In insects, antimicrobial humoral immunity is governed by two distinct gene cascades, IMD pathway mainly targeting Gram-negative bacteria and Toll pathway preferentially targeting Gram-positive bacteria, which are widely conserved among diverse metazoans. However, recent genomic studies uncovered that IMD pathway is exceptionally absent in some hemipteran lineages like aphids and assassin bugs. How the apparently incomplete immune pathways have evolved with functionality is of interest. Here we report the discovery that, in the hemipteran stinkbug Plautia stali, both IMD and Toll pathways are present but their functional differentiation is blurred. Injection of Gram-negative bacteria and Gram-positive bacteria upregulated effector genes of both pathways. Notably, RNAi experiments unveiled significant functional permeation and crosstalk between IMD and Toll pathways: RNAi of IMD pathway genes suppressed upregulation of effector molecules of both pathways, where the suppression was more remarkable for IMD effectors; and RNAi of Toll pathway genes reduced upregulation of effector molecules of both pathways, where the suppression was more conspicuous for Toll effectors. These results suggest the possibility that, in hemipterans and other arthropods, IMD and Toll pathways are intertwined to target wider and overlapping arrays of microbes, which might have predisposed and facilitated the evolution of incomplete immune pathways.
PsDefensin1
Relative expression level to L32 (fig. S10a).
Defensin1.csv
PsDefensin2
Relative expression level to L32 (fig.S10b).
Defensin2.csv
PsHemiptericin
Relative expression level to L32 (fig. S10c).
Hemiptericin.csv
PsPGRP-L1a
Relative expression level to L32 (fig. S8a).
PGRPL1a.csv
PsPGRP-L1b
Relative expression level to L32 (fig. S8b).
PGRPL1b.csv
PsPGRP-L2
Relative expression level to L32 (fig. S8c).
PGRPL2.csv
PslysM
Relative expression level to L32 (fig. S8d).
lysM.csv
PsGNBP1
Relative expression level to L32 (fig. S8e).
GNBP1.csv
PsGNBP2
Relative expression level to L32 (fig. S8f).
GNBP2.csv
PsImd
Relative expression level to L32 (fig. S9a).
IMD.csv
PsMyD88
Relative expression level to L32 (fig. S9b).
MyD88.csv
PsRelish
Relative expression level to L32 (fig. S9c).
Relish.csv
PsDorsalA
Relative expression level to L32 (fig. S9d).
DorsalA.csv
PsDorsalB
Relative expression level to L32 (fig. S9e).
DorsalB.csv
PsLysozyme b1
Relative expression level to L32 (fig.S11a).
Lysb1.csv
PsLysozyme b2
Relative expression level to L32 (fig.S11b).
Lysb2.csv
PsLysozyme c1
Relative expression level to L32 (fig.S11c).
Lysc1.csv
PsLysozyme c2
Relative expression level to L32 (fig.S11d).
Lysc2.csv
PsLysozyme c3
Relative expression level to L32 (fig.S11e).
Lysc3.csv
PsLysozyme i1
Relative expression level to L32 (fig.S11f).
Lysi1.csv
PsDefensin1 pathwayRNAi and Ec
Relative expression level to L32 (fig. 4a).
Def1 pathwayRNAi and Ec.csv
PsDefensin1 pathwayRNAi and Ml
Relative expression level to L32 (fig. 4b).
Def1 pathwayRNAi and Ml.csv
PsDefensin2 pathwayRNAi and Ec
Relative expression level to L32 (fig. 4c).
Def2 pathwayRNAi and Ec.csv
PsDefensin2 pathwayRNAi and Ml
Relative expression level to L32 (fig. 4d).
Def2 pathwayRNAi and Ml.csv
PsHemiptericin pathwayRNAi and Ec
Relative expression level to L32 (fig. 4e).
Hemi pathwayRNAi and Ec.csv
PsHemiptericin pathwayRNAi and Ml
Relative expression level to L32 (fig. 4f).
Hemi pathwayRNAi and Ml.csv
PsLysozyme b1 pathwayRNAi and Ec
Relative expression level to L32 (fig. 4g).
Lysb1 pathwayRNAi and Ec.csv
PsLysozyme b1 pathwayRNAi and Ml
Relative expression level to L32 (fig. 4h).
Lysb1 pathwayRNAi and Ml.csv
PsLysozyme c1 pathwayRNAi and Ec
Relative expression level to L32 (fig. 4i).
Lysc1 pathwayRNAi and Ec.csv
PsLysozyme c1 pathwayRNAi and Ml
Relative expression level to L32 (fig. 4j).
