Abstract
Cyclotides are unique plant cyclic-peptides that can serve as agrochemicals, pharmaceutical scaffolds for drug delivery, and therapeutic agents. Currently, cyclotides are obtained only via direct extraction from limited plants. Hence, they serve as valuable candidates for synthesis via plant cell bioprocesses. In this study, callus lines (47 in total) were successfully induced from the leaf and petiole explants of the Indian medicinal plant, V. odorata, on a solidified woody plant medium (WPM) supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) (4.5 mg/l). Two fast growing callus lines, VOP-4 and VOL-44, were selected for the development of cell suspension cultures having a doubling time of 8 and 6 days, respectively. Further, known (15) and novel (9) cyclotides were identified for the first time in the callus and cell suspension cultures of V. odorata, using liquid chromatography and Fourier transform mass spectrometry. The cyclotides were identified based on their monoisotopic mass (2.5–4 kDa), hydrophobic nature, disulfide bonds, circular structure and amino acid sequence. Some of the cyclotides identified in the study (vodo I96, vodo I97, vodo I98) were exclusively produced in callus/cell suspension cultures and not in the parent plant. The study revealed that besides germplasm conservation, plant cell bioprocessing of V. odorata could be a potential alternative for in vitro production of known and novel cyclotides.
Similar content being viewed by others
References
Burman R, Gunasekera S, Strömstedt AA, Göransson U (2014) Chemistry and biology of cyclotides: circular plant peptides outside the box. J Nat Prod 77:724–736. doi:10.1021/np401055j
Craik DJ, Conibear AC (2011) The chemistry of cyclotides. J Org Chem 76:4805–4817. doi:10.1021/cr60113a001
Craik DJ, Daly NL, Bond T, Waine C (1999) Plant cyclotides: a unique family of cyclic and knotted proteins that defines the cyclic cystine knot structural motif. J Mol Biol 294:1327–1336. doi:10.1006/jmbi.1999.3383
Daly NL, Rosengren KJ, Craik DJ (2009) Discovery, structure and biological activities of cyclotides. Adv Drug Deliv Rev 61:918–930. doi:10.1016/j.addr.2009.05.003
Davazdahemami S (2010) Collection, cultivation and establishment rare and endangered medicinal plants of Iran in order to conservation and reclamation of them. http://agris.fao.org/agris-search/search.do?recordID=IR2012070062
Dörnenburg H (2010) Cyclotide synthesis and supply: from plant to bioprocess. Biopolymers 94:602–610. doi:10.1002/bip.21466
Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50:151–158. doi:10.1016/0014-4827(68)90403-5
Gustafson KR, Walton LK, Sowder RC et al (2000) New circulin macrocyclic polypeptides from Chassalia parvifolia. J Nat Prod 63:176–178. doi:10.1021/np990432r
Huang T-K, McDonald KA (2012) Bioreactor systems for in vitro production of foreign proteins using plant cell cultures. Biotechnol Adv 30:398–409. doi:10.1016/j.biotechadv.2011.07.016
Kedarisetti P, Mizianty MJ, Kaas Q et al (2014) Prediction and characterization of cyclic proteins from sequences in three domains of life. Biochim Biophys Acta J 1844:181–190. doi:10.1016/j.bbapap.2013.05.002
Kintzing JR, Filsinger Interrante MV, Cochran JR (2016) Emerging strategies for developing next-generation protein therapeutics for cancer treatment. Trends Pharmacol Sci 37:993–1008. doi:10.1016/j.tips.2016.10.005
Koehbach J, Attah AF, Berger A et al (2013) Cyclotide discovery in Gentianales revisited: identification and characterization of cyclic cystine-knot peptides and their phylogenetic distribution in rubiaceae plants. Pept Sci 100:438–452. doi:10.1002/bip.22328
Lim T (2014) Edible medicinal and non-medicinal plants. Springer, New York
Lloyd G, McCown B (1980) Commercially feasible micropropagation of mountain laurel Kalmia latifolia, by use of shoot-tip culture Comb. Proc Int Plant Prop Soc 30:421–427
Malik AR, Siddique MAA, Sofi PA, Butola JS (2011) Ethnomedicinal practices and conservation status of medicinal plants of North Kashmir Himalayas. Res J Med Plants 5:515–530. doi:10.3923/rjmp.2011.515.530
Muhammad N, Naveed I, Saqlan Naqvi SM, Mahmood T (2013) Standardization of tissue culture conditions and estimation of free scavenging activity in Viola odorata L. Pak J Bot 45:197–202
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497. doi:10.1111/j.1399-3054.1962.tb08052.x
Nguyen GKT, Zhang S, Nguyen NTK et al (2011) Discovery and characterization of novel cyclotides originated from chimeric precursors consisting of albumin-1 chain a and cyclotide domains in the fabaceae family. J Biol Chem 286:24275–24287. doi:10.1074/jbc.M111.229922
