Calcium-alginate coated synthetic seed production, storage and assessment of genetic stability in Alpinia galanga (L.) Willd

, , , , , ,

Research Articles | Published:

Print ISSN : 0970-4078.
Online ISSN : 2229-4473.
Pub Email:
Doi: 10.1007/s42535-021-00314-x
First Page: 527
Last Page: 533
Views: 600

Keywords: Alpinia galanga , Synthetic seed, Calcium-alginate, RAPD


An efficient encapsulation method was developed for artificial seed production in Alpinia galanga. Shoot buds obtained from in vitro proliferated plantlets on Murashige and Skoog (MS) medium containing 2.5 mg/l 6-benzyladenine (BA) was used for encapsulation. Calcium-alginate matrix composed of 3% sodium (Na) alginate and 100 mM calcium chloride (CaCl2.2H2O) was optimum for uniform, firm synseed preparation. Highest germination and plantlet conversion (87%) was obtained on MS medium containing 2.0 mg/l BA and 0.5 mg/l α-naphthalene acetic acid (NAA) under light/dark (16/8 h) photoperiod. Synseeds stored at 4 °C exhibited 85% and 23% germination after 30 days and 120 days respectively. In vitro grown plantlets were hardened successfully to field condition with 76% survival rate. 8 random amplified polymorphic DNA (RAPD) markers were used to check the genetic stability of regenerated plants. A monomorphic banding profile confirmed the genetic similarity of plants obtained from synseeds.

                Alpinia galanga
              , Synthetic seed, Calcium-alginate, RAPD

*Pdf Download

(*Only SPR Members can download pdf file; #Open Access;)



Adhikari S, Bandyopadhyay TK, Ghosh P (2014) Assessment of genetic stability of Cucumis sativus L. regenerated from encapsulated shoot tips. Sci Horti 170:115–122.

Ahmad Z, Shahzad A (2019) Cash crops: synseed production, propagation, and conservation. In: Faisal M, Alatar A (eds) Synthetic seeds. Springer, Cham.

Asmah HN, Hasnida HN, Zaimah NN, Noraliza A, Salmi NN (2011) Synthetic seed technology for encapsulation and regrowth of in vitro-derived Acacia hyrid shoot and axillary buds. Afr J Biotech 10:7820–7824.

Babu KN, Samsudeen K, Divakaran M, Pillai GS, Sumathi V, Praveen K, Ravindran PN, Peter KV (2016) Protocols for in vitro propagation, conservation, synthetic seed production, embryo rescue, microrhizome production, molecular profiling, and genetic transformation in ginger (Zingiber officinale Roscoe.). Protocols for in vitro cultures and secondary metabolite analysis of aromatic and medicinal plants, 2nd edn. Humana Press, New York, pp 403–426

Banerjee S, Singh S, Pandey H, Pandey P, Rahman L (2012) Conversion and storage of Curcuma amada Roxb. synseeds on Luffa sponge matrix and RAPD analysis of the converted plantlets. Indust Crop Product 36:383–388.

Banjerdpongchai R, Punyati P, Nakrob A, Pompimon W, Kongtawelert P (2011) 4′-Hydroxycinnamaldehyde from Alpinia galanga (Linn.) induces human leukemic cell apoptosis via mitochondrial and endoplasmic reticulum stress pathways. Asian Pac J Cancer Prev 12:593–598.

Benelli C (2016) Encapsulation of shoot tips and nodal segments for in vitro storage of “Kober 5BB” grapevine rootstock. Horticult 2:10.

Ekinci H, Çiftçi YÖ, Nadarajan J (2019) Medium- and long-term conservation of ornamental plants using synthetic seed technology. In: Faisal M, Alatar A (eds) Synthetic seeds. Springer, Cham.

Fonseka DL, Wickramaarachchi WW, Madushani RP (2019) Synthetic seed production as a tool for the conservation and domestication of Celastrus paniculatus: a rare medicinal plant. Ann Res Rev Biol 1:1–8.

