Supplemental Data

Files in this Data Supplement:

• Supplemental Figure 1 - Supplemental Figure 1. The deduced amino acid sequence of the Botrytis induced Kinase 1 (BIK1) protein. The BIK protein conserved sequences containing the kinase subdomains within its catalytic domain are underlined (Hanks and Quinn, 1991). The conserved amino acid triplets DFG (subdomain VII) and APE (subdomain VIII) that define the borders of the activation domain segment of the protein kinases are outlined (Johnson et al., 1996). A potential myristoylation motif at the N-terminus of BIK1 is shown in bold.
• Supplemental Figure 2 - Supplemental Figure 2. Phylogenetic tree showing the relationship between BIK1 and related plant protein kinases.
• Supplemental Figure 3 - Supplemental Figure 3. Multiple sequence alignment used for phylogenetic analysis. Sequences were aligned using ClustalW (Thompson et al., 1994) and adjusted by hand. The multiple sequence alignment was then used to construct the phylogenetic tree using mean character distances with the neighbor-joining method from the PAUP program (Swofford, 2000). Numbers depict bootstrap values (above 50%) derived from 1000 bootstrap resamples. Proteins kinases included in the analysis are AAT40481 (Solanum demissum), MLPKe (Brassica rapa), PTO (Lycopersicum esculentum), XP_493889 and NP_910058 (Oryza sativa) and Arabidopsis protein kinases. Black shading indicates amino acids conserved in all entries, and green shading indicates amino acids identical to the BIK1 sequence.
• Supplemental Figure 4 - Supplemental Figure 4. Expression of BIK1 in Arabidopsis Wild type, ein2, coi1, nahG and pad2 Plants During Botrytis Infection. Ten micrograms of total RNA was loaded in each lane. The RNA blot was hybridized to full length BIK1 cDNA.
• Supplemental Figure 5 - Supplemental Figure 5. Immunocomplex Assays Showing the Activation of MPK3 and MPK6 in Wild-type and bik1 plants in Response to Botrytis Infection. MPK3 and MPK6 were immunoprecipitated from leaf cell extracts of Botrytis infected plants. MPK activity was measured in immunocomplex kinase assays using MBP as substrate, and levels of MPK3 and MPK6 proteins were detected in Western blots.
• Supplemental Figure 6 - Supplemental Figure 6. The BIK1 cDNA Rescues thebik1 Altered Growth Traits to Wild type levels. Complementation of (A) Reduced primary root length, (B) Increased number and length of root hair growth traits to wild type levels. All assays were repeated three times.