The F-box protein CORONATINE INSENSITIVE1 (COI1) plays a central role in jasmonate (JA) signaling and is required for all JA responses in Arabidopsis (Arabidopsis thaliana). To dissect JA signal transduction, we isolated the partially suppressing coi1 (psc1) mutant, which partially suppressed coi1 insensitivity to JA inhibition of root growth. The psc1 mutant partially restored JA sensitivity in coi1-2 background and displayed JA hypersensitivity in wild-type COI1 background. Genetic mapping, sequence analysis, and complementation tests revealed that psc1 is a leaky mutation of DWARF4 (DWF4) that encodes a key enzyme in brassinosteroid (BR) biosynthesis. Physiological analysis showed that an application of exogenous BR eliminated the partial restoration of JA sensitivity by psc1 in coi1-2 background and the JA hypersensitivity of psc1 in wild-type COI1 background. Exogenous BR also attenuated JA inhibition of root growth in the wild type. In addition, the expression of DWF4 was inhibited by JA, and this inhibition was dependent on COI1. These results indicate that (1) BR is involved in JA signaling and negatively regulates JA inhibition of root growth, and (2) the DWF4 is down-regulated by JA and is located downstream of COI1 in the JA-signaling pathway.

Jasmonate (JA) regulates plant development, mediates defense responses, and induces anthocyanin biosynthesis as well. Previously, we isolated the psc1 mutant that partially suppressed coi1 insensitivity to JA, and found that brassinosteroid (BR) was involved in JA signaling and negatively regulated JA inhibition of root growth in Arabidopsis. In this study it was shown that JA-induced anthocyanin accumulation was reduced in BR mutants or in wild type treated with brassinazole, an inhibitor of BR biosynthesis, whereas it was induced by an application of exogenous BR. It was also shown that the 'late' anthocyanin biosynthesis genes including DFR, LDOX, and UF3GT, were induced slightly by JA in the BR mutants relative to wild type. Furthermore, the expression level of JA-induced Myb/bHLH transcription factors such as PAP1, PAP2, and GL3, which are components of the WD-repeat/Myb/bHLH transcriptional complexes that mediate the 'late' anthocyanin biosynthesis genes, was lower in the BR mutants than that in wild type. These results suggested that BR affects JA-induced anthocyanin accumulation by regulating the 'late' anthocyanin biosynthesis genes and this regulation might be mediated by the WD-repeat/Myb/bHLH transcriptional complexes.

Fumarylacetoacetate hydrolase (FAH) hydrolyzes fumarylacetoacetate to fumarate and acetoacetate, the final step in the tyrosine (Tyr) degradation pathway that is essential to animals. Deficiency of FAH in animals results in an inborn lethal disorder. However, the role for the Tyr degradation pathway in plants remains to be elucidated. In this study, we isolated an Arabidopsis (Arabidopsis thaliana) short-day sensitive cell death1 (sscd1) mutant that displays a spontaneous cell death phenotype under short-day conditions. The SSCD1 gene was cloned via a map-based cloning approach and found to encode an Arabidopsis putative FAH. The spontaneous cell death phenotype of the sscd1 mutant was completely eliminated by further knockout of the gene encoding the putative homogentisate dioxygenase, which catalyzes homogentisate into maleylacetoacetate (the antepenultimate step) in the Tyr degradation pathway. Furthermore, treatment of Arabidopsis wild-type seedlings with succinylacetone, an abnormal metabolite caused by loss of FAH in the Tyr degradation pathway, mimicked the sscd1 cell death phenotype. These results demonstrate that disruption of FAH leads to cell death in Arabidopsis and suggest that the Tyr degradation pathway is essential for plant survival under short-day conditions.

X, et al. A leaky mutation in DWARF4 reveals an antagonistic role of brassinosteroid in the inhibition of root growth by jasmonate in Arabidopsis.

Fumarylacetoacetate hydrolase (FAH) catalyzes the final step in Tyr degradation pathway essential to animals but not well understood in plants. Previously, we found that mutation of SSCD1 encoding Arabidopsis FAH causes cell death under short day, which uncovered an important role of Tyr degradation pathway in plants. Since phytohormones salicylic acid (SA) and jasmonate (JA) are involved in programmed cell death, in this study, we investigated whether sscd1 cell death is related to SA and JA, and found that (1) it is accompanied by up-regulation of JA-and SA-inducible genes as well as accumulation of JA but not SA; (2) it is repressed by breakdown of JA signaling but not SA signaling; (3) the up-regulation of reactive oxygen species marker genes in sscd1 is repressed by breakdown of JA signaling; (4) treatment of wild-type Arabidopsis with succinylacetone, an abnormal metabolite caused by loss of FAH, induces expression of JA-inducible genes whereas treatment with JA induces expression of some Tyr degradation genes with dependence of JA signaling. These results demonstrated that cell death resulted from loss of FAH in Arabidopsis is related to JA but not SA, and suggested that JA signaling positively regulates sscd1 cell death by up-regulating Tyr degradation. Programmed cell death (PCD) is a sequence of genetically regulated events resulting in the elimination of specific cells, tissues, or whole organs 1 , which is required both for normal development and to face stress conditions 2-4. In plants, one well-characterized example of PCD is hypersensitive response taking place on incompatible plant-pathogen interactions 3 , which leads to cell death and then forms visible lesions at the site infected by an avirulent pathogen, as a result, limits the pathogen spread 4. Phytohormones including salicylic acid (SA) and jasmonate (JA) appear to be key players for hypersensitive response regulation 5. To date, a large number of mutants displaying spontaneous cell death lesions have been identified in plants including Arabidopsis, rice, barley, maize, and so on 6-9. These mutants have been named as lesion-mimic mutants (LMM) because of the form of lesions in the absence of pathogen infection 10. In some of LMM, the SA or JA signaling has been activated 9,11. By isolating LMM's genes, many of regulators that play important roles in PCD and SA or JA signal defense responses have been identified, including ACCELERATED CELL DEATH11, LESION SIMULATING DISEASE1, and NICOTIANA BENTHAMIANA HOMEOBO1 12-14. SA is involved in plant defense and cell death 15,16. The level of SA correlates with the expression of PATHO-GENESIS-RELATED1 (PR1) gene and resistance to pathogen attack 17,18. The NON-EXPRESSOR OF PATHO-GENESIS-RELATED GENES1 (NPR1) gene is required for SA-induced expression of PR1 gene and resistance in Arabidopsis 19,20 .