Arzu UÇAR TÜRKER, Buhara YÜCESAN, Ekrem GÜREL

Adventitious shoot regeneration from stem internode explants of Verbena officinalis L., a medicinal plant

Verbena officinalis L. (vervain) is a medicinal plant that shows several biological activities, including antimicrobial, antiinflammatory, analgesic, neuroprotective, hypnotic/sedative, gastroprotective, hepatoprotective, anticancer, cicatrizant, and antioxidant. This report describes an efficient plant regeneration system for V. officinalis via adventitious shoot development from stem internode and petiole explants. Stem internode explants were more effective than petiole explants in terms of both the number of shoots per explant and the percentages of explants forming shoots. Increasing benzyladenine (BA) concentrations steadily increased shoot formation up to 13.32 µM, but 22.22 µM BA caused a sharp decrease. The highest number of shoots (17.1 shoots per explant at 100% frequency) was obtained when 13.32 µM BA was combined with 5.71 µM indole-3-acetic acid (IAA). Rooting of the regenerated shoots was readily achieved when multiple shoots were singled out and cultured on medium containing varying concentrations of different auxins [IAA, indole-3-butyric acid (IBA), 2,4-dichlorophenoxyacetic acid (2,4-D), or naphthalene acetic acid (NAA)]. IBA was more effective in terms of the number of roots developed, producing a mean of 7.1 roots per shoot at 4.92 µM, whereas IAA was more effective for the frequency of shoots developing roots: 100% of the shoots rooted at 5.71 µM IAA. More than 90% of the regenerants survived through the hardening off process, when they were transferred to foam cups and kept under growth room conditions for 2 weeks before finally being moved to room conditions at low humidity. Approximately 3 months after the transfer to room conditions, the flowering of the regenerated plants could be observed.

Anahtar Kelimeler:

Key words: Verbena officinalis, in vitro culture, stem internode explant, micropropagation

Adventitious shoot regeneration from stem internode explants of Verbena officinalis L., a medicinal plant

Keywords:

Key words: Verbena officinalis, in vitro culture, stem internode explant, micropropagation,

