A chimeric ecdysone receptor facilitates methoxyfenozide-dependent restoration of male fertility in ms45 maize.
A mutation in the maize Ms45 gene results in abortion of microspore development and a male-sterile phenotype. MS45 protein has been localized to the tapetum and maximally expressed in anthers at the early vacuolate stage of microspore development. Molecular complementation analysis determined that a transformed copy of the gene fully restored fertility to ms45 maize. In this report, using phenotypic complementation as an assay, chimeric transcriptional activators were expressed to regulate a gal:MS45 gene and test the ability of a multi-component system to restore male fertility. A high frequency of phenotypic complementation was observed when either C1-GAL4 or VP16-GAL4 activators were transcribed by promoters that expressed at a stage of anther development that precedes the early vacuolate stage of microsporogenesis. For the conditional regulation of male fertility, these transcriptional activators were modified by the addition of regions that include the ligand-binding domain from the European corn borer ecdysone receptor to generate the nuclear receptors C1-GAL4-EcR (CGEcR) and VP16-GAL4-EcR (VGEcR). These chimeric receptors were introduced with the gal:MS45 gene into ms45 maize, and in the absence of ligand, these plants were male sterile. In contrast, application of the ecdysone agonist, methoxyfenozide, to plants containing either a constitutive (Ubiquitin1) or anther-specific (maize 5126) VGEcR resulted in the restoration of fertility to ms45 plants grown in either the greenhouse or in the field.
Unger
Erica
E
Agronomic Traits Department, Pioneer Hi-Bred International, Johnston, Iowa 50131, USA. erica.unger-wallace@pioneer.com
Cigan
A Mark
AM
Trimnell
Mary
M
Xu
Rui-ji
RJ
Kendall
Tim
T
Roth
Brad
B
Albertsen
Marc
M
D002678:Chimera / Q000235:genetics; D004440:Ecdysone / Q000378:metabolism; D005298:Fertility; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant / Q000235:genetics*; D017930:Genes, Reporter / Q000187:drug effects; D006834:Hydrazines / Q000494:pharmacology*; D007605:Juvenile Hormones / Q000494:pharmacology*; D010641:Phenotype; D011401:Promoter Regions, Genetic; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics / Q000378:metabolism*; D011987:Receptors, Steroid / Q000378:metabolism*; D012098:Reproduction / Q000502:physiology*; D012641:Selection, Genetic; D013170:Spores / Q000235:genetics*; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism; D014170:Transformation, Genetic; D025801:Ubiquitin / Q000235:genetics; D003313:Zea mays / Q000235:genetics* / Q000378:metabolism*
C070633:Gal-VP16; D006834:Hydrazines; D007605:Juvenile Hormones; D018160:Receptors, Cytoplasmic and Nuclear; D011987:Receptors, Steroid; D015534:Trans-Activators; D025801:Ubiquitin; C055469:ecdysone receptor; D004440:Ecdysone; C406712:methoxyfenozide