Figure 1.
Flag leaf sheaths and peduncles of the NILs examined in this study.
NIL designations are indicated beneath each peduncle and genotypes are specified in parentheses. The introduced alleles in the genotypes are underlined. The bar indicates 1 cm.
Figure 2.
Electron micrographs of the cuticle surfaces of flag leaf blades, sheaths, peduncles, and glumes of the NILS.
The tissues are indicated on the top and the NIL designations at the left. The bars indicate 2.5 µm.
Figure 3.
Wax composition of the six NILs.
(a) Total wax load of the flag leaf sheaths was measured by GC-MS. (b) β-diketone, fatty acid, aldehyde, primary (1°) alcohol, alkane, and sterol and triterpene (ST&TP) content. The numbers on the y-axes indicate average content expressed as µg per g dried tissue (dry weight, DW). The bars indicate standard deviation of the mean estimated from six biological replicates. (c) The percentage of wax species in each genotype was calculated from the means.
Figure 4.
Homolog variation of major wax species.
Carbon atom numbers of aldehydes (a) and primary (1°) alcohols (b), and β-diketones (c) are indicated on the x-axes. Their contents are indicated on y-axes as µg per g dried tissue. The bars indicate standard deviation of the mean calculated from six biological replicates. β-D, β-diketone; and OH-β, hydroxyl-β-diketones.
Figure 5.
Expression of cuticular wax genes in the wheat flag leaf sheath of the Iw NILs compared to W1W2.
Genes with two- or higher-fold changes are depicted and expression data for all genes analyzed are listed in Table S2. The bars represent standard deviation of the mean fold-change of mRNA levels. Asterisks indicate that the difference is significant at P<0.05 (*) or at P<0.01 (**).
Figure 6.
CER4-6 expression in sheaths of Iw1iw2, iw1Iw2, and W1W2 at different developmental stages.
(a) Transcription levels of CER4-6 at stage F4.0 compared to that of the reference gene TaRPII36. (b) Fold changes of CER4-6 transcription at stage F9.0 compared to that at stage F4.0. The bars represent standard deviation of the mean fold-change of mRNA levels. Asterisks indicate that the difference is significant at P<0.01 (**).
Figure 7.
The expression of cuticular wax- and cutin-related genes in the wheat flag leaf sheath of W1w2, w1W2, and W1W2 plants compared to w1w2.
Genes with two- or higher-fold changes are depicted and expression data for all genes analyzed are listed in Table S3. The bars represent standard deviation of the mean fold-change of mRNA levels. Asterisks indicate that the difference is significant at P<0.05 (*) or at P<0.01 (**).
Figure 8.
Cuticle permeability of the NILs.
Cuticle permeability was evaluated by air drying at room temperature (a and c) and chlorophyll leaching in 80% ethanol (b and d). The numbers on the x-axes represent hours of treatment. Water loss or chlorophyll leaching at each time point is represented on the y-axes as percentages of the total water content or total chlorophyll content in the tissue. Measurements taken from four individuals were averaged.