Pistil drip following pollination: a simple in planta Agrobacterium-mediated transformation in cotton.
Transgenic cotton plants were developed by pistil drip inoculation in a solution containing Agrobacterium carrying a gene for resistance to the herbicide Basta (bar), 10% (w/v) sucrose, 0.05% (v/v) Silwet L-77 and 40 mg acetosyringone l(-1). Pistil drip during 17:00-19:00 on the first day of flowering resulted in 0.07-0.17% Basta-resistant plants/number of viable seeds generated, and stigma excision prior to pistil drip during this time period gave rise to a transformation efficiency of 0.46-0.93%, in contrast with 0.04-0.06% generated from pistil drip during 9:00-11:00 on the second day of flowering. PCR and Southern blot analysis confirmed the integration of the bar gene into the cotton genome, and a T1 and T2 generation herbicide resistance test consistently revealed expression and stable heritability of the bar gene in the two generations.
Tianzi
Chen
C
National Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095, China.
Shenjie
Wu
W
Jun
Zhao
Z
Wangzhen
Guo
G
Tianzhen
Zhang
Z
D000613:Aminobutyrates / Q000494:pharmacology; D001426:Bacterial Proteins; D015139:Blotting, Southern; D024881:Drug Resistance, Bacterial; D035264:Flowers / Q000502:physiology*; D003368:Gossypium / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D054817:Pollination; D016133:Polymerase Chain Reaction; D012231:Rhizobium / Q000235:genetics*; D013997:Time Factors; D014170:Transformation, Genetic; D019076:Transgenes
D000613:Aminobutyrates; D001426:Bacterial Proteins; C003121:phosphinothricin