makino. J. Photochem Photobiol B, Biol 196:111509–111515

    Article  CAS  Google Scholar 

  • Afsah EM, Fadda AA, Bondock S, Hammouda MM (2015) Synthesis and some reactions of functionalized 11, 12-dihydro-5H-dibenzo [b, g] azonines. Zeitschrift für Naturforschung B 70:385–391

    Article  CAS  Google Scholar 

  • Ain QU, Khan H, Mubarak MS, Pervaiz A (2016) Plant alkaloids as antiplatelet agent: drugs of the future in the light of recent developments. Front. Pharmacol 7:292–300

    Article  PubMed  PubMed Central  Google Scholar 

  • Amirkia V, Heinrich M (2014) Alkaloids as drug leads–a predictive structural and biodiversity-based analysis. Phytochem Lett 10:48–53

    Article  Google Scholar 

  • Andi SA, Gholami M, Ford CM, Maskani F (2019) The effect of light, phenylalanine and methyl jasmonate, alone or in combination, on growth and secondary metabolism in cell suspension cultures of Vitis vinifera. J Photochem Photobiol B, Biol 199:111625–111634

    Article  CAS  Google Scholar 

  • Anita TZ, Mahdi C, Beltran MAG, Aulanni’am A (2019) Identification and molecular docking analysis alkaloids Polyalthia longifolia leaves from Indonesia and the Philippines as anti-inflammatory. J Phys Conf Ser 1374:012049

    Article  CAS  Google Scholar 

  • Asuk AA, Agiang MA, Dasofunjo K, Willie AJ (2015) The biomedical significance of the phytochemical, proximate and mineral compositions of the leaf, stem bark and root of Jatropha curcas. Asian Pac J Trop Biomed 5:650–657

    Article  CAS  Google Scholar 

  • Azliza MA, Ong HC, Vikineswary S, Noorlidah A, Haron NW (2012) Ethno-medicinal resources used by the Temuan in Ulu Kuang Village. Studies on Ethno-Medicine 6:17–22

    Article  Google Scholar 

  • Baek S, Ho TT, Lee H, Jung G, Kim YE, Jeong CS, Park SY (2019) Enhanced biosynthesis of triterpenoids in Centella asiatica hairy root culture by precursor feeding and elicitation. Plant Biotechnol Rep 14:45–53

    Article  Google Scholar 

  • Besher S, Al-Ammouri Y, Murshed R (2014) Production of tropane alkaloids in the in vitro and callus cultures of Hyoscyamus aureus and their genetic stability assessment using ISSR markers. Physiol Mol Biol Plants 20:343–349

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bienaimé C, Melin A, Bensaddek L, Attoumbré J, Nava-Saucedo E, Baltora-Rosset S (2015) Effects of plant growth regulators on cell growth and alkaloids production by cell cultures of Lycopodiella inundata. Plant Cell Tissue Organ Cult (PCTOC) 123:523–533

    Article  Google Scholar 

  • Calixto NO, Pinto ME, Ramalho SD, Burger M, Bobey AF, Young MC, Bolzani VS, Pinto AC (2016) The genus Psychotria: phytochemistry, chemotaxonomy, ethnopharmacology and biological properties. J Braz Chem Soc 27:1355–1378

    CAS  Google Scholar 

  • Chen CY, Wu HM, Chao WY, Lee CH (2013) Review on pharmacological activities of liriodenine. African J Pharm Pharmacol 7:1067–1070

    Article  Google Scholar 

  • Connolly JD, Haque ME, Kadir AA (1996) Two 7, 7′-bisdehydroaporphine alkaloids from Polyalthia bullata. Phytochemistry 43:295–297

    Article  CAS  Google Scholar 

  • Das J, Mao AA, Handique PJ (2013) Callus-mediated organogenesis and effect of growth regulators on production of different valepotriates in Indian valerian (Valeriana jatamansi Jones.). ActaPhysiol Plant 35:55–63

