Antioxidant activity of in vitro propagated Stevia rebaudiana Bertoni plants of different origins

An efficient in vitro protocol for propagation of Stevia rebaudiana Bertoni is described. Multiple shoots were induced in vitro from shoot tip and nodal segments on Murashige and Skoog medium containing 6-benzylaminopurine, zeatin, or thidiazuron alone and in combination with naphthalene acetic acid or indole-3-acetic acid. A high frequency of shoot induction as well as maximum number of shoots per shoot tip explant was observed on Murashige and Skoog medium supplemented with 6-benzylaminopurine (1.0 mg L-1) alone and combined with indole-3-acetic acid (0.1 mg L-1). For root induction, in vitro shoots were transferred to rooting media containing naphthalene acetic acid, indole-3-acetic acid, or indole-3-butyric acid. The highest rooting frequency and the highest number of roots was observed in half-strength Murashige and Skoog medium supplemented with 0.1 mg L-1 indole-3-butyric acid. The rooted in vitro plants were successfully acclimatized in a growth chamber and transferred to the field. Leaf extracts of plants propagated in vitro and adapted to field conditions are characterized by high levels of water-soluble antioxidant capacity (expressed as equivalents of ascorbic acid), phenols, and flavonoids, and therefore by high total antioxidant potential, expressed as DPPH radical scavenging activity.

Antioxidant activity of in vitro propagated Stevia rebaudiana Bertoni plants of different origins

An efficient in vitro protocol for propagation of Stevia rebaudiana Bertoni is described. Multiple shoots were induced in vitro from shoot tip and nodal segments on Murashige and Skoog medium containing 6-benzylaminopurine, zeatin, or thidiazuron alone and in combination with naphthalene acetic acid or indole-3-acetic acid. A high frequency of shoot induction as well as maximum number of shoots per shoot tip explant was observed on Murashige and Skoog medium supplemented with 6-benzylaminopurine (1.0 mg L-1) alone and combined with indole-3-acetic acid (0.1 mg L-1). For root induction, in vitro shoots were transferred to rooting media containing naphthalene acetic acid, indole-3-acetic acid, or indole-3-butyric acid. The highest rooting frequency and the highest number of roots was observed in half-strength Murashige and Skoog medium supplemented with 0.1 mg L-1 indole-3-butyric acid. The rooted in vitro plants were successfully acclimatized in a growth chamber and transferred to the field. Leaf extracts of plants propagated in vitro and adapted to field conditions are characterized by high levels of water-soluble antioxidant capacity (expressed as equivalents of ascorbic acid), phenols, and flavonoids, and therefore by high total antioxidant potential, expressed as DPPH radical scavenging activity.

