The function of chloroplast ferredoxin‐NADP+ oxidoreductase positively regulates the accumulation of bamboo mosaic virus in Nicotiana benthamiana

• Published in: Molecular plant pathology 2022-04, Vol.23 (4), p.503-515
• Main Authors: Chen, I‐Hsuan, Chen, Xiang‐Yu, Chiu, Guan‐Zhi, Huang, Ying‐Ping, Hsu, Yau‐Heiu, Tsai, Ching‐Hsiu
• Format: Article
• Language: English
• Subjects: Accumulation; Bamboo; BaMV; chloroplast; Chloroplasts; Coat protein; Electron transport; Electron transport chain; Experiments; Ferredoxin; ferredoxin NADP+ oxidoreductase; Fluorescence; Gene expression; Gene silencing; Genomes; Genotype & phenotype; Heparin; Immunoprecipitation; in vitro replication assay photosynthesis enzyme; Infections; Inoculation; Mutants; Mutation; NADP; Nicotiana benthamiana; Original; Oxidoreductase; Photosynthesis; Proteins; Protoplasts; Replicase; RNA polymerase; Viral infections; viral RNA accumulation; Viruses; Yeasts
• ISSN: 1464-6722; 1364-3703
• DOI: 10.1111/mpp.13174

A gene down‐regulated in Nicotiana benthamiana after bamboo mosaic virus (BaMV) infection had high identity to the nuclear‐encoded chloroplast ferredoxin NADP+ oxidoreductase gene (NbFNR). NbFNR is a flavoenzyme involved in the photosynthesis electron transport chain, catalysing the conversion of NADP+ into NADPH. To investigate whether NbFNR is involved in BaMV infection, we used virus‐induced gene silencing to reduce the expression of NbFNR in leaves and protoplasts. After BaMV inoculation, the accumulation of BaMV coat protein and RNA was significantly reduced. The transient expression of NbFNR fused with orange fluorescent protein (OFP) localized in the chloroplasts and elevated the level of BaMV coat protein. These results suggest that NbFNR could play a positive role in regulating BaMV accumulation. Expressing a mutant that failed to translocate to the chloroplast did not assist in BaMV accumulation. Another mutant with a catalytic site mutation could support BaMV accumulation to some extent, but accumulation was significantly lower than that of the wild type. In an in vitro replication assay, the replicase complex with FNR inhibitor, heparin, the RdRp activity was reduced. Furthermore, BaMV replicase was revealed to interact with NbFNR in yeast two‐hybrid and co‐immunoprecipitation experiments. Overall, these results suggest that NbFNR localized in the chloroplast with functional activity could efficiently assist BaMV accumulation. The nuclear‐encoded chloroplast ferredoxin NADP+ oxidoreductase gene (NbFNR) could assist bamboo mosaic virus accumulation efficiently when expressed in Nicotiana benthamiana leaves but not the mutants with failure in translocating to the chloroplast (NbFNR/ΔTP) and with a catalytic site mutation (NbFNR/mut).