Abstract
Stevia rebaudiana (Bertoni), commonly known as a natural sweetener plant or sweet herb grabs the economic and scientific interests owing to its sweetness and therapeutic properties present in its leaves mainly due to the presence of steviol glycosides (SGs), with stevioside being the most abundant, followed by rebaudioside. Stevioside, the most abundant active component, is greatly preferred by diabetic patients since it is a non-calorie sweetener and is approved by the Food and Drug Administration as a dietary supplement. This shows the benefits of stevia over other artificial sweeteners as an ingredient for the food business, subsequently, making stevia an increasingly appropriate substitute for saccharine in various drinks, beverages, and bakery products. Stevia also offers therapeutic benefits having anti-hyperglycemic, anti-hypertensive, and immunomodulatory effects. These beneficial effects largely focused on the importance of stevia. Sexually developed plants were not efficient because of low fertility and reduced viability. However, quality planting material can be produced through biotechnological approaches like micropropagation. To date, a great deal of studies has been completed on the tissue culture intervened approaches for the mass propagation of stevia. In this chapter, the above-mentioned approaches and their significance are emphasized for the large-scale production of identical plant materials and for the improvement of important calorie-free SGs.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Abraham J, Thomas TD (2016) Recent advances in Asteraceae tissue culture. In: Plant tissue culture: propagation, conservation and crop improvement. Springer, Singapore, pp 161–195
Aggarwal D, Reddy MS, Kumar A (2016) Biotechnological approaches for the improvement of eucalyptus. In: Plant tissue culture: propagation, conservation and crop improvement. Springer, Singapore, pp 219–244
Ahmad U, Ahmad RS (2018) Antidiabetic property of aqueous extract of Stevia rebaudiana Bertoni leaves in Streptozotocin-induced diabetes in albino rats. BMC Complement Altern Med 18:179
Ahmed MB, Salahin M, Karim R, Razvy MA, Hannan MM, Sultana R, Hossain M, Islam R (2007) An efficient method for in vitro clonal propagation of a newly introduced sweetener plant (Stevia rebaudiana Bertoni.) in Bangladesh. Am-Eur J Sci Res 2:121–125
Alan AR, Zeng H, Assani A, Shi WL, McRae HE, Murch SJ, Saxena PK (2007) Assessment of genetic stability of the germplasm lines of medicinal plant Scutellaria baicalensis Georgi (Huang-qin) in long-term, in vitro maintained cultures. Plant Cell Rep 26:1345–1355
Alhady MR (2011) Micropropagation of Stevia rebaudiana Bertoni. A new sweetening crop in Egypt. Global J Biotechnol Biochem 6:178–182
Ali A, Ahmad T, Abbasi NA, Hafiz IA (2009) Effect of different concentrations of auxins on in vitro rooting of olive cultivar ‘Moraiolo’. Pak J Bot 41:1223–1231
Anbazhagan M, Kalpana M, Rajendran R, Natarajan V, Dhanavel D (2010) In vitro production of Stevia rebaudiana Bertoni. Emir J Food Agric 22:216–222
Atalay E, Erisen S, Yorgancilar M, Tanur M (2011) Micropropagation of Stevia rebaudiana Bertoni. Curr Opin Biotechnol 22:15–152
Aziz R, Khaled K (2017) Stevia Rebaudiana Bertoni; Rapid micropropagation, stevioside accumulation and genetic fidelity using ISSR markers. J Agric Chem Biotechnol 8:295–301
Borah R, Kumaria S, Choudhury H (2017) In vitro plant regeneration of Magnolia punduana: an endemic and threatened plant species. Plant Tissue Cult Biotechnol 27:153–159
Brahmachari G, Mandal LC, Roy R, Mondal S, Brahmachari AK (2011) Stevioside and related compounds–molecules of pharmaceutical promise: a critical overview. Arch Pharm 344:5–19