___

  • Duke J. Stevia rebaudiana. In: Duke J. ed. CRC Handbook of Alternative Cash Crops. CRC Press; 1993: pp. 422–424.
  • Goettemoeller J, Ching A. Seed germination in Stevia rebaudiana. In: Janick J. ed. Perspectives on New Crops and New Uses. ASHS Press; 1999: pp. 510–511.
  • Bhosle S. Commercial cultivation of Stevia rebaudiana. Agrobios Newslett 3: 43–45, 2004.
  • Gujral S. Stevia: 0% calorie, 100% sweet, 100% nature. Sci Tech Entrepreneur 12: 1–7, 2004.
  • Kim IS, Yang M, Lee OH et al. The antioxidant activity and the bioactive compound content of Stevia rebaudiana water extracts. LWT Food Sci Tech 44: 1328–1332, 2011.
  • Komissarenko NF, Derkach AI, Kovalyov IP at al. Diterpene glycosides and phenylpropanoids of Stevia rebaudiana Bertoni. Rast Research 1: 53–64, 1994.
  • Hollman PCH, Hertog MGL, Katan MB. Analysis and health effects of flavonoids. Food Chem 57: 43–46, 1996.
  • Siddhuraju P, Becker K. The antioxidant and free radical scavenging activities of processed cowpea (Vigna unguiculata (L.) Walp.) seed extracts. Food Chem 101: 10–19, 2007.
  • Tadhani MB, Patel VH, Subhash R. In vitro antioxidant activity of Stevia rebaudiana leaves and callus. J Food Comp Anal 20: 323–329, 2007.
  • Megeji NW, Kumar JK, Singh V et al. Introducing Stevia rebaudiana, a natural zero-calorie sweetener. Curr Cell Sci 88: 801–804, 2005.
  • Brandle J. Genetic control of rebaudioside A and C concentration in leaves of the sweet herb Stevia rebaudiana. Can J Plant Sci 79: 85–92, 1999.
  • Kinghorn AD. Biologically active compounds from plants with reputed medicinal and sweetening properties. J Nat Prod 50: 1009–1024, 1999.
  • Felippe GM, Lucas NMC. Estudo da viabilidabe dos frutos de Stevia rebaudiana. Hoehnea 1: 95–105, 1971.
  • Sakaguchi M, Kan T. Japanese researches on Stevia rebaudiana. Ci Cult 34: 235–248, 1982.
  • Carneiro JWP, Muniz AS, Guedes TA. Greenhouse bedding plant production of Stevia rebaudiana (Bert) Bertoni. Can J Plant Sci 77: 473–474, 1997.
  • Sivaram L, Mukundan U. In vitro culture studies on Stevia rebaudiana. In Vitro Cell Dev Biol Plant 39: 520–523, 2003.
  • Slavova Y, Nenkova D, Ivanova I. Study on the influence of the substance of benzymidazol upon Stevia rebaudiana Bertoni, cultivated in vitro. Bulg J Agric Sci 9: 225–228, 2003.
  • Mitra A, Pal A. In vitro regeneration of Stevia rebaudiana (Bert) from nodal explants. J Plant Biochem Biotech 16: 59–62, 200 Ahmed MB, Salahin M, Karim R et al. An efficient method for in vitro clonal propagation of a newly introduced sweetener plant (Stevia rebaudiana) in Bangladesh. Am-Eu J Sci Res 2: 121–125, 2007.
  • Debnath M. Clonal propagation and antimicrobial activity of an endemic medicinal plant Stevia rebaudiana. J Med Plants Res 2: 45–51, 2008.
  • Anbazhagan M, Kalpana M, Rajendran R et al. In vitro production of Stevia rebaudiana Bertoni. Emir J Food Agric 22: 216–222, 2010.
  • Satpathy S, Das M. In vitro shoot multiplication in Stevia rebaudiana Bert., a medicinally important plant. Gen Appl Plant Physiol 36: 167–175, 2010.
  • Shatnawi AM, Shibli RA, Abu-Romman SM et al. Clonal propagation and cryogenic storage of the medicinal plant Stevia rebaudiana. Span J Agric Res 9: 213–220, 2011.
  • Cenkci S, Temel M, Kargıoğlu M et al. Propagation of endangered Thermopsis turcica Tan, Vural & Küçüködük using conventional and in vitro techniques. Turk J Biol 33: 327–333, 200 Nakamura S, Tamura Y. Variation in the main glycosides of stevia. Jpn J Trop Agric 29: 109–116, 1985.
  • Murashige T, Skoog F. A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol Plantarum 15: 473–479, 1962.
  • Prieto P, Pineda M, Aguilar M. Spectrophotometric quantitation of antioxidant capacity through the formation of a phosphomolybdenum complex: specific application to the determination of vitamin E. Anal Biochem 269: 337–341, 1999.
  • Tepe B, Sokmen M, Akpulat HA et al. Screening of the antioxidant potentials of six Salvia species from Turkey. Food Chem 95: 200–204, 2006.
  • Pfeffer H, Dannel F, Römheld V. Are there connections between phenol metabolism, ascorbate metabolism and membrane integrity in leaves of boron-deficient sunflower plants? Physiol Plantarum 104: 479–485, 1998.
  • Zhishen J, Mengcheng T, Jianming W. The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals. Food Chem 64: 555–559, 1999.
  • Rafiq M, Dahot MU, Mangrio SM et al. In vitro clonal propagation and biochemical analysis of field established Stevia rebaudiana Bertoni. Pak J Bot 39: 2467–474, 2007.
  • Hossain MA, Shamim AHM, Jahan KTA et al. Micropropagation of stevia. Int J Sustain Crop Prod 3: 1–9, 2008.
  • Uçar Türker A, Yücesan B, Gürel E. Adventitious shoot regeneration from stem internode explants of Verbena officinalis L., a medicinal plant. Turk J Biol 34: 297–304, 2010.
  • Moncaleán P, Rodrígues A, Fernández B. In vitro response of Actinidia deliciosa explants to different BA incubation periods. Plant Cell Tissue Organ Cult 67: 257–266, 2001.
  • Foyer CH, Souriau N, Perret S et al. Overexpression of glutathione reductase but not glutathione synthetase leads to increases in antioxidant capacity and resistance to photoinhibition in poplar trees. Plant Physiol 109: 1047–1057, 19
  • Benavente-García O, Castillo J, Lorente J et al. Antioxidant activity of phenolics extracted from Olea europaea L. leaves. Food Chem 68: 457–462, 2000.
  • Shukla S, Mehta A, Bajpai VK et al. In vitro antioxidant activity and total phenolic content of ethanolic leaf extract of Stevia rebaudiana Bert. Food Chem Toxicol 47: 2338–2343, 2009.
  • Kahkonen MP, Hopia AI, Vuorela HJ. Antioxidant activity of plant extracts containing phenolic compounds. J Agr Food Chem 47: 3954–3962, 1999.
Turkish Journal of Biology-Cover
  • ISSN: 1300-0152
  • Yayın Aralığı: Yılda 6 Sayı
  • Yayıncı: TÜBİTAK
Sayıdaki Diğer Makaleler

