Rapid and reproducible Agrobacterium-mediated transformation of sorghum

Arlene Howe, Shirley Sato, Ismail Dweikat, Michael E Fromm, Tom Clemente

Research output: Contribution to journalArticle

73 Citations (Scopus)

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™ 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™ 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.

Original languageEnglish (US)
Pages (from-to)784-791
Number of pages8
JournalPlant Cell Reports
Volume25
Issue number8
DOIs
StatePublished - Aug 1 2006

Fingerprint

Agrobacterium
genetically modified organisms
plasmids
geneticin
nopaline
aminoglycosides
genotype
Spermatophytina
seed set
gene transfer
embryo (plant)
transgenes
transgenic plants
immatures
greenhouses
genetic markers

Keywords

  • GUSPlus™
  • Genetic engineering
  • Immature embryo
  • nptII

ASJC Scopus subject areas

  • Agronomy and Crop Science
  • Plant Science

Cite this

Howe, A., Sato, S., Dweikat, I., Fromm, M. E., & Clemente, T. (2006). Rapid and reproducible Agrobacterium-mediated transformation of sorghum. Plant Cell Reports, 25(8), 784-791. https://doi.org/10.1007/s00299-005-0081-6

Rapid and reproducible Agrobacterium-mediated transformation of sorghum. / Howe, Arlene; Sato, Shirley; Dweikat, Ismail; Fromm, Michael E; Clemente, Tom.

In: Plant Cell Reports, Vol. 25, No. 8, 01.08.2006, p. 784-791.

Research output: Contribution to journalArticle

Howe, A, Sato, S, Dweikat, I, Fromm, ME & Clemente, T 2006, 'Rapid and reproducible Agrobacterium-mediated transformation of sorghum', Plant Cell Reports, vol. 25, no. 8, pp. 784-791. https://doi.org/10.1007/s00299-005-0081-6
Howe, Arlene ; Sato, Shirley ; Dweikat, Ismail ; Fromm, Michael E ; Clemente, Tom. / Rapid and reproducible Agrobacterium-mediated transformation of sorghum. In: Plant Cell Reports. 2006 ; Vol. 25, No. 8. pp. 784-791.
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