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Transgenic rice expressing a codon-modified synthetic CP4-EPSPS confers tolerance to broad-spectrum herbicide, glyphosate

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Highly tolerant herbicide-resistant transgenic rice was developed by expressing codon-modified synthetic CP4 - EPSPS . The transformants could tolerate up to 1 % commercial glyphosate and has the potential to be used for DSR (direct-seeded rice).

Abstract

Weed infestation is one of the major biotic stress factors that is responsible for yield loss in direct-seeded rice (DSR). Herbicide-resistant rice has potential to improve the efficiency of weed management under DSR. Hence, the popular indica rice cultivar IR64, was genetically modified using Agrobacterium-mediated transformation with a codon-optimized CP4-EPSPS (5-enolpyruvylshikimate-3-phosphate synthase) gene, with N-terminal chloroplast targeting peptide from Petunia hybrida. Integration of the transgenes in the selected rice plants was confirmed by Southern hybridization and expression by Northern and herbicide tolerance assays. Transgenic plants showed EPSPS enzyme activity even at high concentrations of glyphosate, compared to untransformed control plants. T 0, T 1 and T 2 lines were tested by herbicide bioassay and it was confirmed that the transgenic rice could tolerate up to 1 % of commercial Roundup, which is five times more in dose used to kill weeds under field condition. All together, the transgenic rice plants developed in the present study could be used efficiently to overcome weed menace.

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Acknowledgments

We are grateful to Nuziveedu Seeds Limited (NSL), Hyderabad for providing financial assistance. We acknowledge the help of Dr Vikrant Nain for codon optimization of mCP4-EPSPS gene. We are also thankful to our colleagues Hari Kishore CM and Shruti Yadava for help.

Conflict of interest

The authors do not have any conflict of interest.

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Author notes

  1. Polumetla Ananda Kumar

    Present address: Institute of Biotechnology, Acharya NG Ranga Agricultural University, Rajendranagar, Hyderabad, 500030, Telangana, India

Authors and Affiliations

  1. National Research Centre on Plant Biotechnology, Lal Bahadur Shastri Building, Pusa Campus, New Delhi, 110012, India

    Sushil Chhapekar, Sanagala Raghavendrarao, Gadamchetty Pavan, Chopperla Ramakrishna, Vivek Kumar Singh, Mullapudi Lakshmi Venkata Phanindra, Gurusamy Dhandapani, Rohini Sreevathsa & Polumetla Ananda Kumar

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  1. Sushil Chhapekar
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  2. Sanagala Raghavendrarao
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Correspondence to Polumetla Ananda Kumar.

Additional information

Communicated by Manoj Prasad.

Electronic supplementary material

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299_2014_1732_MOESM1_ESM.jpg

Supplementary Fig. 1: mCP4-EPSPS gene-specific PCR analysis of putative T 0 transgenic rice plant. M: 1-kb ladder, W: Water control; UC: Untransformed control plant; Lanes 1-15: Putative transgenic plants; PC: Binary plasmid control. (JPEG 13 kb)

299_2014_1732_MOESM2_ESM.jpg

Supplementary Fig. 2: Immunostrip analysis of T 0 CP4-EPSPS transgenic rice. UC; Untransformed control plant, Lane 1 to 15; T 0 putative transgenic plants (JPEG 44 kb)

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Chhapekar, S., Raghavendrarao, S., Pavan, G. et al. Transgenic rice expressing a codon-modified synthetic CP4-EPSPS confers tolerance to broad-spectrum herbicide, glyphosate. Plant Cell Rep 34, 721–731 (2015). https://doi.org/10.1007/s00299-014-1732-2

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  • DOI: https://doi.org/10.1007/s00299-014-1732-2

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