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Image Search Results
Journal: Proceedings of the National Academy of Sciences of the United States of America
Article Title: Duplication and neofunctionalization of a horizontally transferred xyloglucanase as a facet of the Red Queen coevolutionary dynamic.
doi: 10.1073/pnas.2218927121
Figure Lengend Snippet: Fig. 1. Evolutionary history of oomycete GH12. (A) Maximum likelihood tree constructed with IQ-Tree v2.0.3 [WAG+R5 model of evolution (51) selected as the best- fit model by ModelFinder (52)]. The tree was constructed from an alignment of 161 sequences comprising 211 amino acids to confirm the evolutionary relationships between oomycete GH12. The final tree was visualized with iTOL (53), and nodes indicate results (%) of nonparametric bootstrap (200 pseudoreplicates) (54). The tree is rooted with a fungal outgroup because fungi were previously identified as the putative donor group of the GH12 HGT into the oomycetes (31, 33). The eleven P. sojae GH12 paralogs are visually identifiable as they have adjacent transcriptome profiles (column B). Symbols indicate the P. sojae paralog with a structurally inferred additional binding site (gray), and paralogs with C-terminal tails 1 and 2 (blue and green, respectively). The numbers of GH12 orthologs across representative Phytophthora spp., and H. arabidopsidis (an obligate pathogen of A. thaliana) genomes are shown to the Left of the phylogenetic tree, indicating lineage-specific putative local duplications or losses. Some are colored black, indicating they are absent in the tree due to exclusion because of gappy gene models, or they formed uninformative branches, potentially affecting bootstrap resolution (i.e., very long branches or very short indicating little protein variation). All P. sojae paralogs were retained. (B) P. sojae life-cycle transcriptome data [FungiDB: (55, 56)] were used to identify how the eleven P. sojae GH12 paralogs were expressed across (column 1) mycelial, (column 2), cyst and (column 3) 3 d postinfection (soybean hypocotyls infected with P. sojae strain P6497)— expressed in FPKM(Log2). C. Xyloglucanase functional data generated during this study from (column 1) cell culture agar plate-based enzyme assay, (column 2) concentrated supernatant agar-plate based enzyme assay, (column 3) concentrated supernatant DNS reducing sugar assay, and (column 4) concentrated supernatant mass spectrometry of xyloglucan breakdown products (black fill = function detected, white fill = no function detected). (D) ROS data in response to P. sojae xyloglucanase variants, presented as percentage ROS generation (%) of the variant that triggered the highest ROS accumulation in N. benthamiana (i.e., P. sojae_482953, which is presented as 100%). (E) FungiDB (55, 56) was used to locate approximate genomic coordinates of each P. sojae GH12 gene—the relative approximate distances between the paralogs are shown in kilobases (kb), providing additional support for the sister paralog relationships in the phylogenetic tree for: P. sojae_559651 PsXEG1 and P. sojae_360375 PsXLP1 (PHYSO_scaffold_4); and P. sojae_482953 (PHYSO_scaffold_2). Scaffold numbers are shown in the figure.
Article Snippet: For
Techniques: Construct, Binding Assay, Infection, Functional Assay, Generated, Cell Culture, Enzymatic Assay, Mass Spectrometry, Variant Assay