Brunner AM, Li J, DiFazio SP, Shevchenko O, Montgomery BE, Mohamed R, Wei H, Ma C, Elias AA, Van Wormer K, Strauss SH (2007) Genetic containment of forest plantations. Tree Gene Genomics 3:75–100
Carakostas MC, Curry LL, Boileau AC, Brusick DJ (2008) Overview: the history, technical function and safety of rebaudioside A, a naturally occurring steviol glycoside, for use in food and beverages. Food Chem Toxicol 46:S1–S10
Chan P, Xu DY, Liu JC, Chen YJ, Tomlinson B, Huang WP, Cheng JT (1998) The effect of stevioside on blood pressure and plasma catecholamines in spontaneously hypertensive rats. Life Sci 63:1679–1684
Chandana BC, Kumari Nagaveni HC, Heena MS, Shashikala SK, Lakshmana D (2018) Role of plant tissue culture in micropropagation, secondary metabolites production and conservation of some endangered medicinal crops. J Pharmacogn Phytochem 3:246–251
Chandra S, Bandopadhyay R, Kumar V, Chandra R (2010) Acclimatization of tissue cultured plantlets: from laboratory to land. Biotechnol Lett 32:1199–1205
D’Amato F, Bayliss MW (1985) Cytogenetics of plant cell and tissue cultures and their regenerates. Crit Rev Plant Sci 3:73–112
Das A, Gantait S, Mandal N (2010) Micropropagation of an elite medicinal plant: Stevia rebaudiana Bert. Int J Agric Res 6:40–48
Deshmukh N, Talkal R, Khan N (2017) In vitro propagation and determination of genetic stability of micropropagated plants of Stevia rebaudiana. Int J Adv Res Innov Ideas Educ 3:884–894
Doležel J, Valárik M, Vrána J, Lysák MA, Hibová E, Bartos J, Gasmanová N, Doleželová M, Safár J, Simková H (2004) Molecular cytogenetics and cytometry of bananas (Musa spp.). In: Banana improvement: cellular molecular biology and induced mutations, vol 1, pp 229–244
Domãnguez F, Chávez M, Garduño-Ramírez ML, Chávez-Avila VM, Mata M, Cruz-Sosa F (2010) Honokiol and magnolol production by in vitro micropropagated plants of Magnolia dealbata, an endangered endemic Mexican species. Nat Prod Commun 5:235–240
Ferreira CM, Handro W (1988) Micropropagation of Stevia rebaudiana through leaf explants from adult plants. Planta Med 54:157–160
Fiuk A, Bednarek PT, Rybczyński JJ (2010) Flow cytometry, HPLC-RP, and metAFLP analyses to assess genetic variability in somatic embryo-derived plantlets of Gentiana pannonica Scop. Plant Mol Biol Rep 28:413–420
Fletcher RA, Gilly A, Sankhla N, Davis TD (2000) Triazoles as plant growth regulators and stress protectants. Hortic Rev 24:55–138
Fujita H, Edahiro T (1978) Safety utilization of stevia sweetener. Food Ind 22:1–8
Gantait S, Mandal N, Bhattacharyya S, Das PK (2009) In vitro mass multiplication with genetic clonality in elephant garlic (Allium ampeloprasum L.). J Crop Weed 5:100–104
Gantait S, Das A, Mandal N (2015) Stevia: a comprehensive review on ethnopharmacological properties and in vitro regeneration. Sugar Tech 17:95–106
Gantait S, Das A, Banerjee J (2018) Geographical distribution, botanical description and self-incompatibility mechanism of genus Stevia—a review. Sugar Tech 20:1–10
George EF, Sherrington PD (1984) Plant propagation by tissue culture. Exegetics Ltd., London
Hossain MA, Siddique AB, Rahman SM, Hossain MA (2010) Chemical composition of the essential oils of Stevia rebaudiana Bertoni leaves. Asian J Trad Med 5:56–61
Hutchinson M, Murch SJ, Saxena PK (1996) Morphoregulatory role of thidiazuron: evidence of the involvement of endogenous auxin in thidiazuron-induced somatic embryogenesis of geranium (Pelargonium·hortorum Bailey). J Plant Physiol 149:573–579
Hwang SJ (2006) Rapid in vitro propagation and enhanced stevioside accumulation in Stevia rebaudiana Bert. J Plant Biol 49:267–270
