Current issue
Online first
Archive
About the Journal
Editorial Office
Editorial Board
Publisher
Reviewers
Editorial policies
Ethical standards and procedures
Abstracting and indexing
Contact
Subscription
Instructions for authors
Article processing charge (APC)
English language editing services
Books and Events
Books
Events
RESEARCH PAPER
In vitro regeneration of Portulaca grandiflora Hook. and analysis of betalain content within in vivo plants
1
Department of Botany, Bhavan’s Sheth R.A. College of Science, Gujarat University, Ahmedabad 380001, Gujarat, India
2
Department of Botany, Faculty of Science, The Maharaja Sayajirao University of Baroda, Vadodara 390002, Gujarat, India
Submission date: 2024-11-25
Final revision date: 2025-06-12
Acceptance date: 2025-08-20
Publication date: 2025-09-22
Corresponding author
Aruna Joshi
Department of Botany, Faculty of Science, The Maharaja Sayajirao University of Baroda, Vadodara 390002, Gujarat, India
Department of Botany, Faculty of Science, The Maharaja Sayajirao University of Baroda, Vadodara 390002, Gujarat, India
BioTechnologia 2025;106(3):291-308
KEYWORDS
TOPICS
ABSTRACT
Background:
Synthetic colors are widely used in the food and cosmetic industries to make products more appealing to consumers. However, the health hazards associated with synthetic colors have prompted their replacement with natural colors. Portulaca grandiflora is a promising candidate for natural color extraction, as it is rich in betalains. This study presents a reliable, reproducible three-step regeneration protocol for this plant and analyzes its betalain content.
Material and methods:
In vitro shoot cultures were established on Murashige and Skoog (MS) medium supplemented with different plant growth regulators. Anatomical studies were conducted to determine the stages of shoot primordium development. Betalains were extracted from in vivo plants using 60% methanol and subjected to spectrophotometric analysis. The effect of sodium ascorbate on betalain stability was also evaluated.
Results:
Juvenile leaf explants regenerated shoots on MS medium supplemented with 10 µM 6-benzyladenine and 5 µM indole-3-acetic acid. Shoots were multiplied with 20 µM BA (6.25 ± 0.85 shoots/explant), and elongation was achieved with 5 µM gibberellic acid (GA3) (8.2 ± 0.37 shoots/explant). Shoot primordia developed from well-organized meristemoid cells. The betalain content in the stem was 26.66 ± 0.19 mg/100 g, but this pigment degraded within 24 h (42.19% degradation). The addition of 50 mM sodium ascorbate prevented betalain degradation, even after 24 h.
Conclusions:
This study reports a regeneration protocol from juvenile leaf explants and demonstrates that betalain stability in the stem can be maintained with 50 mM sodium ascorbate.
Synthetic colors are widely used in the food and cosmetic industries to make products more appealing to consumers. However, the health hazards associated with synthetic colors have prompted their replacement with natural colors. Portulaca grandiflora is a promising candidate for natural color extraction, as it is rich in betalains. This study presents a reliable, reproducible three-step regeneration protocol for this plant and analyzes its betalain content.
Material and methods:
In vitro shoot cultures were established on Murashige and Skoog (MS) medium supplemented with different plant growth regulators. Anatomical studies were conducted to determine the stages of shoot primordium development. Betalains were extracted from in vivo plants using 60% methanol and subjected to spectrophotometric analysis. The effect of sodium ascorbate on betalain stability was also evaluated.
Results:
Juvenile leaf explants regenerated shoots on MS medium supplemented with 10 µM 6-benzyladenine and 5 µM indole-3-acetic acid. Shoots were multiplied with 20 µM BA (6.25 ± 0.85 shoots/explant), and elongation was achieved with 5 µM gibberellic acid (GA3) (8.2 ± 0.37 shoots/explant). Shoot primordia developed from well-organized meristemoid cells. The betalain content in the stem was 26.66 ± 0.19 mg/100 g, but this pigment degraded within 24 h (42.19% degradation). The addition of 50 mM sodium ascorbate prevented betalain degradation, even after 24 h.
Conclusions:
This study reports a regeneration protocol from juvenile leaf explants and demonstrates that betalain stability in the stem can be maintained with 50 mM sodium ascorbate.
REFERENCES (51)
1.
Anghel AI, Olaru OT, Gatea F, Dinu M, Ancuceanu RV, Istudor V. 2013. Preliminary research on Portulaca grandiflora Hook. species (Portulacaceae) for therapeutic use. Farmacia. 61(4): 694–702.
2.
Azni MM, Moradi H, Ghasemi K, Biparva P. 2021. Elicitation of dopamine biosynthesis in common purslane as affected by methyl jasmonate and silicon. J Plant Nutr. 44(20): 3083–3098. https://doi.org/10.1080/019041....
