Plant Genetic Research
Scientific Journal

Construction of Plant Expression Vectors Harboring WRI1 and LPAAT Genes and Its Transformation in Tobacco Plants

Document Type : Original Article

Authors

Sahar Dashchi 1
Hassan Rahnama 2
Kianoosh Cheghamirza 1
Katayun Zamani 2

1 Department of Agronomy and Plant Breeding, Faculty of Agricultural Science and Engineering, Razi University, Kermanshah, Iran

2 Agricultural Biotechnology Research Institute of Iran, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran

http://dx.doi.org/10.52547/pgr.7.2.4
Abstract
In oilseed crops, a number of genes involved in the production of triacylglycerol have been identified that changes in their expression have increase the seed oil content. WRI1 and LPAAT are key genes in this synthetic pathway that their overexpression can increase the oil content. In this study, the expression vectors carrying WRI1 and LPAAT genes were designed and constructed for genetic transformation of tobacco (Nicotiana tabacum) plants. The synthetic WRI1 and LPAAT genes were isolated from the PGH.WRI1 and PGH.LPAAT cloning vector using specific restriction enzymes and then cloned in the PGH.O3.2.2 intermediate vector under the control of SBP and Napin promoters and E9 terminator. Finally, the genetic cassettes were transferred to the plant transformation pBin19 binary vector. The resulting constructs were transferred to Agrobacterium tumefaciens strain EHA105 and were used for genetic transformation of tobacco plants. Molecular analysis of transgenic plants confirmed the presence and activity of WRI1 and LPAAT genes. Seeds from transgenic plants were selected on the medium containing kanamycin and developed strong and healthy seedlings.

Keywords

Agrobacterium tumefaciens
Oil seed
Tobacco
WRI1
LPAAT
Plant Genetic Research
Volume 7, Issue 2
December 2020
Pages 41-54

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