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Expression patterns of cp4-epsps gene in diverse transgenic Saccharum officinarum L. genotypes

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Abstract

Glyphosate, a functional analogue of phosphoenolpyruvate (PEP), blocks the shikimate pathway by inhibiting the enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS; EC 2.5.1.19) through interference with the conversion of (shikimate-3-phosphate) S3P and PEP to 5-enolpyruvylshikimate-3-phosphate (EPSP) and subsequently leads to plant death. This metabolic pathway possesses great potential to be used for development of herbicide resistant transgenic crops and here in this study, we wanted to check the expression potential of CP4-EPSPS gene in various sugarcane genotypes. A synthetic version of CP4-EPSPS gene synthesized commercially, cloned in pGreen0029 vector, was transformed into regenerable embryogenic calli of three different sugarcane cultivars HSF-240, S2003US-778 and S2003US-114 using biolistic gene transfer approach for comparative transcriptional studies. Transgenic lines screened by PCR analysis were subjected to Southern hybridization for checking transgene integration patterns. All the tested lines were found to contain multiple (3–6) insert copies. Putative transgenic plants produced the CP4-EPSPS protein which was detected using immunoblot analysis. The CP4-EPSPS transcript expression detected by qRT-PCR was found to vary from genotype to genotype and is being reported first time. In vitro glyphosate assay showed that transformed plants were conferring herbicide tolerance. It is concluded that different cultivars of sugarcane give variable expression of the same transgene and reasons for this phenomenon needs to be investigated.

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Authors and Affiliations

  1. Agricultural Biotechnology Division, National Institute for Biotechnology and Genetic Engineering, P.O. Box 577, Jhang Road, Faisalabad, Pakistan

    Muhammad Imran, Shaheen Asad & Zahid Mukhtar

  2. Pakistan Institute of Engineering and Applied Sciences, Nilore, Islamabad, Pakistan

    Muhammad Imran, Shaheen Asad & Zahid Mukhtar

  3. Laboratory of Genomics and Molecular Biology, Department of Biological Sciences, Escola Superior Agricultura Luiz De Quiroz, University of Sao Paulo, Piracicaba, SP, Brazil

    Andre Luiz Barboza

  4. Department of Bioinformatics and Biotechnology, International Islamic University, Islamabad, Pakistan

    Zafar M. Khalid

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  1. Muhammad Imran
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  2. Andre Luiz Barboza
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  3. Shaheen Asad
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  4. Zafar M. Khalid
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  5. Zahid Mukhtar
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Imran, M., Barboza, A.L., Asad, S. et al. Expression patterns of cp4-epsps gene in diverse transgenic Saccharum officinarum L. genotypes. Physiol Mol Biol Plants 25, 779–786 (2019). https://doi.org/10.1007/s12298-019-00650-y

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  • DOI: https://doi.org/10.1007/s12298-019-00650-y

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