Lysc1 pathwayRNAi and Ml.csv
PsDefensin1 PGRPRNAi and Ec
Relative expression level to L32 (fig. 5a).
Def1 PGRPRNAi and Ec.csv
PsDefensin1 PGRPRNAi and Ml
Relative expression level to L32 (fig. 5b).
Def1 PGRPRNAi and Ml.csv
PsDefensin2 PGRPRNAi and Ec
Relative expression level to L32 (fig. 5c).
Def2 PGRPRNAi and Ec.csv
PsDefensin2 PGRPRNAi and Ml
Relative expression level to L32 (fig. 5d).
Def2 PGRPRNAi and Ml.csv
PsHemiptericin PGRPRNAi and Ec
Relative expression level to L32 (fig. 5e).
Hemi PGRPRNAi and Ec.csv
PsHemiptericin PGRPRNAi and Ml
Relative expression level to L32 (fig. 5f).
Hemi PGRPRNAi and Ml.csv
PsLysozyme b1 PGRPRNAi and Ec
Relative expression level to L32 (fig. 5g).
Lysb1 PGRPRNAi and Ec.csv
PsLysozyme b1 PGRPRNAi and Ml
Relative expression level to L32 (fig. 5h).
Lysb1 PGRPRNAi and Ml.csv
PsLysozyme c1 PGRPRNAi and Ec
Relative expression level to L32 (fig. 5i).
Lysc1 PGRPRNAi and Ec.csv
PsLysozyme c1 PGRPRNAi and Ml
Relative expression level to L32 (fig. 5j).
Lysc1 PGRPRNAi and Ml.csv
PsDefensin1 GNBPRNAi and Ec
Relative expression level to L32 (fig. 5a).
Def1 GNBPRNAi and Ec.csv
PsDefensin1 GNBPRNAi and Ml
Relative expression level to L32 (fig. 5b).
Def1 GNBPRNAi and Ml.csv
PsDefensin2 GNBPRNAi and Ec
Relative expression level to L32 (fig. 5c).
Def2 GNBPRNAi and Ec.csv
PsDefensin2 GNBPRNAi and Ml
Relative expression level to L32 (fig. 5d).
Def2 GNBPRNAi and Ml.csv
PsHemiptericin GNBPRNAi and Ec
Relative expression level to L32 (fig. 5e).
Hemi GNBPRNAi and Ec.csv
PsHemiptericin GNBPRNAi and Ml
Relative expression level to L32 (fig. 5f).
Hemi GNBPRNAi and Ml.csv
PsLysozyme b1 GNBPRNAi and Ec
Relative expression level to L32 (fig. 5g).
Lysb1 GNBPRNAi and Ec.csv
PsLysozyme b1 GNBPRNAi and Ml
Relative expression level to L32 (fig. 5h).
Lysb1 GNBPRNAi and Ml.csv
PsLysozyme c1 GNBPRNAi and Ec
Relative expression level to L32 (fig. 5i).
Lysc1 GNBPRNAi and Ec.csv
PsLysozyme c1 GNBPRNAi and Ml
Relative expression level to L32 (fig. 5j).
Lysc1 GNBPRNAi and Ml.csv
PsRelish RNAi timelapse
Relative expression level to L32 (fig. S12).
relish rnai timelapse.csv
PsImd RNAi check
Relative expression level to L32 (fig. S13a).
imdrnaicheck.csv
PsMyD RNAi check
Relative expression level to L32 (fig. S13b).
mydrnaicheck.csv
PsRelish RNAi check
Relative expression level to L32 (fig. S14a).
relishRNAi.csv
PsDorsalA RNAi check
Relative expression level to L32 (fig. S14b).
dorsalARNAi.csv
PsDorsalB RNAi check
Relative expression level to L32 (fig. S14c).
dorsalBRNAi.csv
PsPGRP-L1a RNAi check
Relative expression level to L32 (fig. S15a).
PGRPL1a rnai check.csv
PsPGRP-L1b RNAi check
Relative expression level to L32 (fig. S15b).
PGRPL1b rnai check.csv
PsPGRP-L2 RNAi check
Relative expression level to L32 (fig. S15c).
PGRPL2 rnai check.csv
PsLysM RNAi check
Relative expression level to L32 (fig. S15d).
LysM rnai check.csv
PsGNBP1 RNAi check
Relative expression level to L32 (fig. S16a).
gnbp1rnaicheck.csv
PsGNBP2 RNAi check
Relative expression level to L32 (fig. S16b).
gnbp2rnaicheck.csv