| 研究実績の概要 |
I have currently identified more than 20,000 LRR-RK_XIIs from 300 angiosperms. Many of these receptors might be redundant and recognize the same ligand. As a result, it is challenging to characterize receptors individually. Categorizing LRR-RK_XIIs would solve the redundancy problem and greatly reduce the number of candidates to be characterized. Unlike other LRR-RK subgroups, LRR-RK_XIIs exhibit lineage-specific expansion. For example, the, LRR-RK_XII flagellin-sensing 2 (FLS2) is an ancient receptor that recognizes bacterial flagellin and exists in most angiosperms. While another LRR-RK_XII, EF-Tu receptor (EFR), is a recently evolved receptor that recognizes prokaryotic elongation factor (EF-Tu) and exists only in the plant family Brassicaceae. The strategy is to generate a phylogenetic tree according to the LRR domain. The LRR-RK_XIIs will then be split/classified into multiple groups based on their position in the phylogenetic tree. To determine whether the receptors within a group are likely to recognize the same ligand, I will perform LRR-conservation analysis (RCM). In brief, all the amino acid sequences of the inner LRR surface within a group will be aligned and the conservation scores of each amino acid will be calculated. A high proportion of conserved amino acids indicate that most members within the group are likely to recognize the same ligand. Then, a representative member will be selected from these groups, and I will determine the pathogen-recognition capacity of these candidates.
|
| 現在までの達成度 (区分) |
現在までの達成度 (区分)
2: おおむね順調に進展している
理由
Currently I am working with two bioinformaticians (Dr. Marc W Schmid and Mr. Michele Wyler) to identify and classify LRR-RLK-XIIs. In a trial run, we have successfully classified 12,000 LRR-RK_XIIs into more than 100 distinct subgroups. I have also performed structural and ligand-binding predictions on these subgroups and identified promising PRR candidates. These candidates are being cloned into constructs for expression. We are now repeating the LRR-RLK-XII identification and classification again with more species (300 angiosperms). Once this is finished, I will finalize the list of candidates for the screening. In addition, I am currently optimizing and testing the sensitivity of receptors to different pathogen extracts, such as bacteria and nematodes. After optimization, the screening for PRRs that recognize pathogen extracts is ready to be started.
|
| 今後の研究の推進方策 |
The research project is on-going and progress as planned. The bioinformatics work is on-going and expected to be finished within a few months. Cloning of PRR constructs is also on-going and expected to be ready by the end of 2022. Screening of LRR-responsiveness against pathogens is expected to take at least a year. Depending on the number of candidates that response to pathogens, further function characterization, identification of potentially ligands and transferring candidates to other plant species will take up the rest of the time of this project.
|
