Abstract
Napier grass (Cenchrus purpureus Schum.) is one of the highest yielding tropical C4 forage grasses and a promising candidate feedstock for the production of biofuel. Genetic engineering and genome editing have tremendous potential for crop improvement and require the establishment of a transformation protocol. In this study, transgenic Napier grass was generated by introducing a minimal cassette (MC) encoding the antibiotic resistance gene nptII with biolistic gene transfer into embryogenic callus. Paromomycin was superior to geneticin as a selective agent in the selection of transgenic calli. Two high-biomass producing Napier grass accessions were successfully transformed. Southern blot analysis revealed simple transgene integration patterns with 33% of the regenerated transgenic lines having a single insert of the minimal transgene expression cassette. NPTII-ELISA confirmed transgene expression in all transgenic lines. The results of this study will enable the acceleration of Napier grass improvement by genetic transformation or genome editing.
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Abbreviations
- MC:
-
Minimal cassette
- CIM:
-
Callus induction media
- CPA:
-
4-Chlorophenoxyacetic acid
- 2,4-D:
-
2,4-Dichlorophenoxyacetic acid
- MS:
-
Murashige and Skoog
- CaMV:
-
Cauliflower mosaic virus
- Hsp70:
-
Heat shock protein 70
- CIM-S:
-
Callus induction media with selection
- CTAB:
-
Cetyl trimethyl ammonium bromide
- PCR:
-
Polymerase chain reaction
- ELISA:
-
Enzyme-linked immunosorbent assay
- BSA:
-
Bovine serum albumin
- CRISPR:
-
Clustered regularly interspaced short palindromic repeats
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Acknowledgements
We like to thank USDA National Institute of Food and Agriculture for financial support of Hatch project 1020425 and Embrapa for the post-doctoral fellowship for Fabio Faleiro.
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Faleiro, F.G., Kim, J.Y., Parikh, A. et al. Generation of Transgenic Napier Grass (Cenchrus purpureus Schum.) Plants by Biolistic Gene Transfer of a Minimal Expression Cassette. Tropical Plant Biol. 14, 371–380 (2021). https://doi.org/10.1007/s12042-021-09298-z
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DOI: https://doi.org/10.1007/s12042-021-09298-z