A PLANT TRANSFORMATION VECTOR CONTAINING THE GENE dehD FOR THE DEVELOPMENT OF CULTIVARS RESISTANT TO MONOCHLOROACETIC ACID
E. Mohamed1, F. A. Rahiman1, R. A. Wahab2, C. R. C M. Zain3, M. A Javed1 and F. Huyop*1
1Faculty of Biosciences & Medical Engineering, Universiti Teknologi Malaysia, UTM Skudai, 81310 Johor, Malaysia
2Faculty of Science, Universiti Teknologi Malaysia, UTM Skudai, 81310 Johor, Malaysia
3Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600, UKM, Bangi, Malaysia
*Corresponding author Email: email@example.com
A dehalogenase D gene (dehD) capable of degrading monochloroacetic acid (MCA) has been previously isolated from Rhizobium sp. RC1 and characterized. The 804-bp dehD gene was inserted into pCAMBIA under the control of the cauliflower mosaic virus 35S promoter and designated pCAMdehD, with a total size of 10,592 bp. The pCAMdehD was introduced into tobacco (Nicotiana benthamiana) via Agrobacterium-mediated transformation. The integration and expression of dehD in N. benthamiana was confirmed by PCR and reverse transcription PCR, respectively. MCA-resistant transformants were selected in tissue cultures containing 60 µg/L MCA. Analysis of plants using a leaf-painting assay revealed that transgenic N. benthamiana was resistant to4.0 g/L MCAcompared with 2.0 g/L for non-transformed controls. The use of dehD could thus be advantageous for herbicide-resistant plant breeding systems, and it is also a suitable marker gene for plant-transformation studies. To the best of our knowledge, this is the first report detailing transgenic N. benthamiana engineered with dehD from Rhizobium sp. RC1.
Key words: Binary vector, dehD gene, Herbicide resistant, Monochloroacetic acid, Tobacco.