TY - JOUR
T1 - Assessment of ptxD gene as an alternative selectable marker for Agrobacterium-mediated maize transformation
AU - Nahampun, Hartinio N.
AU - López-Arredondo, Damar
AU - Xu, Xing
AU - Herrera-Estrella, Luis
AU - Wang, Kan
N1 - Funding Information:
HN and KW thank Bronwyn Frame for her expertise in maize transformation, scientific discussion and critical review of the manuscript. This work was supported in part by the U.S. Department of Agriculture National Institute of Food and Agriculture (Hatch Project No. IOW05162 to KW), the Iowa State University Crop Bioengineering Consortium (KW), the Howard Hughes Medical Institute, Grant 55005946 (LHE) and Charoen Pokphand Indonesia (HN).
Publisher Copyright:
© 2016, Springer-Verlag Berlin Heidelberg.
PY - 2016/5/1
Y1 - 2016/5/1
N2 - Key message: Bacterial phosphite oxidoreductase gene and chemical phosphite can be used as a selection system forAgrobacterium-mediated maize transformation. Abstract: Application of phosphite (Phi) on plants can interfere the plant metabolic system leading to stunted growth and lethality. On the other hand, ectopic expression of the ptxD gene in tobacco and Arabidopsis allowed plants to grow in media with Phi as the sole phosphorous source. The phosphite oxidoreductase (PTXD) enzyme catalyzes the conversion of Phi into phosphate (Pi) that can then be metabolized by plants and utilized as their essential phosphorous source. Here we assess an alternative selectable marker based on a bacterial ptxD gene for Agrobacterium-mediated maize transformation. We compared the transformation frequencies of maize using either the ptxD/Phi selection system or a standard herbicide bar/bialaphos selection system. Two maize genotypes, a transformation amenable hybrid Hi II and an inbred B104, were tested. Transgene presence, insertion copy numbers, and ptxD transcript levels were analyzed and compared. This work demonstrates that the ptxD/Phi selection system can be used for Agrobacterium-mediated maize transformation of both type I and type II callus culture and achieve a comparable frequency as that of the herbicide bar/bialaphos selection system.
AB - Key message: Bacterial phosphite oxidoreductase gene and chemical phosphite can be used as a selection system forAgrobacterium-mediated maize transformation. Abstract: Application of phosphite (Phi) on plants can interfere the plant metabolic system leading to stunted growth and lethality. On the other hand, ectopic expression of the ptxD gene in tobacco and Arabidopsis allowed plants to grow in media with Phi as the sole phosphorous source. The phosphite oxidoreductase (PTXD) enzyme catalyzes the conversion of Phi into phosphate (Pi) that can then be metabolized by plants and utilized as their essential phosphorous source. Here we assess an alternative selectable marker based on a bacterial ptxD gene for Agrobacterium-mediated maize transformation. We compared the transformation frequencies of maize using either the ptxD/Phi selection system or a standard herbicide bar/bialaphos selection system. Two maize genotypes, a transformation amenable hybrid Hi II and an inbred B104, were tested. Transgene presence, insertion copy numbers, and ptxD transcript levels were analyzed and compared. This work demonstrates that the ptxD/Phi selection system can be used for Agrobacterium-mediated maize transformation of both type I and type II callus culture and achieve a comparable frequency as that of the herbicide bar/bialaphos selection system.
KW - Agrobacterium-mediated transformation
KW - Maize
KW - Phosphite
KW - Phosphite oxidoreductase
KW - ptxD gene
UR - http://www.scopus.com/inward/record.url?scp=84958818964&partnerID=8YFLogxK
U2 - 10.1007/s00299-016-1942-x
DO - 10.1007/s00299-016-1942-x
M3 - Article
C2 - 26883223
AN - SCOPUS:84958818964
SN - 0721-7714
VL - 35
SP - 1121
EP - 1132
JO - Plant Cell Reports
JF - Plant Cell Reports
IS - 5
ER -