Poth AG, Colgrave ML, Philip R et al (2011) Discovery of cyclotides in the Fabaceae plant family provides new insights into the cyclization, evolution, and distribution of circular proteins. ACS Chem Biol 6:345–355. doi:10.1021/cb100388j
Prakash MG, Gurumurthi K (2009) Effects of type of explant and age, plant growth regulators and medium strength on somatic embryogenesis and plant regeneration in Eucalyptus camaldulensis. Plant Cell Tissue Organ Cult 100:13–20. doi:10.1007/s11240-009-9611-1
Saether O, Craik DJ, Campbel ID et al (1995) Elucidation of the primary and three-dimensional structure of the uterotonic polypeptide kalata B1. BioChemistry 34:4147–4158
Singh P (1965) Pharmacognosy of Viola odorata l. and its adulterants. Q J Crude Drug Res 2:712–719
Slazak B, Jacobsson E, Kuta E, Göransson U (2015) Exogenous plant hormones and cyclotide expression in Viola uliginosa (Violaceae). Phytochemistry 117:527–536. doi:10.1016/j.phytochem.2015.07.016
Trabi M, Svangård E, Herrmann A et al (2004) Variations in cyclotide expression in Viola species. J Nat Prod 67:806–810. doi:10.1021/np034068e
Uhlig T, Kyprianou T, Martinelli FG et al (2014) The emergence of peptides in the pharmaceutical business: from exploration to exploitation. EuPA open. Proteomics 4:58–69. doi:10.1016/j.euprot.2014.05.003
Walsh G (2014) Biopharmaceutical benchmarks 2014. Nat Biotechnol 32:992–1000. doi:10.1038/nbt0910-917
Wang CKL, Kaas Q, Chiche L, Craik DJ (2008) CyBase: a database of cyclic protein sequences and structures, with applications in protein discovery and engineering. Nucleic Acids Res 36:206–210. doi:10.1093/nar/gkm953
Wilson SA, Roberts SC (2012) Recent advances towards development and commercialization of plant cell culture processes for synthesis of biomolecules. Plant Biotechnol J 10:249–268. doi:10.1111/j.1467-7652.2011.00664.x.Recent
Yan MM, Xu C, Kim CH, et al (2009) Effects of explant type, culture media and growth regulators on callus induction and plant regeneration of Chinese jiaotou (Allium chinense). Sci Hortic (Amsterdam) 123:124–128. doi:10.1016/j.scienta.2009.07.021
Acknowledgements
The financial support for this research work from the Department of Biotechnology, Ministry of Science and Technology, Government of India is gratefully acknowledged by the authors (Sanction Order No. BT/PR6829/GBD/27/489/2012). The authors thank Dr. Nandita Madhavan for providing access to semi-preparative HPLC facility at IIT Madras (Currently: Associate Professor, Indian Institute of Technology Bombay, Mumbai, India). The authors wish to thank Dr. Suresh Baburaj [Survey Officer, Central Council for Research in Homoeopathy (CCRH)], for providing the Viola odorata plant material required for the study. The authors thank Prof. P. Balaram (Molecular Biophysics Unit, Indian Institute of Science Bangalore, India) for his valuable suggestions for deciphering the sequence of the cyclotide(s) identified in this study. The authors also thank Dr. V. Sabareesh (Advanced Centre for Bio Separation Technology, Vellore Institute of Technology University, Tamil Nadu, India) for initial discussion on de novo sequencing of peptides.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Electronic supplementary material
Below is the link to the electronic supplementary material.
11240_2017_1223_MOESM1_ESM.tif
Supplementary Fig. S1. De novo peptide sequencing of vodo I66 using collision-induced dissociation (CID) based MS/MS spectra of (a) 3643.61 Da; (b) 3428.48 Da; (c) 919.47 Da and (d) 1258.51 Da. The b and y ions are indicated in blue and red font respectively. The peptide sequence deduced from each MS/MS data is indicated in green font in the respective panel. (TIFF 353 KB). (TIF 352 KB)
11240_2017_1223_MOESM2_ESM.tif
Supplementary Fig. S2. De novo peptide sequencing of vodo I98 using collision-induced dissociation (CID) based MS/MS spectra of (a) 3522.54 Da; (b) 2729.15 Da; (c) 1433.62 Da and (d) 1589.70 Da.The b and y ions are indicated in blue and red font respectively. The peptide sequence deduced from each MS/MS data is indicated in green font in the respective panel, where ‘X’ indicates I/L (TIFF 334 KB). (TIF 333 KB)
Rights and permissions
About this article
Cite this article
Narayani, M., Chadha, A. & Srivastava, S. Callus and cell suspension culture of Viola odorata as in vitro production platforms of known and novel cyclotides. Plant Cell Tiss Organ Cult 130, 289–299 (2017). https://doi.org/10.1007/s11240-017-1223-6
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11240-017-1223-6