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Figure 1.

Phylogenetic tree of GT43 proteins from selected species.

A multiple alignment of protein sequences from Arabidopsis (At), rice (Oryza sativa, Os), poplar (Populus trichocarpa, Pt), spikemoss (Selaginella moellendorffii, Sm) and human (Homo sapiens, Sp) was used to generate a neighbor-joining tree using MEGA5. Bootstrap values are indicated and the scale bar shows evolutionary distances in units of the number of amino acid substitutions per site. See Materials and Methods section for sequence IDs.

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Table 1.

Nomenclature used for the mutated versions of IRX9, IRX9L and IRX14.

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Figure 2.

Representative photos of wild type, mutants and transformed plants.

The nomenclature for the constructs used to transform the irx9 and irx14 plants is explained in Table 1. Rosettes were from 4-week-old plants and mature plants were 6-week-old.

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Figure 3.

Rosette diameter of 4-week-old mutant plants transformed with the different constructs.

The nomenclature for the constructs used to transform the irx9 and irx14 plants is explained in Table 1. The bars show average ± SD (n = 6). Averages that are not significantly different (ANOVA, Tukey's test, p>0.05) are indicated with the same letter.

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Figure 4.

Stem height of 6-week-old mutant plants transformed with the different constructs.

The nomenclature for the constructs used to transform the irx9 and irx14 plants is explained in Table 1. The bars show average ± SD (n = 6). Averages that are not significantly different (ANOVA, Tukey's test, p>0.05) are indicated with the same letter.

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Figure 5.

Xylose content in cell wall polysaccharides isolated from stems of 6-week-old mutant plants transformed with the different constructs.

Cell walls were prepared from stems, hydrolyzed in TFA, and the xylose content determined by HPAEC. The nomenclature for the constructs used to transform the irx9 and irx14 plants is explained in Table 1. The bars show average ± SD (n = 4). Averages that are not significantly different (ANOVA, Tukey's test, p>0.05) are indicated with the same letter.

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Figure 6.

Microcopy analysis of stems from 6-week-old plants.

The LM10 anti-xylan monoclonal antibody was used for immunodetection of xylan in transverse stem sections. The nomenclature for the constructs used to transform the irx9 and irx14 plants is explained in Table 1. Irregular xylem phenotype (indicated with arrows) was observed in the irx9 and irx14 mutants, as well as in the irx14 plants transformed with the 35S:IRX14-1 construct. All the other transformants showed normal vessel phenotype.

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