Fatty acid composition of root and shoot samples of some Astragalus L. (Fabaceae) taxa growing in the east and southeast of Turkey

Cumali KESKİN, Semra KAÇAR

Phylogenetic analysis of Rhizoctonia solani AG-4 isolates from common beans in Black Sea coastal region, Turkey, based on ITS-5.8S rDNA

Melike Çebi KILIÇOĞLU, İbrahim ÖZKOÇ

Antimicrobial, antioxidant, and synergistic properties of two nutraceutical plants: Terminalia catappa L. and Colocasia esculenta L.

Sumitra CHANDA, Kalpna RAKHOLIYA, Komal DHOLAKIA, Yogesh BARAVALIA

Using seedlessness-related molecular markers in grapevine breeding for seedlessness via marker-assisted selection into Muscat of Hamburg × Sultani progeny

Murat AKKURT, Atilla ÇAKIR, Mina SHİDFAR, Filiz MUTAF, Gökhan SÖYLEMEZOĞLU

Purification of antioxidant protein isolated from Peganum harmala and its protective effect against CCl4 toxicity in rats

Hanaa AHMED, Helal Abu El ZAHAB, Gamia ALSWIAI

Compositional studies and antioxidant potential of Albizia lebbeck (L.) Benth. pods and seeds

Muhammad ZIA-UL-HAQ, Shakeel AHMAD, Mughal QAYUM, Sezai ERCİŞLİ

Karyotype traits in Romanian selections of edible blue honeysuckle

Elena TRUTA, Gabriela VOCHITA, Craita Maria ROSU, Maria Magdalena ZAMFIRACHE

Salicylic acid delays leaf rolling by inducing antioxidant enzymes and modulating osmoprotectant content in Ctenanthe setosa under osmotic stress

Mehmet DEMİRALAY, Aykut SAĞLAM, Asım KADIOĞLU

Comparison of the volatiles of Daphne pontica L. and D. oleoides Schreber subsp. oleoides isolated by hydro- and microdistillation methods

İlhan GÜRBÜZ, Betül DEMİRCİ, Gerhard FRANZ

Antioxidant activity of in vitro propagated Stevia rebaudiana Bertoni plants of different origins

Ely ZAYOVA, Ira STANCHEVA, Maria GENEVA, Maria PETROVA, Lyudmila DIMITROVA