Effect of Modified Heptamethyltrisiloxane on the Efficiency of Agrobacterium-Mediated Transformation and Expression of Recombinant Structure in Plant Cell and Tissue Culture

Abstract:

Detergents represent a unique class of chemical compounds. They can alter surface and interphase bonds, and form micellar systems. These detergent properties allow to alter wettability of surfaces, stabilize or destabilize dispersed systems, and modify the properties of liquid phases. Therefore, the use of detergents is virtually unlimited in chemical synthesis and processing, medicine, biological systems and agricultural biology. The article includes the studies of the feasibility of application of 1,1,1,3,5,5,5-heptamethyltrisiloxane modified by polyalkylene oxide in combination with allyloxypolyethyleneglycol in the ratio of 10:1 as a detergent for agrobacterial-mediated transformation. Tween 20 detergent was used as a means of control in the concentration of 5%. As a result of histochemical analysis of transformed tissues, a significant difference was determined in expression of beta-glucuronidase reporter gene. The study of gene expression by calculating the relative content of mRNA showed that the initial number of mRNA copies of genetic makers transfected by co-culturing in a liquid medium with addition of Tween 20 is on the average 32% lower than when using modified heptamethyltrisiloxane

• Periodical: Key Engineering Materials (Volume 683)
• Pages: 506-510
• DOI: https://doi.org/10.4028/www.scientific.net/KEM.683.506
• Online since: February 2016
• Authors: Mikhail Skaptsov*, Maxim Kutsev, Maxim Filipenko, Evgeniy Khrapov, Harue Shinoyama
• Keywords: Complementary DNA, Electrophoresis, GUS-Assay, Modified Heptamethyltrisiloxane, Polymerase Chain Reaction, RNA, Surfactant, Tween20

References

[1] M. Mishra , P. Muthuprasanna, K. Surya-prabha, P. Sobhita-rani, I.A. Satish-babu, I. Sarath-chandiran,G. Arunachalam, S. Shalini, Basics and potential applications of surfactants - a review, Int. J. PharmTech Res. 1, 4 (2009) 1354-1365.

[2] R. Azarmi, A. Ashjaran, Type and application of some common surfactants, J. Chem. Pharm. Res. 7, 2 (2015) 632-640.

[3] R.K. Sahoo, N. Biswas, A. Guha, N. Sahoo, K. Kuotsu, Nonionic surfactant vesicles in ocular delivery: innovative approaches and perspectives, BioMed Res. Int. 2014 (2014).

DOI: 10.1155/2014/263604

[4] R.B. Horsch, J.E. Fry, N.L. Hoffmann, M. Wallroth, D. Eichholtz, S.G. Rogers, R.T. Fraley, A simple and general method for transferring genes into plants, Science 227 (1985) 1229-1231.

DOI: 10.1126/science.227.4691.1229

[5] R.T. Fraley, S.G. Rogers, R.B. Horsch, G.F. Barry, Gene transfer in plants: a tool for studying gene expression and plant development, in: L. Bogard (eds. ), Mol. Dev. Biol., A.R. Liss Inc., New York, 1986, pp.15-26.

[6] G.F. Arcos-Ortega, R.A. Chan-Kuuk, W.A. González-Kantún, R. Souza-Perera, Y.E. Nakazawa-Ueji, E. Avilés-Berzunza, G. Godoy-Hernández, M.A. Lawton, J.J. Aguilar,. Agrobacterium tumefaciens-transient genetic transformation of Habanero pepper (Capsicum chinense Jacq. ) leaf explants, Electro. J. Biotech. 13, 4 (2010).

DOI: 10.2225/vol13-issue4-fulltext-10

[7] A. Paul, S. Bakshi, D.P. Sahoo, M.C. Kalita, L. Sahoo, Agrobacterium-mediated genetic transformation of Pogostemon cablin (Blanco) Benth. using leaf explants: bactericidal effect of leaf extracts and counteracting strategies, Appl. Biochem. Biotechnol. 166, 8 (2012).

DOI: 10.1007/s12010-012-9612-0

[8] R. Mora-Lugo, J. Zimmermann, A. M Rizk, M. Fernandez-Lahore, Development of a transformation system for Aspergillus sojae based on the Agrobacterium tumefaciens-mediated approach, BMC Microbiology. 14 (2014) 247-254.

DOI: 10.1186/s12866-014-0247-x

[9] Butenko R.G. Cell biology of higher plants in vitro and biotechnology based on them, FBK-Press, Moscow, (1999).

[10] T. Murashige, F. Skoog, A revised medium for rapid growth and bioassays with tobacco tissue cultures, Plant Physiol. 15, 13 (1962) 473-497.

DOI: 10.1111/j.1399-3054.1962.tb08052.x

[11] M.V. Skaptsov, M.G. Kutsev, Effect of 24-epibrassinolide on duration of sorrel (Rumex acetosa L. ) cultivation in vitro, Tomsk State Univ. J. Biol. 2, 22 (2013) 52-56.

[12] H. Shinoyama, M. Komano, Y. Nomura, T. Nagai, introduction of delta-endotoxin gene of Bacillus thuringiensis to chrysanthemum [Dendranthema×grandiflorum (Ramat. ) Kitamura] for Insect Resistance, Breeding Sci. 52, 1 (2002) 43-50.

DOI: 10.1270/jsbbs.52.43

[13] R.A. Jefferson, T.A. Kavanagh, M.W. Bevan, Gus fusions: ß-glucuronidase as a sensitive and versatile gene marker in higher plants, EMBO J. 6 (1987) 3901-3907.

DOI: 10.1002/j.1460-2075.1987.tb02730.x