FIGURE 4. Effect of AT1G52460 deficiency on the level of transcripts of neighboring genes and the content of polyisoprenoids.
A AT1G52460 gene structure. Exons and introns are indicated by thick and thin lines, respectively. The T-DNA insertion sites in two independent mutant lines: SALK_066806 and GK_823G12 are depicted.
B Levels of AT1G52440, AT1G52450, and AT1G52460 transcripts (qPCR analysis) in the leaves of 3-week-old plants – WT (Col) and AT1G52460-deficient plants were compared. P≤0.0001 (one-way ANOVA followed by Turkey post-tests); ns, non-significant.
C The content of Dols and Prens estimated in leaves of 3-week-old plants using HPLC/UV, shown are means ± SD of five independent biological replicates. The phenotypic appearance of 4-week-old detached leaves of AT1G52460-deficient line (SALK_066806,abh heterozygous mutant) and wild-type (Col-0) plants grown in soil is shown in Figure S6. Seed germination and segregation analysis of F1 progeny of self-pollinated of heterozygous SALK_066806 and GK_823G12 lines is shown in Table S5.
FIGURE 5. Correlations between the content of seven metabolites estimated in the seedlings of Arabidopsis accessions (A) and the EstC mapping population (B). The original distributions (green bars), together with the approximation of the normal distribution of the data (blue curve) with outliers removed, are presented on the diagonal. Correlation patterns for each metabolite pair are presented at the appropriate intersection; please note that outliers (red dots) were not taken into consideration for the analysis. Above each diagonal panel, the Shapiro-Wilk statistics (W, p) for normal distribution is presented, while for out-of-diagonal panels Pearson (P) and Spearman (S) correlation coefficients together with the associated significance levels are shown (please note that ‘0’ means p< 1e-7). Bearing in mind the statistically significant deviations from normal distribution shown in the diagonal panels, the significance of the observed correlations should be interpreted in terms of the Spearman rather than Pearson coefficient (see Materials and methods). Cumulative distribution functions (CDF) of the content of seven studied metabolites analyzed in the seedlings of Arabidopsis accessions and AI-RILs are shown in Figure S7.
FIGURE 6. Biosynthetic routes leading to isoprenoids in a plant cell; the involvement of the genes cis-prenyltransferase 3 (CPT3) and putative role of alpha-beta hydrolase (ABH) is indicated. Depicted are seven metabolites analyzed in this study. Two pathways, the mevalonate (MVA) and methylerythritol phosphate (MEP) pathways, are generating IPP in parallel, both contributing to particular isoprenoids (Hemmerlin, Harwood, & Bach, 2012; Akhtar et al., 2017; Jozwiak et al., 2017). Blue arrows illustrate the exchange of intermediates between the MVA and MEP pathways. Abbreviations: DMAPP-dimethylallyl diphosphate, FPP-farnesyl diphosphate, GPP-geranyl diphosphate, GGPP-geranylgeranyl diphosphate, IPP-isopentenyl diphosphate.