Dual Roles of GSNOR1 in Cell Death and Immunity in Tetraploid Nicotiana tabacum .

S-nitrosoglutathione reductase 1 (GSNOR1) is the key enzyme that regulates cellular homeostasis of S- nitrosylation. Although extensively studied in Arabidopsis , the roles of GSNOR1 in tetraploid Nicotiana species have not been investigated previously. To study the function of Nt GSNOR1, we knocked out two NtGSNOR1 genes simultaneously in Nicotiana tabacum using clustered regularly interspaced short palindromic repeats (CRISPR)/caspase 9 (Cas9) technology. To our surprise, spontaneous cell death occurred on the leaves of the CRISPR/Cas9 lines but not on those of the wild-type (WT) plants, suggesting that Nt GSNOR1 negatively regulates cell death. The natural cell death on the CRISPR/Cas9 lines could be a result from interactions between overaccumulated nitric oxide (NO) and hydrogen peroxide (H ₂ O ₂ ). This spontaneous cell death phenotype was not affected by knocking out two Enhanced disease susceptibility 1 genes (NtEDS11a/1b) and thus was independent of the salicylic acid (SA) pathway. Unexpectedly, we found that the NtGSNOR1a/1b knockout plants displayed a significantly ( p < 0.001) enhanced resistance to paraquat-induced cell death compared to WT plants, suggesting that Nt GSNOR1 functions as a positive regulator of the paraquat-induced cell death. The increased resistance to the paraquat-induced cell death of the NtGSNOR1a/1b knockout plants was correlated with the reduced level of H ₂ O ₂ accumulation. Interestingly, whereas the N gene-mediated resistance to Tobacco mosaic virus (TMV) was significantly enhanced ( p < 0.001), the resistance to Pseudomonas syringae pv. tomato DC3000 was significantly reduced ( p < 0.01) in the NtGSNOR1a/1b knockout lines. In summary, our results indicate that Nt GSNOR1 functions as both positive and negative regulator of cell death under different conditions and displays distinct effects on resistance against viral and bacterial pathogens.
Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
(Copyright © 2021 Li, Ren, Guo, Ran, Ren, Wu, Xu, Liu and Liu.)