FAH catalyzes the final step of Tyr degradation pathway and the deficiency of FAH in animals causes an inborn lethal disease, which was named as HT1 in humans1,3. Phe could be converted to Tyr and then degraded in animals, the dietary restriction of Tyr as well as Phe can improve the condition of HT1 patients4,9. In plants, the SSCD1 gene encodes the Arabidopsis FAH, mutation of SSCD1 results in spontaneous cell death under SD6. Like as in animals, Phe could be also converted into Tyr in plants and then degraded10. However, in our study the death of sscd1 seedlings was not increased but repressed by Phe treatment (Fig. 1). So, why would Phe treatment repress the cell death resulting from loss of FAH in plants?
Previously, we demonstrated that the sscd1 cell death is mediated by Chl biosynthesis8. The inhibition of the ALAD activity by SUAC in the sscd1 mutant influences Chl biosynthesis resulting in impairment of feedback inhibition of Chl biosynthesis from the light–dark transition under SD, which activates Chl biosynthesis and accumulation of Pchlide in the dark, and then upon re-illumination the excessive accumulation of Pchlide induces the mass production of ROS and thereby causes cell death8. Thus, it is the excess ROS that induces the sscd1 cell death. In this study, treatment of sscd1 seedlings with Phe distinctly repressed the up-regulation of ROS marker genes including APX2, OXI1, ZP and BAP1 (Fig. 2), which indicated that ROS is reduced by Phe treatment.
In Chl biosynthetic pathway there are two pivotal control points mainly regulated by HEMA1 and CHLH respectively32,36−38. In our study, after treatment with Phe, both the content of Chl and the expression levels of HEMA1 and CHLH were increased (Fig. 3), suggesting that Phe treatment could promote Chl biosynthesis. The increase in Chl biosynthesis would restore to some extent the feedback inhibition of Chl biosynthesis in the sscd1 mutant from the light–dark transition, as a result, reducing the accumulation of Pchlide in the dark and the production of ROS after subsequent exposure to light.
In plants, Phe could be metabolized through the phenylpropanoid pathway to produce secondary metabolites, which plays an important role in plant against stress including UV-light, drought and pathogen attack10,11,26,39−41. PAL catalyzes the first step of the phenylpropanoid pathway, which is a key step in phenylpropanoid biosynthesis13,34,42. The activity of PAL could be inhibited by AOPP and promoted by MeJA16. Treatment with AOPP prevents the increase in resistance to B. cinerea due to the application of external Phe41. In our study, the repression of sscd1 seedlings death by Phe was reduced by AOPP treatment (Fig. 4), however, it was enhanced by MeJA (Fig. 5), which suggested that the suppression of sscd1 cell death by Phe is related to the phenylpropanoid pathway. Since the secondary metabolites produced by Phe metabolism through the phenylpropanoid pathway have antioxidant function41, ROS could be also reduced by the metabolism of Phe through the phenylpropanoid pathway, which should be another important cause for the repression of sscd1 cell death by Phe.
Recently, we found that JA signaling is involved in the sscd1 cell death31. In the sscd1 mutant, the accumulation of SUAC results in the generation of ROS, which induces cell death as well as JA synthesis31. JA up-regulates Tyr degradation pathway, producing more SUAC, which promotes cell death31. Once JA signaling is broken by mutation of COI1 encoding a JA receptor35, the up-regulation of Tyr degradation pathway by JA is eliminated, reducing the production of SUAC, as a result, the sscd1 cell death is repressed31. In this study, MeJA treatment markedly increased the expression level of PAL1 in wild type but not in the coi1-2 mutant (Fig. 6), indicating that JA signaling up-regulates the phenylpropanoid pathway. The repression of sscd1 cell death by Phe could be enhanced by MeJA treatment in the sscd1 single mutant (Figs. 5 and 7) but not in the sscd1coi1 double mutant (Fig. 7), which suggested that MeJA treatment enhances Phe inhibition of the sscd1 cell death through JA signaling. Therefore, JA has a dual regulatory effect on the sscd1 cell death. On the one hand, JA up-regulates the Tyr degradation pathway, promoting the sscd1 cell death, on the other hand, JA up-regulates the phenylpropanoid pathway, inhibiting the sscd1 cell death. For this reason, the death of sscd1 seedlings was not increased by MeJA treatment, it seemed to decrease slightly (Figs. 5 and 7), which suggested that the effect of MeJA treatment on the sscd1 cell death through phenylpropanoid pathway might be greater than that through Tyr degradation pathway.
In conclusion, although Phe can be degraded through Tyr degradation pathway, unlike in animals, Phe treatment does not increase the cell death resulting from loss of FAH in plants, instead, it represses the cell death. A possible mechanism for the repression of sscd1 cell death by Phe treatment can be described as follows (Fig. 8). Loss of FAH in the sscd1 mutant results in a decline of Chl biosynthesis, which impairs the feedback inhibition of Chl biosynthesis from light–dark transition under SD, leading to the accumulation of ROS and then cell death. Phe treatment, on the one hand, promotes Chl biosynthesis, increasing the feedback inhibition of Chl biosynthesis from light–dark transition under SD, and on the other hand, activates phenylpropanoid pathway, both of which reduce ROS and subsequent cell death. In addition, in the sscd1 mutant ROS induces cell death as well as JA synthesis. JA signaling up-regulates Tyr degradation pathway, promoting the sscd1 cell death, however, it also up-regulates PAL1 that activates phenylpropanoid pathway, repressing the sscd1 cell death. Since the effect of MeJA treatment on the sscd1 cell death through phenylpropanoid pathway might be greater than that through Tyr degradation pathway, the repression of sscd1 cell death by Phe could be enhanced by MeJA treatment.