![]() ![]() (A) Primers used for amplification of the 5′ and 3′ integration sites of the CESA5KO‐npt vector are indicated as black arrows on the diagram showing the PpCESA5KO‐npt vector integrated so as to delete PpCESA5. ![]() Within each ‘test’ set, means with the same letter do not differ significantly (p > .05, Fisher's Exact Test).įigure S2 PCR‐based genotyping of ppcesa5KO lines. As measured by percentages of lines that produced gametophores, all of the mutant proteins fully restored gametophore formation. In each experiment, complementation with PpCESA5 provided a positive control, and an empty vector was tested as a negative control. ![]() ) was complemented with vectors driving expression of PpCESA5 with phenylalanine at position 968 changed to glycine (G), tyrosine (Y), tryptophan (W), aspartate (D), or lysine (K). n > 50 seedlings per line were measured within 2 trials. The restored root elongation at the restrictive temperature showed that the F967G variant of AtCesA1 was fully functional. The lengths of the seedling primary roots of Col‐0 wild type, the rsw1–1 mutant, and F967G plants were measured after 5 days growth at 22 ☌ followed by 2 days growth at restrictive 29 ☌. A For AtCesA1, two independent F967G lines were generated by transforming the rsw1–1 temperature sensitive mutant in AtCesA1 (Arioli et al., 1998, Science 279: 717–720. Methods were described previously (Slabaugh et al., 2016, Glycobiology 26: 509–519. patens PpCesA5 with substitution of the phenylalanine just beyond the acidic motif (corresponding to F913 in AtCesA7). Figure S1 Phenotype complementation assays for Arabidopsis AtCesA1 or P. ![]()
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