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Ecorehabilitation and biochemical studies of Ceropegia bulbosa Roxb.: a threatened medicinal succulent

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Abstract

In the present endeavour, a propagation protocol has been standardized for two varieties of threatened medicinal succulent, Ceropegia bulbosa Roxb. (varieties bulbosa and lushii). The manuscript focuses on morphogenic response of shoot tips when cultured on MS media augmented with growth regulators. Maximum shoot length and highest number of shoots were observed when 6-benzyladenine (8.88 μM) and α-naphthalene acetic acid were used in combination. During multiplication, antioxidant enzymes have shown variation in their activity. Total antioxidant activity and phenolic content were higher in the in vitro plants. Chlorophyll and protein content was higher in in vivo plants while proline was higher in in vitro plants. Superoxide dismutase activity was higher in naturally grown plants while, activities of catalase, glutathione reductase and peroxidase were elevated in the in vitro plants. Isolated cerpegin (pyridine alkaloid) was characterized using thin layer chromatography and attenuated total reflectance spectroscopy.

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Abbreviations

AAE:

Ascorbic acid equivalents

BA:

6-Benzyladenine

CAT:

Catalase

GAE:

Gallic acid equivalents

GR:

Glutathione reductase

IAA:

Indole-3-acetic acid

IBA:

Indole-3-butyric acid

Kin:

Kinetin (N6-furfuryladenine)

MS:

Murashige and Skoog

NAA:

α-Naphthalene acetic acid

PGRs:

Plant growth regulators

POD:

Peroxidase

SOD:

Superoxide dismutase

TPC:

Total phenolic content

References

  • Abbasi BH, Khan M, Guo B, Bokhari SA, Khan MA (2011) Efficient regeneration and antioxidative enzyme activities in Brassica rapa var. turnip. Plant Cell Tiss Organ Cult 105:337–344

    Article  CAS  Google Scholar 

  • Adibatti NA, Thirugnanasambantham P, Kulothungan C, Viswanathan S, Kameswaran L, Balakrishna K, Sukumar E (1991) A Pyridine alkaloid from Ceropegia juncea. Phytochemistry 30:2449–24501

    Article  CAS  Google Scholar 

  • Aebi H (1974) Catalases. In: Bergmeyer HU (ed) Methods of enzymatic analysis. Academic Press, Verlag Chemie, pp 673–684

    Chapter  Google Scholar 

  • Ahmad N, Javed SB, Khan I, Anis M (2013) Rapid plant regeneration and analysis of genetic fidelity in micropropagated plants of Vitex trifolia: an important medicinal plant. Acta Physiol Plant 35:2493–2500

    Article  CAS  Google Scholar 

  • Ananthan R, Latha M, Ramkumar KM, Pari L, Baskar C, Narmatha BV (2004) Modulatory effects of Gymnema montanum leaf extract on alloxan induced oxidative stress in wistar rats. Nutrition 20:280–285

    Article  CAS  PubMed  Google Scholar 

  • Arnon DL (1949) A copper enzymes in isolated chloroplasts: polypheno oxidase in Beta vulgaris. Plant Physiol 24:1–15

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Avetisyan A, Karapetyan L (2010) Convenient synthesis of Cerpegin. Synth Commun 40(17):2607–2609

    Article  CAS  Google Scholar 

  • Bates LS, Wadern RP, Teare ID (1973) Rapid estimation of free proline for water stress determination. Plant Soil 39:205–207

    Article  CAS  Google Scholar 

  • Batkova P, Pospisilova J, Synkova H (2008) Production of reactive oxygen species and development of antioxidative systems during in vitro growth and ex vitro transfer. Biol Plant 52:413–422

    Article  CAS  Google Scholar 

  • Beena MR, Martin KP, Kirti PB, Hariharan M (2003) Rapid in vitro propagation of medicinally important Ceropegia candelabrum. Plant Cell Tiss Organ Cult 72:285–289

    Article  CAS  Google Scholar 

  • Benson EE, Lynch PT, Jones J (1992) Variation in free-radical damage in rice cell-suspensions with different embryogenic potentials. Planta 188:296–305

    Article  CAS  PubMed  Google Scholar 

  • Beuchamp C, Fridovich I (1971) Superoxide dismutase: improved assays and an acrylamide gels. Anal Biochem 44:276–287

    Article  Google Scholar 

  • Borthakur A, Das SC, Kalita MC, Sen P (2011) In vitro plant regeneration from apical buds of Albizzia odoratissima (L.f.) Benth. Adv App Sci Res 2(5):457–464