Ibrahim IA, Nasr MI, Mohammed BR, El-Zefzafi MM (2008) Plant growth regulators affecting in vitro cultivation of Stevia rebaudiana. Sugar Tech 10:254–259
Israeli Y, Reuveni O, Lahav E (1991) Qualitative aspects of somaclonal variations in banana propagated by in vitro techniques. Sci Hortic 48:71–88
Janarthanam B, Gopalakrishnan M, Sai GL, Sekar A (2009) Plant regeneration from leaf derived callus of Stevia rebaudiana Bertoni. Plant Tissue Cult Biotechnol 19:133–141
Jeppesen PB, Gregersen S, Poulsen CR, Hermansen K (2000) Stevioside acts directly on pancreatic β cells to secrete insulin: actions independent of cyclic adenosine monophosphate and adenosine triphosphate- sensitive K+-channel activity. Meta 49:208–214
Kalpana M, Anbazhagan M, Natarajan V, Dhanavel D (2010) Improved micropropagation method for the enhancement of biomass in Stevia rebaudiana Bertoni. Rec Res Sci Tech 2:8–13
Karim Z, Uesugi D, Nakayama N, Hossain MM, Ishihara K, Hamada H (2015) Identification of stevioside using tissue culture-derived stevia (Stevia rebaudiana) leaves. Biochem Insights 8:33–37
Kawiak A, Łojkowska E (2004) Application of RAPD in the determination of genetic fidelity in micropropagated Drosera plantlets. In Vitro Cell Dev Biol Plant 40:592–595
Kazmi A, Khan MA, Mohammad S, Ali A, Ali H (2019) Biotechnological production of natural calorie free steviol glycosides in Stevia rebaudiana: an update on current scenario. Curr Biotechnol 8:70–84
Keshvari T, Najaphy A, Kahrizi D, Zebarjadi A (2018) Callus induction and somatic embryogenesis in Stevia rebaudiana Bertoni as a medicinal plant. Cell Mol Biol 64:46–49
Krikorian AD, Berquam DL (1969) Plant cell and tissue cultures: the role of Haberlandt. Bot Rev 35:59–67
Lassner MW, Orton TJ (2018) Detection of somatic variation. Isozymes in plant genetics and breeding, part A. Elsevier, Amsterdam, pp 209–218
Lata H, Chandra S, Techen N, Wang YH, Khan IA (2013) Molecular analysis of genetic fidelity in micropropagated plants of Stevia rebaudiana Bert. using ISSR marker. Am J Plant Sci 4:964–971
Lemus-Mondaca R, Vega-Gálvez A, Zura-Bravo L, Ah-Hen K (2012) Stevia rebaudiana Bertoni, source of a high-potency natural sweetener: a comprehensive review on the biochemical, nutritional and functional aspects. Food Chem 132:1121–1132
Li J, Baga M, Hucl P, Chibbar RN (2011) Development of microsatellite markers in canary seed (Phalaris canariensis L.). Mol Breed 28:611–621
Lozzi A, Abdelwahd R, Mentag R, Abousalim A (2019) Development of a new culture medium and efficient protocol for in vitro micropropagation of Ceratonia siliqua L. In Vitro Cell Dev Biol Plant 55:615–624
Majumder S, Rahman MM (2016) Micropropagation of Stevia rebaudiana Bertoni. through direct and indirect organogenesis. J Inn Pharma Biol Sci 3:47–56
Martins M, Sarmento D, Oliveira DD (2004) Genetic stability of micropropagated almond plantlets, as assessed by RAPD and ISSR markers. Plant Cell Rep 23:492–496
Meera Manjusha AV, Sathyanarayana BN (2008) Acclimatization studies in stevia (Stevia rebaudiana Bert.). In: Proc. IV International symposium on acclimatization and establishment of micropropagated plants, pp 129–133
Melis MS (1999) Effects of chronic administration of Stevia rebaudiana on fertility in rats. J Ethnopharmcol 67:157–161
Merkle S, Dean JFD (2000) Forest tree biotechnology. Curr Opin Biotechnol 11:298–302
Mitra A, Pal A (2006) In vitro regeneration of Stevia rebaudiana (Bert.) from the nodal explants. J Plant Biochem Biotechnol 16:59–62
Mitra M, Gantait S, Mandal N (2020) Coleus forskohlii: advancements and prospects of in vitro biotechnology. Appl Microbiol Biotechnol 104:2359–2371
