Diverse PGRPs cooperatively maintain homeostasis of facultative symbiont Arsenophonus to promote reproduction of Nilaparvata lugens
Data files
Jan 09, 2026 version files 50.72 KB
-
Rawdata.xlsx
41.66 KB
-
README.md
9.06 KB
Abstract
Peptidoglycan recognition proteins (PGRPs) play a critical role in insect innate immunity in defending against pathogen invasion and regulating the homeostasis of endosymbionts. Arsenophonus, an emerging clade of intracellular symbionts, has been extensively studied for its roles in host reproductive manipulation and environmental adaptation. However, the molecular mechanisms by which host insects maintain Arsenophonus homeostasis through PGRPs remain largely unexplored. Here, we investigated the functional roles of PGRPs in regulating Arsenophonus homeostasis in the brown planthopper, Nilaparvata lugens, a devastating pest of rice crops. While a previous study reported the existence of only two PGRPs (PGRP-LC and PGRP-LB) in N. lugens, we further determined that PGRP-LC produced two functional isoforms, PGRP-LCa and PGRP-LCb, respectively. PGRP-LCa and PGRP-LCb shared identical intracellular domains but possessed different extracellular domains. Simultaneously, Arsenophonus was observed to localize to the gut muscle layer, fat bodies and ovarian tissues by TEM. Functional assays revealed that PGRP-LCa and PGRP-LCb bound not only peptidoglycan but also directly interacted with cultured Arsenophonus. Artificially transfected Arsenophonus upregulated the expression of PGRP-LCa, PGRP-LCb, and Relish, while suppressing PGRP-LB. PGRP-LCa or Relish knockdown increased Arsenophonus density, whereas silencing PGRP-LCb reduced the symbiont load and female fecundity by the non-IMD pathway. Notably, inhibiting PGRP-LB not only elevated Arsenophonus density but also triggered symbiont dispersal from the gut muscle layer into epithelium. Our findings indicate that PGRPs cooperatively maintain Arsenophonus homeostasis to ensure the fecundity in N. lugens. These studies provide insight into the interaction of immunity between host and endosymbiont.
Dataset DOI: 10.5061/dryad.73n5tb3bm
Description of the data and file structure
Overview
This dataset contains the raw experimental data underlying the manuscript: Faliang Qin, Ping Li, Xuan He, Sen Zheng, Lihaotian Gao, Shuo Wang, Xingya Wang. Diverse PGRPs cooperatively maintain homeostasis of facultative symbiont Arsenophonus to promote reproduction of Nilaparvata lugens. Insect Molecular Biology, 2025.
The experiments investigate the role of PGRPs in Arsenophonus homeostasis in N. lugens. The data include: (1) Tissue-specific expression profiles of PGRP-LB, PGRP-LCa, and PGRP-LCb in female adults of N. lugens; (2) Effects of Arsenophonus injection on PGRPs expression; (3) Effects of PGRPs RNAi on the relative abundance of Arsenophonus in whole body or tissues of N .lugens; (4) Effects of PGRPs RNAi on egg production.
All data are provided in one Excel workbook: Rawdata.xlsx
Dryad datasets are treated as independent publications; this README describes the content of each worksheet and the main variables, so that the dataset can be understood and reused without referring to the article.
For full experimental details and interpretation, please refer to the associated article in Insect Molecular Biology.
Files and variables
File: Rawdata.xlsx
Description: Sheet: Fig 1. These data were used to generate: Figure 1C–E: The conservative domains and tissue profiles of PGRPs in N. lugens (C) Tissue-specific expression profiles of PGRP-LB in female adults of N. lugens. (D) Tissue-specific expression profiles of PGRP-LCa in female adults of N. lugens. (E) Tissue-specific expression profiles of PGRP-LCb in female adults of N. lugens. The relative expression of genes was calculated using the 2-ΔΔCt method with N. lugens 18S rRNA gene as an internal standard for normalization. FB, Fat body; SG, Salivary glands; Ov, Ovary. Data are means ± SEMs. Pairwise comparisons were performed using Student’s t- test (*P < 0.05, **P < 0.01, ***P < 0.001).
sheet: Fig 4. These data were used to generate: Figure 4A–D: Arsenophonus could induce the innate immunity of N. lugens. (A) Relative expression level of PGRP-LCa at 3 h, 6 h, and 12 h after the cultured Arsenophonus injection. (B) Relative expression level of PGRP-LCb at 3 h, 6 h, and 12 h after the cultured Arsenophonus injection. (C) Relative expression level of Relsh at 3 h, 6 h and 12 h after the cultured Arsenophonus injection. (D) Relative expression level of PGRP-LB at 3 h, 6 h and 12 h after the cultured Arsenophonus injection. Each treatment was replicated three times with 10 individuals per replication. The relative expression of genes was calculated using the 2-ΔΔCt method with N. lugens 18S rRNA gene as an internal standard for normalization. Data are means ± SEMs. Significant differences between treatments are indicated by asterisks (*P < 0.05; **P < 0.01; ***P < 0.001).