    CAS  Google Scholar 

  • Britto SJ, Natarajan E, Arockiasamy DI (2003) In vitro flowering and shoot multiplication from nodal explants of Ceropegia bulbosa Roxb. var. bulbosa. Taiwania 48(2):106–111

    Google Scholar 

  • Chance B, Maehly AC (1995) Assay of catalases and peroxidises. Methods Enzymol 2:764–775

    Article  Google Scholar 

  • Dhaka N, Kothari SL (2005) Micropropagation of Eliptica alba (L.) Hassk-An important medicinal plant. In Vitro Cell Dev Biol Plant 41:658–661

    Article  Google Scholar 

  • Dhir R, Shekhawa GS (2013) Production, storability and morphogenic response of alginate encapsulated axillary meristems and genetic fidelity evaluation of in vitro regenerated Ceropegia bulbosa: a pharmaceutically important threatened plant species. Ind Crops Prod 47:139–144

    Article  CAS  Google Scholar 

  • Dhir R, Shekhawat GS (2012) Critical review on Tecomella undulata: a medicinally potent endangered plant species of Indian Thar Deseart. Int J Curr Res 4(6):36–44

    Google Scholar 

  • Dutta Gupta S, Datta S (2003) Antioxidant enzyme activities during in vitro morphogenesis of gladiolus and the effect of application of antioxidants on plant regeneration. Biol Plant 47:179–183

    Article  Google Scholar 

  • Faisal M, Anis M (2007) Regeneration of plants from alginate encapsulated shoots of Tylophora indica (Burm.f.) Merrill, an endangered medicinal plant. J Hortic Sci Biotechnol 82:351–354

    CAS  Google Scholar 

  • Giri L, Jugran A, Rawat S, Dhyani P, Andola H, Bhatt ID, Rawal RS, Dhar U (2012) In vitro propagation, genetic and phytochemical assessment of Habenaria edgeworthii: an important Astavarga plant. Acta Physiol Plant 34(3):869–875

    Article  Google Scholar 

  • Gupta SD, Ibaraki Y, (2006) Evaluation of photosynthetic capacity in micropropagated plants by image analysis. In: Gupta and Ibaraki (ed) Plant tissue culture engineering, Springer, Netherlands, pp 15–29

  • Hong H, Comins DL (1996) A three-step synthesis of cerpegin. J Org Chem 61:391–392

    Article  CAS  Google Scholar 

  • Huang CL, Hsieh MT, Hsieh WC, Sagare AP, Tsay HS (2000) In vitro propagation of Limonium wrightii (Hance) Ktze. (Plumbaginaceae), an ethnomedicinal plant from shoot tip, leaf and inflorescence-node explants. In Vitro Cell Dev Biol Plant 36:220–224

    Article  CAS  Google Scholar 

  • Jain M, Bhandari A, Bhandari A, Patel P (2012) Characterization and in vitro antiurolithiatic activity of cerpegin alkaloid from Ceropegia bulbosa var. Lushii root. Int J Drug Dev Res 4(4):154–160

    CAS  Google Scholar 

  • Jana S, Shekhawat GS (2012) In vitro regeneration of Anethum graveolens, antioxidative enzymes during organogenesis and RAPD analysis for clonal fidelity. Biol Plant 56(1):9–14

    Article  CAS  Google Scholar 

  • Khan MA, Pradhan D (2012) Antiurolithc activity of Ceropegia bulbosa extract in rats. Der Pharmacia Sinica 3(1):148–152

    CAS  Google Scholar 

  • Kirtikar KR, Basu BD (1975) Indian medicinal plants. Bishen Singh Mahendra Pal Singh, Dehradun

    Google Scholar 

  • Kumaran A, Karunakaran JR (2007) In vitro antioxidant activities of methanol extracts of five Phyllanthus species from India. Food Sci Technol 40(2):344–352

    CAS  Google Scholar 

  • Kwon YI, Vattem DA, Shetty K (2006) Clonal herbs of lamiaceae species against diabetes and hypertension. Asia Pac J Clin Nut 15:107–118

    Google Scholar 

  • Lowry OH, Rosenberg NJ, Farr AL, Randall RJ (1951) Protein measurement with folin phenol reagent. J Biol Chem 193:265–275

    CAS  PubMed  Google Scholar 

  • Mabberley DJ (1978) The plant book. Cambridge University Press, Cambridge

    Google Scholar 

  • Malik SK, Chaudhury R, Kalia RK (2005) Rapid in vitro multiplication and conservation of Garcinia indica: a tropical medicinal tree species. Sci Hortic 106:539–553

