Entomopathogenic fungi, often acknowledged primarily for their insecticidal properties, fulfill diverse roles within ecosystems. These roles encompass endophytism, antagonism against plant diseases, promotion of the growth of plants, and inhabitation of the rhizosphere, occurring both naturally and upon artificial inoculation, as substantiated by a growing body of contemporary research. Numerous studies have highlighted the beneficial aspects of endophytic colonization. This review aims to systematically organize information concerning the direct (nutrient acquisition and production of phytohormones) and indirect (resistance induction, antibiotic and secondary metabolite production, siderophore production, and mitigation of abiotic and biotic stresses) implications of endophytic colonization. Furthermore, a thorough discussion of these mechanisms is provided. Several challenges, including isolation complexities, classification of novel strains, and the impact of terrestrial location, vegetation type, and anthropogenic reluctance to use fungal entomopathogens, have been recognized as hurdles. However, recent advancements in biotechnology within microbial research hold promising solutions to many of these challenges. Ultimately, the current constraints delineate potential future avenues for leveraging endophytic fungal entomopathogens as dual microbial control agents.

Xenorhabdus hominickii, an entomopathogenic bacterium, inhibits eicosanoid biosynthesis of target insects to suppress their immune responses by inhibiting phospholipase A₂ (PLA₂) through binding to a damage-associated molecular pattern (DAMP) molecule called dorsal switch protein 1 (DSP1) from Spodoptera exigua, a lepidopteran insect. However, the signalling pathway between DSP1 and PLA₂ remains unknown. The objective of this study was to determine whether DSP1 could activate Toll immune signalling pathway to activate PLA₂ activation and whether X. hominickii metabolites could inhibit DSP1 to shutdown eicosanoid biosynthesis. Toll-Spätzle (Spz) signalling pathway includes two Spz (SeSpz1 and SeSpz2) and 10 Toll receptors (SeToll1-10) in S. exigua. Loss-of-function approach using RNA interference showed that SeSpz1 and SeToll9 played crucial roles in connecting DSP1 mediation to activate PLA₂. Furthermore, a deletion mutant against SeToll9 using CRISPR/Cas9 abolished DSP1 mediation and induced significant immunosuppression. Organic extracts of X. hominickii culture broth could bind to DSP1 at a low micromolar range. Subsequent sequential fractionations along with binding assays led to the identification of seven potent compounds including 3-ethoxy-4-methoxyphenol (EMP). EMP could bind to DSP1 and prevent its translocation to plasma in response to bacterial challenge and suppress the up-regulation of PLA₂ activity. These results suggest that X. hominickii inhibits DSP1 and prevents its DAMP role in activating Toll immune signalling pathway including PLA₂ activation, leading to significant immunosuppression of target insects.

Dorsal switch protein 1 (DSP1) acts as a damage-associated molecular pattern (DAMP) molecule to activate immune responses inTenebrio molitor. From a previous study inSpodoptera exigua, we found that DSP1 activates Toll immune signalling pathway to induce immune responses by melanisation, PLA₂activity and AMP synthesis. However, the target site of DSP1 in this pathway remains unknown. The objective of this study was to determine the role of spätzle processing enzyme in the DSP1 induced toll immune signalling pathway. To address this, we analyzed spätzle processing enzyme (Tm-SPE) of the three-step serine protease cascade ofT.molitorToll pathway.Tm-SPEexpressed in all developmental stages and larval tissues. Upon immune challenge, its expression levels were upregulated but significantly reduced after RNA interference (RNAi). In addition, the induction of immune responses upon immune challenge or recombinant DSP1 injection was significantly increased. Loss of function using RNA interference revealed that theTm-SPEis involved in connecting DSP1 induced immune responses like hemocyte nodule formation, phenoloxidase (PO) activity, phospholipase A₂(PLA₂) activity and antimicrobial peptide (AMP) synthesis. These suggest thatTm-SPEcontrols the DSP1 induced activation of Toll immune signalling pathway required for both cellular and humoral immune responses. However, to confirm the target molecule of DSP1 in three-step proteolytic cascade, we have to check other upstream serine proteases like Spatzle activating enzyme (SAE) or modular serine protease (MSP).

We know that heat shock can activate the functional components in soybeans, but we don't know the type, level, and duration of heat shock for maximum activation. To address this, the present study investigated the changes in functional components like polyphenols, antioxidants, and isoflavones in soybeans at various temperature levels and durations with their respective functionality or health benefits. For this, treated seed samples were extracted with 70 % ethanol. Heat shock at 60 °C for 2 h increased polyphenol content (60.67 % of control) and antioxidant activity for both ABTS (41.14 % of control) and DPPH (217.72 % of control). This also increased the beneficial aglycone form of isoflavones that includes daidzein (8.36-fold of control), glycitein (3.85-fold of control) and genistein (20.50-fold of control) but decreased the harmful β-glucoside form (3.65-fold) including daiazin (1.84-fold of control); glycitin (1.45-fold of control) and genistin (23.88-fold of control) over untreated dry seed. This may happen because of the conversion of conjugated β-glucoside isoflavones to their aglycone forms that have various health benefits. Maximum inhibition of NO production in RAW 264.7 cells was achieved by samples elicited for 2 h with 300 μg/mL concentration. This sample also confirmed the maximum anti-obesity activity treated against 3-T-3L1 cells. This study summarized that heat shock at 60 °C for 2 h increased polyphenols, antioxidants, and aglycon isoflavone in soybeans resulting in increased anti-inflammatory and anti-obesity activity.