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Chevallier A. The Encyclopedia of Medicinal Plants. Dorling Kindersley Limited. London; 1996. 2. Grieve M. A Modern Herbal, vol. 2. Dover Publications. New York; 1982. 3. Baytop T. Türkiye’de Bitkiler ile Tedavi. Nobel Tip Kitabevleri. Istanbul; 1999.
Guarrera PM, Forti G, Marignoli S. Ethnobotanical and
ethnomedicinal uses of plants in the district of Acquapendente
(Latium, Central Italy). J Ethnopharmacol 96: 429-44, 2005. 5.
Hernández NE, Tereschuk ML, Abdala LR. Antimicrobial activity
of flavonoids in medicinal plants from Tafí del Valle (Tucumán,
Lai SW, Yu MS, Yuen WH et al. Novel neuroprotective effects of the aqueous extracts from Verbena officinalis Neuropharmacology 50: 641-50, 2006. L.
Lai SW, Yu MS, Yuen WH et al. Potential neuroprotective agent from botanical extract: an experience of using Verbena officinalis against beta-amyloid peptide neurotoxicity. Neurosignals 15: 146-47, 2006.
Carlini EA. Plants and the central nervous system. Pharmacol Biochem Behav 75: 501-12, 2003.
Singh B, Saxena A, Chandan BK et al. Hepatoprotective activity of verbenalin on experimental liver damage in rodents. Fitoterapia 69: 135-40, 1998.
Dudai N, Weinstein Y, Krup M et al. Citral is a new inducer of caspase-3 in tumor cell lines. Planta Med 71: 484-88, 2005.
Rimpler H, Schafer B. Hastatoside, new iridoid from Verbena officinalis and Verbena hastata (Verbenaceae). Tetrahedron Lett 17: 1463-64, 1973.
Rimpler H, Schafer B. Hastatoside, a new iridoid from Verbena hastata L. and Verbena officinalis L. Z Naturforsch C 34: 311-18, 1979.
Damtoft S, Godthjælpsen L, Jensen SR et al. Age-dependent variations of the efficiency of iridoid biosynthesis in Verbena officinalis. Phytochemistry 22: 2614-15, 1983.
Makboul AM. Chemical constituents of Verbena officinalis. Fitoterapia 57: 50-51, 1986.
Calvo MI, San Julian A, Fernandez M. Identification of the major compounds in extracts of Verbena officinalis L. (Verbenaceae) by HPLC with post-column derivatization. Chromatographia 46: 241-44, 1997.
Deepak M, Handa SS. 3 alpha,24-dihydroxy-urs-12-en-28-oic acid from Verbena officinalis. Phytochemistry 49: 269-71, 1998.
Muller A, Ganzera M, Stuppner H. Analysis of the aerial parts of Verbena officinalis L by micellar electrokinetic capillary chromatography. Chromatographia 60: 193-97, 2004.
Reynaud J, Couble A, Raynaud J. Flavonoids of Verbena officinalis L. J Plant Physiol 135: 380-81, 1989.
Carnat A, Carnat AP, Chavignon O et al. Luteolin 7- diglucuronide, the major flavonoid compound from Aloysia triphylla and Verbena officinalis. Planta Med 61: 490, 1995.
Hansel R, Kallmann S. Verbascoside, a main constituent of Verbena officinalis. Arch Pharm 319: 227-30, 1986.
Woodword FI. Life at the edge: a 14-year study of a Verbena officinalis population’s interactions with climate. J Ecol 85: 899- 906, 1997.
Brandel M, Schütz W. Seasonal dormancy patterns and stratification requirements in seeds of Verbena officinalis L. Basic and App Eco 4: 329-37, 2003.
Rout GR, Samantaray S, Das P. In vitro manipulation and propagation of medicinal plants. Biotechnol Adv 18: 91-120, 2000.
Naika HR, Krishna V. Plant regeneration from callus culture of Clematis gouriana Roxb. – a rare medicinal plant. Turk J Biol 32: 99-103, 2008.
Davis PH. Flora of Turkey and the East Aegean Islands, vol. 7. Edinburgh University Press. Edinburgh; 1982.
Murashige T, Skoog F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473-97, 1962.
Eisvand HR, Arefi HM, Tavakol-Afshari R. Effects of various treatments on breaking seed dormancy of Astragalus siliquosus. Seed Sci Technol 34: 747-52, 2006.
Olmez Z, Temel F, Gokturk A et al. Effect of cold stratification treatments on germination of drought tolerant shrubs seeds. J Environ Biol 28: 447-53, 2007.
Vandelook F, Bolle N, Van Assche JA. Seed dormancy and germination of the European Chaerophyllum temulum (Apiaceae), a member of a trans-Atlantic genus. Ann Bot 100: 233-39, 2007.
Trigiano RN, Gray DJ. Plant Tissue Culture: Concepts and Laboratory Exercises, 2nd edition. CRC Press. Boca Raton; 2000.
Yucesan B, Turker AU, Gurel E. TDZ-induced high frequency plant regeneration through multiple shoot formation in witloof chicory (Cichorium intybus L.). Plant Cell Tiss Org Cult 91: 243- 50, 2007.
Tamura M, Togami J, Ishiguro K et al. Regeneration of transformed verbena (Verbena × hybrida) by Agrobacterium tumefaciens. Plant Cell Rep 21: 459-66, 2003.
Watad AA, Ahroni A, Zuker A et al. Adventitious shoot formation from carnation stem segments: a comparison of different culture procedures. Sci Hortic 65: 313-20, 1996.
Ahroni A, Zuker A, Rozen Y et al. An efficient method for adventitious shoot regeneration from stem-segment explants of gypsophila. Plant Cell Tissue Organ Cult 49: 101-106, 1997.
Chen CC, Chen SJ, Sagare AP et al. Adventitious shoot regeneration from stem internode explants of Adenophora triphylla (Thunb.) A. DC. (Campanulaceae) - an important medicinal herb. Bot Bull Acad Sinica 42: 1-7, 2001.
Gurel E, Wren MJ. In vitro development from leaf explants of sugar beet (Beta vulgaris L.): rhizogenesis and the effect of sequential exposure to auxin and cytokinin. Ann Bot 75: 31-38, 1995. Hosoki T, Katahira S. Micropropagation of Verbena tenera by node culture. Plant Cell Tissue Organ Cult 36: 373-375, 1994.
Charrière F, Sotta B, Miginiac E et al. Induction of adventitious shoots or somatic embryos on in vitro cultured zygotic embryos of Helianthus annuus: variation of endogenous hormone levels. Plant Physiol Biochem 37: 751-57, 1999.