Embryogenic cell suspensions for high-capacity genetic transformation and regeneration of switchgrass (Panicum virgatum L.)

Ondzighi-Assoume, Christine A.; Willis, Jonathan D.; Ouma, Wilson K.; Allen, Sara M.; King, Zachary; Parrott, Wayne A.; Liu, Wusheng; Burris, Jason N.; Lenaghan, Scott C.; Stewart, Jr., C. Neal

doi:10.1186/s13068-019-1632-3

Embryogenic cell suspensions for high-capacity genetic transformation and regeneration of switchgrass (Panicum virgatum L.)

Journal Article · Mon Dec 16 00:00:00 EST 2019 · Biotechnology for Biofuels

DOI:https://doi.org/10.1186/s13068-019-1632-3· OSTI ID:1618774

Ondzighi-Assoume, Christine A.; Willis, Jonathan D.; Ouma, Wilson K.; Allen, Sara M.; King, Zachary; Parrott, Wayne A.; Liu, Wusheng; Burris, Jason N.; Lenaghan, Scott C.;

Abstract Background

Switchgrass ( Panicum virgatum L.), a North American prairie grassland species, is a potential lignocellulosic biofuel feedstock owing to its wide adaptability and biomass production. Production and genetic manipulation of switchgrass should be useful to improve its biomass composition and production for bioenergy applications. The goal of this project was to develop a high-throughput stable switchgrass transformation method using Agrobacterium tumefaciens with subsequent plant regeneration.

Results

Regenerable embryogenic cell suspension cultures were established from friable type II callus-derived inflorescences using two genotypes selected from the synthetic switchgrass variety ‘Performer’ tissue culture lines 32 and 605. The cell suspension cultures were composed of a heterogeneous fine mixture culture of single cells and aggregates. Agrobacterium tumefaciens strain GV3101 was optimum to transfer into cells the pANIC-10A vector with a hygromycin-selectable marker gene and a pporRFP orange fluorescent protein marker gene at an 85% transformation efficiency. Liquid cultures gave rise to embryogenic callus and then shoots, of which up to 94% formed roots. The resulting transgenic plants were phenotypically indistinguishable from the non-transgenic parent lines.

Conclusion

The new cell suspension-based protocol enables high-throughput Agrobacterium -mediated transformation and regeneration of switchgrass in which plants are recovered within 6–7 months from culture establishment.

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Sponsoring Organization:: USDOE Advanced Research Projects Agency - Energy (ARPA-E)

Grant/Contract Number:: AR0000313

OSTI ID:: 1618774

Journal Information:: Biotechnology for Biofuels, Journal Name: Biotechnology for Biofuels Journal Issue: 1 Vol. 12; ISSN 1754-6834

Publisher:: Springer Science + Business MediaCopyright Statement

Country of Publication:: Netherlands

Language:: English

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