Study creating and cloning hairy root lines in Talinum paniculatum plants
Main Article Content
Abstract
Talinum paniculatum Gaertn. contains flavonoid and saponins that have strong antioxidant properties, which are used to treat a number of conditions such as inflammation, allergies, stomach ulcers, ... However, the amount of flavonoid synthesized naturally in Talinum paniculatum plants is very low (about 0.897 mg /g fresh leaves). Therefore a method has been proposed for enhancing flavonoid content in Talinum paniculatum plants by applying tissue culture technique to produce the hairy roots to enhance biomass. This study present the results of optimization of the gene transfer process to create the hairy root lines through Agrobacterium rhizogenes (A. rhizogenes) in Talinum paniculatum plants. Of the three types of materials that infect by A. rhizogenes (cotyledon, stem, leaf tissue), leaf tissue is a suitable material for transforming and inducing hairy roots. Density of bacteria corresponding to OD600 value=0.6; concentration AS 100 μmol/l; Infection time of 10 minutes; 2 days of co-culture; cefotaxime concentrations of 500 mg/l are suitable conditions for inducing hairy roots from leaf tissue. In state of the liquid MS medium without growth regulator, shaking culture conditions are suitable for hairy roots growth. The test result of presence of rolC gene by PCR and the absence of the virD2 gene asserted that 5 lines of hairy roots were produced from Talinum paniculatum plants.
References
[2] Shimoda H., Nishida N., Ninomiya K., Matsuda H., Yoshikawa M. and Javaberine A., New TNF-alpha and nitric oxide production inhibitor, from the roots of Talinum paniculatum, Heterocycle, 55 (2001): 2043-2050.
[3] Winarni D., Efek ekstrak akar ginseng Jawa dan Korea terhadap libido mencit jantan pada prakondisi testosteron rendah, Berkala Penelitian Hayati, 12(2) (2007): 153-159.
[4] Afolabi O. B., Oloyede O. I., Antioxidant Properties of the Extracts of Talinum Triangulare and its Effect on Antioxidant enzymes in Tissue Homogenate of Swiss Albino Rat, Toxicol Int, 21(3) (2014): 307–313.
[5] Gupta S. K., Liu R. B., Liaw S. Y., Chan H., Tsay H. S., Enhanced tanshinone production in hairy roots of ‘Salvia miltiorrhiza Bunge’ under the influence of plant growth regulators in liquid culture, Botanical Studies, 52 (2011): 435-443.
[6] Kai G., Xu H., Zhou C., Liao P., Xiao J., Luo X., You L., Zhang L., Metabolic engineering tanshinone biosynthetic pathway in Salvia miltiorrhiza hairy root cultures. Metabolic Enginerring, 13(3) (2011): 319-327.
[7] Zhao J., Zhou L. and Wu J., Promotion of Salvia miltiorrhiza hairy root growth and tanshinone production by polysaccharide–protein fractions of plant growth-promoting rhizobacterium Bacillus cereus, Process Biochemistry, 45 (2010): 1517-1522.
[8] Mehrotra S., Kukreja A.K., Khanuja S.P.S., Mishra B.N., Genetic transformation studies and scale up of hairy root culture of Glycyrrhiza glabra in bioreactor, Elec J Biotechnol, 11(2) (2008): 1–7.
[9] Yogananth N., Jothi Basu M., TLC method for determination of plumbagin in hairy root culture of Plumbago rosea L, Glob J Biotechnol Biochem, 4(1) (2009), pp. 66–69.
[10] Majumdar S., Garai S., Jha S., Genetic transformation of Bacopa monnieri by wild type strains of Agrobacterium rhizogenes stimulates production of bacopa saponins in transformed calli and plants, Plant Cell Rep, 30(5) (2011), pp. 941–954.
