Biofilm formation in Burkholderia thailandensis, a model for the causative agent of melioidosis, is critically regulated by N-acyl homoserine lactone (AHL)-mediated quorum sensing (QS), contributing to its pathogenicity and treatment failure. This study investigates a novel combinatorial strategy to disrupt this process by employing methyl gallate, a plant-derived biofilm inhibitor, together with the AHL-lactonase YtnP. Our results demonstrate that methyl gallate not only attenuated B. thailandensis virulence in a Galleria mellonella infection model but also showed a markedly slower development of resistance compared to conventional antibiotics like tetracycline and meropenem. Strikingly, the combination of sub-inhibitory concentrations of methyl gallate and YtnP exhibited a synergistic effect, significantly enhancing the susceptibility of the bacterium to methyl gallate. This synergy was underpinned by a profound suppression of AHL signal molecules (C8-HSL, 3-OH-C8-HSL, and 3-OH-C10-HSL) and a concurrent downregulation of key QS regulatory genes (btaI1-3, btaR1-3). Consequently, the combination treatment robustly impaired biofilm formation and disrupted its structural integrity, as visualized by confocal microscopy. Molecular docking analyses provided a structural basis for this synergy, revealing stable binding of C8-HSL within the catalytic pocket of YtnP. Our findings posit the methyl gallate-YtnP combination as a potent and resistance-retarding anti-virulence approach against B. thailandensis biofilms.

BACKGROUND:The brown planthopper (Nilaparvata lugens, BPH) is a notorious sap-sucking pest on rice (Oryza sativa) causing substantial yield losses. During BPH oviposition, its egg-associated secretions trigger rice defense responses; however, the role of their specific components in plant immune modulation remains poorly understood. This study aimed to investigate the effect of the oviduct-secreted protein 1 (NlOdsp1) from BPH egg-associated secretions on plant defense. RESULTS:NlOdsp1, a planthopper-specific gene, is predominantly expressed in the female adults and oviduct of BPH. NlOdsp1 activates cell death and reactive oxygen species production in Nicotiana benthamiana and impairs the performance of the whitefly and aphid. Moreover, its 39 amino acid peptide is recognized by plants as a minimal immunogenic epitope. NlOdsp1 significantly induces H2O2 accumulation in rice and enhances rice resistance to BPH. CONCLUSION:NlOdsp1 from BPH egg-associated secretions serves as a critical and unavoidable elicitor, triggering plant resistance to various insects. The essential role of this protein for BPH reproduction may preclude evolutionary adaptations to evade detection by rice. © 2025 Society of Chemical Industry.

Plants activate defense machinery when infested by herbivorous insects but avoid such costs in the absence of herbivory. However, the key signaling pathway regulators underlying such flexibility and the mechanisms that insects exploit these components to disarm plant defense systems remain elusive. Here, it is reported that immune repressor 14-3-3e in rice Oryza sativa (OsGF14e) regulates immune homeostasis. Infestation with brown planthopper (BPH) Nilaparvata lugens decreased OsGF14e expression; however, the level of downregulation is limited both by the short duration and the specific feeding location. OsGF14e interacts with Enhanced Disease Resistance 1-like (OsEDR1l), a Raf-like MAP kinase kinase kinase (MAPKKK), and repressed jasmonic acid, jasmonic acid-isoleucine, and H2O2 accumulation by enhancing OsEDR1l abundance and signaling ability. OsGF14e and OsEDR1l overexpression renders rice susceptible to BPH, whereas their knockout increases plant resistance but compromises rice growth and grain yield. Intriguingly, BPH 14-3-3e protein (Nl14) that shares high sequence homology and structural similarity with OsGF14e is identified from BPH saliva and egg-associated secretions. Mediated through BPH feeding and oviposition, Nl14, similar to OsGF14e, interacts with OsEDR1l and triggers the OsEDR1l signaling, thereby suppressing plant defenses and facilitating BPH infestation. Apparently, structural and functional mimicry makes it possible for this newly discovered BPH effector to exploit rice OsGF14e-EDR1l immune suppression module. The results reveal a novel mechanism deployed by herbivorous insects, in a manner similar to certain pathogen effectors, to evade host plant defenses by mimicking host immune regulators.

