Abstract
Genus Ophiorrhiza has recently emerged as one of the promising sources of Camptothecin (CPT), an antitumour monoterpene indole alkaloid. It possesses CPT in its every part and has a relatively short life span. To determine whether differentiation plays any role in the synthesis and/or accumulation of CPT, the concentration of CPT was analyzed across various tissues of Ophiorrhiza rugosa var. decumbens obtained through both direct as well as indirect modes of regeneration. The results revealed that the plants obtained from both types of regeneration showed similar levels of CPT. It was also observed that with differentiation, the accumulation of CPT increases, as the callus, being an undifferentiated mass of cells, had only traces of CPT. In contrast, the completely differentiated in-vitro plant obtained from it showed a significantly higher percentage of CPT in shoots (0.22% dry weight) and roots (0.247% dw). The CPT when analyzed after hardening, varied among different organs of the plant. It was also observed that the inflorescence accumulated the highest concentration of CPT (0.348% dw) once the flowering began, accompanied by a decrease in remaining organs. This decrease may result from CPT being mobilized to the inflorescence as a chemical defense mechanism. These findings allowed us to determine the ideal plant harvesting age for CPT extraction. The findings could be used to decide the right stage of plant harvest, which is just before the onset of blooming.
This is a preview of subscription content, log in via an institution to check access.
Data availability
Not applicable.
Code availability
Not applicable.
References
Cui L, Ni X, Ji Q, Teng X, Yang Y, Wu C, Kai G (2015) Co-overexpression of geraniol-10-hydroxylase and strictosidine synthase improves anticancer drug camptothecin accumulation in Ophiorrhiza pumila. Sci Rep 5(1):8227
Deb DB, Mondal DC (1997) Taxonomic revision of the genus Ophiorrhiza L. (Rubiaceae) in Indian subcontinent. Nelumbo 39(1–4):1–148. https://doi.org/10.20324/nelumbo/v39/1997/74298
Degambada KD, Peramune Arachchilage ASPK, Salim N, Chandrika UG, Abeysekera AM (2023) Cellular localisation and quantification of Camptothecin in different plant parts of Nothapodytes nimmoniana (J. Graham) Mabberley of Sri Lankan origin. Phytochem Anal 34(4):453–460. https://doi.org/10.1002/pca.3226
Flores HE, Hoy MW, Pickard JJ (1987) Secondary metabolites from root cultures. Trends Biotechnol 5(3):64–69. https://doi.org/10.1016/S0167-7799(87)80013-6
Fulzele DP, Satdive RK (2005a) Distribution of anticancer drug camptothecin in Nothapodytes foetida. Fitoterapia 76(7–8):643–648. https://doi.org/10.1016/j.fitote.2005.07.005
Fulzele DP, Satdive RK (2005b) Comparison of techniques for the extraction of the anticancer drug camptothecin from Nothapodytes foetida. J Chromatogr A 1063(1–2):9–13. https://doi.org/10.1016/j.chroma.2004.11.020
Hao X, Wang C, Zhou W, Ruan Q, Xie C, Yang Y, Kai G (2023) OpNAC1 transcription factor regulates the biosynthesis of the anticancer drug camptothecin by targeting loganic acid O-methyltransferase in Ophiorrhiza pumila. J Integr Plant Biol 65(1):133–149. https://doi.org/10.1111/jipb.13377
Kamble S, Gopalakrishnan R, Eapen S (2013) High-frequency plant regeneration and accumulation of the anticancer alkaloid camptothecin in Ophiorrhiza rugosa var. decumbens Deb & Mondal. J Herbs Spices Med Plants 19(4):321–328. https://doi.org/10.1080/10496475.2013.794182
Kaplan I, Halitschke R, Kessler A, Sardanelli S, Denno RF (2008) Constitutive and induced defenses to herbivory in above-and belowground plant tissues. Ecol 89(2):392–406. https://doi.org/10.1890/07-0471.1
Krishnakumar G, Dintu KP, Varghese SC, Nair DS, Gopinath G, Rameshkumar KB, Krishnan PN (2020) Ophiorrhiza, a promising herbaceous source of the anticancer compound camptothecin. Plant Sci Today 7(2):240–250. https://doi.org/10.14719/pst.2020.7.2.660
Lee JY, Goto E, Yoshida H, Hikosaka S (2022) Optimal harvest-time to maximize the annual camptothecin production by Ophiorrhiza pumila in a plant factory with artificial light. J Nat Med 76(4):865–872. https://doi.org/10.1007/s11418-022-01634-1
Linsmaier EM, Skoog F (1965) Organic growth factor requirements of tobacco tissue cultures. Physiol Plant 18:100–127. https://doi.org/10.1111/j.1399-3054.1965.tb06874.x
Lorence A, Nessler CL (2004) Camptothecin, over four decades of surprising findings. Phytochem 65(20):2735–2749. https://doi.org/10.1016/j.phytochem.2004.09.001
Mingzhang A, Jing W, Yue S, Wentao G, Longjiang Y (2011) Camptothecin distribution and content in Nothapodytes nimmoniana. Nat Prod Commun 6(2):197–200. https://doi.org/10.1177/1934578X1100600210
