l-cysteine desulfhydrase-related H2 S production is involved in OsSE5-promoted ammonium tolerance in roots of Oryza sativa.

Previous studies revealed that rice heme oxygenase PHOTOPERIOD SENSITIVITY 5 (OsSE5) is involved in the regulation of tolerance to excess ammonium by enhancing antioxidant defence. In this study, the relationship between OsSE5 and hydrogen sulfide (H2 S), a well-known signalling molecule, was investigated. Results showed that NH4 Cl triggered the induction of l-cysteine desulfhydrase (l-DES)-related H2 S production in rice seedling roots. A H2 S donor not only alleviated the excess ammonium-triggered inhibition of root growth but also reduced endogenous ammonium, both of which were aggravated by hypotaurine (HT, a H2 S scavenger) or dl-propargylglycine (PAG, a l-DES inhibitor). Nitrogen metabolism-related enzymes were activated by H2 S, thus resulting in the induction of amino acid synthesis and total nitrogen content. Interestingly, the activity of l-DES, as well as the enzymes involved in nitrogen metabolism, was significantly increased in the OsSE5-overexpression line (35S:OsSE5), whereas it impaired in the OsSE5-knockdown mutant (OsSE5-RNAi). The application of the HT/PAG or H2 S donor could differentially block or rescue NH4 Cl-hyposensitivity or hypersensitivity phenotypes in 35S:OsSE5-1 or OsSE5-RNAi-1 plants, with a concomitant modulation of nitrogen assimilation. Taken together, these results illustrated that H2 S function as an indispensable positive regulator participated in OsSE5-promoted ammonium tolerance, in which nitrogen metabolism was facilitated.

Heme-heme oxygenase 1 system is involved in ammonium tolerance by regulating antioxidant defence in Oryza sativa.

Despite substantial evidence showing the ammonium-altered redox homeostasis in plants, the involvement and molecular mechanism of heme-heme oxygenase 1 (heme-HO1), a novel antioxidant system, in the regulation of ammonium tolerance remain elusive. To fill in these gaps, the biological function of rice HO1 (OsSE5) was investigated. Results showed that NH4 Cl up-regulated rice OsSE5 expression. Oxidative stress and subsequent growth inhibition induced by excess NH4 Cl was partly mitigated by pretreatment with carbon monoxide (CO, a by-product of HO1 activity) or intensified by zinc protoporphyrin (ZnPP, a potent inhibitor of HO1 activity). Pretreatment with HO1 inducer hemin, not only up-regulated OsSE5 expression and HO activity, but also rescued the down-regulation of antioxidant transcripts, total and related isozymatic activities, thus significantly counteracting the excess NH4 Cl-triggered reactive oxygen species overproduction, lipid peroxidation and growth inhibition. OsSE5 RNAi transgenic rice plants revealed NH4 Cl-hypersensitive phenotype with impaired antioxidant defence, both of which could be rescued by CO but not hemin. Transgenic Arabidopsis plants over-expressing OsSE5 also exhibited enhanced tolerance to NH4 Cl, which might be attributed to the up-regulation of several antioxidant transcripts. Altogether, these results illustrated the involvement of heme-HO1 system in ammonium tolerance by enhancing antioxidant defence, which may improve plant tolerance to excess ammonium fertilizer.