sheet: Fig 5. These data were used to generate: Figure 5A–I: Arsenophonus density was regulated by the Imd pathway of N. lugens. (A) Arsenophonus density in whole body of female adults (0-6 h after eclosion) after microinjection dsPGRP-LCa and dsGFP (n = 24). (B) Arsenophonus density in the gut and carcass of female adults (0-6 h after eclosion) after microinjection dsPGRP-LCa and dsGFP (Each treatment was replicated four times with 10 individuals per replication). (C) The number eggs in dsPGRP-LCa and dsGFP-injected female adults within 10 d after eclosion (n = 15). (D) Arsenophonus density in whole body of female adults (0-6 h after eclosion) after dsPGRP-LCb and dsGFP injection (n = 24). (E) Arsenophonus density in the gut and carcass of female adults (0-6 h after eclosion) after dsPGRP-LCb and dsGFP injection (Each treatment was replicated four times with 10 individuals per replication). (F) The number eggs in dsPGRP-LCa and dsGFP-injected female adults within 10 d after eclosion (n = 15). (G) Arsenophonus density in whole body of female adults (0-6 h after eclosion) after dsGFP and dsRelish injection (n = 24). (H) Arsenophonus density in the gut and carcass of female adults (0-6 h after eclosion) after dsGFP and dsRelish injection (Each treatment was replicated four times with 10 individuals per replication). (I) The number eggs in dsRelish and dsGFP-injected female adults within 10 d after eclosion (n = 15). The relative symbiont density was calculated using the 2-ΔCt method with N. lugens 18S rRNA gene as an internal standard for normalization. Data are means ± SEMs. Significant differences between treatments are indicated by asterisks (*P < 0.05; **P < 0.01; ***P < 0.001).
sheet: Fig 6. These data were used to generate: Figure 6A-B, and C: PGRP-LB regulates Arsenophonus density and infection in the gut of N. lugens. (A) Arsenophonus density in whole body of female adults (0-6 h after eclosion) after microinjection dsPGRP-LCa and dsGFP (n = 24). (B) Arsenophonus density in the gut and carcass of female adults (0-6 h after eclosion) after microinjection dsPGRP-LCa dsGFP (Each treatment was replicated four times with 10 individuals per replication). (C) The number eggs in dsPGRP-LB and dsGFP-injected female adults within 10 d after eclosion (n = 15). The relative symbiont density was calculated using the 2-ΔCt method with N. lugens 18S rRNA gene as an internal standard for normalization. Data are means ± SEMs. Significant differences between treatments are indicated by asterisks (**P < 0.01; ***P < 0.001).
sheet: Fig S2. These data were used to generate Figure S2: The abundance of Arsenophonus in the gut of N.lugens. The different gut parts of female adult (2 d after eclosion) were dissected and used to extract the DNA for qPCR. Each treatment was replicated three times with 10 individuals per replication. The relative symbiont density was calculated using the 2-ΔCt method with N. lugens 18S rRNA gene as an internal standard for normalization. Data are means ± SEMs. n = 3. Significant differences between treatments are indicated by asterisks (**P < 0.01).
sheet: Fig S5. These data were used to generate Figure S5: The binding affinities of recombinant PGRP-LCa and PGRP-LCb to different type PGN. The relative band intensities in Western blot analysis were quantified using ImageJ software.
sheet: Fig S6. These data were used to generate: Figure S6: Heated-Arsenophonus could induce the innate immunity of N. lugens. The heat-killed Arsenophonus were microinjected into fifth-instar nymphs of N. lugens to quantify the gene responses associated with the IMD pathway. (A) Relative expression level of PGRP-LCa at 3 h, 6 h, and 12 h after the heat-killed Arsenophonus injection. (B) Relative expression level of PGRP-LCb at 3 h, 6 h, and 12 h after the heat-killed Arsenophonus injection. (C) Relative expression level of Relish at 3 h, 6 h and 12 h after the heat-killed Arsenophonus injection. (D) Relative expression level of PGRP-LB at 3 h, 6 h and 12 h after the heat-killed Arsenophonus injection. Each treatment was replicated three times with 10 individuals per replication. The relative expression of genes was calculated using the 2-ΔΔCt method with N. lugens 18S rRNA gene as an internal standard for normalization. Data are means ± SEMs. Significant differences between treatments are indicated by asterisks (*P < 0.05; ***P < 0.001).
sheet: Fig S7.These data were used to generate Figure S7A-E: Relative expression of genes after dsRNA microinjection. (A) Relative expression of PGRP-LCa in dsGFP-injected and dsPGRP-LCa-injected female adult (0-6 h after eclosion) of N.lugens (n = 3). (B) Relative expression of PGRP-LCb and Relish in dsGFP-injected and dsPGRP-LCb-injected female adult (0-6 h after eclosion) of N.lugens (n = 3). (C) Relative expression of Relish in dsGFP-injected and dsRelish-injected female adult (0-6 h after eclosion) of N.lugens (n = 3). (D) Relative expression of PGRP-LB and Relish in dsGFP-injected and dsPGRP-LB-injected female adult (0-6 h after eclosion) of N.lugens (n = 3). (E) Relative expression of PGRP-LCb and Relish in dsGFP-injected and dsPGRP-LCb + Relish-injected female adult (0-6 h after eclosion) of N.lugens (n = 3). (F) Relative abundance of Arsenophonus in the gut of N. lugens after dsPGRP-LCb+Relish injection. The relative expression of genes was calculated using the 2-ΔΔCt method with N. lugens 18S rRNA gene as an internal standard for normalization. Data are means ± SEMs. Significant differences between treatments are indicated by asterisks (*P < 0.05; ***P < 0.001).
Nymphs of N. lugens were injected with dsRNAs targeting PGRP-LCa, PGRP-LCb, PGRP-LB, Relish, or control dsGFP. qRT-PCR was used to quantify transcript levels of PGRP-LCa, PGRP-LCb, and PGRP-LB in whole bodies or selected tissues. Relative expression was calculated using the 2 −ΔΔCt or 2 −ΔCt method with appropriate reference genes. Statistical comparisons between treatments were performed using Student’s t-test in GraphPad Prism, and the corresponding P values and test statistics are included in the tables where applicable. For full experimental details and interpretation, please refer to the associated article in Insect Molecular Biology.