A plant transformation vector containing the gene dehd for the development of cultivars resistant to monochloroacetic acid

• Main Authors: Mohamed, E., Rahiman, F. A., Wahab, R. A., Zain, C. R. C. M., Javed, M. A., Huyop, F.
• Format: Article
• Language: English
• Published: Pakistan Agricultural Scientists Forum 2016
• Subjects: QH301 Biology
• Online Access: http://eprints.utm.my/id/eprint/72227/1/RAWahab2016_APlantTransformationVectorContaining.pdf; http://eprints.utm.my/id/eprint/72227/; https://www.scopus.com/inward/record.uri?eid=2-s2.0-84984908570&partnerID=40&md5=6c2bf9f5330fa5856afb0e5b372af4f3
• Institution: Universiti Teknologi Malaysia

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 to 4.0 g/L MCA compared 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.