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Figure 4. In planta characterization of the indole-specific glucosinolate transporter GTR3.(A–B) Indole glucosinolate content in (A) root and (B) rosette of non-grafted hydroponically grown wildtype, gtr3, gtr1 gtr2 dko and gtr1 gtr2 gtr3 tko plants. The box is determined by the 25th and 75th percentiles. The whiskers are determined by the 5th and 95th percentiles. Median and mean are shown as line and square. Groups in subfigures are determined by one-way ANOVA followed by Tukey HSD Calculator multiple comparison post-hoc analysis (p<0.05). Data presented is one of two individual experiments, each containing 8–12 repeats (n) (see Figure 4—source data 1–2 for individual glucosinolate data points and individual n; error bars and parentheses are s.d. (C) Indole glucosinolate concentrations of micro-grafted 3-week-old plate-grown Arabidopsis wildtype (Col-0) and mutants. Rosettes and roots from wild type (wt), the glucosinolate biosynthesis null mutant myb28 myb29 cyp79b2 cyp79b3 (qko) and the gtr1 gtr2 gtr3 mutant (tko) were reciprocally grafted using 4-day-old seedlings. Glucosinolate content in the rosette and roots was quantified by LC-MS in 3-week-old plants. Data presented is one of two individual experiments, each containing 8–16 repeats (n) (see Figure 4—source data 3–4 for individual glucosinolate data points and individual n; error bars and parentheses are s.d. Groups in subfigures are determined by one-way ANOVA (p<0.05). n.d. none detected.DOI:
http://dx.doi.org/10.7554/eLife.19466.00910.7554/eLife.19466.010Figure 4—source data 1. Glucosinolate content in rosettes of hydroponically grown wildtype, gtr3, gtr1 gtr2 dko and gtr1 gtr2 gtr3 tko plants.Glucosinolate content in rosettes of three-week-old hydroponically grown plants determined by LC-MS. Data presented is one of two individual experiments. Data are given as means and standard deviation (SD) for individual glucosinolates (nmoles/mg FW), total short-chained aliphatic glucosinolates (SC), total long-chained aliphatic glucosinolates (LC), total aliphatic glucosinolates (AG) and total indole glucosinolates (IG). Differences were tested by ANOVA followed by Post-hoc Tukey HSD Calculator multiple comparison (3mtp, 3-methylthiobutylglucosinolate; 3msp, 3-methylsulfinylpropylglucosinolate; 4mtb, 4-methylthiobutylglucosinolate; 4msb, 4-methylsulfinylbutylglucosinolate;5msp,5-methylsulfinylpentylglucosinolate;7mth,7-(methylthio)heptylglucosinolate,7msh,7-ethylsulfinylheptylglucosinolate;8mso, 8-methylsulfinyloctylglucosinolate; I3M, indol-3-ylmethylglucosinolate; 4MOI3M, 4-methoxy-indol-3-ylmethylglucosinolate; NMOI3M, n,-methoxyindol-3-ylmethylglucosinolate).DOI:
http://dx.doi.org/10.7554/eLife.19466.010Figure 4—source data 2. Glucosinolate content in roots of hydroponically grown wildtype, gtr3, gtr1 gtr2 dko and gtr1 gtr2 gtr3 tko plants.Glucosinolate content in roots of three-week-old hydroponically grown plants determined by LC-MS. Data presented is one of two individual experiments. Data are given as means and standard deviation (SD) for individual glucosinolates (nmoles/mg FW), total short-chained aliphatic glucosinolates (SC), total long-chained aliphatic glucosinolates (LC), total aliphatic glucosinolates (AG) and total indole glucosinolates (IG). Differences were tested by ANOVA followed by Post-hoc Tukey HSD Calculator multiple comparison (3mtp, 3-methylthiobutylglucosinolate; 3msp, 3-methylsulfinylpropylglucosinolate; 4mtb, 4-methylthiobutylglucosinolate; 4msb, 4-methylsulfinylbutylglucosinolate;5msp,5-methylsulfinylpentylglucosinolate;7mth,7-(methylthio)heptylglucosinolate,7msh,7-ethylsulfinylheptylglucosinolate;8mso, 8-methylsulfinyloctylglucosinolate; I3M, indol-3-ylmethylglucosinolate; 4MOI3M, 4-methoxy-indol-3-ylmethylglucosinolate; NMOI3M, n,-methoxyindol-3-ylmethylglucosinolate).DOI:
