Sorghum genetic transformation by particle bombardment

Liu, Guoquan, Campbell, Bradley C. and Godwin, Ian D. (2014). Sorghum genetic transformation by particle bombardment. In Robert J. Henry and Agnelo Furtado (Ed.), Cereal genomics: methods and protocols (pp. 219-234) New York, NY, United States: Humana Press. doi:10.1007/978-1-62703-715-0-18

Author Liu, Guoquan
Campbell, Bradley C.
Godwin, Ian D.
Title of chapter Sorghum genetic transformation by particle bombardment
Title of book Cereal genomics: methods and protocols
Place of Publication New York, NY, United States
Publisher Humana Press
Publication Year 2014
Sub-type Research book chapter (original research)
DOI 10.1007/978-1-62703-715-0-18
Open Access Status
Series Methods in Molecular Biology
ISBN 9781627037143
ISSN 1064-3745
Editor Robert J. Henry
Agnelo Furtado
Volume number 1099
Chapter number 18
Start page 219
End page 234
Total pages 16
Total chapters 24
Collection year 2015
Subjects 1312 Molecular Biology
1311 Genetics
Abstract/Summary Particle bombardment transformation describes the acceleration of high-velocity microparticles coated with exotic genes through the plant-protective cell walls, in order for the introduced genes to be integrated into the host genome. This technique has proven to be an effective and versatile approach towards plant genetic modification in preceding decades. Particle bombardment has been successfully applied to cereals including rice, maize, wheat, barley, and sorghum. Historically, sorghum has been considered as one of the most recalcitrant major crops with regard to successful genetic transformation; however, tremendous progress has been made in recent years. Transformation efficiency by particle bombardment has now improved from approximately 1 % to in excess of 20 % utilizing an optimized tissue culture and DNA delivery system. The protocol described in this chapter routinely generates transformants at 10-25 % efficiency within sorghum genotype Tx430. The process generally takes 11-16 weeks from initiation of immature embryos to planting of transformants. This protocol covers the operation of both the Bio-Rad PDS-1000/He System and particle inflow gun. Three factors are crucial to an efficient particle bombardment transformation system: (1) an efficient tissue culture system, (2) a highly efficient DNA delivery system, and (3) an effective selection strategy.
Keyword Biotechnology
Genetic transformation
Immature embryo
Particle bombardment
Reporter gene
Selective marker
Tissue culture
Q-Index Code BX
Q-Index Status Confirmed Code
Institutional Status UQ

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