Rapid and reproducible Agrobacterium-mediated transformation of sorghum

Arlene Howe, Shirley Sato, Ismail Dweikat, Mike Fromm, Tom Clemente

Research output: Contribution to journalArticlepeer-review

81 Scopus citations

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 2006

Keywords

  • GUSPlus™
  • Genetic engineering
  • Immature embryo
  • nptII

ASJC Scopus subject areas

  • Agronomy and Crop Science
  • Plant Science

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