Galactoglucans exhibit diverse properties, yet their broad application is limited by production challenges. Herein, we isolated a high-polysaccharide-producing strain, Sinorhizobium sp. ACT002. In shake flasks, ACT002 achieved a productivity of 4.21 ± 0.16 g/L/d, surpassing reported values for Rhizobium/Sinorhizobium species. The purified polysaccharide, G2, had a weight-average molecular weight of 2.23 × 106 Da and displayed a smooth surface as observed by SEM. Structural analyses, including FT-IR, methylation-GC/MS, and 1D/2D NMR, identified G2 as a linear galactoglucan composed of the disaccharide repeating unit →3)-β-D-Glcp-(1→3)-(4,6-Pyr)-α-D-Galp-(1→, closely resembling the EPS II produced by S. meliloti. The 4,6-positions of galactosides were substituted with equimolar pyruvyl groups, while its acetyl group content was lower than that of EPS II. Rheological characterization revealed a critical concentration of 3.23 g/L for pseudoplastic behavior. Among various salts, KCl exerted more pronounced effects on the Carreau-Yasuda flow properties, viscoelasticity, and thermal stability of G2. Moreover, G2 possessed moisture absorption/retention capacities comparable to hyaluronan. It also exhibited notable antioxidant activity, UV absorption, and minimal hemolytic effects. Topical application of G2 effectively mitigated UV-B-induced skin damage in mice, promoting restoration of epidermal integrity and tissue organization. These findings highlight the potential application of G2 in rheology modification, moisture retention, and UV protection.

Herbivore infestation triggers complex defense mechanisms in rice, primarily regulated by phytohormones. However, the traditional approach of analyzing plant hormone profiles as an indicator of plant responses to herbivore stress suffers from time-consuming detection, high costs, and destructive sampling of plant tissues. In this study, we propose and validate a novel approach using plant electrophysiological parameters—capacitance, resistance, impedance, and reactance—as a rapid, non-invasive biomarker of rice responses to two major herbivorous pests: the brown planthopper (Nilaparvata lugens, BPH) and the striped stem borer (Chilo suppressalis, SSB). Systematic modelling revealed herbivore-specific temporal electrophysiological response dynamics, reflecting perturbations in water and nutrient transport, dielectric substance movement, and metabolic energy flux in infested rice plants. Based on these dynamics, we developed predictive models for rice yield potential, drought resistance, and metabolic adaptability under biotic stress. In parallel, we analyzed phytohormone levels and found that changes in electrophysiological traits were strongly associated with jasmonic acid and jasmonic acid-isoleucine accumulation, indicating that these electrical signals capture key aspects of the plant’s immune status. Beyond detecting herbivore-induced defenses, electrical patterns were also mirrored in transgenic rice overexpressing planthopper-derived elicitors (e.g. Myosin/PDI1), indicating electrophysiological signatures act as defense activation markers, thus enabling high-throughput screening of herbivore elicitors. Together, our results highlight the potential of plant bioelectrical signals as fast, integrative, and scalable indicators of stress responses, with applications in crop protection and elicitor-assisted resistance breeding.