    Article  Google Scholar 

  • Marquele FD, Di Mambro VM, Georgetti SR, Casagrande R, Yara ML, Valim YML, Fonseca MJV (2005) Assessment of the antioxidant activities of Brazilian extracts of propolis alone and in topical pharmaceutical formulations. J Pharm Biomed Anal 39:455–462

    Article  CAS  PubMed  Google Scholar 

  • Meratan AA, Ghaffari SM, Niknam V (2009) In vitro organogenesis and antioxidant enzymes activity in Acanthophyllum sordidum. Biol Plant 53:5–10

    Article  CAS  Google Scholar 

  • Mishra J, Rawat B, Agnihotri RK, Chandra A, Nautiyal S (2012) In vitro propagation, genetic and secondary metabolite analysis of Aconitum violaceum Jacq. Acta physiol Plant 35(8):2589–2599

    Google Scholar 

  • Mitrovic A, Janosevic D, Budimir S, Pristov JB (2012) Changes in antioxidative enzymes activities during Tacitus bellus direct shoot organogenesis. Biol Plant 56(2):357–361

    Article  CAS  Google Scholar 

  • Molassiotis AN, Dimassi K, Diamantidis G, Therios I (2004) Changes in peroxidases and catalase activity during in vitro rooting. Biol Plant 48:1–5

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Nadkarni KM (1976) Indian materia medica. Popular Prakashan, Bombay

    Google Scholar 

  • Nakanishi K (1962) Infrared absorption spectroscopy. Holden-Day, San Francisco

    Google Scholar 

  • Obert B, Benson EE, Millam S, Pretovb A, Bremner DH (2005) Moderation of morphogenetic and oxidative stress responses in flax in vitro cultures by hydroxynonenal and desferrioxamine. J Plant Physiol 162:537–547

    Article  CAS  PubMed  Google Scholar 

  • Patil VM (1998) Micropropagation of Ceropegia spp. In Vitro Cell Dev Biol Plant 34:240–243

    Article  Google Scholar 

  • Phulwaria M, Shekhawat NS, Rathore JS, Singh RP (2013) An efficient in vitro regeneration and ex vitro rooting of Ceropegia bulbosa Roxb.—A threatened and pharmaceutical important plant of Indian Thar Desert. Ind Crops Prod 42:25–29

    Article  CAS  Google Scholar 

  • Piovan A, Caniato R, Mariella Cappelletti E, Filippini R (2010) Organogenesis from shoot segments and via callus of endangered Kosteletzkya pentacarpos (L.) Ledeb. Plant Cell Tiss Organ Cult 100:309–315

    Article  Google Scholar 

  • Raja B, Pugalendi KV (2010) Evaluation of antioxidant activity of Melothria maderaspatana in vitro. Cent Eur J Biol 5(2):224–230

    Article  Google Scholar 

  • Rathore MS, Chikara J, Shekhawat NS (2011) Plantlet regeneration from callus cultures of selected genotype of Aloe vera L.—An ancient plant for modern herbal industries. Appl Biochem Biotechnol 163:860–868

    Article  CAS  PubMed  Google Scholar 

  • Ravishankara MN, Shrivastava N, Padh H, Rajani M (2002) Evaluation of antioxidant properties of root bark of Hemidesmus indicus R. Br. (Anantmul). Phytomedicine 9:153–160

    Article  CAS  PubMed  Google Scholar 

  • Roy S, Sehgal R, Padhy BM, Kumar VL (2005) Antioxidant and protective effect of latex of Calotropis procera against alloxan-induced diabetes in rats. J Ethnopharmacol 102:470–473

    Article  CAS  PubMed  Google Scholar 

  • Sahai A, Shahzad A (2013) High frequency in vitro regeneration system for conservation of Coleus forskohlii: a threatened medicinal herb. Acta Physiol Plant 35:473–481

    Article  CAS  Google Scholar 

  • Sahai A, Shahzad A, Anis M (2010) High frequency plant production via shoot organogenesis and somatic embryogenesis from callus in Tylophora indica (Burm.f.) Merr., an endangered plant species. Turk J Bot 34:11–20

    CAS  Google Scholar 

  • Sarasan V, Cripps R, Ramsay MM, Atherton C, McMichen M, Prendergast G, Rowntree JK (2006) Conservation in vitro of threatened plants—progress in the past decade. In Vitro Cell Dev Biol Plant 42:206–214