    CAS  Google Scholar 

  • Desgagné-Penix I, Facchini PJ (2012) Systematic silencing of benzylisoquinoline alkaloid biosynthetic genes reveals the major route to papaverine in Opium poppy. Plant J 72:331–344

    Article  PubMed  Google Scholar 

  • Dias DA, Urban S, Roessner U (2012) A historical overview of natural products in drug discovery. Metabolites 2:303–336

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. ExpCell Res 50:151–158

    CAS  Google Scholar 

  • Guo P, Wang Z, Li G, Liu Y, Xie Y, Wang Q (2016) First discovery of polycarpine, polycarpaurines A and C, and their derivatives as novel antiviral and antiphytopathogenic fungus agents. J Agric Food Chem 64:4264–4272

    Article  CAS  PubMed  Google Scholar 

  • Gupta P, Sharma S, Saxena S (2015) Biomass yield and steviol glycoside production in callus and suspension culture of Stevia rebaudiana treated with proline and polyethylene glycol. Appl Biochem Biotechnol 176:863–874

    Article  CAS  PubMed  Google Scholar 

  • Hosseini NS, Hagh ZG, Khoshghalb H (2020) Morphological, antioxidant enzyme activity and secondary metabolites accumulation in response of polyethylene glycol-induced osmotic stress in embryo-derived plantlets and callus cultures of Salvia leriifolia. Plant Cell Tissue Organ Cult (PCTOC) 140:143–155

    Article  CAS  Google Scholar 

  • Iriti M, Faoro F (2009) Chitosan as a MAMP, searching for a PRR. Plant Signal Behav 4:66–68

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Isah T (2016) Anticancer alkaloids from trees: development into drugs. Pharmacogn Rev 10:90–99

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Isah T, Umar S, Mujib A, Sharma MP, Rajasekharan PE, Zafar N, Frukh A (2018) Secondary metabolism of pharmaceuticals in the plant in vitro cultures: strategies, approaches, and limitations to achieving higher yield. Plant Cell. Tissue Organ Cult (PCTOC) 132:239–265

    Article  CAS  Google Scholar 

  • Jawahar G, Punita DL, Rajasheker G, Manoharachary C, Venkatachalam P, Kishor PK (2018) Feeding elicitors and precursors enhance colchicine accumulation in morphogenic cultures of Gloriosa superba L. Plant Cell Tissue Organ Cult (PCTOC) 135:235–245

    Article  CAS  Google Scholar 

  • Jeet A, Singh Y, Singh P, Nimoriya R, Bilung CJ, Kanojiya S, Tripathi V, Mishra DK (2020) Strategies for indole alkaloids enrichment through callus culture from Alstonia scholaris (L.) R. Br. Plant Growth Regul 90:383–392

    Article  CAS  Google Scholar 

  • John BIJU, Sulaiman CT, George S, Reddy VRK (2014) Spectrophotometric estimation of total alkaloids in selected Justicia species. Int J Pharm Pharm Sci 6:647–648

    Google Scholar 

  • Kamarul Zaman MA, Azzeme AM, Ramli SN, Shaharuddin NA, Ahmad S, Abdullah SNA (2020a) Solvent extraction and its effect on phytochemical yield and antioxidant capacity of woody medicinal plant, Polyalthia bullata. BioRes 15:9555–9568

    Article  Google Scholar 

  • Kamarul Zaman MA, Azzeme AM, Ramli SN, Shaharuddin NA, Ahmad S, Abdullah SNA (2020b) Induction, multiplication, and evaluation of antioxidant activity of Polyalthia bullata callus, a woody medicinal plant. Plants 9:1772

    Article  PubMed Central  Google Scholar 

  • Kaur J, Singh A, Pathak T, Kumar K (2017) Role of PGRs in anticancer alkaloids (vincristine and vinblastine) production. In: Naeem M, Aftab T, Khan M (eds) Catharanthus roseus. Springer, Cham, pp 309–319

    Chapter  Google Scholar 

  • Kuo CI, Chao CH, Lu MK (2011) Effects of auxins on the production of steroidal alkaloids in rapidly proliferating tissue and cell cultures of Solanum lyratum. Phytochem Anal 23:400–404

