Trichoderma-secreted anthranilic acid promotes lateral root development via auxin signaling and RBOHF-induced endodermal cell wall remodeling.

Trichoderma spp. have evolved the capacity to communicate with plants by producing various secondary metabolites (SMs). Nonhormonal SMs play important roles in plant root development, while specific SMs from rhizosphere microbes and their underlying mechanisms to control plant root branching are still largely unknown. In this study, a compound, anthranilic acid (2-AA), is identified from T. guizhouense NJAU4742 to promote lateral root development. Further studies demonstrate that 2-AA positively regulates auxin signaling and transport in the canonical auxin pathway. 2-AA also partly rescues the lateral root numbers of CASP1pro:shy2-2, which regulates endodermal cell wall remodeling via an RBOHF-induced reactive oxygen species burst. In addition, our work reports another role for microbial 2-AA in the regulation of lateral root development, which is different from its better-known role in plant indole-3-acetic acid biosynthesis. In summary, this study identifies 2-AA from T. guizhouense NJAU4742, which plays versatile roles in regulating plant root development.

Clustered surface amino acid residues modulate the acid stability of GH10 xylanase in fungi.

Acidic xylanases are widely used in industries such as biofuels, animal feeding, and fruit juice clarification due to their tolerance to acidic environments. However, the factors controlling their acid stability, especially in GH10 xylanases, are only partially understood. In this study, we identified a series of thermostable GH10 xylanases with optimal temperatures ranging from 70 to 90 °C, and among these, five enzymes (Xyn10C, Xyn10RE, Xyn10TC, Xyn10BS, and Xyn10PC) exhibited remarkable stability at pH 2.0. Our statistical analysis highlighted several factors contributing to the acid stability of GH10 xylanases, including electrostatic repulsion, π-π stacking, ionic bonds, hydrogen bonds, and Van der Waals interactions. Furthermore, through mutagenesis studies, we uncovered that acid stability is influenced by a complex interplay of amino acid residues. The key amino acid sites determining the acid stability of GH10 xylanases were thus elucidated, mainly concentrated in two surface regions behind the enzyme active center. Notably, the critical residues associated with acid stability markedly enhanced Xyn10RE's thermostability by more than sixfold, indicating a potential acid-thermal interplay in GH10 xylanases. This study not only reported a series of valuable genes but also provided a range of modification targets for enhancing the acid stability of GH10 xylanases. KEY POINTS: • Five acid stable and thermostable GH10 xylanases were reported. • The key amino acid sites, mainly forming two enriched surface regions behind the enzyme active center, were identified responsible for acid stability of GH10 xylanases. • The finding revealed interactive amino acid sites, offering a pathway for synergistic enhancement of both acid stability and thermostability in GH10 xylanase modifications.

Repellent effect of two anthranilates against the tropical bed bug Cimex hemipterus (Hemiptera: Cimicidae) demonstrated in laboratory assays.

BACKGROUND: The bites and blood sucking of bed bugs (Cimex spp.) (Hemiptera: Cimicidae) pose a serious threat to human physical and mental health. Application of an effective repellent can prevent or reduce bed bug bites. Previous studies on repellent screening mainly focused on Cimex lectularius L. In this study, we investigated the repellent effect of two safe food additives, ethyl anthranilate (EA) and butyl anthranilate (BA), against Cimex hemipterus (F.), and also explored the role of antennae and mouthparts on C. hemipterus perception of repellents. RESULTS: Both EA and BA had a strong repellent effect against tropical bed bugs and their repellency was similar or lower than that of N,N-diethyl-3-methyl benzoyl amide, depending on whether or not a CO2 source was present. EA had higher repellency than BA and exhibited repellency to C. hemipterus for 3 days when a CO2 source was present. C. hemipterus avoided resting on 20% EA- and BA-treated harborages. Applying 20% EA and BA on rabbit skin significantly reduced the blood intake of C. hemipterus within 2 h. C. hemipterus could perceive EA and BA after their antennae or mouthparts or both antennae and mouthparts were removed. CONCLUSION: Both EA and BA had strong repellency against C. hemipterus, with EA being more repellent. Ablation of antennae and mouthparts did not affect the perception of EA and BA by C. hemipterus. © 2023 Society of Chemical Industry.

A Case Study on Tropical Bed Bug, Cimex hemipterus (Hemiptera: Cimicidae) Infestation and Management in Dormitories.

Numerous bed bug research papers have been published in the past 20 yr as a result of bed bug (Cimex spp.) (Hemiptera: Cimicidae) resurgence in the world. Yet, few of them focused on the management of the tropical bed bug, C. hemipterus (F.). Here, we describe a case of tropical bed bug infestation in two dormitory buildings and effectiveness of a tropical bed bug treatment program. The study site consisted of 125 dormitories in two buildings. An initial building-wide monitoring with ClimbUp interceptors revealed 25 infestations. The spatial distribution of bed bug infested rooms showed a significant aggregated distribution pattern with same infestation status for neighboring units sharing walls. All infested rooms were monitored every 2 wk and treated using a combination of steam and diatomaceous earth (DE) dust application if bed bugs were still found. For the 25 initially identified infested rooms, after 14 wk treatment, 44% of them no longer had bed bugs, and the mean number of bed bugs captured per room decreased by 94.1%. A combination of steam and DE dust treatment is an effective strategy for suppressing tropical bed bug infestations in dormitory environment.

