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

Abstract

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.

Authors:
; ; ; ; ; ; ; ; ; ORCiD logo
Publication Date:
Sponsoring Org.:
USDOE Advanced Research Projects Agency - Energy (ARPA-E)
OSTI Identifier:
1618774
Grant/Contract Number:  
AR0000313
Resource Type:
Published Article
Journal Name:
Biotechnology for Biofuels
Additional Journal Information:
Journal Name: Biotechnology for Biofuels Journal Volume: 12 Journal Issue: 1; Journal ID: ISSN 1754-6834
Publisher:
Springer Science + Business Media
Country of Publication:
Netherlands
Language:
English

Citation Formats

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., and Stewart, Jr., C. Neal. Embryogenic cell suspensions for high-capacity genetic transformation and regeneration of switchgrass (Panicum virgatum L.). Netherlands: N. p., 2019. Web. https://doi.org/10.1186/s13068-019-1632-3.
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. Embryogenic cell suspensions for high-capacity genetic transformation and regeneration of switchgrass (Panicum virgatum L.). Netherlands. https://doi.org/10.1186/s13068-019-1632-3
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., and Stewart, Jr., C. Neal. Mon . "Embryogenic cell suspensions for high-capacity genetic transformation and regeneration of switchgrass (Panicum virgatum L.)". Netherlands. https://doi.org/10.1186/s13068-019-1632-3.
@article{osti_1618774,
title = {Embryogenic cell suspensions for high-capacity genetic transformation and regeneration of switchgrass (Panicum virgatum L.)},
author = {Ondzighi-Assoume, Christine A. and Willis, Jonathan D. and Ouma, Wilson K. and Allen, Sara M. and King, Zachary and Parrott, Wayne A. and Liu, Wusheng and Burris, Jason N. and Lenaghan, Scott C. and Stewart, Jr., C. Neal},
abstractNote = {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.},
doi = {10.1186/s13068-019-1632-3},
journal = {Biotechnology for Biofuels},
number = 1,
volume = 12,
place = {Netherlands},
year = {2019},
month = {12}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record
https://doi.org/10.1186/s13068-019-1632-3

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