    Article  Google Scholar 

  • Schaedle M, Bassham JA (1997) Chloroplast glutathione reductase. Plant Physiol 59:1011–1012

    Article  Google Scholar 

  • Sen A, Batra A (2013) The study of in vitro and in vivo antioxidant activity and total phenolic content of Phyllanthus amarus schum. & thonn.: a medicinally important plant. Int. J Pharm Pharm Sci 5(3):942–947

    Google Scholar 

  • Shekhawat MS, Shekhawat NS (2011) Micropropagation of Arnebia hispidissima (Lehm). DC. and production of alkannin from callus and cell suspension culture. Acta Physiol Plant 33:1445–1450

    Article  CAS  Google Scholar 

  • Shekhawat GS, Mathur S, Batra A (2009) Role of phytohormones and nitrogen in somatic embryogenesis induction in cell culture derived from leaflets of Azadirachta indica. Biol Plant 53(4):707–710

    Article  CAS  Google Scholar 

  • Skálová D, Navrátilová B, Richterová L, Knitl M, Sochor M, Vašut RJ (2012) Biotechnological methods of in vitro propagation in willows (Salix spp.). Cent Eur J Biol 7(5):931–940

    Article  Google Scholar 

  • Sukumar E, Hamsaveni GR, Bhima RR (1996) Pharmacological actions of cerpegin, a novel pyridine alkaloid from Ceropegia juncea. Fitoterapia 66:403–406

    Google Scholar 

  • Swarna J, Ravindhran R (2012) In vitro propagation and assessment of genetic integrity of Talinum triangulare (Jacq.) Willd: a valuable medicinal herb. Acta Physiol plant 34:1987–1996

    Article  CAS  Google Scholar 

  • Thakar J, Bhargava S (1999) Seasonal variation in antioxidant enzymes and the sprouting response of Gmelina arborea Roxb. nodal sectors cultured in vitro. Plant Cell Tiss Organ Cult 59:181–187

    Article  Google Scholar 

  • Tripathi M, Kumari N (2010) Micropropagation of a tropical fruit tree Spondias mangifera Willd. through direct organogenesis. Acta Physiol Plant 32:1011–1015

    Article  Google Scholar 

  • Villemin D, Cheikh N, Liao L, Bar N, Lohier JF, Sopkova J, Choukchou-Braham N, Mostefa-Kara B (2012) Two versatile routes towards cerpegin and analogues: applications of a one pot reaction to new analogues of cerpegin. Tetrahedron 68:4906–4918

    Article  CAS  Google Scholar 

  • Walter SK, Gillet J (1998) 1997 IUCN Red list of threatened plants. World conservation monitring centre, Cambridge, UK

  • Wang J, Gao W, Zhang L, Huang L (2013) Establishment and quality assessment of tissue cultures in Glycyrrhiza uralensis Fisch. Appl Biochem Biotechnol 169:588–594

    Article  CAS  PubMed  Google Scholar 

  • Yang YL, Zhang YY, Lu J, Zhang H, Liu Y, Jiang Y, Shi RX (2012) Exogenous H2O2 increased catalase and peroxidase activities and proline content in Nitraria tangutorum callus. Biol Plant 56(2):330–336

    Article  CAS  Google Scholar 

  • Zayova E, Stancheva I, Geneva M, Petrova M, Vasilevska-Ivanova R (2012) Morphological evaluation and antioxidant activity of in vitro- and in vivo-derived Echinacea purpurea plants. Cent Eur J Biol 7(4):698–707

    Article  CAS  Google Scholar 

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Author contribution

G. S. Shekhawat conceptualized the study and Richa Dhir designed and executed the experiments.

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Authors and Affiliations

  1. Department of Bioscience and Biotechnology, Banasthali University, 304022, Rajasthan, India

    Richa Dhir

  2. Department of Botany, Jai Narain Vyas University, Jodhpur, 342001, Rajasthan, India

    G. S. Shekhawat

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  1. Richa Dhir
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  2. G. S. Shekhawat
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Corresponding author

Correspondence to G. S. Shekhawat.

Additional information

Communicated by A. Krolicka.

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Dhir, R., Shekhawat, G.S. Ecorehabilitation and biochemical studies of Ceropegia bulbosa Roxb.: a threatened medicinal succulent. Acta Physiol Plant 36, 1335–1343 (2014). https://doi.org/10.1007/s11738-014-1512-1

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  • DOI: https://doi.org/10.1007/s11738-014-1512-1

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