RLM3 Antibody

FAQs for RLM3-Related Research in Plant Immunity

Advanced Research Questions

• How to resolve contradictions in RLM3’s role across necrotrophic vs. biotrophic pathogens?

  • Hypothesis testing:

    • RLM3 may regulate distinct defense pathways. For example, it influences callose deposition (anti-necrotrophic) but not JA/ET signaling .

    • Test cross-talk with salicylic acid (SA) pathways using SA biosynthesis mutants.

    • Leverage transcriptomics to identify RLM3-dependent genes (e.g., WRKY60 repression) .

• What structural features of RLM3 enable broad-spectrum recognition of fungal effectors?

  • Findings:

    • RLM3 recognizes LARS-family effectors (e.g., AvrLm3, Ecp11-1) via conserved motifs .

    • Critical residues for effector binding (e.g., AvrLm3 I58, G131, F134) were identified through allele mining .

  • Experimental design:

    • Use site-directed mutagenesis on AvrLm3 homologs (e.g., F. oxysporum effectors) and test recognition in RLM3-transgenic plants .

    • Employ cryoEM or AlphaFold2 to model RLM3-effector complexes .

Methodological Challenges & Solutions

• How to address variability in RLM3 expression across plant tissues?

  • Solutions:

    • Quantify splice variants via RT-PCR and Nanopore sequencing .

    • Normalize expression data using housekeeping genes validated for Brassica napus (e.g., ACTIN2).

• What bioinformatics tools are optimal for analyzing RLM3-linked immune networks?

  • Tools:

    • ExpoSeq: For antibody-like sequence analysis in effector recognition .

    • ModelAngelo: Integrates cryoEM density maps to guide antibody/effector-binding site modeling .

    • Antibody Analytics: Predicts developability of synthetic antibodies for functional studies .

Data Contradiction Analysis

• Why does RLM3 knockout increase susceptibility to some pathogens but not others?

  • Resolution:

    • Pathogen-specific effector recognition: RLM3 targets LARS-family effectors but not others (e.g., B. cinerea effectors) .

    • Redundant pathways: RLM1/RLM2 may compensate in certain contexts .

Key Research Gaps

• Role of RLM3 in transcriptional regulation (e.g., nuclear RCC1 proteins) .

• Engineering RLM3 homologs in non-host species for broad-spectrum resistance .