Lysine malonylation (Kmal), an evolutionarily conserved post-translational modification, serves as a critical regulator of cellular processes including transcriptional control, metabolic coordination, and enzyme activation. While Kmal sites have been mapped in rice (Oryza sativa L.) seeds, their dynamic regulation in rice responses to biotic stresses remains poorly characterized. Here, we reported a global profiling of lysine-malonylated proteins in rice leaf sheaths, and the changes in these proteins under herbivore (Nilaparvata lugens/Chilo suppressalis) or viral (rice stripe virus/rice black-streaked dwarf virus) stresses. Using affinity enrichment and proteomics, we identified 3,113 Kmal sites across 1,324 proteins in wild-type rice leaf sheaths, these data demonstrated that lysine-malonylated proteins are involved in diverse biological processes. Kmal levels were significantly upregulated following herbivore infestation or viral infection, with two herbivores inducing more pronounced changes than viruses infection, revealing stress-specific malonylation landscapes. Kmal preferentially targeted highly expressed proteins in energy metabolism (e.g., glycolysis and the tricarboxylic acid cycle) and photosynthesis, exhibiting an inverse correlation between hypermalonylation and protein abundance. Cleavage under targets and tagmentation analyses revealed Kmal-mediated chromatin remodeling through promoter occupancy at defense-related genes. In addition, the histone deacetylases OsHDA702-704, OsHDA711-713 were functionally characterized as key regulators mediating the erasure of specific malonylation marks in rice plants, with OsHDA711 knockout lines exhibiting enhanced resistance against both herbivore and virus infection. Our work establishes Kmal as a regulator in rice biotic immunity, uncovering novel insights into Kmal-mediated plant defense responses against herbivorous pests and viral pathogens. These findings identify potential genetic targets for developing rice varieties with broad-spectrum immunity to biotic stresses, enhancing crop resilience.

Solid-phase extraction ultrahigh-performance liquid chromatography-tandem mass spectrometry (SPE-UHPLC-MS/MS) was used to evaluate the contamination of ochratoxin A (OTA) and assessed the human exposure risk of OTA in Rasa roxburghii. A more suitable method for OTA extraction and purification of R. roxburghii was obtained. Treated 25 mL of R. roxburghii juice with enzymatic hydrolysis at a concentration of 0.06 mg/mL, filtered the resulting mixture and concentrated the filtrate to dry, then redissolved with 0.2 mL of methanol and diluted with 0.4 mL of ultra-pure water. Added sample solution to the activated hydrophilic-lipophilic balance (HLB) column, washed with 6 mL of ultra-pure water and purified by eluting with 6 mL of methanol. The eluent was collected and dried using nitrogen at 40 degrees C, then redissolved in 1 mL of methanol and filtered for detection. The hazard quotient (HQ) values of all R. roxburghii fruit and juice, which were storage at room temperature, 4 and -20 degrees C from 0 day to 63 days, ranged from 0.077% to 35.792%, which were within the allowable limits for human consumption. From the perspective of OTA contamination, the results indicated that the maximal storage time of R. roxburghii fruit were 14 days at room temperature, 35 days at 4 degrees C and 63 days at -20 degrees C. And the maximal storage time of R. roxburghii juice were 7 days for sealed storage and the same-day for open storage at room temperature, 14 days for sealed storage and 7 days for open storage at 4 degrees C, and 63 days for sealed storage and 56 days for open storage at -20 degrees C during the experiment period. And all thirty samples randomly sampled from the market were OTA negative. The results of this study can lay a foundation for the formulation of OTA limit standards in fruits and juice in the future, and provide a reference for consumers to consume R. roxburghii more healthily.

OBJECTIVE:To explore the taste-related quality markers of Qingbanxia, the alum-processed Pinellia ternata tuber. METHODS:Eighteen samples of Banxia and Qingbanxia were analyzed by the Ultra-High Performance Liquid Chromatography coupled with Q-Exactive Orbitrap Mass Spectrometry. Data of all samples were pre-processed by Compound Discoverer 3.3 Software. The discrimination was analyzed by Principal Component Aanalysis, and Orthogonal Partial Least-square Discriminant Analysis. The chemical markers were identified by MS/MS fragments based on the fragment rules. The electronic tongue was utilized to determine the taste traits of Banxia and Qingbanxia. Furthermore, the taste-related material basis was discovered according to correlation analysis and molecular docking. RESULTS:Sixteen potential chemical markers of Banxia and Qingbanxia were identified. Lauryldiethanolamine is a unique bitter component. The taste spectrum of bitterness, sourness and umami changes significantly during the processing of Banxia, with sourness increasing and bitterness and umami decreasing. CONCLUSION:A new approach to explore the taste-related quality markers in alum-processed Banxia was established for the first time based on the Orbitrap MS technology and electronic tongue technology. The bitterness chemical markers were identified for the first time. The mechanism of the sourness of Qingbanxia was clarified. The identification of taste-related quality markers and the generation of comprehensive taste profiles offer an objective and reproducible method for assessing processing efficacy, overcoming the limitations of traditional subjective taste tests. These findings have significant implications for the quality control of Banxia and other traditional Chinese medicine.