Modi AR, Patil G, Kumar N, Singh AS, Subhash N (2012) A simple and efficient in vitro mass multiplication procedure for Stevia rebaudiana Bertoni and analysis of genetic fidelity of in vitro raised plants through RAPD. Sugar Tech 14:391–397
Moktaduzzaman M, Rahman SMM (2009) Regeneration of Stevia rebaudiana and analysis of somaclonal variation by RAPD. Biotechnology 8:449–455
Mundhara R, Rashid A (2006) TDZ-induced triple-response and shoot formation on intact seedlings of Linum, putative role of ethylene in regeneration. Plant Sci 170:185–190
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497
Murch SJ, Campbell SSB, Saxena PK (2001) The role of serotonin and melatonin in plant morphogenesis: regulation of auxin-induced root organogenesis in in vitro-cultured explants of St. John's Wort (Hypericum perforatum L.). In Vitro Cell Dev Biol Plant 37:786–793
Murthy BNS, Murch SJ, Saxena PK (1995) Thidiazuron-induced somatic embryogenesis in intact seedlings of peanut (Arachis hypogea): Endogenous growth regulator levels and significance of cotyledons. Physiol Plant 94:268–276
Nehra NS, Kartha KK, Stushnott C, Giles KL (1992) The influence of plant growth regulator concentrations and callus age on somaclonal variation in callus culture regenerants of strawberry. Plant Cell Tissue Org Cult 29:257–268
Ojha A, Sharma VN, Sharma V (2010) An efficient protocol for in vitro clonal propagation of natural sweetener plant (Stevia rebaudiana Bertoni). Afr J Plant Sci 4:319–321
Rademacher W (2000) Growth retardants: effects on gibberellin biosynthesis and other metabolic pathways. Ann Rev Plant Physiol Plant Mol Biol 51:501–531
Ramírez-Mosqueda MA, Iglesias-Andreu LG, Ramírez-Madero G, Hernández-Rincón EU (2016) Micropropagation of Stevia rebaudiana Bert. in temporary immersion systems and evaluation of genetic fidelity. S Afr J Bot 106:238–243
Rani V, Raina SN (1996) PCR technology. BOTA 46:78–81
Rani V, Raina SN (2000) Genetic fidelity of organized meristem-derived micropropagated plants: a critical reappraisal. In Vitro Cell Dev Biol Plant 36:319–330
Rani V, Parida A, Raina SN (1995) Random amplified polymorphic DNA (RAPD) markers for genetic analysis in micropropagated plants of Populus deltoides Marsh. Plant Cell Rep 14:459–462
Razak UN, Ong CB, Yu TS, Lau LK (2014) In vitro micropropagation of Stevia rebaudiana Bertoni in Malaysia. Braz Arc Biol Technol 57:23–28
Ryynänen L, Aronen T (2005) Genome fidelity during short-and long-term tissue culture and differentially cryostored meristems of silver birch (Betula pendula). Plant Cell Tissue Org Cult 83:21–32
Sahoo Y, Pattnaik SK, Chand PK (1997) Plant regeneration from callus cultures of Morus indica L. derived from seedlings and mature plants. Sci Hortic 69:85–98
Sairkar P, Chandravanshi MK, Shukla NP, Mehrotra NN (2009) Mass production of an economically important medicinal plant Stevia rebaudiana using in vitro propagation techniques. J Med Plants Res 3:266–270
Shahzad A, Parveen S (2013) In vitro conservation protocols for some commercially important medicinal plants. In: Recent trends in biotechnology and therapeutic applications of medicinal plants. Springer, Dordrecht, pp 323–347
Shenoy VB, Vasil IK (1992) Biochemical and molecular analysis of plants derived from embryogenic cultures of Napier grass (Pennisetum purpureum K. Schum.). Theor Appl Genet 83:947–955
Singh P, Dwivedi P, Atri N (2014) In vitro shoot multiplication of Stevia and assessment of stevioside content and genetic fidelity of the regenerants. Sugar Tech 16:430–439
Singh M, Saharan V, Dayma J, Rajpurohit D, Sen Y, Sharma A (2017a) In vitro propagation of Stevia rebaudiana (Bertoni): an overview. Int J Curr Microbiol App Sci 6:1010–1022
