• Jaspreet K. Sembi Department of Botany, Panjab University, Chandigarh
  • Parvinderdeep S. Kahlon Institute of Cellular and Molecular Botany, University of Bonn, Bonn
  • Tania Kalsotra
  • Sukhjeet Kaur Sidhu SUS College of Engineering and Technology, Tangori, Mohali, Punjab, India



In the present study, effect of ascorbic acid, a known growth adjuvant on encapsulated protocorm-like-bodies (PLBs) of Rhynchostylis retusa Bl. was investigated. PLBs were encapsulated in calcium alginate (3.5% sodium alginate and 100mM calcium chloride) prepared in Mitra et al. (1976) basal medium and supplemented with different concentration of ascorbic acid (5, 10, 15, 20mM). The encapsulated PLBs were stored at 25°C. Their germination response and germination potential was evaluated after every 4 weeks on basal media. Control set of encapsulated PLBs, failed to germinate after 32 weeks. However, PLBs with 15mM ascorbic acid in the encapsulated matrix showed the best response; nearly 90% germinated even after 32 weeks of storage. The survival and germination frequency was directly proportional to the level of ascorbic acid in the alginate mix upto 15mM level but declined on further increase. Differentiation of PLBs into plantlet was better in synthetic seeds containing lower concentration of ascorbic acid (5mM) as compared to higher levels (15, 20mM) whereas multiplication of secondary PLBs was more pronounced at higher levels. Chlorophyll content was inversely proportional to the level of ascorbic acid in the nutrient mix; lush green PLBs were observed at low concentration of ascorbic acid (5mM). This study highlights the potential of ascorbic acid as an aid to growth and survival of encapsulated PLBs upon storage.


Download data is not yet available.


Abdelwahd RN, Hakam M, Labhilili MU and Sripada. Use of an adsorbent and antioxidants to reduce the effects of leached phenolics in in vitro plantlet regeneration of faba bean. African Journal of Biotechnology 2008; 7(8): 997-1002

Babbar N, Oberoi HS, Uppal DS and Patil RT. Total phenolic content and antioxidant capacity of extracts obtained from six important fruit residues. Food Research International 2011; 44(1): 391-396

Britt J. The status of the commercial production of potted orchid around the world’, Horticulture Technology 2000; 10(3): 435-436

Bybordi A. Effect of Ascorbic Acid and Silicium on Photosynthesis, Antioxidant Enzyme Activity, and Fatty Acid Contents in Canola Exposure to Salt Stress. Journal of Integrative Agriculture 2012; 11(10): 1610-1620

Foyer CH. Ascorbic acid. In: Alscher, RG and Hess, JL, editors. Antioxidants in Higher Plants. Boca Raton: CRC Press; 1993. p. 31-58.

Gantait S and Sinniah UR. Storability, post-storage conversion and genetic stability assessment of alginate-encapsulated shoot tips of monopodial orchid hybrid Aranda Wan Chark Kuan ‘Blue’ × Vanda coerulea Griff. ex. Lindl. Plant Biotechnology Reports 2013; 7(3): 257-266

Jalal JS, Kumar P and Pangtey YPS. Ethnomedicinal Orchids of Uttarakhand, Western Himalaya. Ethnobotanical Leaflets 2008; 12: 1227-1230

Ko WH, Su CC, Chen CL and Chao CP. Control of lethal browning of tissue culture planlets of Cavendish banana cv. Formosana with ascorbic acid. Plant Cell Tissue Organ Culture 2009; 96: 137-141

Kumar A, Nandi SK, Bag N and Palni LMS. Tissue culture studies in two important orchid taxa: Rhynchostylis retusa (L.) Bl. and Cymbidium elegans Lindl. In: Gyanodaya, P, editor. Role of plant tissue culture in biodiversity conservation and economic development. Nainital; 2002. p. 113-124.

Mitra GC, Prasad RN and Chowdhary AR. Inorganic salts and differentiation of protocorms in seed callus of an orchid and correlated changes in its free amino acid content. Indian Journal of Experimental Biology 1976; 14: 350-351

Miyake C and Asada K. Thylakoid-bound ascorbate peroxidase in spinach chloroplasts and photoreduction of its primary oxidation product, monodehydroaseorbate radicals in the thylakoids. Plant Cell Physiology 1992; 33(5): 541-553

Mohanty P, Nongkling P, Das MC, Kumaria S and Tandon P. Short-term storage of alginate-encapsulated protocorm-like bodies of Dendrobium nobile Lindl,: an endangered medicinal orchid from North-east India. Biotech 2013; 3(3): 235-239

Nisyawati and Kariyana K. Effect of ascorbic acid, activated charcoal and light duration on shoot regeneration of banana cultivar barangan (Musa acuminata l.) in vitro culture. IJRRAS 2013; 15(1): 13-17

Noctor G and Foyer CH. Ascorbate and glutathione: keeping active oxygen under control. Annual Revision of Plant Physiol Plant Molecular Biology 1998; 49: 249-279

Pradhan S, Tiruwa B, Subedee BR and Pant B. In vitro germination and propagation of a threatened medicinal orchid, Cymbidium aloifolium (L.) Sw. through artificial seed. Asian Pacific Journal of Tropical Biomedicine 2014; 4(12): 971-976.

Radhika B and Murthy N. Preliminary phytochemical analysis and in vitro bioactivity against clinical pathogens of medicinally important orchid of Rhynchostylis retusa Bl. American Journal of PharmTech Research 2013; 3(4) 510-520

Richard W, Joy IV, Kamlesh RP and Trevor AT. Ascorbic acid enhancement of organogenesis in tobacco callus. Plant Cell, Tissue and Organ Culture 1988; 13(3): 219-228

Sembi JK, Kahlon PK, Kalsotra T and Sidhu S. Effect of ascorbic acid, proline and mannitol on the desiccation tolerance of synthetic seeds in Rhynchostylis Retusa Bl. American Journal of Life Science Researches 2014; 2(4): 520-533

Shawky AB. A synthetic seed method through encapsulation of in vitro proliferated bulblets of garlic (Allium sativum L.). Arab Journal of Biotechnology 2006; 9(3): 415-426

Siew WL, Kwok MY, Ong YM, Liew HP and Yew BK. Effective Use of Synthetic Seed Technology in the Regeneration of Dendrobiumm White Fairy Orchid. Journal of Ornamental Plants 2014; 4: 1-7

Smirnoff N. The Function and Metabolism of Ascorbic Acid in Plant’, Annals of Botany 1996; 78: 661-669

Stasolla C and Yeung EC. Ascorbic acid improves conversion of of white spruce somatic embryo. In Vitro CeIl Develpomental Biology Plant 1999; 35(4): 316-319

Stasolla C and Yeung EC. Ascorbic acid metabolism during white spruce somatic embryo maturation and germination’, Physiologia Plantarum Vol. 111, No. 2, pp.196–205.

Vij, S.P., Sood, A. and Plaha, K.K. (1984) ‘Propagation of Rhynchostylis retusa BL. (Orchidaceae) by direct organogenesis from leaf cultures. Botanical Gazette 2001; 145(2): 210–214

Yagya PA and Fischer A. Distribution pattern of the epiphytic orchid Rhynchostylis retusa under strong human influence in Kathmandu valley, Nepal. Botanica Orientalis Journal of Plant Science 2011; 8: 90–99

Zhou B, Xinfang W, Rongting W and Jingming J. Quantification of the enzymatic browning and secondary metabolites in the callus culture system of Nigella glandulifera Freynet Sint. Asian Journal of traditional medicines 2010; 5(3): 109-116



How to Cite




Regular Articles