3.
Becerra DC, Forero AP, Gongora GA. 2004. Age and physiological condition of donor plants affect in vitro morphogenesis in leaf explants of Passiflora edulis f. flavicarpa. Plant Cell Tiss Organ Cult. 79: 87–90. https://doi.org/10.1023/B:TICU....
4.
Bhuiyan MNH, Adachi T. 2002. Efficient regeneration from hypocotyl cultures of betalain-forming plant Portulaca sp. cv. ‘Jewel’. Stimulatory effect of thidiazuron. Plant Biotechnol. 19(1): 57–61.
5.
Calvi P, Terzo S, Amato A. 2022. Betalains: colours for human health. Nat Prod Res. 1–20. https://doi.org/10.1080/147864....
6.
Chen S, Xiong Y, Yu X, Pang J, Zhang T, Wu K, Ren H, Jian S, Teixeira da Silva JA, Xiong Y, et al. 2020. Adventitious shoot organogenesis from leaf explants of Portulaca pilosa L. Sci Rep. 10: 3675. https://doi.org/10.1038/s41598....
7.
Chung HH, Schwinn KE, Ngo HM, Lewis DH, Massey B, Calcott KE, Crowhurst R, Joyce DC, Gould KS, Davies KM, Harrison DK. 2015. Characterization of betalain biosynthesis in Parakeelya flowers identifies the key biosynthetic gene DOD as belonging to an expanded LigB gene family that is conserved in betalain-producing species. Front Plant Sci. 6: 499. https://doi.org/10.3389/fpls.2....
8.
Cruz CF, Santos WF, Souza CS, Machado MD, Carvalho IF, Rocha DI, Silva ML. 2019. In vitro regeneration and flowering of Portulaca grandiflora Hook. Ornamental Hortic. 25(4): 443–449. https://doi.org/10.1590/2447-5....
9.
El-Ashry AAE, El-Bahr MK, Gabr AMM. 2020. Effect of light quality on betalain content of red beet (Beta vulgaris L.) cultured in vitro. Egypt Pharm J. 19(2): 143–148. https://doi.org/10.4103/epj.ep....
10.
Escribano J, Cabanes J, Jiménez-Atiénzara M, Ibañez-Tremolada M, Gómez-Pando LR, García-Carmona F, Gandía-Herrero F. 2017. Characterization of betalains, saponins and antioxidant power in differently colored quinoa (Chenopodium quinoa) varieties. Food Chem. 234: 285–291.https://doi.org/10.1016/j.food....
11.
Gandía-Herrero F, Escribano J, García-Carmona F. 2005. Betaxanthins as pigments responsible for visible fluorescence in flowers. Planta. 222(4): 586–593. https://doi.org/10.1007/s00425....
12.
Geng F, Moran R, Day M, Halteman W, Zhang D. 2016. Increasing in vitro shoot elongation and proliferation of ‘G.30’ and ‘G.41’ apple by chilling explants and plant growth regulators. HortScience. 51(7): 899–904. https://doi.org/10.21273/HORTS....
13.
Georgiev V, Ilieva M, Bley T, Pavlov A. 2008. Betalain production in plant in vitro systems. Acta Physiol Plant. 30: 581–593. https://doi.org/10.1007/s11738....
14.
Gilbert MG, Phillips SM. 2000. A review of the opposite leaved species of Portulaca in Africa and Arabia. Kew Bull. 55: 769–802. https://doi.org/10.2307/411362....
15.
Guerrero-Rubio MA, Hernández-García S, García-Carmona F, Gandía-Herrero F. 2019. Extension of life-span using a RNAi model and in vivo antioxidant effect of Opuntia fruit extracts and pure betalains in Caenorhabditis elegans. Food Chem. 274: 840–847. https://doi.org/10.1016/j.food....
16.
Guralnick LJ, Gilbert K, Denio D, Antico N. 2020. The development of Crassulacean acid metabolism (CAM) photosynthesis in cotyledons of the C4 species, Portulaca grandiflora (Portulacaceae). Plants. 9(1): 55. https://doi.org/10.3390/plants....
17.
Johansen DA. 1940. A plant microtechnique. London: McGraw-Hill Book Company, Inc.
18.
Kalyan K, Sil S. 2015. Proto corm-like bodies and plant regeneration from foliar explants of Coelogyne flaccida, a horticulturally and medicinally important endangered orchid of Eastern Himalaya. Lankesteriana. 15: 151–158. https://doi.org/10.15517/lank.....
19.
