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Berry skin colour is a crucial determinant of red/black grape berry quality. We investigated the effects of combination treatments with amino acids and a low concentration of ABA on anthocyanin accumulation in grapes. Among the amino acids tested, isoleucine and phenylalanine resulted in high anthocyanin contents in grape cell cultures. The combination treatments with isoleucine or phenylalanine, and a low concentration of ABA enhanced anthocyanin accumulation in grape cells and detached grape berries. The combination treatment with isoleucine, but not with phenylalanine, and ABA upregulated MybA1 expression. Field-grown grapevines received combination treatments with isoleucine or phenylalanine, and ABA in two growing seasons. In the 2015 growing season, the combination treatments with isoleucine or phenylalanine, and a low concentration of ABA accelerated anthocyanin accumulation in grape berry skins of field-grown grapevines on days 10 and 31 post treatment. The effects on anthocyanin accumulation became negligible at harvest. The effect of the combination treatment with phenylalanine and a low concentration of ABA on anthocyanin accumulation was masked in the 2017 growing season due to the unexpected stimulation of anthocyanin accumulation by the low concentration of ABA, although the combination treatment accelerated anthocyanin accumulation on days 3 and 10 post treatment. Taken together, the results suggested that exogenous isoleucine and phenylalanine interacted with ABA-mediated anthocyanin accumulation in grape berry skins of field-grown grapevines when the activity of ABA used to treat the grapevines was inadequate.

Network robustness is a crucial property of the plant immune signaling network because pathogens are under a strong selection pressure to perturb plant network components to dampen plant immune responses. Nevertheless, modulation of network robustness is an area of network biology that has rarely been explored. While two modes of plant immunity, Effector-Triggered Immunity (ETI) and Pattern-Triggered Immunity (PTI), extensively share signaling machinery, the network output is much more robust against perturbations during ETI than PTI, suggesting modulation of network robustness. Here, we report a molecular mechanism underlying the modulation of the network robustness in Arabidopsis thaliana. The salicylic acid (SA) signaling sector regulates a major portion of the plant immune response and is important in immunity against biotrophic and hemibiotrophic pathogens. In Arabidopsis, SA signaling was required for the proper regulation of the vast majority of SA-responsive genes during PTI. However, during ETI, regulation of most SA-responsive genes, including the canonical SA marker gene PR1, could be controlled by SA-independent mechanisms as well as by SA. The activation of the two immune-related MAPKs, MPK3 and MPK6, persisted for several hours during ETI but less than one hour during PTI. Sustained MAPK activation was sufficient to confer SA-independent regulation of most SA-responsive genes. Furthermore, the MPK3 and SA signaling sectors were compensatory to each other for inhibition of bacterial growth as well as for PR1 expression during ETI. These results indicate that the duration of the MAPK activation is a critical determinant for modulation of robustness of the immune signaling network. Our findings with the plant immune signaling network imply that the robustness level of a biological network can be modulated by the activities of network components. [ABSTRACT FROM AUTHOR]

To study the signal transduction of cytokinins, we characterized cytokinin-binding proteins (CBPs) isolated from tobacco callus Nicotiana tabacum. Two high-affinity CBPs, CBP1 and CBP2, were isolated from the soluble fraction of tobacco callus BY-2 cells by anion exchange chromatography on a DEAE-cellulose column and affinity chromatography on a benzyladenine (BA)-linked Sepharose 4B column. Cytokinin-binding activity was determined by the equilibrium dialysis method. The degree of purification of CBP1 and CBP2 was 270 and 600-fold, respectively. These proteins had molecular masses of 34 kDa and 26 kDa, and to bind benzyladenine (BA) with dissociation constants (Kd) of 8.9 × 10−6 M and 1.1 × 10−6 M, respectively. Binding of BA to CBP2 was inhibited by zeatin and kinetin but not by adenine, adenosine, ATP or IAA. The optimum pH for binding of BA to CBP1 and CBP2 was approximately pH 6.5 and 7.5, respectively. CBP1 showed significant homology (90%) with endochitinase and CBP2 with osmotin-like protein (OLP). These findings and the results of immunoblotting analysis and cytokininbinding assay of recombinant OLP indicated that CBP2 is OLP, a stress protein. [ABSTRACT FROM AUTHOR]

