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Receptor kinase (RK) families process information from small molecules, short peptides, or glycan ligands to regulate core cellular pathways in plants. To date, whether individual plant RKs are capable of processing signals from distinct types of ligands remains largely unexplored. Addressing this requires the discovery of structurally unrelated ligands that engage the same receptor. Here, we focus on FLAGELLIN-SENSING 2 (FLS2), an RK that senses a peptide of bacterial flagellin to activate antibacterial immunity in Arabidopsis. We interrogate >20,000 potential interactions between small molecules and the sensory domain of FLS2 using a large-scale reverse chemical screen. We discover two small molecules that interact with FLS2 in atypical ways. The surrogate ligands weakly activate the receptor to drive a functional antibacterial response channeled via unusual gene expression programs. Thus, chemical probes acting as biased ligands can be exploited to discover unexpected levels of output flexibility in RKs signal transduction., Competing Interests: Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

Flagellin, the protein subunit of the bacterial flagellum, stimulates the innate immune receptor Toll-like receptor 5 (TLR5) after pattern recognition or evades TLR5 through lack of recognition. This binary response fails to explain the weak agonism of flagellins from commensal bacteria, raising the question of how TLR5 response is tuned. Here, we screened abundant flagellins present in metagenomes from human gut for both TLR5 recognition and activation and uncovered a class of flagellin-TLR5 interaction termed silent recognition. Silent flagellins were weak TLR5 agonists despite pattern recognition. Receptor activity was tuned by a TLR5-flagellin interaction distal to the site of pattern recognition that was present in Salmonella flagellin but absent in silent flagellins. This interaction enabled flagellin binding to preformed TLR5 dimers and increased TLR5 signaling by several orders of magnitude. Silent recognition by TLR5 occurred in human organoids and mice, and silent flagellin proteins were present in human stool. These flagellins were produced primarily by the abundant gut bacteria Lachnospiraceae and were enriched in nonindustrialized populations. Our findings provide a mechanism for the innate immune system to tolerate commensal-derived flagellins while remaining vigilant to the presence of flagellins produced by pathogens.

This is the first study that examined and compared the survival, LC50, and RR50 estimates of Megacopta cribraria F. (Hemiptera: Plataspidae) nymphs and adults that were exposed to two native Beauveria bassiana isolates (Previously codified as NI8 and KUDSC strains) at four concentrations. The greatest reduction in survival and mortality was observed primarily on or after 10 d post-exposure to B. bassiana isolates. Survival of early instars (2nd, 3rd) were not affected by either strains or concentration at 3 d and 5 d post-exposure. Survival of later instars (5th) and adults was significantly reduced when exposed to the KUDSC strain at all concentrations. Comparison of dose−mortality values (LC50) using resistance ratios (RR50) were significantly different between life stages of the kudzu bug for both strains of B. bassiana. The LC50 values showed that kudzu bug adults are more susceptible than any other life stage when exposed to either strain. The KUDSC strain was more pathogenic than NI8 10 d after exposure, but NI8 exhibited significantly higher pathogenicity than KUDSC 20 d after exposure. Our results suggest potential field application of B. bassiana for kudzu bug control and their integration into pest management strategies to suppress them before they cause economic damage to soybean crops.