Gantait S, Kundu S, Ali N, Sahu NC (2015) Synthetic seed production of medicinal plants: a review on influence of explants, encapsulation agent and matrix. Acta Physiol Plant 37(5):98.

Haque SK, Ghosh B (2014) Somatic embryogenesis and synthetic seed production—a biotechnological approach for true-to-type propagation and in vitro conservation of an ornamental bulbaceous plant drimiopsis kirkii baker. Appl Biochem Biotechnol 172:4013–4024.

Hsu WY, Simonne A, Weissman A, Kim JM (2010) Antimicrobial activity of greater galangal [Alpinia galanga (Linn.) Swartz.] flowers. Food Sci Biotech 19:873–880.

Indrayan AK, Agrawal P, Rathi AK, Shutru A, Agrawal NK, Tyagi DK (2009) Nutritive value of some indigenous plant rhizomes resembling ginger. Nat Prod Rad 8:507–513

Jaju SB, Indurwade NH, Sakarkar DM, Fuloria NK, Ali MD, Das S, Basu SP (2009) Galango flavonoid isolated from rhizome of Alpinia galanga (L.) Sw. (Zingiberaceae). Trop J Pharm Res.

Jantan I, Rafi IA, Jalil J (2005) Platelet-activating factor (PAF) receptor-binding antagonist activity of Malaysian medicinal plants. Phytomed 12:88–92.

Javed SB, Alatar AA, Anis M, Faisal M (2017) Synthetic seed production and germination studies, for short term storage and long distance transport of Erythrina variegate L.: a multipurpose tree legume. Indust Crop Product 105:41–46.

Khan MI, Ahmad N, Anis M, Alatar AA, Faisal M (2018) In vitro conservation strategies for the Indian willow (Salix tetrasperma Roxb.), a vulnerable tree species via propagation trough synthetic seeds. Biocat Agri Biotech 16:17–21.

Latha C, Shriram VD, Jahagirdar SS, Dhakephalkar PK, Rojatkar SR (2009) Antiplasmid activity of 1′-acetoxychavicol acetate from Alpinia galanga against multi-drug resistant bacteria. J Ethnopharm 123:522–525.

Maqsood M, Khusrau M, Mujib A, Kaloo ZA (2021) Synthetic seed technology in some ornamental and medicinal plants: an overview. In: Siddique I (ed) Propagation and genetic manipulation of plants. Springer, Singapore.

Micheli M, Standardi A (2016) From somatic embryo to synthetic seed in citrus spp. through the encapsulation technology. In: Germana M, Lambardi M (eds) In vitro embryogenesis in higher plants methods in molecular biology, vol 1359. Humana Press, New York

Murashige T (1978) The impact of plant tissue culture on agriculture. In: Thorpe T (ed) Frontiers of plant tissue culture. International Association for Plant Tissue Culture. Univ Calgary, Alberta, pp 15–26

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

Negahdar N, Hashemabadi D, Kaviani B (2021) In vitro conservation and cryopreservation of Buxus sempervirens L., a critically endangered ornamental shrub. Russ J Plant Physiol 68(4):661–8.

Oonmetta-aree J, Suzuki T, Gasaluck P, Eumkeb G (2006) Antimicrobial properties and action of galangal (Alpinia galanga Linn.) on Staphylococcus aureus. LWT-Food Sci Tech 39:1214–1220.

Panich U, Kongtaphan K, Onkoksoong T, Jaemsak K, Phadungrakwittaya R, Thaworn A, Akarasereenont P, Wongkajornsilp A (2010) Modulation of antioxidant defense by Alpinia galanga and Curcuma aromatica extracts correlates with their inhibition of UVA-induced melanogenesis. Cell Biol Toxicol 26:103–116.

Priyadharshini S, Manokari M, Shekhawat MS (2020) In vitro conservation strategies for the critically endangered Malabar river lily (Crinum malabaricum Lekhak & Yadav) using somatic embryogenesis and synthetic seed production. S Afr J Bot 135:172–180.

