Background: Extensive studies have demonstrated that the COBRA gene is critical for biosynthesis of cell wall constituents comprising structural tissues of roots, stalks, leaves and other vegetative organs, however, its role in fruit development and ripening remains largely unknown. Results: We identified a tomato gene (SlCOBRA-like) homologous to Arabidopsis COBRA, and determined its role in fleshy fruit biology. The SlCOBRA-like gene is highly expressed in vegetative organs and in early fruit development, but its expression in fruit declines dramatically during ripening stages, implying a primary role in early fruit development. Fruit-specific suppression of SlCOBRA-like resulted in impaired cell wall integrity and up-regulation of genes encoding proteins involved in cell wall degradation during early fruit development. In contrast, fruit-specific overexpression of SlCOBRA-like resulted in increased wall thickness of fruit epidermal cells, more collenchymatous cells beneath the epidermis, elevated levels of cellulose and reduced pectin solubilization in the pericarp cells of red ripe fruits. Moreover, transgenic tomato fruits overexpressing SlCOBRA-like exhibited desirable early development phenotypes including enhanced firmness and a prolonged shelf life. Conclusions: Our results suggest that SlCOBRA-like plays an important role in fruit cell wall architecture and provides a potential genetic tool for extending the shelf life of tomato and potentially additional fruits.

Plant pathogens, such as bacteria, fungi, oomycetes and nematodes, rely on wide range of virulent effectors delivered into host cells to suppress plant immunity. Although phytobacterial effectors have been intensively investigated, little is known about the function of effectors of plant-parasitic nematodes, such as Globodera pallida, a cyst nematode responsible for vast losses in the potato and tomato industries. Here, we demonstrate using in vivo and in vitro ubiquitination assays the potato cyst nematode (Globodera pallida) effector RHA1B is an E3 ubiquitin ligase that employs multiple host plant E2 ubiquitin conjugation enzymes to catalyze ubiquitination. RHA1B was able to suppress effector-triggered immunity (ETI), as manifested by suppression of hypersensitive response (HR) mediated by a broad range of nucleotide-binding leucine-rich repeat (NB-LRR) immune receptors, presumably via E3-dependent degradation of the NB-LRR receptors. RHA1B also blocked the flg22-triggered expression of Acre31 and WRKY22, marker genes of pathogen‐associated molecular pattern (PAMP)‐triggered immunity (PTI), but this did not require the E3 activity of RHA1B. Moreover, transgenic potato overexpressing the RHA1B transgene exhibited enhanced susceptibility to G. pallida. Thus, our data suggest RHA1B facilitates nematode parasitism not only by triggering degradation of NB-LRR immune receptors to block ETI signaling but also by suppressing PTI signaling via an as yet unknown E3-independent mechanism. Globodera pallida is a plant-parasitic cyst nematode that causes vast losses in economically important crops such as potato and tomato. To successfully parasitize host plants, G. pallida produces proteins called effectors to overcome plant defenses. Here, we report identification of a novel G. pallida effector RHA1B as an E3 ubiquitin ligase, which is responsible for ubiquitin-proteasome-mediated protein degradation in general. We found that RHA1B can suppress plant defense signaling via both E3-dependent and -independent manners. In particular, it promotes degradation of a broad range of NB-LRR immune receptors. In addition, expression of RHA1B in potato plants made the plants more susceptible to G. pallida infection, indicating that RHA1B acts as an effector that aids parasitism. Overall, we found RHA1B as the first effector with ubiquitin ligase activity identified from eukaryotic pathogen infecting plants or animals. Our data suggest nematode uses RHA1B as a powerful weapon to manipulate host cellular signaling pathways, thereby interfering with plant immunity for successful parasitism.

Summary: The plant‐specific NAC ( NAM, ATAF1,2, CUC2) transcription factors play significant roles in diverse physiological processes. In this study, we determined the regulation of a stress‐related tomato (Solanum lycopersicum) NAC1 (SlNAC1) transcription factor at both the transcriptional and the post‐translational level. The SlNAC1 protein was found to be stable in the presence of proteasome‐specific inhibitor MG132 or MG115 and ubiquitinated in plant cells, suggesting that the SlNAC1 is subject to the ubiquitin–proteasome system‐mediated degradation. Deletion analysis identified a short segment of 10 amino acids (aa261–270) that was required for ubiquitin–proteasome system‐mediated degradation, among which two leucine residues (L268 and L269) were critical for the protein instability of SlNAC1. Fusion of the degron (SlNAC1_191–270) containing these 10 amino acids to green fluorescent protein was found to be sufficient to trigger the degradation of the fusion protein. In addition, the SlNAC1 gene is strongly upregulated during Pseudomonas infection, while repression of the NAC1 ortholog in Nicotiana benthamiana resulted in enhanced susceptibility to Pseudomonas bacteria. These results suggest that rapid upregulation of the NAC1 gene resulting in more protein production is likely one of the strategies plants use to defend themselves against pathogen infection.

BAK1 (brassinosteroid-insensitive 1 (BRI1) associated receptor kinase 1) plays major roles in multiple signaling pathways as a coreceptor to regulate plant growth and development and stress response. However, the role of BAK1 in high light signaling is still poorly understood. Here we observed that overexpression of BAK1 in Arabidopsis interferes with the function of high light in promoting plant growth and development, which is independent of the brassinosteroid (BR) signaling pathway. Further investigation shows that high light enhances the phosphorylation of BAK1 and catalase activity, thereby reducing hydrogen peroxide (H₂O₂) accumulation. Catalase3 (CAT3) is identified as a BAK1-interacting protein by affinity purification and LC-MS/MS analysis. Biochemical analysis confirms that BAK1 interacts with and phosphorylates all three catalases (CAT1, CAT2, and CAT3) of the Arabidopsis genome, and the trans-phosphorylation sites of three catalases with BAK1-CD are identified by LC-MS/MS in vitro. Genetic analyses reveal that the BAK1 overexpression plants knocked out all the three CAT genes completely abolishing the effect of BAK1 on suppression of high light-promoted growth. This study first unravels the role of BAK1 in mediating high light-triggered activation of CATs, thereby degrading H₂O₂ and regulating plant growth and development in Arabidopsis.