Immune systems respond to "non-self" molecules termed microbe-associated molecular patterns (MAMPs). Microbial genes encoding MAMPs have adaptive functions and are thus evolutionarily conserved. In the presence of a host, these genes are maladaptive and drive antagonistic pleiotropy (AP) because they promote microbe elimination by activating immune responses. The role AP plays in balancing the functionality of MAMP-coding genes against their immunogenicity is unknown. To address this, we focused on an epitope of flagellin that triggers antibacterial immunity in plants. Flagellin is conserved because it enables motility. Here, we decode the immunogenic and motility profiles of this flagellin epitope and determine the spectrum of amino acid mutations that drives AP. We discover two synthetic mutational tracks that undermine the detection activities of a plant flagellin receptor. These tracks generate epitopes with either antagonist or weaker agonist activities. Finally, we find signatures of these tracks layered atop each other in natural Pseudomonads., Competing Interests: Declaration of interests J.L.D. is a co-founder of, and shareholder in, AgBiome LLC, a corporation whose goal is to use plant-associated microbes to improve plant productivity., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Immune systems restrict microbial pathogens by identifying "non-self" molecules called microbe-associated molecular patterns (MAMPs). It is unclear how immune responses are tuned to or by MAMP diversity present in commensal microbiota. We systematically studied the variability of commensal peptide derivatives of flagellin (flg22), a MAMP detected by plants. We define substantial functional diversity. Most flg22 peptides evade recognition, while others contribute to evasion by manipulating immunity through antagonism and signal modulation. We establish a paradigm of signal integration, wherein the sequential signaling outputs of the flagellin receptor are separable and allow for reprogramming by commensal-derived flg22 epitope variants. Plant-associated communities are enriched for immune evading flg22 epitopes, but upon physiological stress that represses the immune system, immune-activating flg22 epitopes become enriched. The existence of immune-manipulating epitopes suggests that they evolved to either communicate or utilize the immune system for host colonization and thus can influence commensal microbiota community composition., Competing Interests: Declaration of interests J.L.D. is a co-founder of, and shareholder in, AgBiome LLC, a corporation whose goal is to use plant-associated microbes to improve plant productivity., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Methyl benzoate (MB) is a component of bee semiochemicals. Recent discovery of insecticidal activity of MB against insect pests provides a potential alternative to chemical insecticides. The aim of this study was to examine any potential adverse impact of MB on honey bees. By using two different methods, a spray for contact and feeding for oral toxicity, LC 50 s were 236.61 and 824.99 g a.i./L, respectively. The spray toxicity was 2002-fold and 173,163-fold lower than that of imidacloprid and abamectin. Piperonyl butoxide (PBO, inhibiting P450 oxidases [P450]) significantly synergized MB toxicity in honey bees, indicating P450s are the major MB-detoxification enzymes for bees. Assessing additive/synergistic interactions indicated that MB synergistically or additively aggravated the toxicity of all four insecticides (representing four different classes) in honey bees. Another adverse effect of MB in honey bees was the significant decrease of orientation and flight ability by approximately 53%. Other influences of MB included minor decrease of sucrose consumption, minor increase of P450 enzymatic activity, and little to no effect on esterase and glutathione S-transferase (GST) activities. By providing data from multiple experiments, we have substantially better understanding how important the P450s are in detoxifying MB in honey bees. MB could adversely affect feeding and flight in honey bees, and may interact with many conventional insecticides to aggravate toxicity to bees. However, MB is a relatively safe chemical to bees. Proper formulation and optimizing proportion of MB in mixtures may be achievable to enhance efficacy against pests and minimize adverse impact of MB on honey bees.

Receptor kinases of the Catharanthus roseus RLK1-like (CrRLK1L) family have emerged as important regulators of plant reproduction, growth and responses to the environment 1 . Endogenous RAPID ALKALINIZATION FACTOR (RALF) peptides 2 have previously been proposed as ligands for several members of the CrRLK1L family 1 . However, the mechanistic basis of this perception is unknown. Here we report that RALF23 induces a complex between the CrRLK1L FERONIA (FER) and LORELEI (LRE)-LIKE GLYCOSYLPHOSPHATIDYLINOSITOL (GPI)-ANCHORED PROTEIN 1 (LLG1) to regulate immune signalling. Structural and biochemical data indicate that LLG1 (which is genetically important for RALF23 responses) and the related LLG2 directly bind RALF23 to nucleate the assembly of RALF23-LLG1-FER and RALF23-LLG2-FER heterocomplexes, respectively. A conserved N-terminal region of RALF23 is sufficient for the biochemical recognition of RALF23 by LLG1, LLG2 or LLG3, and binding assays suggest that other RALF peptides that share this conserved N-terminal region may be perceived by LLG proteins in a similar manner. Structural data also show that RALF23 recognition is governed by the conformationally flexible C-terminal sides of LLG1, LLG2 and LLG3. Our work reveals a mechanism of peptide perception in plants by GPI-anchored proteins that act together with a phylogenetically unrelated receptor kinase. This provides a molecular framework for understanding how diverse RALF peptides may regulate multiple processes, through perception by distinct heterocomplexes of CrRLK1L receptor kinases and GPI-anchored proteins of the LRE and LLG family.