During herbivore feeding, plants can recognize herbivore-associated molecular patterns (HAMPs) present in saliva and trigger pattern-triggered immunity (PTI). Piercing-sucking insects secrete gel saliva, forming salivary sheaths that aid in feeding. However, the role of proteins within these salivary sheaths in modulating plant defences remains poorly understood. In this study, we identified a novel HAMP, Nlsp5, from the salivary sheath of the brown planthopper (Nilaparvata lugens, BPH). Nlsp5 is a planthopper-specific protein and acts as an elicitor of BAK1-dependent PTI responses in both tobacco and rice plants. Moreover, the 19-amino-acid peptide (NP19) within Nlsp5 functions as a minimal immunogenic epitope, which is specifically recognized by plants, stimulating jasmonic acid and hydrogen peroxide pathways. Through exogenous treatment with synthetic NP19 and overexpressing Nlsp5 in rice, we further found that the induced defence responses not only impaired planthopper performance directly but also triggered the emission of volatile compounds that attract a common parasitoid. Additionally, NP19 treatment enhanced the resistance of rice, tobacco, and cotton to several chewing and sap-sucking insects. However, silencing Nlsp5 in BPH disrupted salivary sheath formation, reducing insect feeding efficiency. This study demonstrates that Nlsp5 from the BPH salivary sheath acts as an unavoidable HAMP, triggering resistance in multiple plants to various insect pests. The critical role of this protein in insect feeding precludes evolutionary adaptations to evade detection by plants.

Storability is a critical quality trait of wax gourd, yet the role of magnesium (Mg) in enhancing its storage potential remains underexplored. This study demonstrated that the application of 60 kg MgO hm⁻2 significantly improved fruit firmness by 18.2 % at harvest and 16.3 % after 95 days of storage, compared with control (CK). Mg treatment predominantly increased the levels of ethanol-soluble Mg (MgEth) and pectate-/protein-bound Mg (MgNaCl), which together accounted for 88.2–96.9 % of total Mg, while enhancing total pectin, chelate-soluble pectin (CSP), and sodium carbonate-soluble pectin (SSP). Upregulated pectin methylesterase (PME) activity (28.2 % increase) catalyzes pectin demethylation, facilitating CSP formation and cell wall stabilization. Atomic force microscopy (AFM) revealed the proportion of CSP and SSP with a chain width greater than 120 nm increased significantly, contributing to improved cell wall elasticity and delayed softening. Correlation analysis highlighted strong positive relationships between Mg forms (e.g., MgNaCl) and pectin fractions, whereas transcriptomic analysis identified key glycometabolic and cell wall remodeling pathways. Notably, hub genes (Bhi04G001183, Bhi11G000824, Bhi12G001032, and Bhi11G000479) strongly correlated with PME activity and CSP content. These findings provide molecular and physiological insights into the role of Mg in enhancing wax gourd storability, thereby offering practical solutions for reducing postharvest losses, improving shelf life, and supporting sustainable agricultural practices.

