23325) in B73 and also other maize lines with low constitutive HDMBOA-Glc accumulation.
23325) in B73 along with other maize lines with low constitutive HDMBOA-Glc accumulation. R. maidis progeny production is negatively correlated with DIMBOA-Glc and positively correlated with HDMBOA-Glc production. In the existing study, we use near-isogenic lines (NILs) derived from B73 and CML277 to further characterize the effects of Bx10c inactivation on benzoxazinoid accumulation and aphid resistance in maize. Genotyped RILs in the NAM population are on typical 3.6 heterozygous,four generating it feasible to seek out person lines that are nonetheless heterozygous for practically any desired genomic interval. Such heterozygotes is often self-pollinated and genotyped to create heterogeneous inbred households (HIFs).15 5 RILs derived from a B73 CML277 cross, Z005E0109, Z005E0153, Z005E0174, Z005E0176, and Z005E018, had been predicted to become heterozygous in a region containing the Bx10a,b, c genes (Fig. 1). To Genotype the Bx10c alleles, genomic DNA was amplified by PCR making use of the following 3 primers: forward, 5-AGCACGGCAA CAACCTTGG-3; reverse for the transposon insertion allele 5-CGCGGTGGTG AGAACCGTTT -3; and reverse for the functional gene, 5-AAGTGCACGT TGCCATCAGA TGGAG -3, as described previously..six Heterozygosity at the Bx10c gene may be verified only for Z005E0109 and Z005E0174, as well as the latter was chosen for additional experiments. As Brd Inhibitor Source Z005E0174 was predicted to*Correspondence to: Vered Tzin; E mail: [email protected] Submitted: 08/10/2013; Revised: 10/09/2013; Accepted: 10/10/2013 Citation: Mijares V, Meihls L, Jander G, Tzin V. Near-isogenic lines for measuring phenotypic effects of DIMBOA-Glc methyltransferase activity in maize. Plant Signaling Behavior 2013; 8:e26779; PMID: 24168845; dx.doi.org/10.4161/psb.26779 landesbioscience.com Plant signaling Behavior e26779-Figure 1. Generation of near-isogenic lines for the Bx10a, b, c locus. (A) Genotype information for Z005E0174, a recombinant inbred line derived from B73 cmL277, are shown (marker data had been downloaded from panzea.org). (B) Genotype of Bx10a, b, c near-isogenic lines derived from Z005E0174, with the location on the genome that differs among the 2 near-isogenic lines, Bx10cniL-B73 and Bx10cniL-cmL277, marked in green. the approximate Bx10a, b, c position on chromosome 1 is marked with an arrow.be heterozygous for regions of chromosome 1, two, three, and four (Fig. 1A), it was selfed for three generations, prior to choosing sibling lines using the B73 or CML277 alleles, respectively, at the Bx10c locus. Single nucleotide polymorphism (SNP) markers within the other H4 Receptor Antagonist supplier previously heterozygous regions of chromosomes 1 (Fig. 1A) have been genotyped by PCR amplification and Sanger sequencing (Table 1) to confirm that these regions of the genome are homozygous for the B73 or CML277 alleles in the NILs (illustrated graphically in Figure 1B). As well as the predicted SNPs from panzea.org, our DNA sequencing identified 5 other SNPs in the amplified regions which are polymorphic among B73 and CMLTable 1. Genotyping snPs of near-isogenic lines derived from Z005E0174 on chromosome 1 to 4. SNP NameSNP locationPCR amplification primer sequence (5′ 3′) aGccatcGat aGtcGaaacG taGGcacGaat ttGcttGaa tccaaGcGta tGctcaaGaa GcacaataGc GaGatGGtca taatcctGGcG atGaGatcc tttGccaaGc tcctccatac GaacctcGca tGGttGaGat aGGGcacttG GttccaGata GctcatcaGG cttttGGaaG GcacaaGcca cccactattt GctcatcaGG cttttGGaaG GcacaaGcca cccactattt ccacGGttac aGcactaGca GtccaacccG aaactGaaaa ccacGGttac aGcactaGca GtccaacccG aaactGaaaa caacacGcGttattGGacac GccaGaactGGacGaGtaGc ccaaa.