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Identifying Mu Insertions in Nutrient Transport-related Genes in the UniformMu Maize Population. 40. CesA11. 30. 20. 10. Relative mRNA expression (RQ). 20. 16. CesA12. 12. 8. 4. 0. pith. silk. ligule. stem. husk. pollen. tassel top. leaf blade. cob 7 dbp. cob 0 dbp.

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  1. Identifying Mu Insertions in Nutrient Transport-related Genes in the UniformMu Maize Population 40 CesA11 30 20 10 Relative mRNA expression (RQ) 20 16 CesA12 12 8 4 0 pith silk ligule stem husk pollen tassel top leaf blade cob 7 dbp cob 0 dbp leaf sheath lat root tip floret 0 dbp prop root tip tassel bottom mature anthers mature lat root mature prop root developing prop root lat. root initiation zone Male reproductive Female reproductive Vegetative Root Results: 65DAG Coordinate 7 CesA7 5 3 1 Relative mRNA expression (RQ) 10 8 CesA7-P 6 4 2 0 Stephanie M. Locke, Brent. A. O’Brien, Ryan Gannon, Johnathan Saunders, Laura Morales, Solute carrier family 2, facilitated glucose transporter member 8 (SLC2A8) Phloem protein 2A13 (PP2 A13) Solute carrier family 23 member 2 (SCF23) Plant neutral invertase family protein • Primarily functions in carbohydrate binding • In A. Thaliana, this gene was found to be a N-terminal protein involved in myristoylation and control responses to phloem damage. • GRMZM2G010855 • Hydrolyzes sucrose to fructose and glucose in the cytoplasm • GRMZM2G084940 • SCF23 represents integral membrane proteins that transport organic molecules and inorganic ions. • GRMZM2G437859 • This gene functions in carbohydrate transmembrane transporter activity. • GRMZM2G160504 • GRMZM2G160460 Wayne T. Avigne, Donald McCarty, Charles Hunter, and Karen E. Koch ABC-type transporter • ABC (ATP binding cassette) transporters are transmembrane proteins that utilize the energy of ATP hydrolysis to pump substrate across the membrane against a concentration gradient. • GRMZM2G156630 Plant Molecular and Cellular Biology Program, University of Florida, Gainesville, FL, 32611 Overview: Research presented here tests the hypotheses that insertion of a Mu transposable element into any of the selected genes will cause a loss of function that results in a distinct mutant phenotype. The UniformMu maize population is a resource developed by introgressing the highly transposon-mutagenic line Robertson’s Mutator into the phenotypically-uniform inbred W22. This population provides the ability to identify and map Mu insertions, allowing researchers to select and characterize lines harboring mutations in genes of interest. Here we use both a forward and reverse genetics approach to identify insertions in genes potentially affecting nutrient transport. To this end, we attempt to associate insertions with mutant phenotypes through co-segregationalanalyses. Plasmodesmatacallose binding protein 4 (PDCB4) • Localizes to the plasmodesmata and is predicted to bind callose, which is deposited at the sieve plate in response to injury and pathogen infection. • GRMZM2G125241 Reverse Genetics Segregation/genotypes Prominent Phenotypes Insertions Tested Empty pericarp (ep) Embryo lethal (emb) Leaf discoloration PCR inconclusive to date ? Defective kernel (dek), Rough kernel (rgh), ep, and emb All Het or WT (GRMZM2G160504) NO (other effects possible) All Het (GRMZM2G160460) Small kernel (smk), ep, and emb NO (other effects possible) WT, Het, and Homozygous Phenotype Examples NO (other effects possible) smk and emb All Het NO (other effects possible) WT, Het, and Homozygous ep and dek NO (other effects possible) smk WT, Het, and Homozygous smk Leaf discoloration Forward Genetics Insertions Tested Family 011S-2189 Phenotype = wilty Segregation/genotypes WT, Het, and Homozygous Cellulose synthase-like D3 NO WT, and Homozygous NO Peptidase M1 family protein ep emb WT, Het, and Homozygous Hercules receptor kinase 2 NO Co-segregation of Co-segregation Long-chain acyl-CoAsynthetase 7 -- All WT with main phenotypes KO with phenotype References: McCarty, D.R., Settles, A.M., Suzuki, M., Tan, B.C., and Latshaw, S. et al. (2005). Steady-state transposon mutagenesis in inbred maize. Plant J. 44: 52-61. Mitochondrial transcription termination factor family protein WT, Het, and Homozygous NO WT, Het, and Homozygous PapD-like superfamily protein NO WT, Het, and Homozygous Protein of unknown function (DUF3537) NO WT, Het, and Homozygous Sec-independent periplasmic protein translocase NO WT, Het, and Homozygous NO Succinatedehydrogenase 1-1

  2. Wild type

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