Polar auxin transport (PAT) is facilitated by polar localization of PIN-FORMED (PIN) efflux carriers, which direct auxin flow and regulate developmental events. Brassinosteroids (BRs) and auxin work synergistically to promote growth, and in root geotropisms this cross-talk involves BR-directed polarization of PIN through the mobilization of F-actin. However, the role of BR in PAT during shoot growth, hair formation, and embryogenesis has not been well studied. Orchid seed are mature at a point in development that is analogous to the globular-stage of embryogenesis in typical angiosperms. Thus, this system provided a unique opportunity to study the effects of BR on PAT during embryogenesis-like events, including meristem/first leaf formation and protocorm/stem development, which is followed by protocorm hair formation. In this work, the degree to which BRs rescued embryo-like protocorms from the impact of PAT-disrupting agents, such as PAT inhibitors or high auxin levels, was determined based on growth responses. This study first established that auxin and BRs work together synergistically to promote seedling elongation in Spathoglottis. Repressed seedling growth caused by the PAT-disrupting agents was alleviated with eBL, suggesting that BRs enhance PAT in embryogenesis-like stages of young protocorms. However, similar responses were not evident in seed embryos. Results from this study also suggested that BRs may enhance orchid protocorm elongation by regulating auxin transport through an F-actin-mediated mechanism. With regard to protocorm hairs, increased eBL levels inhibited formation, whereas reduced BR biosynthesis altered hair patterning, and prevented outgrowth of auxin-stimulated hairs. Moreover, PAT inhibitors and repression of BR biosynthesis caused hair bud formation without hair outgrowth, suggesting a role for BR in PAT during protocorm hair development.

The ecology and physiology of ectomycorrhizal (EcM) symbiosis with conifer trees are well documented. In comparison, however, very little is known about the molecular regulation of these associations. In an earlier study, we identified three EcM-regulated Pinus expressed sequence tags (EST), two of which were identified as homologous to the Medicago truncatula nodulin MtN21. The third EST was a homologue to the receptor-like kinase Clavata1. We have characterized the expression patterns of these genes and of auxin- and mycorrhiza-regulated genes after induction with indole-3-butyric acid in Pinus sylvestris and in a time course experiment during ectomycorrhizal initiation with the co-inoculation of 2,3,5-triiodobenzoic acid, an auxin transport inhibitor. Our results suggest that different P. sylvestris nodulin homologues are associated with diverse processes in the root. The results also suggest a potential role of the Clv1-like gene in lateral root initiation by the ectomycorrhizal fungus.

Little is known about the roles of peroxisomes in the necrotrophic fungal plant pathogens. In the present study, a Pex6 gene encoding an ATPase-associated protein was characterized by analysis of functional mutations in the tangerine pathotype of Alternaria alternata, which produces a host-selective toxin. Peroxisomes were observed in fungal cells by expressing a mCherry fluorescent protein tagging with conserved tripeptides serine-lysing-leucine and transmission electron microscopy. The results indicated that Pex6 plays no roles in peroxisomal biogenesis but impacts protein import into peroxisomes. The number of peroxisomes was affected by nutritional conditions and H2O2, and their degradation was mediated by an autophagy-related machinery termed pexophagy. Pex6 was shown to be required for the formation of Woronin bodies, the biosynthesis of biotin, siderophores, and toxin, the uptake and accumulation of H2O2, growth, and virulence, as well as the Slt2 MAP kinase-mediated maintenance of cell wall integrity. Adding biotin, oleate, and iron in combination fully restored the growth of the pex6-deficient mutant (Δpex6), but failed to restore Δpex6 virulence to citrus. Adding purified toxin could only partially restore Δpex6 virulence even in the presence of biotin, oleate, and iron. Sensitivity assays revealed that Pex6 plays no roles in resistance to H2O2 and superoxide, but plays a negative role in resistance to 2-chloro-5-hydroxypyridine (a hydroxyl radical-generating compound), eosin Y and rose Bengal (singlet oxygen-generating compounds), and 2,3,5-triiodobenzoic acid (an auxin transport inhibitor). The diverse functions of Pex6 underscore the importance of peroxisomes in physiology, pathogenesis, and development in A. alternata.

Plants, like other multicellular organisms, survive through a delicate balance between growth and defense against pathogens. Salicylic acid (SA) is a major defense signal in plants, and the perception mechanism as well as downstream signaling activating the immune response are known. Here, we identify a parallel SA signaling that mediates growth attenuation. SA directly binds to A subunits of protein phosphatase 2A (PP2A), inhibiting activity of this complex. Among PP2A targets, the PIN2 auxin transporter is hyperphosphorylated in response to SA, leading to changed activity of this important growth regulator. Accordingly, auxin transport and auxin-mediated root development, including growth, gravitropic response, and lateral root organogenesis, are inhibited. This study reveals how SA, besides activating immunity, concomitantly attenuates growth through crosstalk with the auxin distribution network. Further analysis of this dual role of SA and characterization of additional SA-regulated PP2A targets will provide further insights into mechanisms maintaining a balance between growth and defense.