Regeneration of Ruta graveolens Transgenic Plants

Year 2017, Volume: 3 Issue: 1, 66 - 71, 31.01.2017

Nadiia A. Matvieieva

Abstract

Here we report the construction of transgenic Ruta graveolens L. plants. The plants were obtained via direct shoot
regeneration from leaves and petioles on Murashige and Skoog (MS) solidified medium supplemented with 6-benzylaminopurine
(1.0 mg L-1) and α-naphthylacetic acid (0,05 mg L-1) in case of A. tumefaciens-mediated transformation as
well as via direct shoot regeneration from rue “hairy” root culture on hormone free half-strength MS medium in case
of A. rhizogenes-mediated transformation. Frequency of “hairy” root formation was up to 30%. All transgenic root
lines were able to form shoots. Using of A. tumefaciens allowed to obtain transgenic rue plants with high frequency
up to 100%. Indolil butyric acid addition to the medium in concentration 0.5 mg L-1 enabled transgenic plant rooting.
The shoots regenerated from rue “hairy” root culture were more sensitive to the growth regulator addition and rooted
in 2 weeks. Rooting of the shoots obtained after A. tumefaciens-mediated transformation was observed only in 6 weeks
under these cultivation conditions.

Keywords

Ruta graveolens L., A. tumefaciens, A. rhizogenes, genetic transformation, plant regeneration, “hairy” root culture

References

• Ahmad N, Faisal M, Anis M, Aref IM (2010) In vitro callus induction and plant regeneration from leaf explants of Ruta graveolens L. South African J of Bot 76:597-600 Cunha MR, Melo TS, Magri FMM, Delorenzi JC (2015) Analgesic activity of Ruta graveolens L. (Rue) extracts. African J. of Pharmacy and Pharmacol 9:1-5. DOI: 10.5897/AJPP2014.4152 Diwan R, Malpathak N (2008) Novel technique for scaling up of micropropagated Ruta graveolens shoots using liquid culture systems: A step towards commercialization. N Biotechnol 5:85-91. DOI: 10.1016/j.nbt.2008.02.002 © Plant Breeders Union of Turkey (BİSAB) 71 Draper J, Scott R, Armitage P, Walden R (1988) Plant Genetic Transformation and Gene Expression. A Laboratory Manual. Oxford; Boston: Blackwell Scientific Publications. Ekiert H, Abou-Mandour AA, Czygan FC (2005) Accumulation of biologically active furanocoumarins in Ruta graveolens ssp. divaricata (Tenore) gams in vitro culture. Pharmazie 60:66-68 França Orlanda JF, Nascimento AR (2015) Chemical composition and antibacterial activity of Ruta graveolens L. (Rutaceae) volatile oils, from São Luís, Maranhão, Brazil South African J of Bot 99:103-106. DOI:10.1016/j.sajb.2015.03. 198 Ghosh S, Sikdar S, Mukherjee A, Khuda-Bukhsh AR (2015) Evaluation of chemopreventive potentials of ethanolic extract of Ruta graveolens against A375 skin melanoma cells in vitro and induced skin cancer in mice in vivo. J Integr Med 13:34- 44. DOI: 10.1016/S2095-4964(15)60156-X Ivanova A, Mikhova B, Najdenski H, Tsvetkova I, Kostova I (2005) Antimicrobial and cytotoxic activity of Ruta graveolens. Fitoterapia 76:344- 347. DOI:10.1016/j.fitote.2005.02.008 Lièvre K, Tran TL, Doerper S, Hehn A, Lacoste P, Thomasset B, Bourgaud F, Gontier E (2009) Agrobacterium-mediated transformation of Ruta graveolens L. Methods Mol Biol 547:235-248. DOI: 10.1007/978-1-60327-287-2 Luchakivskaya Yu, Kishchenko O, Gerasymenko I, Olevinskaya Z, Simonenko Yu, Spivak M, Kuchuk M (2011) High-level expression of human interferon alpha-2b in transgenic carrot (Daucus carota L.) plants. Plant Cell Rep 30:407-415. DOI:10.1007/s00299-010-0942-5 Majumdar S, Garai S, Jha S (2012) Use of the cryptogein gene to stimulate the accumulation of Bacopa saponins in transgenic Bacopa monnieri plants. Plant Cell Rep 31:1899-1909. DOI:10.1007/ s00299-012-1303-3 Matvieieva NA, Drobot, KO, Pasichnyk L.A. N.V. Zhytkevych An efficient regeneration system and antimicrobial activity of Ruta graveolens L. plants/ Ecological Engineering and Environment Protection.-2015, № 1.-Р. 33-39 Meepagala KM, Schrader KK, Wedge DE, Duke SO (2005). Algicidal and antifungal compounds from the roots of Ruta graveolens and synthesis of their analogs. Phytochem 66:2689-2695. DOI:10.1016/j.phytochem.2005.09.019 Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plantarum 15: 473-497 Orlita A, Sidwa-Gorycka M, Kumirska J, Malinski E, Siedlecka EM, Gajdus J, Lojkowska E, Stepnowski P (2008) Identification of Ruta graveolens L. metabolites accumulated in the presence of abiotic elicitors. Biotechnol Prog 24:128-133. DOI: 10.1021/bp070261d Porter J.R. (1991) Host range and implications of plant infection by Agrobacterium rhizogenes. Crit Rev Plant Sci 10:387-421. DOI:10.1080/07352689109382318 Preethi KC, Kuttan G, Kuttan R (2006) Anti-tumour activity of Ruta graveolens extract. Asian Pac J Cancer Prev 7:439-443 Raghav SK, Gupta B, Agrawal C, Goswami K, Das H (2006) Anti-inflammatory effect of Ruta graveolens L. in murine macrophage cells. J. Ethnopharmacol 104:234-239 Sharafi A, Hashemi Sohi H, Mousavi A, Azadi P, Dehsara B, Hosseini Khalifani B (2013) Enhanced morphinan alkaloid production in hairy root cultures of Papaver bracteatum by over-expression of salutaridinol 7-o-acetyltransferase gene via Agrobacterium rhizogenes mediated transformation. World J Microbiol Biotechnol 29:2125 -2131 Sidwa-Gorycka M, Krolicka A, Orlita A, Malinski E, Golebiowski M, Kumirska J, Chromik A, Biskup E, Stepnowski P, Lojkowska E (2009) Genetic transformation of Ruta graveolens L. by Agrobacterium rhizogenes: hairy root cultures a promising approach for production of coumarins and furanocoumarins. Plant Cell Tiss Org Cult 97:59-69. DOI: 10.1007/s11240-009-9498-x Tepfer D (1990) Genetic transformation using Agrobacterium rhizogenes Physiol. Plant 79:140-146 Thiruvengadam M, Praveen N, Kim EH, Kim SH, Chung IM (2014) Production of anthraquinones, phenolic compounds and biological activities from hairy root cultures of Polygonum multiflorum Thunb. Protoplasma 251:555-66. DOI:10.1007/s00709-013-0554-3 Zhang Y, Yan YP, Wu YC, Hua WP, Chen C, Ge Q, Wang ZZ (2014) Pathway engineering for phenolic acid accumulations in Salvia miltiorrhiza by combinational genetic manipulation. Metab Eng 21:71-80. DOI:10.1016/j.ymben.2013.10.009