TY - JOUR
T1 - Transgenic Paulownia elongata S. Y. Hu plants using biolistic-mediated transformation
AU - Castellanos-Hernández, Osvaldo A.
AU - Rodríguez-Sahagún, Araceli
AU - Acevedo-Hernández, Gustavo J.
AU - Rodríguez-Garay, Benjamín
AU - Cabrera-Ponce, José Luis
AU - Herrera-Estrella, Luis Rafael
PY - 2009/9
Y1 - 2009/9
N2 - A biolistic protocol for the stable genetic transformation of the hardwood tree Paulownia elongata was developed. Leaf explants were bombarded using the PDS-1000/He system with plasmid pBI121. The introduced DNA contained the β-glucuronidase (GUS) reporter gene and neomycin phosphotransferase (nptII) as a selection marker. Transformed calli were induced and selected on medium supplemented with 50 mg L-1 kanamycin, and transgenic plants were regenerated through indirect organogenesis. Complete plants were successfully transferred to soil and established under greenhouse conditions. Different helium pressures and explant positions were used and the transformation frequency was calculated. Optimal conditions for genetic transformation were bombardment of the abaxial leaf surface at a pressure of 450 psi. The integration of the transgenes in the plant genome and their stable expression was demonstrated by fluorometric GUS assay, determination of NPTII activity and PCR analysis. This method allows the production of transgenic trees of P. elongata in a relatively short time.
AB - A biolistic protocol for the stable genetic transformation of the hardwood tree Paulownia elongata was developed. Leaf explants were bombarded using the PDS-1000/He system with plasmid pBI121. The introduced DNA contained the β-glucuronidase (GUS) reporter gene and neomycin phosphotransferase (nptII) as a selection marker. Transformed calli were induced and selected on medium supplemented with 50 mg L-1 kanamycin, and transgenic plants were regenerated through indirect organogenesis. Complete plants were successfully transferred to soil and established under greenhouse conditions. Different helium pressures and explant positions were used and the transformation frequency was calculated. Optimal conditions for genetic transformation were bombardment of the abaxial leaf surface at a pressure of 450 psi. The integration of the transgenes in the plant genome and their stable expression was demonstrated by fluorometric GUS assay, determination of NPTII activity and PCR analysis. This method allows the production of transgenic trees of P. elongata in a relatively short time.
KW - Biolistics
KW - NptII
KW - Organogenesis
KW - Transformation
KW - Tree
UR - http://www.scopus.com/inward/record.url?scp=70350554408&partnerID=8YFLogxK
U2 - 10.1007/s11240-009-9590-2
DO - 10.1007/s11240-009-9590-2
M3 - Article
AN - SCOPUS:70350554408
VL - 99
SP - 175
EP - 181
JO - Plant Cell, Tissue and Organ Culture
JF - Plant Cell, Tissue and Organ Culture
SN - 0167-6857
IS - 2
ER -