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Abstract Background Wheat (Triticum aestivum L.) O-methyltransferase (TaOMT2) catalyzes the sequential methylation of the flavone, tricetin, to its 3'-methyl- (selgin), 3',5'-dimethyl- (tricin) and 3',4',5'-trimethyl ether derivatives. Tricin, a potential multifunctional nutraceutical, is the major enzyme reaction product. These successive methylations raised the question as to whether they take place in one, or different active sites. We constructed a 3-D model of this protein using the crystal structure of the highly homologous Medicago sativa caffeic acid/5-hydroxyferulic acid O-methyltransferase (MsCOMT) as a template with the aim of proposing a mechanism for multiple methyl transfer reactions in wheat. Results This model revealed unique structural features of TaOMT2 which permit the stepwise methylation of tricetin. Substrate binding is mediated by an extensive network of H-bonds and van der Waals interactions. Mutational analysis of structurally guided active site residues identified those involved in binding and catalysis. The partly buried tricetin active site, as well as proximity and orientation effects ensured sequential methylation of the substrate within the same pocket. Stepwise methylation of tricetin involves deprotonation of its hydroxyl groups by a His262-Asp263 pair followed by nucleophilic attack of SAM-methyl groups. We also demonstrate that Val309, which is conserved in a number of graminaceous flavone OMTs, defines the preference of TaOMT2 for tricetin as the substrate. Conclusions We propose a mechanism for the sequential methylation of tricetin, and discuss the potential application of TaOMT2 to increase the production of tricin as a nutraceutical. The single amino acid residue in TaOMT2, Val309, determines its preference for tricetin as the substrate, and may define the evolutionary differences between the two closely related proteins, COMT and flavone OMT.

Rackwitz–Fiessler (RF) method is well accepted as an efficient way to solve the uncorrelated non-Normal reliability problems by transforming original non-Normal variables into equivalent Normal variables based on the equivalent Normal conditions. However, this traditional RF method is often abandoned when correlated reliability problems are involved, because the point-by-point implementation property of equivalent Normal conditions makes the RF method hard to clearly describe the correlations of transformed variables. To this end, some improvements on the traditional RF method are presented from the isoprobabilistic transformation and copula theory viewpoints. First of all, the forward transformation process of RF method from the original space to the standard Normal space is interpreted as the isoprobabilistic transformation from the geometric point of view. This viewpoint makes us reasonably describe the stochastic dependence of transformed variables same as that in Nataf transformation (NATAF). Thus, a corresponding enhanced RF (EnRF) method is proposed to deal with the correlated reliability problems described by Pearson linear correlation. Further, we uncover the implicit Gaussian copula hypothesis of RF method according to the invariant theorem of copula and the strictly increasing isoprobabilistic transformation. Meanwhile, based on the copula-only rank correlations such as the Spearman and Kendall correlations, two improved RF (IRF) methods are introduced to overcome the potential pitfalls of Pearson correlation in EnRF. Later, taking NATAF as a reference, the computational cost and efficiency of above three proposed RF methods are also discussed in Hasofer–Lind reliability algorithm. Finally, four illustrative structure reliability examples are demonstrated to validate the availability and advantages of the new proposed RF methods. [ABSTRACT FROM AUTHOR]

Plants employ numerous cell-surface and intracellular immune receptors to perceive a variety of immunogenic signals associated with pathogen infection and subsequently activate defenses. Immune signaling is potentiated by the major defense hormone salicylic acid (SA), which reprograms the transcriptome for defense. Here we highlight recent advances in understanding the mechanisms underlying activation of the main classes of immune receptors, summarize the current understanding of their signaling mechanisms, and discuss an updated model for SA perception and signaling. In addition, we discuss how different receptors are organized into networks and the implications of such networks in the integration of complex danger signals for appropriate defense outputs. Plants possess a sophisticated and complex immune signaling system, many components or which are either conserved or have analogous counterparts in animals. Jian-Min Zhou and Yuelin Zhang review plant immune signaling mechanisms, from sensing mechanisms to transcriptional responses, noting key parallels to animal immunity. [ABSTRACT FROM AUTHOR]

Plants utilize nucleotide-binding, leucine-rich repeat receptors (NLRs) to detect pathogen effectors, leading to effector-triggered immunity. The NLR ZAR1 indirectly recognizes the Xanthomonas campestris pv. campestris effector AvrAC and Pseudomonas syringae effector HopZ1a by associating with closely related receptor-like cytoplasmic kinase subfamily XII-2 (RLCK XII-2) members RKS1 and ZED1, respectively. ZAR1, RKS1, and the AvrAC-modified decoy PBL2UMP form a pentameric resistosome in vitro , and the ability of resistosome formation is required for AvrAC-triggered cell death and disease resistance. However, it remains unknown whether the effectors induce ZAR1 oligomerization in the plant cell. In this study, we show that both AvrAC and HopZ1a can induce oligomerization of ZAR1 in Arabidopsis protoplasts. Residues mediating ZAR1–ZED1 interaction are indispensable for HopZ1a-induced ZAR1 oligomerization in vivo and disease resistance. In addition, ZAR1 residues required for the assembly of ZAR1 resistosome in vitro are also essential for HopZ1a-induced ZAR1 oligomerization in vivo and disease resistance. Our study provides evidence that pathogen effectors induce ZAR1 resistosome formation in the plant cell and that the resistosome formation triggers disease resistance. This study shows that the bacterial pathogen effectors AvrAC and HopZ1a induce oligomerization of the NLR protein ZAR1 in Arabidopsis protoplasts. Structural requirements for ZAR1 resistosome assembly in vitro are reveavled to be essential for HopZ1a-induced ZAR1 oligomerization in vivo and disease resistance in plants, suggesting that the ZAR1 resistosome forms in vivo during immune activation. [ABSTRACT FROM AUTHOR]