    Article  PubMed  Google Scholar 

  • Liu Y, Liang Z, Zhang Y (2010) Induction and in vitro alkaloid yield of calluses and protocorm-like bodies (PLBs) from Pinellia ternata. In Vitro Cell Dev Biol - Plant 46:239–245

    Article  CAS  Google Scholar 

  • Machado MW, Stern Neto C, Salgado J, Zaffari G, Barison A, Campos FR, Corilo YE, Eberlin MN, Biavatti MW (2010) Search for alkaloids on callus culture of Passiflora alata. Braz Arch Biol Technol 53:901–910

    Article  CAS  Google Scholar 

  • Mahood HE, Alwash BM, Ibrahim KM (2018) Improvement of alkaloids yield using phenylalanine as a precursor supplemented to Moringa oleifera L. callus cultures. Biochem Cell Arch 18:913–919

    Google Scholar 

  • Maqbool S, Younus I, Nesar RSS (2019) A short review on medicinal uses and pharmacological activities of Cocculus laurifolius leaves. International Journal of Advanced Research and Review. 4:14–20

    Google Scholar 

  • McCarthy-Suárez I (2017) Role of reactive oxygen species in auxin herbicide phytotoxicity: current information and hormonal implications—are gibberellins, cytokinins, and polyamines involved? Botany 95:369–385

    Article  Google Scholar 

  • McCown BH, Lloyd G (1981) Woody plant medium (WPM) – a mineral nutrient formulation for microculture of woody plant species. HortScience 16:453

    Google Scholar 

  • Mishra MR, Srivastava RK, Akhtar N (2018) Enhanced alkaloid production from cell culture system of Catharanthus roseus in combined effect of nutrient salts, sucrose and plant growth regulators. J Biomed Biotechnol 4:14–34

    Article  Google Scholar 

  • Mohamad M, Mohsin HF, Singh GKS (2017) A study on the effect of Eurycoma longifolia and Polyalthia bullata on reproductive organs and androgens of male Sprague-Dawley rats. J Eng Appl Sci 12:6994–6999

    CAS  Google Scholar 

  • Mohammed A, Chiruvella KK, Rao YK, Geethangili M, Raghavan SC, Ghanta RG (2015) In vitro production of echioidinin, 7-o-methywogonin from callus cultures of Andrographis lineata and their cytotoxicity on cancer cells. PLoS One 10:e0141154–e0141168

    Article  PubMed  PubMed Central  Google Scholar 

  • Moreira R, Pereira DM, Valentão P, Andrade PB (2018) Pyrrolizidine alkaloids: chemistry, pharmacology, toxicology and food safety. Int J Mol Sci 19:1668–1690

    Article  PubMed Central  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Naik PM, Al-Khayri JM (2016) Impact of abiotic elicitors on in vitro production of plant secondary metabolites: a review. Int J Adv Biotechnol Res 1:1–7

    Article  Google Scholar 

  • Nantapap SS, Punyanitya N, Nuntasaen W, Pompimon W, Meepowpan P (2017) Flavones from aerial parts of Polyalthia bullata and cytotoxicity against cancer cell lines. Chem Nat Compd 53:762–763

    Article  CAS  Google Scholar 

  • Paarakh PM, Khosa RL (2009) Phytoconstituents from the genus Polyalthia- a review. J Pharm Res 2:594–605

    CAS  Google Scholar 

  • Perviz S, Khan H, Pervaiz A (2016) Plant alkaloids as an emerging therapeutic alternative for the treatment of depression. Front Pharmacol 7:1–7

    Article  Google Scholar 

  • Pillai SK, Siril EA (2019) Enhanced production of berberine through callus culture of Tinospora cordifolia (Willd.) Miers ex Hook F. and Thoms. Proc Natl Acad Sci India B 1-9

