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Metabolic engineering of isoflavonoid genistein in indica rice by expressing Isoflavone Synthase from Glycine max

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Abstract

Isoflavonoids are a diverse group of secondary metabolites derived from the phenylpropanoid pathway distributed mostly in leguminous plants and possess numerous health benefits. Metabolic engineering is a necessary approach for the bioproduction of various secondary metabolites including isoflavonoids. The present study was carried out to engineer the Glycine max Isoflavone Synthase (GmIFS) in indica rice towards the production of the isoflavone genistein. The Agrobacterium-mediated genetic transformation was carried out and transgenic plants were recovered after three rounds of selection and subsequent regeneration and rooting. The presence and stable integration of the transgene were confirmed by PCR and Southern hybridization in both T0 and T1 generations. Biochemical analysis indicated that the transgenic plants recorded higher phenolic, flavonoid, DPPH radical scavenging activity and total antioxidant potential as compared to the wild type (WT). The relative gene expression analysis revealed that the transgenic plants showed higher levels of gene expression as compared to the WT in both T0 and T1 generations. In addition, HPLC analysis revealed enhanced production of the isoflavonoid genistein with transgenic T0 plant IFS7 and T1 plant IFS 7 − 1 recording the maximum genistein content (11.0 µg/g and 8.0 µg/g). Results of the present study indicate that the stable integration of the GmIFS resulted in the enhanced production of isoflavonoid genistein in rice. These results provide evidence that genetic manipulation of isoflavonoid biosynthesis genes could be a viable strategy to enhance the bioproduction of isoflavonoids in non-leguminous plants like rice which could contribute significantly in eliminating nutritional inadequacies.

Key message

• Attempt to engineer the isoflavonoid pathway in rice was found to be successful.

• Transgenic rice plants expressing Glycine max Isoflavone Synthase demonstrated the accumulation of genistein.

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Data availability

The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

Safia N thanks University Grants Commission-Basic Science Research, New Delhi for fellowship support (UGC BSR No. F.25.1/2014-15). Author Sathish S acknowledges Indian Council for Medical Research (ICMR), New Delhi (No.3/1/2/102/2018-Nut.) for fellowship support. We acknowledge the University Grants Commission – Special Assistance Programme (UGC-SAP-II: F-3-20/2013) and the Department of Science and Technology -Fund for Improvement of S&T Infrastructure (DST-FIST: SR/FST/LSI-618/2014) for providing infrastructure to carry out this research.

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Authors and Affiliations

  1. Plant Genetic Engineering Laboratory, Department of Biotechnology, Bharathiar University, Coimbatore, 641 046, India

    Safia Nayeem, Sathish Sundararajan, Venkatesh Rajendran, Ashwini Malla & Sathishkumar Ramalingam

  2. Technologico de Monterrey, Centre of Bioengineering, Epigmenio Gonzalez #500, Fracc. San Pablo, Campus Queretaro, Santiago de Querétaro, Queretaro, Mexico

    Ashutosh Sharma

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  1. Safia Nayeem
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  2. Sathish Sundararajan
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  3. Venkatesh Rajendran
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  4. Ashwini Malla
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  5. Ashutosh Sharma
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  6. Sathishkumar Ramalingam
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Contributions

SN carried out the experiments and prepared the manuscript, SS contributed to experimental analysis, manuscript preparation, and data representations. VR contributed to data analysis and representations, AM carried out the initial cloning experiments in IFS, AS contributed to experimental analysis, manuscript evaluation and SR conceptualized, supervised the research, and critically evaluated the manuscript.

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Correspondence to Sathishkumar Ramalingam.

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All authors read, approved the manuscript, and declare that there is no conflict of interest.

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Communicated by Manoj Prasad.

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Nayeem, S., Sundararajan, S., Rajendran, V. et al. Metabolic engineering of isoflavonoid genistein in indica rice by expressing Isoflavone Synthase from Glycine max. Plant Cell Tiss Organ Cult 155, 243–253 (2023). https://doi.org/10.1007/s11240-023-02577-5

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  • DOI: https://doi.org/10.1007/s11240-023-02577-5

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