title: Assessment of ptxD gene as an alternative selectable marker for Agrobacterium-mediated maize transformation.
abstract: Bacterial phosphite oxidoreductase gene and chemical phosphite can be used as a selection system for Agrobacterium -mediated maize transformation. 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.
authors.affiliation: Interdepartmental Plant Biology Major, Iowa State University, Ames, IA, 50011-1010, USA.|Department of Agronomy, Iowa State University, Ames, IA, 50011-1010, USA.|PT.BISI International, Tbk., Jl. Raya Pare-Wates Km 9, Ds. Sumberagung, Kab, Kediri, East Java, 64175, Indonesia.
authors.forename: Hartinio N
authors.initials: HN
authors.lastname: Nahampun
authors.affiliation: StelaGenomics México, S de RL de CV, Av. Camino Real de Guanajuato s/n, 36821, Irapuato, Guanajuato, Mexico.
authors.forename: Damar
authors.initials: D
authors.lastname: López-Arredondo
authors.affiliation: Interdepartmental Plant Biology Major, Iowa State University, Ames, IA, 50011-1010, USA.|Department of Agronomy, Iowa State University, Ames, IA, 50011-1010, USA.|Zhejiang University, Hangzhou, China.
authors.forename: Xing
authors.initials: X
authors.lastname: Xu
authors.affiliation: Laboratorio Nacional de Genómica para la Biodiversidad, Unidad de Genómica Avanzada del Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional, Km 9.6 carretera Irapuato-León, 36500, Irapuato, Guanajuato, Mexico.
authors.forename: Luis
authors.initials: L
authors.lastname: Herrera-Estrella
authors.affiliation: Department of Agronomy, Iowa State University, Ames, IA, 50011-1010, USA. kanwang@iastate.edu.
authors.forename: Kan
authors.initials: K
authors.lastname: Wang
mesh_terms: D060054:Agrobacterium / Q000502:physiology*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D005819:Genetic Markers / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D005838:Genotype; D009247:NADH, NADPH Oxidoreductases / Q000235:genetics* / Q000378:metabolism; D017905:Phosphites / Q000378:metabolism*; D030821:Plants, Genetically Modified; D014170:Transformation, Genetic; D003313:Zea mays / Q000235:genetics* / Q000378:metabolism / Q000382:microbiology
chemical_list: D001426:Bacterial Proteins; D005819:Genetic Markers; D017905:Phosphites; D009247:NADH, NADPH Oxidoreductases; C429539:NAD phosphite oxidoreductase
keywords: Agrobacterium-mediated transformation; Maize; Phosphite; Phosphite oxidoreductase; ptxD gene