  • Pliankong P, Suksa-Ard P, Wannakrairoj S (2018) Chitosan elicitation for enhancing of vincristine and vinblastine accumulation in cell culture of Catharanthus roseus (L.) G. Don. J Agric Sci 10:287–293

    Google Scholar 

  • Ptak A, Simlat M, Morańska E, Skrzypek E, Warchoł M, Tarakemeh A, Laurain-Mattar D (2019) Exogenous melatonin stimulated Amaryllidaceae alkaloid biosynthesis in in vitro cultures of Leucojum aestivum L. Ind Crops Prod 138:111458–111466

    Article  CAS  Google Scholar 

  • Ptak A, Tahchy A, Skrzypek E, Wójtowicz T, Laurain-Mattar D (2013) Influence of auxins on somatic embryogenesis and alkaloid accumulation in Leucojum aestivum callus. Open Life Sci 8:591–599

    Article  CAS  Google Scholar 

  • Radchenko OS, Novikov VL, Willis RH, Murphy PT, Elyakov GB (1997) Synthesis of polycarpine, a cytotoxic sulfur-containing alkaloid from the ascidian Polycarpa aurata, and related compounds. Tetrahedron Lett 38:3581–3584

    Article  CAS  Google Scholar 

  • Raj D, Kokotkiewicz A, Drys A, Luczkiewicz M (2015) Effect of plant growth regulators on the accumulation of indolizidine alkaloids in Securinega suffruticosa callus cultures. Plant Cell Tissue Organ Cult (PCTOC). 123:39–45

    Article  CAS  Google Scholar 

  • Ramakrishna A, Ravishankar GA (2011) Influence of abiotic stress signals on secondary metabolites in plants. Plant Signal Behav 6:1720–1731

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ramirez-Estrada K, Vidal-Limon H, Hidalgo D, Moyano E, Golenioswki M, Cusidó RM, Palazon J (2016) Elicitation, an effective strategy for the biotechnological production of bioactive high-added value compounds in plant cell factories. Molecules 21:182–206

    Article  PubMed  PubMed Central  Google Scholar 

  • Rao S, Usha K, Arjun (2015) Production of secondary metabolites from callus cultures of Centella asiatica (L.) Urban. Ann Phytomed 4:74–78

    CAS  Google Scholar 

  • Ratnadewi D (2017) Alkaloids in plant cell cultures. In: Georgiev V, Pavlov A (eds) Alkaloids-alternatives in synthesis, modification and application. IntechOpen, UK, pp 1–23

    Google Scholar 

  • Sayhan H, Beyaz SG, Çeliktaş A (2017) The local anesthetic and pain relief activity of alkaloids. In: Georgiv V (ed) Alkaloids: alternatives in synthesis, modification and application. IntechOpen, United Kingdom, pp 57–84

    Google Scholar 

  • Sharma A, Mathur AK, Ganpathy J, Joshi B, Patel P (2019) Effect of abiotic elicitation and pathway precursors feeding over terpenoid indole alkaloids production in multiple shoot and callus cultures of Catharanthus roseus. Biologia 74:543–553

    Article  CAS  Google Scholar 

  • Singh T, Yadav R, Agrawal V (2020) Effective protocol for isolation and marked enhancement of psoralen, daidzein and genistein in the cotyledon callus cultures of Cullen corylifolium (L.) Medik. Ind Crops Prod 143:111905–111915

    Article  CAS  Google Scholar 

  • Sivanandhan G, Selvaraj N, Ganapathi A, Manickavasagam M (2014) Enhanced biosynthesis of withanolides by elicitation and precursor feeding in cell suspension culture of Withania somnifera (L.) Dunal in shake-flask culture and bioreactor. PLoS One 9:e104005–e104016

    Article  PubMed  PubMed Central  Google Scholar 

  • Song Y (2014) Insight into the mode of action of 2, 4-dichlorophenoxyacetic acid (2,4-D) as an herbicide. J Integr Plant Biol 56:106–113

