Suppression of Nodule Formation by RNAi Knock-Down of Bax inhibitor-1a in Lotus japonicus

Genes (Basel). 2025 Jan 6;16(1):58. doi: 10.3390/genes16010058.

Abstract

Background/objectives: The balanced regulation of innate immunity plays essential roles in rhizobial infection and the establishment and maintenance of symbiosis. The evolutionarily conserved cell death suppressor Bax inhibitor-1 plays dual roles in nodule symbiosis, providing a valuable clue in balancing immunity and symbiosis, while it remains largely unexplored in the legume Lotus japonicus.

Methods/results: In the present report, the BI-1 gene family of L. japonicus was identified and characterized. We identified 6 BI-1 genes that translate into peptides containing 240-255 amino acids with different structural characteristics and isoelectric points. We performed phylogenetic analyses and detected evolutionary conservation and divergence among BI-1 proteins from L. japonicus, Glycine max, Medicago truncatula, Arabidopsis thaliana, and Oryza sativa. Expression profiles among different roots indicated that the inoculation of MAFF303099 significantly increased the expression of most of the L. japonicus BI-1 family genes. We down-regulated the transcripts of LjBI-1a by RNA interference and observed that LjBI-1a promotes nodulation and nodule formation.

Conclusions: These discoveries shed light on the functions of BI-1 genes in L. japonicus, and simultaneously emphasize the potential application of LjBI-1a in enhancing the symbiotic nitrogen fixation ability of legumes.

Keywords: Bax inhibitor-1; RNA interference; infection thread; nodulation; nodule formation; root nodule symbiosis.

MeSH terms

  • Gene Expression Regulation, Plant*
  • Gene Knockdown Techniques
  • Lotus* / genetics
  • Lotus* / metabolism
  • Lotus* / microbiology
  • Phylogeny*
  • Plant Proteins* / genetics
  • Plant Proteins* / metabolism
  • Plant Root Nodulation / genetics
  • RNA Interference*
  • Root Nodules, Plant* / genetics
  • Root Nodules, Plant* / metabolism
  • Root Nodules, Plant* / microbiology
  • Symbiosis* / genetics

Substances

  • Plant Proteins

Grants and funding

This work was supported by funds from the National Natural Science Foundation of China (grant no. 32071964), the Key Research and Development Plan Projects of Hubei Province (2022BBA0036), the Agricultural Science and Technology Innovation Program of CAAS (CAAS-0CRI-ZDRW-202402), and the National Natural Science Foundation of China (grant number: U1904102). The funding body played no role in the study design, sample collection, data analysis, data interpretation, and manuscript drafting.