The rice immune receptor XA21 is activated by the sulfated microbial peptide required for activation of XA21-mediated immunity X (RaxX) produced by Xanthomonas oryzae pv. oryzae ( Xoo ). Mutational studies and targeted proteomics revealed that the RaxX precursor peptide (proRaxX) is processed and secreted by the protease/transporter RaxB, the function of which can be partially fulfilled by a noncognate peptidase-containing transporter component B (PctB). proRaxX is cleaved at a Gly-Gly motif, yielding a mature peptide that retains the necessary elements for RaxX function as an immunogen and host peptide hormone mimic. These results indicate that RaxX is a prokaryotic member of a previously unclassified and understudied group of eukaryotic tyrosine sulfated ribosomally synthesized, posttranslationally modified peptides (RiPPs). We further demonstrate that sulfated RaxX directly binds XA21 with high affinity. This work reveals a complete, previously uncharacterized biological process: bacterial RiPP biosynthesis, secretion, binding to a eukaryotic receptor, and triggering of a robust host immune response., Competing Interests: The authors declare no conflict of interest.

In this Letter, an incorrect version of the Supplementary Information file was inadvertently used, which contained several errors. The details of references 59-65 were missing from the end of the Supplementary Discussion section on page 4. In addition, the section 'Text 3. Y2H on ICD interactions' incorrectly referred to 'Extended Data Fig. 4d' instead of 'Extended Data Fig. 3d' on page 3. Finally, the section 'Text 4. Interaction network analysis' incorrectly referred to 'Fig. 1b and Extended Data Fig. 6' instead of 'Fig. 2b and Extended Data Fig. 7' on page 3. These errors have all been corrected in the Supplementary Information.

Background: The native bee fauna of Mississippi, USA has been historically poorly sampled, but is of particular relevance to determine range limits for species that occur in the southern United States. Currently published literature includes 184 species of bees that occur within the state of Mississippi. Additions to the list of native bees known for Mississippi are reported with notes on range, ecology and resources for identification., New Information: The geographic ranges of seven additional species are extended into the state of Mississippi: Andrena (Melandrena) obscuripennis Smith, 1853, Anthemurgus passiflorae Robertson, 1902, Dieunomia bolliana (Cockerell 1910), Diadasia (Diadasia) enavata (Cresson 1872), Peponapis crassidentata (Cockerell 1949), Triepeolus subnitens Cockerell and Timberlake, 1929 and Brachynomada nimia (Snelling and Rozen 1987). These records raise the total number of published species known from the state to 191. Anthemurgus and Brachynomada are also genera new to Mississippi.

Mitochondria are multifunctional organelles that play a central role in energy metabolism. Owing to the life-essential functions of these organelles, mitochondrial content, quality and dynamics are tightly controlled. Across the species, highly conserved ATP-dependent proteases prevent malfunction of mitochondria through versatile activities. This study focuses on a molecular function of the plant mitochondrial inner membrane-embedded AAA protease (denoted i -AAA) FTSH4, providing its first bona fide substrate. Here, we report that the abundance of the Tim17-2 protein, an essential component of the TIM17:23 translocase (Tim17-2 together with Tim50 and Tim23), is directly controlled by the proteolytic activity of FTSH4. Plants that are lacking functional FTSH4 protease are characterized by significantly enhanced capacity of preprotein import through the TIM17:23-dependent pathway. Taken together, with the observation that FTSH4 prevents accumulation of Tim17-2, our data point towards the role of this i -AAA protease in the regulation of mitochondrial biogenesis in plants., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2018. Published by The Company of Biologists Ltd.)

