Abstract
Zea mays transformants produced by particle bombardment of embryogenic suspension culture cells of the genotype A188 x B73 and selected on kanamycin or bialaphos were characterized with respect to transgene integration, expression, and inheritance. Selection on bialaphos, mediated by the bar or pat genes, was more efficient than selection on kanamycin, mediated by the nptII gene. Most transformants contained multicopy, single locus, transgene insertion events. A transgene expression cassette was more likely to be rearranged if expression of that gene was not selected for during callus growth. Not all plants regenerated from calli representing single transformation events expressed the transgenes, and a non-selectable gene (uidA) was expressed in fewer plants than was the selectable transgene. Mendelian inheritance of transgenes consistent with transgene insertion at a single locus was observed for approximately two thirds of the transformants assessed. Transgene expression was typically, but not always, predictable in progeny plants--transgene silencing, as well as poor transgene transmission to progeny was observed in some plant lines in which the parent plants had expressed the transgene.
MeSH Terms
Cells, Cultured
DNA/genetics
Genes, Plant
Genotype
Kanamycin/pharmacology
Plants, Genetically Modified
Transfection/methods
Transformation, Genetic
Zea mays/genetics
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Register J C
ICI Seeds Research Department, Slater, IA 50244.
Peterson D J
Bell P J
Bullock W P
Evans I J
Frame B
Greenland A J
Higgs N S
Jepson I
Jiao S
References (21)
21 references, click to expand
-
Transformation and inheritance of a hygromycin phosphotransferase gene in maize plants.
Plant Mol Biol. 1992 Jan;18(2):189-200
PMID: 1310057
-
Evaluation of selectable markers for obtaining stable transformants in the gramineae.
Plant Physiol. 1988 Feb;86(2):602-6
PMID: 16665953
-
Establishment and maintenance of friable, embryogenic maize callus and the involvement of L-proline.
Planta. 1985 May;164(2):207-14
PMID: 24249562
-
A mutant neomycin phosphotransferase II gene reduces the resistance of transformants to antibiotic selection pressure.
Proc Natl Acad Sci U S A. 1990 May;87(9):3435-9
PMID: 2159150
-
Stable transformation of tobacco by electroporation: evidence for plasmid concatenation.
Proc Natl Acad Sci U S A. 1986 Aug;83(15):5602-6
PMID: 3016708
-
Molecular and general genetics of a hybrid foreign gene introduced into tobacco by direct gene transfer.
Mol Gen Genet. 1985;199(2):169-77
PMID: 3860712
-
Transformation of Maize Cells and Regeneration of Fertile Transgenic Plants.
Plant Cell. 1990 Jul;2(7):603-618
PMID: 12354967
-
Isolation and characterization of male flower cDNAs from maize.
Plant J. 1993 Jan;3(1):41-9
PMID: 8401606
-
Inactivation of the maize transposable element Activator (Ac) is associated with its DNA modification.
EMBO J. 1987 Feb;6(2):295-302
PMID: 3034583
-
Genetic transformation of maize cells by particle bombardment.
Plant Physiol. 1989 Sep;91(1):440-4
PMID: 16667039
-
A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity.
Anal Biochem. 1983 Jul 1;132(1):6-13
PMID: 6312838
-
Nucleotide sequence of the phosphinothricin N-acetyltransferase gene from Streptomyces viridochromogenes Tü494 and its expression in Nicotiana tabacum.
Gene. 1988 Oct 15;70(1):25-37
PMID: 3240868
-
Transgenic tobacco plants and their progeny derived by microprojectile bombardment of tobacco leaves.
Plant Mol Biol. 1990 Feb;14(2):261-8
PMID: 1966275
-
Activity of a chimeric promoter with the doubled CaMV 35S enhancer element in protoplast-derived cells and transgenic plants in maize.
Plant Mol Biol. 1993 Feb;21(3):415-28
PMID: 8443339
-
Expression and inheritance of kanamycin resistance in a large number of transgenic petunias generated by Agrobacterium-mediated transformation.
Plant Mol Biol. 1988 May;11(3):355-64
PMID: 24272348
-
Dissection of a pollen-specific promoter from maize by transient transformation assays.
Plant Mol Biol. 1992 Jan;18(2):211-8
PMID: 1731984
-
Transgenic maize plants by tissue electroporation.
Plant Cell. 1992 Dec;4(12):1495-505
PMID: 1334743
-
Inheritance and expression of chimeric genes in the progeny of transgenic maize plants.
Biotechnology (N Y). 1990 Sep;8(9):833-9
PMID: 1366794
-
Analysis of the T-DNA structure in a large number of transgenic petunias generated by Agrobacterium-mediated transformation.
Plant Mol Biol. 1988 May;11(3):365-77
PMID: 24272349
-
Segregation of transgenes in maize.
Plant Mol Biol. 1992 Jan;18(2):201-10
PMID: 1731983
-
A simple and rapid method for the preparation of plant genomic DNA for PCR analysis.
Nucleic Acids Res. 1991 Mar 25;19(6):1349
PMID: 2030957