Kugler F, Stintzing FC, Carle R. 2004. Identification of betalains from petioles of differently colored Swiss chard (Beta vulgaris L. ssp. cicla [L.] Alef. cv. Bright Lights) by high-performance liquid chromatography-electrospray ionization mass spectrometry. J Agric Food Chem. 52(10): 2975–2981. https://doi.org/10.1021/jf0359....
20.
Little CHA, Macdonald JE. 2003. Effects of exogenous gibberellin and auxin on shoot elongation and vegetative bud development in seedlings of Pinus sylvestris and Picea glauca. Tree Physiol. 23: 73–83. https://doi.org/10.1093/treeph....
21.
Long Y, Yang Y, Pan G, Shen Y. 2022. New insights into tissue culture plant-regeneration mechanisms. Front Plant Sci. 13: 926752. https://doi.org/10.3389/fpls.2....
22.
McIlvaine T. 1921. A buffer solution for colorimetric comparison. J Biol Chem. 49: 183–186. https://doi.org/10.1016/S0021-....
23.
Moghadam YA, Piri Kh, Bahramnejad B, Ghiasvand T. 2014. Dopamine production in hairy root cultures of Portulaca oleracea (purslane) using Agrobacterium rhizogenes. J Agric Sci Technol. 16: 409–420.
24.
Moreno Sanz P, D’Amato D, Nebish A, Costantini L, Grando MS. 2020. An optimized histological procedure to study the female gametophyte development in grapevine. Plant Methods. 16: 61. https://doi.org/10.1186/s13007....
25.
Mousa AM, Abd-ElShafy E, Bedair R, Khafagi OMA, Abou Elkhier ZA. 2022. Callus induction and influence of biotic and abiotic elicitation on active constituents of Portulaca oleracea L. calli induced in vitro. Egypt J Chem. 65(11): 29–40. https://doi.org/10.21608/ejche....
26.
Murashige T, Skoog F. 1962. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant. 15: 473–497.
27.
Nurcholis W, Aisyah SI, Saraswati ARM, Yudha YS. 2023. Total phenolics, flavonoid contents, and antioxidant activity of three selected Portulaca grandiflora mutants in MV8 generations as a result of recurrent irradiation technique. J Appl Biol Biotechnol. 11(3): 245–249. https://doi.org/10.7324/JABB.2....
28.
Polturak G, Aharoni A. 2018. “La Vie en Rose”: Biosynthesis, source, and applications of betalain pigments. Mol Plant. 11(1): 7–22. https://doi.org/10.1016/j.molp....
29.
Rossi-Hassani BD, Zryd JP. 1995. In vitro culture and plant regeneration of large flowered purslane. Plant Cell Tiss Organ Cult. 41: 281–283. https://doi.org/10.1007/BF000....
30.
Safdari Y, Kazemitabar SK. 2009. Plant tissue culture study on two different races of purslane (Portulaca oleracea L.). Afr J Biotechnol. 8(21): 5906–5912.
31.
Safdari Y, Kazemitabar SK. 2010. Direct shoot regeneration, callus induction and plant regeneration from callus tissue in moss rose (Portulaca grandiflora L.). Plant Omics J. 3(2): 45–51.
32.
Santos-Diaz MS, Velásquez-García Y, González-Chávez MM. 2005. Pigment production by callus of Mammillaria candida Scheidweiler (Cactaceae). Agrociencia. 39: 619–626.
33.
Schaller GC, Street IH, Kieber JJ. 2014. Cytokinin and the cell cycle. Curr Opin Plant Biol. 21: 7–15. https://doi.org/10.1016/j.pbi.....
34.
Schuchovski C, Sant’Anna-Santos BF, Marra RC, Biasi LA. 2020. Morphological and anatomical insights into de novo shoot organogenesis of in vitro ‘Delite’ rabbiteye blueberries. Heliyon. 6(11): e05468. https://doi.org/10.1016/j.heli....
35.
Sedaghati B, Haddad R, Bandehpour M. 2019. Efficient plant regeneration and Agrobacterium-mediated transformation via somatic embryogenesis in purslane (Portulaca oleracea L.), an important medicinal plant. Plant Cell Tiss Organ Cult. 136: 231–245. https://doi.org/10.1007/s11240....
36.
Silva JPP, Bolanho BC, Stevanato N, Massa TB, da Silva C. 2020. Ultrasound-assisted extraction of red beet pigments (Beta vulgaris L.): Influence of operational parameters and kinetic modelling. J Food Process Preserv. 44: e14762. https://doi.org/10.1111/jfpp.1....
37.
Spórna-Kucab A, Tekieli A, Grzegorczyk A, Œwi¹tek £, Rajtar B, Skalicka-WoŸniak K, Starzak K, Nemzer B, Pietrzkowski Z, Wybraniec S. 2022. Metabolite profiling analysis and the correlation with biological activity of betalain-rich Portulaca grandiflora Hook. extracts. Antioxidants. 11: 1654. https://doi.org/10.3390/antiox....
