Figure 1.
Timeline of H. annuus plant growth and treatment.
Table 1.
Peaks detected in extracts of H. annuus glandular trichomes.
Figure 2.
Glandular trichome count per 3 mm×3 mm grid of H. annuus abaxial leaf surface.
‘LEAF2’ was removed from the plant prior to the first methyl jasmonate treatment; ‘LEAF5’ was removed two weeks later, after two treatments with 0.1 mM methyl jasmonate (MeJA) or control spray spaced at one-week intervals. Accession is indicated by marker shape (▴ = ANN1238, • = HA89) and treatment is indicated by filled (control) versus open (MeJA) markers. Data points represent the mean value from 6 plants; error bars show one standard deviation. Where data points from both treatments are not clearly visible (e.g. HA89 EDGE), they are overlapping.
Figure 3.
Representative chromatograms of methylene chloride extracts from the leaf surfaces of H. annuus accessions ANN1238 and HA89.
The horizontal axis displays retention time in minutes. Vertical axes are scaled identically and display total mass counts. Labeled peaks A-K were selected for further analysis and comparison between accessions, and correspond to similarly-labeled peaks in Table 1 and Figures 4 and 5.
Figure 4.
Relative percentages of LC-MS peak area accounted for by 10 peaks shared between glandular trichome extract profiles derived from floral (anther appendage) versus leaf (abaxial surface) samples.
Relative peak percentage is determined as (peak area)/(sum of areas for peaks A-J). Statistical significance of differences among sample sources are marked in the legend: * = p<0.05, x = p<0.1. Peaks are marked in Figure 3; retention time and neutral mass for each peak are given in Table 1.
Figure 5.
Comparison of common peaks from H. annuus foliar glandular trichome LC-MS profiles.
Samples are grouped by accession (ANN1238, HA89) and treatment (C = control, J = 100 µM MeJA) along the horizontal axis. The vertical axis shows the peak area (mass counts) standardized by the estimated number of glandular trichomes extracted per sample (cm2 leaf area *average glandular trichomes/cm2). Each colored section within the columns corresponds to a specific peak, indicated in the legend and corresponding to Table 1. Values shown represent the mean peak area for 5 samples per accession x treatment; statistically significant differences between HA89 and ANN1238-derived samples are marked in the legend: * = p<0.05, ** = p<0.01, *** = p<0.001).
Figure 6.
Heatmap showing normalized transcript accumulation of putative jasmonate-responsive transcripts in H. annuus samples: ‘H’ = HA89, ‘A’ = ANN1238, ‘J’ = MeJA treatment group, ‘C’ = control group, ‘pre’ = pre-treatment, ‘post’ = post-treatment.
Fifty H. annuus transcripts showing significant positive response to exogenous MeJA across multiple comparisons are arrayed in columns with color intensity indicating transcript abundance. A list of transcripts and associated annotation by best blast hit is provided in Table S1. The dendrogram on the left indicates clustering by similarity of transcript patterns among samples.