Herbivore-associated molecular patterns (HAMPs) in saliva enable plants to detect herbivores and activate pattern-triggered immunity (PTI). Piercing-sucking herbivores secrete gel saliva, forming salivary sheaths that assist in feeding, however, the role of proteins within these sheaths in modulation of plant defenses remains poorly understood. Here, a thermostable HAMP, myosin light chain 1-like (myosin) is identified, from the salivary sheath of the small brown planthopper (SBPH) Laodelphax striatellus. Myosin is a widely conserved arthropod protein, and acts as an elicitor of BAK1-dependent PTI responses in several plant species. Plants are able to specifically recognize the myosin 41-amino-acid peptide (MP41), which acts as a minimal immunogenic epitope. Furthermore, myosin and MP41 stimulate jasmonic acid and H2O2 production in rice. The resulting defenses not only diminish planthopper performance directly but also induce volatile emissions, attracting a common parasitoid. Additionally, expression of myosin in rice increased plant resistance to a chewing insect as well as to viral and fungal pathogens. However, silencing myosin in SBPH resulted in disruption of salivary sheath formation, reducing insect feeding efficiency. This study demonstrates that myosin from the SBPH salivary sheath serves as a critical and unavoidable HAMP, triggering broad-spectrum plant resistance to various insects and pathogens.

Herbivore-associated molecular patterns (HAMPs) in saliva enable plants to detect herbivores and activate pattern-triggered immunity (PTI). Piercing-sucking herbivores secrete gel-like saliva that forms salivary sheaths, which aid in feeding. However, the role of proteins within these sheaths in modulating plant defenses remains poorly understood. In this study, we identified a thermostable HAMP, actin-related protein 1 (Lsactin), from the salivary sheath of the small brown planthopper (SBPH) Laodelphax striatellus. Lsactin is a widely conserved arthropod protein that serves as an inducer of the BAK1-dependent PTI response in Nicotiana benthamiana, thereby enhancing its resistance to both Bemisia tabaci and Spodoptera frugiperda. Moreover, this HAMP can also induce cell death in plants such as eggplant, cotton, pepper, and corn. Plants were able to specifically recognize the Lsactin 41-amino-acid peptide (LP41), which functions as a minimal immunogenic epitope. Furthermore, LP41 stimulates the production of jasmonic acid and hydrogen peroxide in rice, enhancing rice's resistance to the SBPH. However, silencing Lsactin in SBPH led to the disruption of salivary sheath formation, which reduced insect feeding efficiency. This study demonstrates that Lsactin from the SBPH salivary sheath serves as a crucial HAMP, triggering plants resistance to various insect species.

Rhodotorula toruloides, an oleaginous yeast known for its high lipid productivity, produces lipids with low very-long-chain fatty acid (VLCFA) content. Meanwhile, the roles of enzymes, particularly the condensing enzymes, involved in VLCFA biosynthesis in R. toruloides remained unclear. In this study, two elongases, RtELO1 and RtELO2, were identified from R. toruloides U13N3 and their tertiary structure and catalytic mechanism were investigated using molecular dynamic methods. Both enzymes exhibited typical ELO-like characteristics, with active sites located within cavities formed by seven transmembrane helixes. RtELO2 displayed higher binding affinity to acyl-CoAs compared to RtELO1, and at least seven amino acid residues, including two crucial histidines in the "HXXHH" box, were identified as important for the condensation reaction. To enhance VLCFA production, an internal ribosome entry site (IRES)-mediated bicistronic strategy was developed to integrate multiple genes into the R. toruloides genome. The efficiency of IRES-mediated translation initiation reached 85.4% of cap-dependent upstream translation, based on EGFP fluorescent intensity. Using this strategy, four genes encoding enzymes involved in the VLCFA biosynthesis cycle (Rtelo2, RtKCR, RtHCD, and RtECR) were introduced into the U13N3 genome in various combinations. The results indicated that the expression of a single elongase had a modest effect on VLCFA production, but the simultaneous expression of multiple genes resulted in cumulative effects. Notably, the transformant harboring four genes exhibited a remarkable 436.8% increase in C22 and C24 VLCFA yield compared to the original strain.

