Brassinosteroids (BRs) are a unique class of plant steroid hormones that orchestrate myriad growth and developmental processes. Although BRs have long been known to protect plants from a suite of biotic and abiotic stresses, our understanding of the underlying molecular mechanisms is still rudimentary. Aiming to further decipher the molecular logic of BR-modulated immunity, we have examined the dynamics and impact of BRs during infection of rice (Oryza sativa) with the root oomycete Pythium graminicola. Challenging the prevailing view that BRs positively regulate plant innate immunity, we show that P. graminicola exploits BRs as virulence factors and hijacks the rice BR machinery to inflict disease. Moreover, we demonstrate that this immunesuppressive effect of BRs is due, at least in part, to negative cross talk with salicylic acid (SA) and gibberellic acid (GA) pathways. BR-mediated suppression of SA defenses occurred downstream of SA biosynthesis, but upstream of the master defense regulators NONEXPRESSOR OF PATHOGENESIS-RELATED GENES1 and OsWRKY45. In contrast, BR alleviated GA-directed immune responses by interfering at multiple levels with GA metabolism, resulting in indirect stabilization of the DELLA protein and central GA repressor SLENDER RICE1 (SLR1). Collectively, these data favor a model whereby P. graminicola coopts the plant BR pathway as a decoy to antagonize effectual SA-and GA-mediated defenses. Our results highlight the importance of BRs in modulating plant immunity and uncover pathogen-mediated manipulation of plant steroid homeostasis as a core virulence strategy.

BackgroundProgressive yield decline in Philippine aerobic rice fields has been recently associated with three closely related Pythium spp., P. arrhenomanes, P. graminicola and P. inflatum. To understand their differential virulence towards rice seedlings, we conducted a comparative survey in which three isolates each of P. arrhenomanes, P. graminicola and P. inflatum were selected to investigate host colonization, host responses and carbon utilization profiles using histopathological analyses, phenoarrays, DNA quantifications and gene expression studies.ResultsThe isolate of the most virulent species, P. arrhenomanes, quickly colonized the outer and inner root tissues of rice seedlings, including the xylem, by which it possibly blocked the water transport and induced severe stunting, wilting and seedling death. The lower virulence of the tested P. graminicola and P. inflatum isolates seemed to be reflected in slower colonization processes, limited invasion of the vascular stele and less systemic spread, in which cell wall fortification appeared to play a role. Progressive hyphal invasions triggered the production of reactive oxygen species (ROS) and phenolic compounds, which was the strongest for the P. arrhenomanes isolate and was delayed or much weaker upon inoculation with the P. inflatum isolate. The necrosis marker OsJamyb seemed faster and stronger induced by the most virulent isolates. Although the isolate of P. inflatum was nutritionally the most versatile, the most virulent Pythium isolate appeared physiologically more adapted to its host, evidenced by its broad amino acid utilization profile, including D-amino acids, L-threonine and hydroxyl-L-proline. The latter two compounds have been implicated in plant defense and their use by P. arrhenomanes could therefore represent a part of its virulence strategy.ConclusionsThis study illustrates that the differential virulence of rice-pathogenic P. arrhenomanes, P. graminicola and P. inflatum isolates is related to their root colonization capacity, the intensity of induced root responses and their ability to utilize amino acids in their colonization niche. Accordingly, this paper presents important knowledge concerning rice root infections by oomycetes, which could be helpful to further disentangle virulence tactics of soil-borne pathogens.

The cultivation of aerobic rice in the tropics enables farmers to save water without lowering productivity. Unfortunately, this system suffers from declining yields due to a disease complex involving nematodes, pathogenic Pythium spp. and nutrient deficiencies. Assessing the impact of each underlying factor can contribute to efficient disease control measures. This study therefore investigated pathogenic and genotypic variability among Pythium species from affected aerobic rice fields in the Philippines using pathogenicity assays and sequence information from the internal transcribed spacer (ITS) region and b-tubulin gene. Three closely related Pythium spp., P. arrhenomanes, P. graminicola and P. inflatum, were recovered from affected aerobic rice fields. All P. arrhenomanes isolates reduced rice seedling growth, whereas only a few P. graminicola isolates and no P. inflatum isolates were pathogenic, indicating that P. arrhenomanes is probably the most important species affecting rice. Both P. arrhenomanes and P. graminicola isolates showed little genetic variation, despite the observed pathogenic variation within P. graminicola. Intraspecific variation was higher among P. inflatum isolates, but again no correlation was observed with phenotype. When screening P. arrhenomanes isolates from other hosts such as sugarcane, maize and several grasses, a link between pathogenic and genetic variability was detected. However, rice and maize isolates seemed to lack host specificity, and therefore crop rotation with maize might be a risky strategy to manage yield decline in Philippine aerobic rice fields.

Next to their essential roles in plant growth and development, phytohormones play a central role in plant immunity against pathogens. In this study we studied the previously reported antagonism between the plant-pathogenic oomycete Pythium arrhenomanes and the root-knot nematode Meloidogyne graminicola, two root pathogens that co-occur in aerobic rice fields. In this manuscript, we investigated if the antagonism is related to imbalances in plant hormone levels, which could be involved in activation of plant defense. Hormone measurements and gene expression analyses showed that the jasmonate (JA) pathway is induced early upon P. arrhenomanes infection. Exogenous application of methyl-jasmonate (MeJA) on the plant confirmed that JA is needed for basal defense against both P. arrhenomanes and M. graminicola in rice. Whereas M. graminicola suppresses root JA levels to increase host susceptibility, Pythium inoculation boosts JA in a manner that prohibits JA repression by the nematode in double-inoculated plants. Exogenous MeJA supply phenocopied the defense-inducing capacity of Pythium against the root-knot nematode, whereas the antagonism was weakened in JA-insensitive mutants. Transcriptome analysis confirmed upregulation of JA biosynthesis and signaling genes upon P. arrhenomanes infection, and additionally revealed induction of genes involved in biosynthesis of diterpenoid phytoalexins, consistent with strong activation of the gene encoding the JA-inducible transcriptional regulator DITERPENOID PHYTOALEXIN FACTOR. Altogether, the here-reported data indicate an important role for JA-induced defense mechanisms in this antagonistic interaction. Next to that, our results provide evidence for induced expression of genes encoding ERF83, and related PR proteins, as well as auxin depletion in P. arrhenomanes infected rice roots, which potentially further contribute to the reduced nematode susceptibility seen in double-infected plants.