Montoro P, Maldini M, Piacente S, Macchia M, Pizza C (2010) Metabolite fingerprinting of Camptotheca acuminata and the HPLC–ESI-MS/MS analysis of camptothecin and related alkaloids. J Pharm Biomed Anal 51(2):405–415. https://doi.org/10.1016/j.jpba.2009.05.013
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
Namdeo AG, Sharma A (2012) HPLC analysis of camptothecin content in various parts of Nothapodytes foetida collected on different periods. Asian Pa J Trop Biomed 2(5):389–393. https://doi.org/10.1016/S2221-1691(12)60062-8
Rajan R, Varghese SC, Kurup R, Gopalakrishnan R, Venkataraman R, Satheeshkumar K, Baby S (2016) HPTLC-based quantification of camptothecin in Ophiorrhiza species of the southern Western Ghats in India. Cogent Chem 2(1):1275408. https://doi.org/10.1080/23312009.2016.1275408
Raveendran VV (2015) Camptothecin-discovery, clinical perspectives and biotechnology. Nat Prod Chem Res 3:175. https://doi.org/10.4172/2329-6836.1000175
Sankar-Thomas YD, Lieberei R (2011) Camptothecin accumulation in various organ cultures of Camptotheca acuminata Decne grown in different culture systems. Plant Cell Tissue Organ Cult 106(3):445–454. https://doi.org/10.1007/s11240-011-9942-6
Sibi CV, Dintu PK, Renjith R, Krishnaraj MV, Roja G, Satheeshkumar K (2012) A new record of Ophiorrhiza trichocarpon Blume (Rubiaceae: Ophiorrhizeae) from Western Ghats, India: Another source plant of camptothecin. J Sci Res 4(2):529. https://doi.org/10.3329/jsr.v4i2.9378
Singh S, Kamble SN, Satdive RK, Fulzele DP (2020) Heterologous overexpression of Nothapodytes foetida strictosidine synthase enhances levels of anti-cancer compound camptothecin in Ophiorrhiza rugosa. Plant Cell Tissue Organ Cult 141:67–76. https://doi.org/10.1007/s11240-020-01767-9
Thiers BM (updated continuously) (2024) Index Herbariorum. https://sweetgum.nybg.org/science/ih/
Venditto VJ, Simanek EE (2010) Cancer therapies utilizing the camptothecin: a review of the in vivo literature. Mol Pharm 7:307–349. https://doi.org/10.1021/mp900243b
Wang X, Tanaka M, Krstin S, Peixoto HS, de Melo Moura CC, Wink M (2016) Cytoskeletal interference–a new mode of action for the anticancer drugs camptothecin and topotecan. Eur J Pharmacol 789:265–274. https://doi.org/10.1016/j.ejphar.2016.07.044
Wen-Zhe LIU (2004) Secretory structures and their relationship to accumulation of camptothecin in Camptotheca acuminata (Nyssaceae). J Integr Plant Biol 46(10):1242
Wetterauer B, Wildi E, Wink M (2018) Production of the anticancer compound camptothecin in root and hairy root cultures of Ophiorrhiza mungos L. Biotechnological Approaches for Medicinal and Aromatic Plants. Springer, Singapore. pp 303–341. doi: https://doi.org/10.1007/978-981-13-0535-1_14
Wink M (1987) Why do lupin cell cultures fail to produce alkaloids in large quantities? Plant Cell Tissue Organ Cult 8:103–111. https://doi.org/10.1007/BF00043147
Yamazaki Y, Urano A, Sudo H, Kitajima M, Takayama H, Yamazaki M, Saito K (2003) Metabolite profiling of alkaloids and strictosidine synthase activity in camptothecin-producing plants. Phytochem 62(3):461–470. https://doi.org/10.1016/S0031-9422(02)00543-5
Yan XF, Wang Y, Yu T, Zhang YH, Dai SJ (2003) Variation in camptothecin content in Camptotheca acuminata leaves. Bot Bull Acad Sin 44:99–105
Acknowledgements
Authors acknowledge the assistance from Mr. Subham Bhakta for HPLC analysis and Head, Nuclear Agriculture & Biotechnology Division for his constant encouragement and support.
Funding
Work communicated in the present manuscript is funded by Department of Atomic Energy, Government of India.
Author information
Authors and Affiliations
Contributions
KS: Methodology, Data curation, Formal analysis, Original draft; RKS: Methodology, Formal analysis, Original draft; SS: Conceptualization, Methodology, Data curation, Formal analysis, Supervision, Original draft and Review.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest in the publication.
Ethical approval
This article does not contain any studies with human participants or animals performed by any of the authors.
Informed consent
All the authors gave their consent for publication of the results.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Sharma, K., Satdive, R.K. & Singh, S. Exploring the influence of organ differentiation on biosynthesis and accumulation of camptothecin in Ophiorrhiza rugosa var. decumbens. 3 Biotech 14, 153 (2024). https://doi.org/10.1007/s13205-024-03999-4
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s13205-024-03999-4