In this study, the properties of remaining starch granules obtained with different degrees of exfoliation were explored by removing the outer layers of A- and B-type wheat starch (AWS and BWS) granules with chemical surface gelatinization. SEM images revealed significant morphological variations with increasing exfoliation. CLSM and amylose content analysis indicated a predominance of lipid complexes in the outer granule layers, particularly in BWS. The structural characteristics of AWS and BWS were analyzed using PLM, XRD, FT-IR and DSC, verifying the conclusion of the alternation of starch crystalline and amorphous zone. And the amorphous regions are proportionally higher in the inner starch layer. Moreover, raw AWS and BWS granules were more easily digested from the outside in, with the RS content decreasing from 80.65 % to 66.92 % and 49.06 % to 45.01 %, respectively. The RS content of cooked WS were affected by the internally structures, particularly lipid content (11.46 % - 19.09 %) in BWS outer layers and amylose content (13.59 % - 19.43 %) in the inner layers. These results revealed the internal radial structural differences and digestibility patterns of AWS and BWS granules.

Prokaryotes have evolved intricate innate immune systems against phage infection1-7. Gabija is a highly widespread prokaryotic defence system that consists of two components, GajA and GajB8. GajA functions as a DNA endonuclease that is inactive in the presence of ATP9. Here, to explore how the Gabija system is activated for anti-phage defence, we report its cryo-electron microscopy structures in five states, including apo GajA, GajA in complex with DNA, GajA bound by ATP, apo GajA-GajB, and GajA-GajB in complex with ATP and Mg2+. GajA is a rhombus-shaped tetramer with its ATPase domain clustered at the centre and the topoisomerase-primase (Toprim) domain located peripherally. ATP binding at the ATPase domain stabilizes the insertion region within the ATPase domain, keeping the Toprim domain in a closed state. Upon ATP depletion by phages, the Toprim domain opens to bind and cleave the DNA substrate. GajB, which docks on GajA, is activated by the cleaved DNA, ultimately leading to prokaryotic cell death. Our study presents a mechanistic landscape of Gabija activation.

BACKGROUND The striped stem borer (SSB, Chilo suppressalis) is one of the most destructive insect pests on rice. As a chewing insect, SSB larval feeding causes a dramatic increase in rice defense responses. However, the effects of oral secretions (OSs) during SSB feeding on rice defense remain largely unexplored. RESULTS In this study, based on transcriptome analysis results, treatment with SSB OSs regulated the expression of genes involved in the plant defense-related pathways of calcium, mitogen-activated protein kinases, reactive oxygen species, jasmonic acid (JA), herbivore-induced plant volatiles (HIPVs), and protease inhibitors. Unsurprisingly, treatment with SSB OSs elicited the accumulation of JA and JA-isoleucine in rice. The defense mechanisms activated by the cascade not only induced the expression of trypsin inhibitors, inhibiting the normal growth of SSB larvae but also induced HIPVs emission, rendering rice attractive to a common larval parasitoid. High-throughput proteome sequencing of SSB OSs led to 534 proteins being identified and 343 proteins with two or more unique peptides being detected. CONCLUSION The study demonstrates that SSB OSs trigger both direct and indirect defense mechanisms in rice, akin to the effects of SSB feeding. It identifies specific proteins in SSB OSs that may influence the interactions between SSB and rice during feeding, providing valuable insights for effectors research. (c) 2024 Society of Chemical Industry.

