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A simplified protocol for genetic transformation of switchgrass (Panicum virgatum L.)

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Abstract

The increasing interest in renewable energy has attracted more research attention on biofuels. In order to generate sustainable amount of biomass feedstock from dedicated biofuel crops such as switchgrass they need to be genetically improved. Genetic transformation is one of the techniques to achieve this goal. The aim of our study was to devise a simplified protocol for switchgrass genetic transformation. We have used NB0 as the basal medium and mature seeds of the cultivar Alamo as the starting material. The nutrient medium used and scutellum-derived callus are fashioned after rice genetic transformation protocols. We obtained friable calluses, which were similar to the type II calluses in other monocotyledonous species. Calluses were amenable for Agrobacterium-mediated genetic transformation with at least 6 % transformation efficiency. The concentration of hygromycin was optimized for successful selection of transgenic calluses. The Green Fluorescent Protein gene was used to monitor and demonstrate successful genetic transformation. Compared to the previously published methods for genetic transformation of switchgrass, our protocol is simpler and equally efficient.

Key message

An efficient, simplified switchgrass genetic transformation method with NB0 basal medium and mature seeds as inoculum was developed. The appropriate concentrations of hormones and selection agent are described.

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Abbreviations

2,4-D :

2,4-Dichlorophenoxyacetic acid

BA:

N 6-Benzyladenine

sGFP:

Synthetic green fluorescent protein

HPT:

Hygromycin phosphotransferase

KOH:

Potassium hydroxide

MS:

Murashige and Skoog

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Acknowledgments

This work was funded by the Science and Engineering Research Council (SERC Grant No.: 0921390036) of the Agency for Science Technology and Research, Singapore; and the National University of Singapore.

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Authors and Affiliations

  1. Department of Biological Sciences, Faculty of Science, National University of Singapore, 10 Science Drive 4, Singapore, 117543, Singapore

    Rengasamy Ramamoorthy & Prakash P. Kumar

  2. Temasek Life Sciences Laboratory, 1 Research Link, National University of Singapore, Singapore, 117604, Singapore

    Prakash P. Kumar

Authors
  1. Rengasamy Ramamoorthy
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  2. Prakash P. Kumar
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Correspondence to Prakash P. Kumar.

Additional information

Communicated by P. Lakshmanan.

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Ramamoorthy, R., Kumar, P.P. A simplified protocol for genetic transformation of switchgrass (Panicum virgatum L.). Plant Cell Rep 31, 1923–1931 (2012). https://doi.org/10.1007/s00299-012-1305-1

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  • DOI: https://doi.org/10.1007/s00299-012-1305-1

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