[11] Thiruvengadam M., Praveen N., Kim E.H., Kim S.H., Chung I.M., Production of anthraquinones, phenolic compounds and biological activities from hairy root cultures of Polygonum multiflorum Thunb, Protoplasma, 251(3) (2014), pp. 555–566.
[12] Thiruvengadam M., Praveen N., Maria John K.M., Yang Y.S., S Kim.H., Chung I.M., Establishment of Momordica charantia hairy root cultures for the production of phenolic compounds and determination of their biological activities, Plant Cell Tissue Organ Cult, 118(3) (2014), pp. 545–557.
[13] Manuhara Y. S. W., Kristanti A. N., Utami E. S. W., Yachya A., Effect of Aeration and Inoculum Density on Biomass and Saponin Content of Talinum Paniculatum Gaertn. Hairy Roots in Balloon-Type Bubble Bioreactor, Journal of Pharmaceutical and Biomedical Science, 2(4) (2012): 47-52.
[14] Murashige T., Skoog F. (1962). A revised medium growth and biosynthesis with tobacco tissue culture. Physiol Plant 15, pp. 473-497.
[15] Lièvre K., Hehn A., Tran T. L. M., Gravot A., Thomasset B., Bourgaud F., Gontier E., Genetic transformation of the medicinal plant Ruta graveolens L. by an Agrobacterium tumefaciens - mediated method. Plant Sci., 168(2005): 883–888.
[16] Veena V., Taylor C. G., Agrobacterium rhizogenes: recent developments and promising applications, In Vitro Cell. Dev. Biol. Plant, 43 (2007): 383-403.
[17] Kiana P., Khosro P., Taiebeh G., Hairy root induction from Portulaca oleracea using Agrobacterium rhizogenes to Noradrenaline’s production, International Research Journal of Applied and Basic Sciences, 3(3) (2012): 642-649..
[18] Sinkar V. P., White F. F., Gordon M. P., Molecular biology of Ri-plasmid, J. Biosci. -Indian Acad.Sci.,11 (1987): 47-57.
[19] Vanhala L., Hiltunen R., Oksman-Caldentey K. M., Virulence of different Agrobacterium strains on hairy root formation of Hyoscyamus muticus, Plan Cell Rep., 14 (1995): 236-240.
[20] Shaghai-Maroof M.A., Soliman K.M., Jorgensen R.A., Allard R.W., Ribosomal DNAsepacer-length polymorphism in barley: mendelian inheritance, chromosomal location, and population dynamics, Proc Natl Acad Sci, 81 (1984): 8014–8019.
[21] Thwe A., Arasu M. V., Li X., Park C. H., Kim S. J., Al-Dhabi N. A., Park S. U., Effect of Different Agrobacterium rhizogenes Strains on Hairy Root Induction and Phenylpropanoid Biosynthesis in Tartary Buckwheat (Fagopyrum tataricum Gaertn), Front Microbiol, 7 (2016): 318.
[22] Diof M. F., Hehn A., Ptak A., Chrétien F., Doerper S., Gontier E., Bourgaud F., Henry M., Chapleur Y., Laurain-Mattar D., Hairy root and tissue cultures of Leucojum aestivum L. - Relationships to galanthamine content, Phytochem.Rev., 6 (2006): 137-141.
[23] Ge X., Wu J., Tanshinone production and isoprenoid pathways in Salvia miltiorrhiza hairy roots induced by Ag+ and yeast elicitor, Plant Science, 168(2) (2005): 487-491.
[24] Chu Hoàng Mậu, Phương pháp phân tích di truyền hiện đại trong chọn giống cây trồng (2008), Nxb Đại học Thái Nguyên.
[25] Manuhara Y. S. W., Kristanti A. N., Utami E. S. W., Yachya A., Effect of sucrose and potassium nitrate on biomass and saponin content of Talinum paniculatum Gaertn. hairy root in balloon-type bubble bioreactor, Asian Pacific Journal of Tropical Biomedicine, Volume 5, Issue 12 ( 2015): p.p 1027-1032.