GENRATING GLUTAMINE SYNTHETASE ENCODED GS1 TRANSGENIC MELIA AZEDARACH L. PLANTS ENHANCED NITROGEN

Ong Xuân Phong; Lý Khánh Linh; La Việt Hồng; Nguyễn Văn Phong; Phạm Bích Ngọc

doi:10.34238/tnu-jst.6382

GENRATING GLUTAMINE SYNTHETASE ENCODED GS1 TRANSGENIC MELIA AZEDARACH L. PLANTS ENHANCED NITROGEN

About this article

Received: 18/08/22 Revised: 26/09/22 Published: 07/10/22

Authors

1. Ong Xuan Phong, Graduate University of Sciences and Technology – VAST; Hanoi Pedagogical University 2
2. Ly Khanh Linh, Institute of Biotechnology - VAST
3. La Viet Hong, Hanoi Pedagogical University 2
4. Nguyen Van Phong, Vietnam National University of Forestry
5. Pham Bich Ngoc , Institute of Biotechnology - VAST

Abstract

Glutamine synthetase encoded by the GS1 gen is a nitrogen-assimilating enzyme that uses ammonia as a substrate for the ATP-dependent conversion of glutamic acid to glutamine. Based on the enzyme pair XmaI and SacI, we have successfully engineered pBI121/GmPrP2:GS1_cmyc:NOS and transformed it into Agrobacterium tumefaciens. Carrying out the gen transformation experiment on more than 700 gen recipients that are stem segments in vitro, 33 regenrated and rooted shoots were obtained after 3 screenings on selective culture medium containing 50 - 150 mg /l kanamycin. The results of checking transgenic lines by PCR technique showed that 25/33 lines appeared with a bar with a size of ̴ 1.1 kb, equivalent to the size of the GS1 gen. The obtained results are the basis for the selection and breeding of transgenic species with fast growth gens.

Keywords

GS1; Melia azedarach L.; Nitrogen; Promoter; Transgenic

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References

[1] X. D. Do, V. T. Bui, V. G. Ho, V. S. Le, and H. H. Chu, " Transformation of Gibberellin 20-oxidase encoded gene to paradise tree (Melia azedarach L.) via Agrobacterium tumefaciens," Journal of Biotechnology, vol. 9, no. 2, pp. 217-222, 2011.

[2] S. Huang, A. S. Raman, J. E. Ream, H. Fujiwara, R. E. Cerny, and S. M. Brown, "Overexpression of 20-oxidase confers a gibberellin-overproduction phenotype in Arabidopsis," Plant physiology, vol. 118, no. 3, pp. 773-781, 1998.

[3] L. K. Ly, T. P. Bui, P. T. Do, A. V. T. Le, P. Van Nguyen, and N. B. Pham, "Demonstration of improved plant growth and biomass production by At1g73160 promoter-controlled root-preferential expression of the AtGA20ox gene," Research Square, 2020, doi: 10.21203/rs.3.rs-19585/v42020.

[4] Y. Wang, B. Fu, L. Pan, L. Chen, X. Fu, and K. Li, "Overexpression of Arabidopsis Dof1, GS1 and GS2 enhanced nitrogen assimilation in transgenic tobacco grown under low-nitrogen conditions," Plant Molecular Biology Reporter, vol. 31, no. 4, pp. 886-900, 2013.

[5] E. Nemeth, Z. Nagy, and A. Pecsvaradi, "Chloroplast Glutamine Synthetase, the Key Regulator of Nitrogen Metabolism in Wheat, Performs Its Role by Fine Regulation of Enzyme Activity via Negative Cooperativity of Its Subunits," Front Plant Sci, vol. 9, p. 191, 2018, doi: 10.3389/fpls.2018.00191.

[6] Z. Zhang, S. Xiong, Y. Wei, X. Meng, X. Wang, and X. Ma, "The role of glutamine synthetase isozymes in enhancing nitrogen use efficiency of N-efficient winter wheat," Sci Rep, vol. 7, no. 1, p. 1000, Apr 20, 2017, doi: 10.1038/s41598-017-01071-1.

[7] T. W. Becker, M. Caboche, E. Carrayol, and B. Hirel, "Nucleotide sequence of a tobacco cDNA encoding plastidic glutamine synthetase and light inducibility, organ specificity and diurnal rhythmicity in the expression of the corresponding genes of tobacco and tomato," Plant Molecular Biology, vol. 19, no. 3, pp. 367-379, 1992.

[8] F. R. Cantón, M.-F. Suárez, M. Jose-Estanyol, and F. M. Cánovas, "Expression analysis of a cytosolic glutamine synthetase gene in cotyledons of Scots pine seedlings: developmental, light regulation and spatial distribution of specific transcripts," Plant molecular biology, vol. 40, no. 4, pp. 623-634, 1999.

[9] F. Gallardo, J. Fu, F. R. Cantón, A. García-Gutiérrez, F. M. Cánovas, and E. G. Kirby, "Expression of a conifer glutamine synthetase gene in transgenic poplar," Planta, vol. 210, no. 1, pp. 19-26, 1999.

[10] V. M. Nguyen, V. H. La, and X. P. Ong, Methods in plant physiology. Hanoi: Hanoi National University Publishing House, 2013.

[11] L. Chen, B. Jiang, C. Wu, S. Sun, W. Hou, and T. Han, "GmPRP2 promoter drives root-preferential expression in transgenic Arabidopsis and soybean hairy roots," BMC plant biology, vol. 14, p. 245, September 16, 2014, doi: 10.1186/s12870-014-0245-z.

[12] T. B. Le, V. C. Phan, V. H. Nong, N. H. Truong, and Q. H. Le, Application of molecular techniques in the study of biological resources in Vietnam. Ha Noi: Science and Technology Publishing House, 2003.

[13] V. T. Bui, X. D. Do, V. S. Le, and H. H. Chu, "The high frequency of transformation procedure for expression of gus gene in chinaberry tree (melia azedarach L.) using agrobacterium," Journal of Biotechnology, vol. 35, no. 2, pp. 227-233, 2013.

DOI: https://doi.org/10.34238/tnu-jst.6382

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