Enhancing the lipid production of oleaginous yeasts is conducive to cutting the cost of feedstock for biodiesel. To increase the lipid productivity of Rhodotorula sp. U13N3, genes involving lipid degradation were knocked out and fermentation conditions were investigated. Results of transcription analysis demonstrated that genes encoding the ATG15-like lipase (ATG15) and peroxisomal acyl-CoA oxidase (ACOX2) were upregulated significantly at the lipogenesis stage. When ATG15 and ACOX2 were knocked out separately from the genome by the CRISPR/Cas9 method, both ΔATG15 and ΔACOX2 mutants showed better lipid production ability than the parent strain. Flow cytometry and confocal microscopic analyses indicated that simultaneous the knockout of ATG15 and ACOX2 did not impact the cell viability, whereas the lipid production was enhanced markedly as the lipid yield increased by 67.03% in shake flasks. Afterward, the ΔATG15ΔACOX2 transformant (TO2) was cultivated in shake flasks in the fed-batch mode; the highest biomass and lipid yield reached 45.76 g/L and 27.14 g/L at 216 h, respectively. Better performance was achieved when TO2 was cultivated in the 1-L bioreactor. At the end of fermentation (180 h), lipid content, yield, yield coefficient, and productivity reached 65.53%, 27.35 g/L, 0.277 g/g glycerol, and 0.152 g/L/h, respectively. These values were at the high level in comparison with Rhodotorula strains cultivated in glycerol media. Besides, fermentation modes did not affect the fatty acid composition of TO2 significantly. In conclusion, blocking the lipid degradation was an applicable strategy to increase the lipid production of Rhodotorula strains without compromising their cell viability. • ATG15-like lipase and acyl-CoA oxidase (ACOX2) participated in lipid degradation. • Knockout of ATG15 and ACOX2 increased lipid productivity, and lipid yield coefficient. • Cell viability maintained at high level in the knockout mutants during fermentation.

Rhodotorula toruloides can utilize crude glycerol as the low-cost carbon source for lipid production, but its growth is subjected to inhibition by methanol in crude glycerol. Here, transcriptome profiling demonstrated that 1004 genes were significantly regulated in the strain R. toruloides TO2 under methanol stress. Methanol impaired the function of membrane transport and subsequently weakened the utilization of glycerol, activities of the primary metabolism and functions of nucleus and ribosome. Afterwards the tolerance of TO2 to methanol was improved by using two-round adaptive laboratory evolution (ALE). The final strain M2-ale had tolerance up to 3.5% of methanol. 1 H NMR-based metabolome analysis indicated that ALE not only improved the tolerance of M2-ale to methanol but also tuned the carbon flux towards the biosynthesis of glycerolipid-related metabolites. The biomass and lipid titer of M2-ale reached 14.63 ± 0.45 g L-1 and 7.06 ± 0.44 g L-1 at 96 h in the crude glycerol medium, which increased up to 17.69% and 31.39%, respectively, comparing with TO2. Afterwards, an effective method for cell lysis was developed by combining sonication and enzymatic hydrolysis (So-EnH). The lytic effect of So-EnH was validated by using confocal imaging and flow cytometry. At last, lipid recovery rate reached 95.4 ± 2.7% at the optimized condition.

The brown planthopper (BPH, Nilaparvata lugens) is a notorious sap-sucking insect pest that damages rice (Oryza sativa) plants throughout Asia. During BPH feeding, saliva enters rice plant tissues, whereas during oviposition egg-associated secretions (EAS) are deposited in damaged plant tissue. Dynamic changes in rice to planthopper salivary effectors have been widely reported. However, the effects of EAS from planthopper on rice immunity remains largely unexplored. In this study, we found that both infestation of rice by gravid BPH female adults and treatment with the EAS elicited a strong and rapid accumulation of jasmonic acid (JA), JA-isoleucine, and hydrogen peroxide in rice. EAS enhanced plant defenses not only in rice but also in tobacco, and these impaired the performance of BPH on rice, as well as the performance of aphids and whiteflies on tobacco. High-throughput proteome sequencing of EAS led to 110 proteins being identified and 53 proteins with 2 or more unique peptides being detected. Some proteins from BPH EAS were also found in the salivary proteome from herbivores, suggesting potential evolutionary conservation of effector functions across feeding and oviposition; however, others were only identified in EAS, and these are likely specifically related to oviposition. These findings point to novel proteins affecting interactions between planthoppers and rice during oviposition, providing an additional source of information for effector studies.