Singh M, Saharan V, Rajpurohit D, Sen Y, Joshi A, Sharma A (2017b) Thidiazuron induced direct shoot organogenesis in Stevia rebaudiana and assessment of clonal fidelity of regenerated plants by RAPD and ISSR. Int J Curr Microbiol Appl Sci 6:1690–1702
Sinta MM, Amanah DM (2019) Acclimatization and early growth of tissue culture-derived Stevia rebaudiana at low altitude area in Bogor, Indonesia. Menara Perkebunan 87:68–76
Sivaram L, Mukundan U (2003) In vitro culture studies on Stevia rebaudiana. In Vitro Cell Dev Biol Plant 39:520–523
Soliman HI, Metwali EM, Almaghrabi OA (2014) Micropropagation of Stevia rebaudiana Betroni and assessment of genetic stability of in vitro regenerated plants using inter simple sequence repeat (ISSR) marker. Plant Biotechnol 37:786–793
Sreedhar RV, Venkatachalam L, Thimmaraju R, Bhagyalalakhmi N, Naeayan MS, Ravishankar GA (2008) Direct organogenesis from leaf explants of Stevia rebaudiana and cultivation in bioreactor. Biol Plant 52:355–360
Swanson SM, Mahady GB, Beecher CWW (1992) Stevioside biosynthesis by callus, root, shoot and rooted-shoot cultures in vitro. Plant Cell Tissue Org Cult 28:151–157
Tefera W, Wannakrairoj S (2006) Synergistic effects of some plant growth regulators on in vitro shoot proliferation of Korarima [Aframonum corrorima (Braun) Jansen]. Afr J Biotechnol 5:1894–1901
Thilakarathne PL, Peiris SE, Lankika PC (2019) Efficient low cost seesap (CSUP) technique for micropropagation of newly introduced sweetener plant, Stevia rebaudiana Bertoni. Afr J Biotechnol 18:540–547
Thiyagarajan M, Venkatachalam P (2012) Large scale in vitro propagation of Stevia rebaudiana (Bert) for commercial application: pharmaceutically important and antidiabetic medicinal herb. Ind Crop Prod 37:111–117
Uddin MS, Chowdhury MSH, Khan MMMH, Uddin MB, Ahmed R, Baten MA (2006) In vitro propagation of Stevia rebaudiana Bert in Bangladesh. Afr J Biotechnol 5:1238–1240
Venkatachalam L, Sreedhar RV, Bhagyalakshmi N (2007) Genetic analyses of micropropagated and regenerated plantlets of banana as assessed by RAPD and ISSR markers. In Vitro Cell Dev Biol Plant 43:267–274
Wang SY, Steffens GL, Faust M (1986) Breaking bud dormancy in apple with a plant bioregulator, thidiazuron. Phytochemistry 25:311–317
Yadav A, Kajla S, Poonia AK, Yadav IS, Yadav RC (2016) An efficient micropropagation protocol for Stevia rebaudiana. Med Plants 8:65–73
Yesmin S (2019) In vitro micropropagation of Stevia rebaudiana Bertoni. Plant Tissue Cult Biotechnol 29:277–284
Yücesan B, Mohammed A, Büyükgöçmen R, Altuğ C, Kavas Ö, Gürel S, Gürel E (2016) In vitro and ex vitro propagation of Stevia rebaudiana Bertoni with high Rebaudioside-A content—a commercial scale application. Sci Hortic 203:20–28
Ziv M (1990) The effect of growth retardants on shoot proliferation and morphogenesis in liquid cultured gladiolus plants. Acta Hortic 280:207–214
Ziv M, Ronen G, Raviv M (1998) Proliferation of meristematic clusters in disposable pre sterilized plastic bioreactors for the large-scale micropropagation of plants. In Vitro Cell Dev Biol Plant 34:152–158
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2021 The Author(s), under exclusive license to Springer Nature Singapore Pte Ltd.
About this chapter
Cite this chapter
Sivasankarreddy, K., Abha Manohar, K., Shukla, G., Vineeta, Rather, M.M., Chakravarty, S. (2021). In Vitro Approaches for Mass Propagation of Stevia rebaudiana. In: Gantait, S., Verma, S.K., Sharangi, A.B. (eds) Biotechnology of Anti-diabetic Medicinal Plants. Springer, Singapore. https://doi.org/10.1007/978-981-16-3529-8_5
Download citation
DOI: https://doi.org/10.1007/978-981-16-3529-8_5
Published:
Publisher Name: Springer, Singapore
Print ISBN: 978-981-16-3528-1
Online ISBN: 978-981-16-3529-8
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)