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Rapid and reproducible Agrobacterium-mediated transformation of sorghum

  • Genetic Transformation and Hybridization
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Abstract

A rapid and reproducible Agrobacterium-mediated transformation protocol for sorghum has been developed. The protocol uses the nptII selectable marker gene with either of the aminoglycosides geneticin or paromomycin. A screen of various A. tumefaciens strains revealed that a novel C58 nopaline chromosomal background carrying the chrysanthopine disarmed Ti plasmid pTiKPSF2, designated NTL4/Chry5, was most efficient for gene transfer to sorghum immature embryos. A NTL4/Chry5 transconjugant harboring the pPTN290 binary plasmid, which carries nptII and GUSPlus TM expression cassettes, was used in a series of stable transformation experiments with Tx430 and C2-97 sorghum genotypes and approximately 80% of these transformation experiments resulted in the recovery of at least one transgenic event. The transformation frequencies among the successful experiments ranged from 0.3 to 4.5%, with the average transformation frequency being approximately 1% for both genotypes. Over 97% of the transgenic events were successfully established in the greenhouse and were fully fertile. Co-expression of GUSPlus TM occurred in 89% of the transgenic T0 events. Seed set for the primary transgenic plants ranged from 145 to 1400 seed/plant. Analysis of T1 progeny demonstrated transmission of the transgenes in a simple Mendelian fashion in the majority of events.

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Acknowledgments

The authors would like to thank Stephen Farrand for A. tumefaciens strains. This work was support by the Nebraska Research Initiative, University of Nebraska's Agricultural Research Division, Nebraska Grain Sorghum Board, University of Nebraska's Plant Science Initiative, and the University of Nebraska's Center for Biotechnology. This is journal series paper number 14586 from the Nebraska Agriculture Research Division.

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Authors and Affiliations

  1. Center for Biotechnology, University of Nebraska, N300 Beadle Center, Lincoln, NE, 68588, USA

    Arlene Howe, Shirley Sato, Mike Fromm & Tom Clemente

  2. Department of Agronomy and Horticulture, University of Nebraska, Lincoln, NE, 68583, USA

    Ismail Dweikat, Mike Fromm & Tom Clemente

  3. Plant Science Initiative, University of Nebraska, N300 Beadle Center, Lincoln, NE, 68588, USA

    Mike Fromm & Tom Clemente

Authors
  1. Arlene Howe
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  2. Shirley Sato
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  3. Ismail Dweikat
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  4. Mike Fromm
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  5. Tom Clemente
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Corresponding author

Correspondence to Tom Clemente.

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Communicated by P.Ozias-Akins

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Howe, A., Sato, S., Dweikat, I. et al. Rapid and reproducible Agrobacterium-mediated transformation of sorghum. Plant Cell Rep 25, 784–791 (2006). https://doi.org/10.1007/s00299-005-0081-6

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  • DOI: https://doi.org/10.1007/s00299-005-0081-6

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