http://dx.doi.org/10.7554/eLife.19466.011Figure 4—source data 3. Glucosinolate content in rosettes of micro-grafted plants.Glucosinolate content in rosettes of three-week-old micro-grafted plants determined by LC-MS. Data presented is one of two individual experiments. Data are given as means and standard deviation (SD) for individual glucosinolates (nmoles/mg FW), total short-chained aliphatic glucosinolates (SC), total long-chained aliphatic glucosinolates (LC), total aliphatic glucosinolates (AG) and total indole glucosinolates (IG). Differences were tested by ANOVA followed by Post-hoc Tukey HSD Calculator multiple comparison (3mtp, 3-methylthiobutylglucosinolate; 3msp, 3-methylsulfinylpropylglucosinolate; 4mtb, 4-methylthiobutylglucosinolate; 4msb, 4-methylsulfinylbutylglucosinolate;5msp,5-methylsulfinylpentylglucosinolate;7mth,7-(methylthio)heptylglucosinolate,7msh,7-ethylsulfinylheptylglucosinolate;8mso, 8-methylsulfinyloctylglucosinolate; I3M, indol-3-ylmethylglucosinolate; 4MOI3M, 4-methoxy-indol-3-ylmethylglucosinolate; NMOI3M, n,-methoxyindol-3-ylmethylglucosinolate).DOI:
http://dx.doi.org/10.7554/eLife.19466.012Figure 4—source data 4. Glucosinolate content in roots of micro-grafted plants.Glucosinolate content in roots of three-week-old micro-grafted plants determined by LC-MS. Data presented is one of two individual experiments. Data are given as means and standard deviation (SD) for individual glucosinolates (nmoles/mg FW). total short-chained aliphatic glucosinolates (SC). Total long-chained aliphatic glucosinolates (LC). total aliphatic glucosinolates (AG) and total indole glucosinolates (IG). Differences were tested by ANOVA followed by Post-hoc Tukey HSD Calculator multiple comparison (3mtp. 3-methylthiobutylglucosinolate; 3msp. 3-methylsulfinylpropylglucosinolate; 4mtb. 4-methylthiobutylglucosinolate; 4msb. 4-methylsulfinylbutylglucosinolate;5msp.5-methylsulfinylpentylglucosinolate;7mth.7-(methylthio)heptylglucosinolate.7msh.7-ethylsulfinylheptylglucosinolate;8mso. 8-methylsulfinyloctylglucosinolate; I3M. indol-3-ylmethylglucosinolate; 4MOI3M. 4-methoxy-indol-3-ylmethylglucosinolate; NMOI3M. n.-methoxyindol-3-ylmethylglucosinolate).DOI:
http://dx.doi.org/10.7554/eLife.19466.013Figure 4—figure supplement 1. In silico expression analysis of the indole-specific glucosinolate transporter GTR3 and the broad-specificity GTR1 and GTR2 transporters.(A–B) Cell type-specific expression of GTR2 and GTR3 relative to GTR1 expression in (A) shoot and (B) root tissue of 7-day-old Arabidopsis seedlings. Data was retrieved from microarray studies of RNA bound to ribosomes that were immunoprecipitated by use of epitope-tagged ribosomal protein from seedlings (Mustroph et al., 2009). Cell type-specific expression of GTR1, GTR2 and GTR3 was inferred based on co-expression with the genes whose promoters were used for driving the ribosomal affinity tag (Mustroph et al., 2009).DOI:
http://dx.doi.org/10.7554/eLife.19466.014 Image published in: Jørgensen ME et al. (2017) © 2017, Jørgensen et al. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View |