The cells of multicellular organisms receive extracellular signals using surface receptors. The extracellular domains (ECDs) of cell surface receptors function as interaction platforms, and as regulatory modules of receptor activation. Understanding how interactions between ECDs produce signal-competent receptor complexes is challenging because of their low biochemical tractability. In plants, the discovery of ECD interactions is complicated by the massive expansion of receptor families, which creates tremendous potential for changeover in receptor interactions. The largest of these families in Arabidopsis thaliana consists of 225 evolutionarily related leucine-rich repeat receptor kinases (LRR-RKs), which function in the sensing of microorganisms, cell expansion, stomata development and stem-cell maintenance. Although the principles that govern LRR-RK signalling activation are emerging, the systems-level organization of this family of proteins is unknown. Here, to address this, we investigated 40,000 potential ECD interactions using a sensitized high-throughput interaction assay, and produced an LRR-based cell surface interaction network (CSI LRR ) that consists of 567 interactions. To demonstrate the power of CSI LRR for detecting biologically relevant interactions, we predicted and validated the functions of uncharacterized LRR-RKs in plant growth and immunity. In addition, we show that CSI LRR operates as a unified regulatory network in which the LRR-RKs most crucial for its overall structure are required to prevent the aberrant signalling of receptors that are several network-steps away. Thus, plants have evolved LRR-RK networks to process extracellular signals into carefully balanced responses.

Maintenance of functional mitochondria is vital for optimal cell performance and survival. This is accomplished by distinct mechanisms, of which preservation of mitochondrial protein homeostasis fulfills a pivotal role. In plants, inner membrane-embedded i -AAA protease, FTSH4, contributes to the mitochondrial proteome surveillance. Owing to the limited knowledge of FTSH4's in vivo substrates, very little is known about the pathways and mechanisms directly controlled by this protease. Here, we applied substrate trapping coupled with mass spectrometry-based peptide identification in order to extend the list of FTSH4's physiological substrates and interaction partners. Our analyses revealed, among several putative targets of FTSH4, novel (mitochondrial pyruvate carrier 4 (MPC4) and Pam18-2) and known (Tim17-2) substrates of this protease. Furthermore, we demonstrate that FTSH4 degrades oxidatively damaged proteins in mitochondria. Our report provides new insights into the function of FTSH4 in the maintenance of plant mitochondrial proteome., Competing Interests: The authors declare no competing financial interests.

FTSH4 is one of the inner membrane-embedded ATP-dependent metalloproteases in mitochondria of Arabidopsis (Arabidopsis thaliana). In mutants impaired to express FTSH4, carbonylated proteins accumulated and leaf morphology was altered when grown under a short-day photoperiod, at 22°C, and a long-day photoperiod, at 30°C. To provide better insight into the function of FTSH4, we compared the mitochondrial proteomes and oxyproteomes of two ftsh4 mutants and wild-type plants grown under conditions inducing the phenotypic alterations. Numerous proteins from various submitochondrial compartments were observed to be carbonylated in the ftsh4 mutants, indicating a widespread oxidative stress. One of the reasons for the accumulation of carbonylated proteins in ftsh4 was the limited ATP-dependent proteolytic capacity of ftsh4 mitochondria, arising from insufficient ATP amount, probably as a result of an impaired oxidative phosphorylation (OXPHOS), especially complex V. In ftsh4, we further observed giant, spherical mitochondria coexisting among normal ones. Both effects, the increased number of abnormal mitochondria and the decreased stability/activity of the OXPHOS complexes, were probably caused by the lower amount of the mitochondrial membrane phospholipid cardiolipin. We postulate that the reduced cardiolipin content in ftsh4 mitochondria leads to perturbations within the OXPHOS complexes, generating more reactive oxygen species and less ATP, and to the deregulation of mitochondrial dynamics, causing in consequence the accumulation of oxidative damage., (© 2016 American Society of Plant Biologists. All Rights Reserved.)

The aim of this study was to examine the potential relationship between the occurrence of orthodontic root resorption and presence of dental anomalies such as tooth agenesis and pipette-shaped roots. Dental anomalies and root resorption were assessed on dental panoramic tomographs (DPT) of 88 subjects, 27 males and 61 females, mean age 28.4 (SD = 11.3 years), selected from orthodontic patients on the basis of the following exclusion criteria: previous fixed appliance treatment, bad quality of the DPTs and no visibility of the periodontal ligament of every tooth, and younger than 15 years of age at the onset of treatment with fixed edgewise appliance lasting at least 18 months. A pipette-shaped root was identified as defined by a drawing. Tooth agenesis was assessed on DPTs and from subjects' dental history. Root resorption was calculated as the difference between the root length before and after treatment, with and without a correction factor (crown length post-treatment/crown length pre-treatment). If one of the four upper incisors showed root resorption of ≥2.3 mm with both formulas, the patient was scored as having root resorption. Chi-square tests indicated that there was no relationship between orthodontic root resorption and agenesis (P = 0.885) nor between orthodontic root resorption and pipette-shaped roots (P = 0.800). There was no relationship between having one of the anomalies and root resorption either (P = 0.750). In the present study, it was not possible to confirm on DPTs a relationship between orthodontic root resorption and dental anomalies, such as agenesis and pipette-shaped roots.

