Fig 1.

(A) The expression cassette of the vector developed for Arabidopsis transformation. Abbreviations: CaMV 35S –Tetramer of Cauliflower Mosaic virus 35S RNA Promoter, YFP, yellow fluorescent protein coding sequence. (B) PCR analysis of genomic DNA from transgenic Arabidopsis lines transformed with microbial A.nidulans α-fucosidase expression cassette and wild type plants. Herbicide resistant lines were confirmed to harbor the full construct using four pairs of primers (see S1 Table for sequences). Lane 1—amplification of AnF genes from corresponding transgenic lines; Lane 5—amplification of the same genes from Col-0 wild type plant; Lane 2—amplification of hybrid fragment containing AnF gene linked to the YFP from mutant lines; Lane 6—amplification of the AnF-YFP fragment from Col-0 wild type plant; Lane 3—amplification of YFP gene from the AnF line; Lane 7 –amplification of YFP from Col-0 wild type plant; Lane 4—amplification of A. thaliana ACTIN-2 gene fragment from AnF line, and Lane 8 –amplification of ACTIN-2 from Col-0 wild type plant. Analysis was done for three independent transgenic lines for each construction; picture shows results of PCR for single plant of each mutant line. (C) Western blot analysis of total proteins from apoplast of Arabidopsis AnF transgenic and wild type plants. The corresponding microbial fucosidase fused with YFP (116kDa) were found in transgenic lines and not in Col-0 control plants. Blots were produced using GFP monoclonal antibodies (1:5000 dilution).