Rao K, Ch B, Narasu LM, Giri A (2010) Antibacterial activity of Alpinia galanga (L) Willd crude extracts. Appl Biochem Biotech 162:871–884.

Saeed T, Shahzad A, Ahmad N, Parveen S (2018) High frequency conversion of non-embryogenic synseeds and assessment of genetic stability through ISSR markers in Gymnema sylvestre. Plant Cell Tiss Org Cult 134:163–168.

Saha S, Sengupta C, Ghosh P (2014) Molecular and phytochemical analysis to assess genetic stability in alginate-encapsulated microshoots of Ocimum gratissimum L. following in vitro storage. Nucl 57:33–43.

Sawangjaroen N, Phongpaichit S, Subhadhirasakul S, Visutthi M, Srisuwan N, Thammapalerd N (2006) The anti-amoebic activity of some medicinal plants used by AIDS patients in southern Thailand. Parasit Res 98:588–592.

Sharma TR, Singh BM, Chauhan RS (1994) Production of disease-free encapsulated buds of Zingiber officinale Rosc. Plant Cell Rep 13:300–302.

Sharma N, Gowthami R, Pandey R (2019) Synthetic seeds: a valuable adjunct for conservation of medicinal plants. In: Faisal M, Alatar A (eds) Synthetic Seeds. Springer, Cham.

Smith RM (1990) Alpinia (Zingiberaceae): a proposed new infrageneric classification. Edinburgh J Bot 47:1–75.

Soundar RC, Aslam A, Shajahan A (2016) Germination and storability of calcium-alginate coated somatic embryos of mango ginger (Curcuma amada Roxb.). Horticul Environ Biotech 57:88–96.

Standardi A, Piccioni E (1998) Recent perspectives on synthetic seed technology using nonembryogenic in vitro–derived explants. Int J Plant Sci 159:968–978.

Sundararaj SG, Agrawal A, Tyagi RK (2010) Encapsulation for in vitro short-term storage and exchange of ginger (Zingiber officinale Rosc.) germplasm. Sci Horticul 125:761–766.

Trakranrungsie N, Chatchawanchonteera A, Khunkitti W (2008) Ethnoveterinary study for antidermatophytic activity of Piper betle, Alpinia galanga and Allium ascalonicum extracts in vitro. Res Vet Sci 84:80–84.

Varshney A, Anis M (2014) Synseed conception for short-term storage, germplasm exchange and potentialities of regeneration genetically stable plantlets of desert date tree (Balanites aegyptiaca Del.). Agroforest Syst 88:321–329.

Vinutha B, Prashanth D, Salma K, Sreeja SL, Pratiti D, Padmaja R, Radhika S, Amit A, Venkateshwarlu K, Deepak M (2007) Screening of selected Indian medicinal plants for acetylcholinesterase inhibitory activity. J Ethnopharm 109:359–363.



Author Information

Shamsudheen Kizhakke Modongal
Plant Molecular Biology Laboratory, Department of Botany, Jamal Mohamed College, Tiruchirappalli, India
Mehaboob Valiyaparambath Musfir
Department of Botany, PSMO College, Tirurangadi, India

Faizal Kunnampalli
Plant Molecular Biology Laboratory, Department of Botany, Jamal Mohamed College, Tiruchirappalli, India

Raja Palusamy
Plant Molecular Biology Laboratory, Department of Botany, Jamal Mohamed College, Tiruchirappalli, India

Thiagu Ganesan
Plant Molecular Biology Laboratory, Department of Botany, Jamal Mohamed College, Tiruchirappalli, India

Raju Chellappan Soundar
Department of Botany, Vivekananda College, Madurai, India

Shajahan Appakan
Plant Molecular Biology Laboratory, Department of Botany, Jamal Mohamed College, Tiruchirappalli, India