Network robustness is a crucial property of the plant immune signaling network because pathogens are under a strong selection pressure to perturb plant network components to dampen plant immune responses. Nevertheless, modulation of network robustness is an area of network biology that has rarely been explored. While two modes of plant immunity, Effector-Triggered Immunity (ETI) and Pattern-Triggered Immunity (PTI), extensively share signaling machinery, the network output is much more robust against perturbations during ETI than PTI, suggesting modulation of network robustness. Here, we report a molecular mechanism underlying the modulation of the network robustness in Arabidopsis thaliana. The salicylic acid (SA) signaling sector regulates a major portion of the plant immune response and is important in immunity against biotrophic and hemibiotrophic pathogens. In Arabidopsis, SA signaling was required for the proper regulation of the vast majority of SA-responsive genes during PTI. However, during ETI, regulation of most SA-responsive genes, including the canonical SA marker gene PR1, could be controlled by SA-independent mechanisms as well as by SA. The activation of the two immune-related MAPKs, MPK3 and MPK6, persisted for several hours during ETI but less than one hour during PTI. Sustained MAPK activation was sufficient to confer SA-independent regulation of most SA-responsive genes. Furthermore, the MPK3 and SA signaling sectors were compensatory to each other for inhibition of bacterial growth as well as for PR1 expression during ETI. These results indicate that the duration of the MAPK activation is a critical determinant for modulation of robustness of the immune signaling network. Our findings with the plant immune signaling network imply that the robustness level of a biological network can be modulated by the activities of network components. [ABSTRACT FROM AUTHOR]

Objectives: We aim to evaluate the risk of recurrence after neoadjuvant chemotherapy followed by radical cystectomy, particularly in ypT2 disease in patients with urothelial carcinoma, because it is not clear if all eligible patients with high-risk muscle-invasive urothelial carcinoma should be treated with adjuvant nivolumab., Materials and Methods: We analysed the radiological and clinicopathological features, including cT and ypT stages, of 197 patients who had undergone two to four cycles of cisplatin-based neoadjuvant chemotherapy and radical cystectomy without adjuvant chemotherapy. We stratified the risk of postoperative recurrence by these factors., Results: The median observation period was 29.6 (interquartile range, 11.4-71.7) months, and disease recurrence was observed in 58 patients. Multivariate analysis revealed that ypT stage ( P  = 0.019) and lymphovascular invasion ( P  = 0.015) were independent risk factors for postoperative recurrence. The ypT2 group ( n  = 38) had significantly better recurrence-free survival than the ypT3 group ( n  = 41) (median recurrence-free survival: not reached vs. 13.4 months, respectively, P  = 0.005). In ypT2 disease, the cT2 and ypT2 group ( n  = 15), which was diagnosed as cT2 preoperatively and then diagnosed as ypT2 postoperatively, had significantly better recurrence-free survival than the cT3/4 and ypT2 group ( n  = 23) (median recurrence-free survival: not reached vs. 63.1 months, respectively, P  = 0.034). There was no significant difference in recurrence-free survival between the ypT ≤ 1 ( n  = 106) and the cT2 and ypT2 groups (median recurrence-free survival: not reached in both, P  = 0.962)., Conclusion: Patients with cT2 and ypT2 stage have a relatively low risk of recurrence and thus have a lower need for adjuvant nivolumab, particularly those with ypT2., Competing Interests: All authors have no conflict of interest to disclose., (© 2023 The Authors. BJUI Compass published by John Wiley & Sons Ltd on behalf of BJU International Company.)