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. 2016 Aug 8;7:1140. doi: 10.3389/fpls.2016.01140

FIGURE 2.

FIGURE 2

Identification of AvrPi54 gene in Magnaporthe oryzae genome which encodes a secreted protein, that interacts with blast resistance protein Pi54. 3-D models of Pi54 protein (A) and candidate AvrPi54 proteins (B) generated by MODELER software inbuilt in Accelrys Discovery Studio 2.1. In silico interaction study between Pi54 protein and the four selected candidate AvrPi54 proteins by docking analysis (carried out in ZDOCK software inbuilt in Accelrys Discovery Studio 2.1) revealed, Mo-01947_9 protein is the best candidate to be qualified as AvrPi54 protein as it showed best interaction with Pi54 protein. The Pi54 protein is shown in blue in all the cases and the interacting amino acids of both Pi54 and candidate AvrPi54 proteins are shown in orange (C). In silico mapping showed that the candidate AvrPi54 (Mo-01947_9) is a unique gene located at chromosome 4. It is a 1258 bp long gene containing a single unfragmented ORF of 462 bp. It encodes a protein consisting of 153 amino acids among which first 19 amino acids from the N-terminal end encodes for a signal peptide (SP) needed for secretion (D). Yeast-2-Hybrid analysis confirmed the interaction between Pi54 protein and candidate AvrPi54 protein (Mo-01947_9) in vitro. Interaction potential was checked for both full length (Mo-01947_9_comp) as well as SP truncated version (Mo-01947_9_trun) of candidate AvrPi54 protein with the Pi54 protein. Better growth of yeast cells clearly suggested better interaction potential of SP truncated version (Mo-01947_9_trun) of candidate AvrPi54 protein with Pi54 protein compared to its full length (Mo-01947_9_comp) version (E). In silico docking analysis also supported this fact as the pose and strength of interaction was better in case of SP truncated version of AvrPi54 protein (Mo-01947_9_trun) compared to the full length version of AvrPi54 protein (Mo-01947_9_comp) (F).