Functional analysis of the OsNPF4.5 nitrate transporter reveals a conserved mycorrhizal pathway of nitrogen acquisition in plants

Proc Natl Acad Sci U S A. 2020 Jul 14;117(28):16649-16659. doi: 10.1073/pnas.2000926117. Epub 2020 Jun 25.

Abstract

Low availability of nitrogen (N) is often a major limiting factor to crop yield in most nutrient-poor soils. Arbuscular mycorrhizal (AM) fungi are beneficial symbionts of most land plants that enhance plant nutrient uptake, particularly of phosphate. A growing number of reports point to the substantially increased N accumulation in many mycorrhizal plants; however, the contribution of AM symbiosis to plant N nutrition and the mechanisms underlying the AM-mediated N acquisition are still in the early stages of being understood. Here, we report that inoculation with AM fungus Rhizophagus irregularis remarkably promoted rice (Oryza sativa) growth and N acquisition, and about 42% of the overall N acquired by rice roots could be delivered via the symbiotic route under N-NO3- supply condition. Mycorrhizal colonization strongly induced expression of the putative nitrate transporter gene OsNPF4.5 in rice roots, and its orthologs ZmNPF4.5 in Zea mays and SbNPF4.5 in Sorghum bicolor OsNPF4.5 is exclusively expressed in the cells containing arbuscules and displayed a low-affinity NO3- transport activity when expressed in Xenopus laevis oocytes. Moreover, knockout of OsNPF4.5 resulted in a 45% decrease in symbiotic N uptake and a significant reduction in arbuscule incidence when NO3- was supplied as an N source. Based on our results, we propose that the NPF4.5 plays a key role in mycorrhizal NO3- acquisition, a symbiotic N uptake route that might be highly conserved in gramineous species.

Keywords: OsNPF4.5; RNA sequencing; arbuscular mycorrhiza; nitrate transporter; nitrogen uptake.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anion Transport Proteins / genetics
  • Anion Transport Proteins / metabolism*
  • Gene Expression Regulation, Plant
  • Glomeromycota / physiology*
  • Mycorrhizae / physiology*
  • Nitrate Transporters
  • Nitrates / metabolism
  • Nitrogen / metabolism*
  • Oryza / genetics
  • Oryza / growth & development
  • Oryza / metabolism*
  • Oryza / microbiology
  • Plant Proteins / genetics
  • Plant Proteins / metabolism*
  • Plant Roots / genetics
  • Plant Roots / growth & development
  • Plant Roots / metabolism
  • Plant Roots / microbiology
  • Sorghum / genetics
  • Sorghum / metabolism
  • Sorghum / microbiology
  • Zea mays / genetics
  • Zea mays / metabolism
  • Zea mays / microbiology

Substances

  • Anion Transport Proteins
  • Nitrate Transporters
  • Nitrates
  • Plant Proteins
  • Nitrogen