Development of plant regeneration and transformation protocols for the desiccation-sensitive weeping lovegrass Eragrostis curvula.
A tissue culture protocol, suitable for transformation, was established for the pasture grass Eragrostis curvula. Callus was generated in the dark from leaf and seed tissues on a medium comprising MS salts supplemented with 2 mg/l 2,4-D, 0.01 mg/l BAP and 2% sucrose. Plant regeneration occurred via organogenesis on the same medium with 6% and 3% sucrose for shoot and root formation, respectively. Optimal regeneration (50 plantlets per callus) occurred when light of 45 micromol/m2 per s was used. The yeast Saccharomyces cerevisiae Hsp12 gene was cloned into the Sac1 of the pCAMBIAUbeeQ vector and callus was transformed by biolistic bombardment. Best transformation (30%) occurred when the target tissue was bombarded twice at a distance of 70 mm using a bombardment pressure of 9,100 kPa. Although successful transformation and transcription of the Hsp12 gene occurred, no Hsp12 protein was found present in tissue extracts of transformed grass.
Ncanana
Sandile
S
Department of Molecular and Cellular Biology, University of Cape Town, Private Bag, Rondebosch, 7700, South Africa.
Brandt
Wolf
W
Lindsey
George
G
Farrant
Jill
J
D000383:Agriculture / Q000379:methods; D019470:Biolistics / Q000379:methods; D004190:Disasters; D031750:Eragrostis / Q000235:genetics / Q000254:growth & development*; D018517:Plant Roots / Q000235:genetics / Q000254:growth & development; D018520:Plant Shoots / Q000235:genetics / Q000254:growth & development; D030821:Plants, Genetically Modified / Q000235:genetics / Q000254:growth & development*; D012038:Regeneration / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics; D046509:Tissue Culture Techniques / Q000379:methods*; D014170:Transformation, Genetic / Q000235:genetics*