Genetic and Physical Maps of W1/Iw1. Genetic and Physical Maps of W1/Iw1.(A) Location of the locus responsible for the dominant glaucous phenotype (W1) based on analysis of 9550 gametes from the AUS2499 × Kofa+Lr19 cross. W1 is contained in a 0.16-cM interval on wheat chromosome 2BS flanked by markers JIC009 and JIC010(B) The locus responsible for the dominant glossy phenotype (Iw1) on wheat chromosome 2BS is found in a similar location based on analysis of 598 gametes from the Svevo × TTD140 cross. Iw1 is completely linked to JIC009 and to the three candidate wheat genes DMP, DMH, and DMC. Underlined markers represent dominant markers present in Svevo and absent in TTD140. (C) Wild emmer Zavitan (glaucous) and TTD140 (glossy) physical maps. Gene content of Zavitan across the W1 genetic interval is shown. Out of 22 genes, 15 correspond to either cytochrome P450s (P450; pink), type-III PKS (blue), or hydrolase (HYD; green) genes (including partial genes and pseudogenes). Genes are ordered based on the Zavitan genome and are numbered accordingly on the Zavitan physical map (middle). Duplicated genes with >99% sequence identity are denoted with the same color and with a different numerical suffix. Pseudogenes are denoted with a psi symbol. Partial genes are denoted with a triangle. The Zavitan best homologs for DMP (two duplicated genes DMP-1.1 and DMP-1.2), DMH (two duplicated genes DMH-1.1 and DMH-1.3), and DMC are shown. Among the 22 Zavitan genes, WES and P450-3.1 were annotated as wax ester synthase from the MBOAT family (in turquoise) and CYP96B30 (in dark pink), correspondingly, suggesting a possible involvement in wax biosynthesis. The TTD140 physical map (top) shows a common haplotype with Zavitan in the regions surrounding flanking markers JIC036 and JIC032 (dashed black line). However, apart from the multiple local duplications of the NB-Arc-like genes (genes 1 and 2, duplications are indicated in black asterisks), all other sequences and genes from DUF4220 to Ank_PGG are unique to the Zavitan W1 haplotype (orange line) compared with the TTD140 Iw1 haplotype (light green line). Black vertical lines in the TTD140 physical map are genes exclusive to TTD140. Numbers in purple indicate the number of observed recombinations. A detailed description of the region is provided in Supplemental Text 2 and 3. (D) Phylogenetic tree of wheat DMC (purple) identified from BL and its homologs from Zavitan accession and from barley, with additional P450s from related clades. TaCYP709C1 and AtCYP96A15 (MAH1) are known to catalyze in-chain hydroxylation and are related by sequence to TaDMC, Zavitan P450(1-3), barley MLOC_12151, and MLOC_71974. Proteins related to TaCYP709C1 and AtCYP96A15 are shown in pink and blue, respectively. Species abbreviations: At, Arabidopsis thaliana; Ph, Petunia hybrida; Sb, Sorghum bicolor; St, Solanum tuberosum; Vs, Vicia sativa; Nt, Nicotiana tabacum; Ta, Triticum aestivum; Hv, Hordeum vulgare; Ht, Helianthus tuberosus; Os, Oryza sativa; El, Euphorbia lagascae; Zm, Zea mays. Known enzymatic activities are shown: ωh, ω-hydroxylation; ich, in-chain hydroxylation; df, diacid formation; ep, epoxidation. Physiological role associated with fatty acid biosynthesis is shown: cutin, cutin synthesis, estolide, estolide synthesis; suberin, suberin synthesis; sporopollenin, sporopollenin synthesis; seed oil, seed oil synthesis; wax, wax synthesis; FA, fatty acid hydroxylation. Genes discussed in this study are underlined. Shelly Hen-Avivi et al. Plant Cell 2016;28:1440-1460 ©2016 by American Society of Plant Biologists