The presence of dental anomalies has been rated radiographically in a number of studies. However, since the reliability of the assessment of these anomalies has rarely been investigated, the aim of this study was to examine inter- and intraexaminer agreement in identifying morphological dental anomalies, such as tooth agenesis, dilacerated, pipette-shaped, blunt, pointed, and short roots. Pre-treatment panoramic radiographs of 40 patients (13 males and 27 females; mean age 27.7 ± 10.8 years) treated between 1983 and 2008 were selected. Four examiners independently assessed the radiographs twice. For a dilacerated root and agenesis, a definition was given. For pipette-shaped, blunt, or pointed roots, a drawing was shown, and for a short root, a ratio was used to identify the anomaly. Intraexaminer agreement of the assessments of the dental anomalies was presented by Cohen's Kappa and varied between -0.01 for short roots and 1.00 for agenesis. With respect to short roots, three of the examiners did not rate them to be present on at least one measurement occasion. This implies that intraexaminer agreement could not be calculated for these three examiners. Interexaminer agreement for dilacerated roots varied between 0.14 and 0.50, for pipette-shaped roots between -0.01 and 0.33, for blunt roots between 0.05 and 0.32, and for pointed roots between 0.17 and 0.37. All values for agenesis were 1.00. It can be concluded that assessing agenesis on panoramic radiographs is reliable. Rating the presence of dilacerated, pipette-shaped, blunt, pointed, and short roots on panoramic radiographs, however, does not result in a reliable assessment.

Study Objective: Automatic titration using the forced oscillation technique (FOT) has recently been developed for the treatment of obstructive sleep apnea syndrome (OSAS). So far, it is not known if therapy with automatic nasal continuous positive airway pressure (nCPAP) using a preset upper pressure limitation or a free range (which might lead to higher mean pressure) is preferable with regard to obstructive events, sleep stages, and pressure characteristics., Design: After diagnostic polysomnography, patients were randomly assigned to two settings with the self-adjusting nCPAP (APAP) device based on the FOT. In mode 1, the pressure variation ranged from 4 to 15.5 cm H(2)O, and in mode 2, the pressure variation ranged from 4 cm H(2)O to an individual upper pressure limit., Patients: Eleven men, aged 53.0 +/- 6.8 years with a body mass index of 32.4 +/- 5.1 kg/m(2) and an apnea-hypopnea index (AHI) of 31.6 +/- 26.6/h., Measurements and Results: Manually titrated pressure was at 9.3 +/- 2.1 cm H(2)O, the mean pressure in mode 1 was 5.4 +/- 1.0 cm H(2)O (p < 0.01), and the mean pressure in mode 2 was 5.1 +/- 0.7 cm H(2)O (p < 0.01). A reduction of respiratory events (baseline AHI, 31.6 +/- 26.6/h; AHI in mode 1, 3.4 +/- 4.5; AHI in mode 2, 5.0 +/- 7.2; each with p < 0.001) and an increase in the "rapid eye movement" stage of sleep (baseline, 13.0 +/- 5.5%; mode 1, 22.0 +/- 7.7 [p < 0. 05]; mode 2, 23.0 +/- 7.9 [p < 0.01]) were achieved. In mode 1, the mean pressure was below the manual pressure 91.7 +/- 9.3% of the time, and in mode 2, the mean pressure was below the manual pressure 90.4 +/- 6.3% of the time. The manual pressure was exceeded by 5.5 +/- 7.4% (mode 1) and by 5.2 +/- 3.1% (mode 2)., Conclusion: We conclude that nCPAP therapy based on the FOT permits the adequate treatment of OSAS with significantly lower pressure than manually titrated nCPAP therapy does. A presetting of an upper pressure limit has no advantage compared to free range.