TY - JOUR
T1 - Rapid production of transgenic sugarcane with the introduction of simple loci following biolistic transfer of a minimal expression cassette and direct embryogenesis
AU - Taparia, Yogesh
AU - Fouad, Walid M.
AU - Gallo, Maria
AU - Altpeter, Fredy
N1 - Funding Information:
Acknowledgments We would like to thank USDA-CSREES and Syngenta Biotechnology, Inc. for their financial support of this work; Dr. Robert Gilbert, EREC, Belle Glade, FL, for providing donor plants of sugarcane cultivar CP88-1762; Jeff Seib for training Yogesh Taparia in the safe handling of radio isotopes; and Conrad Fafard Inc., Apopka, FL, for donation of plant-growing media.
PY - 2012/2/1
Y1 - 2012/2/1
N2 - A protocol is described that supports the production of transgenic sugarcane plants ready for transfer to soil within 3 mo from culture initiation. Biolistic gene transfer into cross-sections of immature leaf whorl explants followed by direct somatic embryogenesis resulted in the stable genetic transformation of the commercially important sugarcane cultivar CP 88-1762. Accelerating the production of transgenic sugarcane plants not only saves time and effort but will likely also minimize somaclonal variation. Southern blot analysis revealed simple transgene integration patterns ranging from one to five hybridization products. NPTII-ELISA confirmed that most of the transgenic plants expressed the transgene stably in vegetative progeny. Using a minimal, linear expression cassette (MC) without vector backbone sequences for the biolistic gene transfer and reducing the amount of MC to 10 ng per shot may have led to simple transgene integration and stable transgene expression. Therefore, this protocol has great potential for the generation of commercial transgenic sugarcane events.
AB - A protocol is described that supports the production of transgenic sugarcane plants ready for transfer to soil within 3 mo from culture initiation. Biolistic gene transfer into cross-sections of immature leaf whorl explants followed by direct somatic embryogenesis resulted in the stable genetic transformation of the commercially important sugarcane cultivar CP 88-1762. Accelerating the production of transgenic sugarcane plants not only saves time and effort but will likely also minimize somaclonal variation. Southern blot analysis revealed simple transgene integration patterns ranging from one to five hybridization products. NPTII-ELISA confirmed that most of the transgenic plants expressed the transgene stably in vegetative progeny. Using a minimal, linear expression cassette (MC) without vector backbone sequences for the biolistic gene transfer and reducing the amount of MC to 10 ng per shot may have led to simple transgene integration and stable transgene expression. Therefore, this protocol has great potential for the generation of commercial transgenic sugarcane events.
KW - Biolistic gene transfer
KW - Direct somatic embryogenesis
KW - Minimal expression cassette
KW - Sugarcane cultivar CP-88-1762
UR - http://www.scopus.com/inward/record.url?scp=84857039992&partnerID=8YFLogxK
U2 - 10.1007/s11627-011-9389-9
DO - 10.1007/s11627-011-9389-9
M3 - Article
AN - SCOPUS:84857039992
SN - 0883-8364
VL - 48
SP - 15
EP - 22
JO - In Vitro Cellular and Developmental Biology - Plant
JF - In Vitro Cellular and Developmental Biology - Plant
IS - 1
ER -