38.
Srivastava A, Joshi A. 2009. In vitro behavior of nodal explants of Portulaca grandiflora under the influence of cytokinins. Acta Univ Latviensis. 753: 43–48.
39.
Srivastava A, Joshi A. 2021. Fatty acid analysis of in vitro shoot cultures of Portulaca oleracea Linn. Plant Physiol Rep. 26: 321–328. https://doi.org/10.1007/s40502....
40.
Srivastava A, Joshi A. 2024. Optimization of callus induction protocol from leaf explants of Portulaca oleracea and assessment of fatty acid profiles. Environ Exp Biol. 22: 157–166. https://doi.org/10.22364/eeb.2....
41.
Swamy GJ, Sangamithra A, Chandrasekar V. 2014. Response surface modeling and process optimization of aqueous extraction of natural pigments from Beta vulgaris using Box–Behnken design of experiments. Dyes Pigments. 111: 64–74. https://doi.org/10.1016/j.dyep....
42.
Timoneda A, Feng T, Sheehan H, Walker-Hale N, Pucker B, Lopez-Nieves S, Guo R, Brockington S. 2019. The evolution of betalain synthesis in Caryophyllales. New Phytol. 224: 71–85. https://doi.org/10.1111/nph.15....
43.
Trezzine GF, Zryd JP. 1991. Two betalains from Portulaca grandiflora. Phytochemistry. 30(6): 1897–1899. https://doi.org/10.1016/0031-9....
44.
Trezzini GF, Zryd JP. 1990. Portulaca grandiflora: A model system for the study of the biochemistry and genetics of betalain synthesis. Acta Hortic. 280: 581–585. https://doi.org/10.17660/ActaH....
45.
Vieira Teixeira da Silva D, Dos Santos Baião D, de Oliveira Silva F, Alves G, Perrone D, Mere Del Aguila E, Flosi Paschoalin VM. 2019. Betanin, a natural food additive: Stability, bioavailability, antioxidant and preservative ability assessments. Molecules. 24: 458. https://doi.org/10.3390/molecu....
46.
Voznesenskaya EV, Koteyeva NK, Edwards GE, Ocampo G. 2010. Revealing diversity in structural and biochemical forms of C4 photosynthesis and a C3–C4 intermediate in genus Portulaca L. (Portulacaceae). J Exp Bot. 61(13): 3647–3662. https://doi.org/10.1093/jxb/er....
47.
Wang C, Yu J, Gallagher DL, Byrd J, Yoo W, Wang Q, Guo O, Dietrich AM, Yang M. 2020. Pyrazines: A diverse class of earthy-musty odorants impacting drinking water quality and consumer satisfaction. Water Res. 182: 115971. https://doi.org/10.1016/j.watr....
48.
Wang T, Liu L, Rakhmanova A, Wang X, Shan Y, Yi Y, Liu B, Zhou Y, Lu X. 2020. Stability of bioactive compounds and in vitro gastrointestinal digestion of red beetroot jam: effect of processing and storage. Food Biosci. 38: 100788. https://doi.org/10.1016/j.fbio....
49.
Winson KWS, Chew BL, Sathasivam K, Subramaniam S. 2021. Effect of amino acid supplementation, elicitation and LEDs on Hylocereus costaricensis callus cultures for the enhancement of betalain pigments. Sci Hortic. 289: 110459. https://doi.org/10.1016/j.scie....
50.
Woo S, Wetzstein HY. 2008. Morphological and histological evaluation of in vitro regeneration in Elliottia racemosa leaf explants induced on media with thidiazuron. J Am Soc Hortic Sci. 133(2): 167–172. https://doi.org/10.21273/JASHS....
51.
Xu M, Zhao X, Fang J, Yang Q, Li J, Yan J. 2024. An effective somatic-cell regeneration and genetic transformation method mediated by Agrobacterium tumefaciens for Portulaca oleracea L. Plants. 13: 2390. https://doi.org/10.3390/plants....
Share
RELATED ARTICLE
| eISSN: | 2353-9461 |
| ISSN: | 0860-7796 |
We process personal data collected when visiting the website. The function of obtaining information about users and their behavior is carried out by voluntarily entered information in forms and saving cookies in end devices. Data, including cookies, are used to provide services, improve the user experience and to analyze the traffic in accordance with the Privacy policy. Data are also collected and processed by Google Analytics tool (more).
You can change cookies settings in your browser. Restricted use of cookies in the browser configuration may affect some functionalities of the website.
You can change cookies settings in your browser. Restricted use of cookies in the browser configuration may affect some functionalities of the website.