    Article  CAS  PubMed  Google Scholar 

  • Tekuri SK, Pasupuleti SK, Konidala KK, Pabbaraju N (2019) Pharmacological effects of Polyalthia cerasoides (Roxb.) Bedd.: a brief review. J Complement Med Res 10:38–49

    Article  CAS  Google Scholar 

  • Truong DH, Nguyen DH, Ta NTA, Bui AV, Do TH, Nguyen HC (2019) Evaluation of the use of different solvents for phytochemical constituents, antioxidants, and in vitro anti-inflammatory activities of Severinia buxifolia. J Food Qual 2019:1–9

    Article  Google Scholar 

  • Verma AK, Singh RR, Singh S (2012) Improved alkaloid content in callus culture of Catharanthus roseus. Bot Serb 36:123–130

    Google Scholar 

  • Xie Y, Feng Y, Di Capua A, Mak T, Buchko GW, Myler PJ, Liu M, Quinn RJ (2020) A phenotarget approach for identifying an alkaloid interacting with the tuberculosis protein Rv1466. Mar Drugs 18:149

    Article  CAS  PubMed Central  Google Scholar 

  • Yang L, Zhu J, Sun C, Deng Z, Qu X (2020) Biosynthesis of plant tetrahydroisoquinoline alkaloids through an imine reductase route. Chem Sci 11:364–371

    Article  CAS  PubMed  Google Scholar 

  • Zhu J, Wang M, Wen W, Yu R (2015) Biosynthesis and regulation of terpenoid indole alkaloids in Catharanthus roseus. Pharmacogn Rev 9:24–28

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zuldin M, Nahazima N, Said IM, Mohd Noor N, Zainal Z, Jin Kiat C, Ismail I (2013) Induction and analysis of the alkaloid mitragynine content of a Mitragyna speciosa suspension culture system upon elicitation and precursor feeding. The Scientific World Journal 2013:1–11

    Article  Google Scholar 

Download references

Acknowledgements

The authors wish to thank Universiti Putra Malaysia for funding the Master of Science Degree for the principle author under Graduate Research Fellowship (GRF) scheme.

Funding

This study was supported by the Putra Graduate Initiative (IPS) grant (GP-IPS/2018/9630400).

Author information

Authors and Affiliations

  1. Department of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400, UPM Serdang, Selangor, Malaysia

    Munirah Adibah Kamarul Zaman, Azzreena Mohamad Azzeme, Illy Kamaliah Ramle, Nurfazlinyana Normanshah, Noor Azmi Shaharuddin & Syahida Ahmad

  2. Institute of Plantation Studies, Universiti Putra Malaysia, 43400, UPM Serdang, Selangor, Malaysia

    Noor Azmi Shaharuddin & Siti Nor Akmar Abdullah

  3. Department of Agriculture Technology, Faculty of Agriculture, Universiti Putra Malaysia, 43400, UPM Serdang, Selangor, Malaysia

    Siti Nor Akmar Abdullah

Authors
  1. Munirah Adibah Kamarul Zaman
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Azzreena Mohamad Azzeme
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Illy Kamaliah Ramle
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Nurfazlinyana Normanshah
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Noor Azmi Shaharuddin
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Syahida Ahmad
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Siti Nor Akmar Abdullah
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

MAKZ carried out the research as part of her postgraduate project. IKR and NN helped in some parts of the research. AMA is the project leader and main supervisor. AMA also reviewed the manuscript. NAS, SA, and SNAA are co-supervisors.

Corresponding author

Correspondence to Azzreena Mohamad Azzeme.

Ethics declarations

Competing interests

The authors declare no competing interests.

Additional information

Editor: Yong Eui Choi

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Zaman, M.A.K., Azzeme, A.M., Ramle, I.K. et al. Prolonged incubation of callus on auxin herbicide 2,4-D displayed significant effect on alkaloid production in callus of the woody medicinal plant Polyalthia bullata. In Vitro Cell.Dev.Biol.-Plant 57, 749–759 (2021). https://doi.org/10.1007/s11627-021-10194-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11627-021-10194-0

Keywords

Search

Navigation