The effector protein AvrP is secreted by the flax rust fungal pathogen (Melampsora lini) and recognized specifically by the flax (Linum usitatissimum) P disease resistance protein, leading to effector-triggered immunity. To investigate the biological function of this effector and the mechanisms of specific recognition by the P resistance protein, we determined the crystal structure of AvrP. The structure reveals an elongated zinc-finger-like structure with a novel interleaved zinc-binding topology. The residues responsible for zinc binding are conserved in AvrP effector variants and mutations of these motifs result in a loss of P-mediated recognition. The first zinc-coordinating region of the structure displays a positively charged surface and shows some limited similarities to nucleic acid-binding and chromatin-associated proteins. We show that the majority of the AvrP protein accumulates in the plant nucleus when transiently expressed in Nicotiana benthamiana cells, suggesting a nuclear pathogenic function. Polymorphic residues in AvrP and its allelic variants map to the protein surface and could be associated with differences in recognition specificity. Several point mutations of residues on the non-conserved surface patch result in a loss of recognition by P, suggesting that these residues are required for recognition.

Microbes have been the preferred hosts for producing high-value chemicals from cheap raw materials. However, metabolic flux imbalance, the presence of competing pathways, and toxic intermediates often lead to low production efficiency. The spatial organization of the substrates, intermediates, and enzymes is critical to ensuring efficient metabolic activity by microorganisms. One of the most common approaches for bringing the key components of biosynthetic pathways together is through molecular scaffolds, which involves the clustering of pathway enzymes on engineered molecules via different interacting mechanisms. In particular, synthetic scaffold systems have been applied to improve the efficiency of various heterologous and synthetic pathways in Escherichia coli and Saccharomyces cerevisiae, with varying degrees of success. Herein, we review the recent developments and applications of protein-based and nucleic acid-based scaffold systems and discuss current challenges and future directions in the use of such approaches.

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Secreted fungal effector proteins and their host targets are good examples to understand the mechanism of host-pathogen co-evolution with genes involved in the interaction undergoing positive selection. SIX genes (secreted in xylem) are obtained via horizontal transfer and can be found within the formae speciales of Fusarium oxysporum. SIX6 and SIX9 of F. oxysporum f. spp. cubense (Foc) are predicted to play a role as effectors. However, their involvement in the pathogenicity of Foc in banana plants has not been determined yet. In the susceptible banana cultivar, we found that the SIX6 and SIX9 genes of Foc TR4 were highly expressed in roots, but not in corms or leaves. The host, however, expressed the pathogenesis-related (PR) genes, PR-1 and PR-3, in corms earlier than in the roots. Phylogenetic analysis on SIX6 and SIX9 genes of F. oxysporum has revealed the separation of SIX6 and SIX9 of Foc from other formae speciales. This leads to detecting genes under positive selection using the ratio nonsynonymous to synonymous substitution rates (Ka/Ks). SIX6 of Foc showed an increase in diversity, but insufficient to drive positive selection. Conversely, SIX9 of Foc showed no divergence in the dN/dS ratio distribution, indicating purifying selection. © 2021 Friends Science Publishers