Exploring the multi-level regulation of lignocellulases in the filamentous fungus Trichoderma guizhouense NJAU4742 from an omics perspective.

BACKGROUND: Filamentous fungi are highly efficient at deconstructing plant biomass by secreting a variety of enzymes, but the complex enzymatic regulation underlying this process is not conserved and remains unclear. RESULTS: In this study, cellulases and xylanases could specifically respond to Avicel- and xylan-induction, respectively, in lignocellulose-degrading strain Trichoderma guizhouense NJAU4742, however, the differentially regulated cellulases and xylanases were both under the absolute control of the same TgXyr1-mediated pathway. Further analysis showed that Avicel could specifically induce cellulase expression, which supported the existence of an unknown specific regulator of cellulases in strain NJAU4742. The xylanase secretion is very complex, GH10 endoxylanases could only be induced by Avicel, while, other major xylanases were significantly induced by both Avicel and xylan. For GH10 xylanases, an unknown specific regulator was also deduced to exist. Meanwhile, the post-transcriptional inhibition was subsequently suggested to stop the Avicel-induced xylanases secretion, which explained the specifically high xylanase activities when induced by xylan in strain NJAU4742. Additionally, an economical strategy used by strain NJAU4742 was proposed to sense the environmental lignocellulose under the carbon starvation condition, that only slightly activating 4 lignocellulose-degrading genes before largely secreting all 33 TgXyr1-controlled lignocellulases if confirming the existence of lignocellulose components. CONCLUSIONS: This study, aiming to explore the unknown mechanisms of plant biomass-degrading enzymes regulation through the combined omics analysis, will open directions for in-depth understanding the complex carbon utilization in filamentous fungi.

Behavioral Response of the Tropical Bed Bug, Cimex hemipterus (Hemiptera: Cimicidae) to Carbon Dioxide.

Bed bugs (Cimex spp.) (Hemiptera: Cimicidae) are obligatory blood-sucking insects. Research on their behavioral response to host-derived olfactory cues will be helpful for designing more effective management tools. However, previous studies mainly focused on Cimex lectularius L., while the related studies on Cimex hemipterus (F.) have been rarely reported. In this study, we evaluated the behavioral responses of C. hemipterus to one of the crucial host-derived cues - carbon dioxide (CO2), at various concentrations. Pitfall traps baited with CO2 captured significantly more C. hemipterus (72%) than unbaited ones (28%). In the Y-tube test, bed bugs' exploratory activity (response rate) gradually decreased (from 70 to 27%) as the CO2 concentration increased (from 0.9 to 14.5%); and bed bugs exhibited a significant attraction to CO2 when the tested CO2 concentrations were between 0.9 and 7.3%. Cimex hemipterus were no longer attracted to CO2 after their antennae or mouthparts or both antennae and mouthparts were removed. In conclusion, C. hemipterus are attracted to CO2, and both antennae and mouthparts are important for CO2 perception.

Overcoming diverse homologous recombinations and single chimeric guide RNA competitive inhibition enhances Cas9-based cyclical multiple genes coediting in filamentous fungi.

Deciphering the complex cellular behaviours and advancing the biotechnology applications of filamentous fungi increase the requirement for genetically manipulating a large number of target genes. The current strategies cannot cyclically coedit multiple genes simultaneously. In this study, we firstly revealed the existence of diverse homologous recombination (HR) types in marker-free editing of filamentous fungi, and then, demonstrated that sgRNA efficiency-mediated competitive inhibition resulted in the low integration of multiple genetic sites during coediting, which are the two major obstacles to limit the efficiency of cyclically coediting of multiple genes. To overcome these obstacles, we developed a biased cutting strategy by Cas9 to greatly enhance the desired HR type and applied a new selection marker labelling strategy for multiple donor DNAs, in which only the donor DNA with the lowest sgRNA efficiency was labelled. Combined with these strategies, we successfully developed a convenient method for cyclically coediting multiple genes in different filamentous fungi. In addition, diverse HRs resulted in a useful and convenient one-step approach for gene functional study combining both gene disruption and complementation. This research provided both a useful one-step approach for gene functional study and an efficient strategy for cyclically coediting multiple genes in filamentous fungi.

Morphological and Molecular Identification of Tropical Bed Bugs From Two Cities of the Pearl River Delta in China.

From the 1960s to the 1980s, with the implementation of nationwide 'Four-Pest Elimination' campaigns (bed bugs, Cimex lectularius L. and Cimex hemipterus (F.) (Hemiptera: Cimicidae), were listed as one of the targeted pests), bed bug infestations were gradually eliminated in most provinces in China. However, during the last two decades, reports of bed bug infestations in the Pearl River Delta of China have shown an upward trend. Up to now, the bed bug species occurring in this area was much less frequently reported. In this study, we used both morphological and molecular methods to accurately identify the species of bed bugs collected from the cities of Guangzhou and Foshan, China. Results indicated that no significant difference was observed in the mean pronotum width-to-length ratio of Guangzhou (2.6) and Foshan (2.4) specimens; however, both were significantly lower than that of a laboratory strain C. lectularius (3.1). The genetic distances of our specimens with C. hemipterus and C. lectularius were 0-0.2% and 22.2-22.6%, respectively. On the basis of the morphological characteristics and mitochondrial DNA sequence data, it can be affirmed that bed bugs collected from Guangzhou and Foshan were C. hemipterus.