Hemiptera

As elytra is used in Coleoptera, we used the term ‘tegmina’ (singular: tegmen) as a synonym to mention the more or less sclerified mesothoracic forewings, a convention in most of Hemiptera; they are usually covering the membranous metathoracic hind wings at repose.
The general venation schema for planthoppers is here provided based on a fulgoromorphan ground plan slightly modified from the one proposed by Shcherbakov (1996 ). Terminology is completed according to Bourgoin (1997 ) who recommended the use of areas (nodal cells, major vein areas) for the interpretation of veins and updated from Bourgoin and Szwedo (2008 ) and Szwedo and Żyła (2009 ), including the recent proposal of the CuA zigzag vein (=arculus auctorum, Emeljanov 1987 ) as autapomorphic for Paraneoptera (Nel et al. 2012 (link)).
The standardized terminology proposed is built upon the various major vein nomenclature systems used and upon homology-driven morphological interpretations concerning both extant and extinct taxa samples according to all major authors in these topics (Metcalf 1913 (link); Muir 1913 , 1923 ; Melichar 1923 ; Fennah 1944 ; Hamilton 1972 ; Emeljanov 1977 , 1987 ; Shcherbakov 1981 , 1996 ; Zelazny 1981 ; Kukalová-Peck 1983 (link); Chou et al. 1985 ; Anufriev and Emeljanov 1988 ; Dworakowska 1988 ; Bourgoin 1997 ; Zelazny and Webb 2011 ; Ding 2006 ; Nel et al. 2012 (link), 2013 (link); Gnezdilov 2013 (link)).
A corresponding terminology between these major systems is proposed (Table 1), and a definition is provided for each structure.Table 1

Corresponding terminologies between main vein system interpretations since Metcalf (1913 (link)), with the recommended standardized one (in bold)

Metcalf (1913 (link))

C

Sc + R + M

Sc + R

Sc (two branches)

R

M (typically 4 branched)

Cu

A1

A2

A3

A3 s branch

Melichar (1923 )

Costal vein

Subcostal vein

Radius 1

Radius 2

Median vein

Cubitus

Clavus suture

Claval vein external branch

Claval vein internal branch

Scutellar + clavus sutural margins

Fennah (1944 )

C

Sc + R + M

Sc + R

Sc

R1, Rs

MP

Cu1

Cu2

Pcu

A1

A2

Hamilton (1972 )

C

Sc

S + M

S

SA

SP

M (forks first in MA and MP)

Cu

P + E

1A

2A

Margin

Emeljanov (1977 )

C

Sc

S + M

Sc + R

R1Sc + R2

R3

M

CuA

CuP

Pcu

A1

A2

Shcherbakov (1981 )

C

Sc + R + M

(Sc +) R

R1

R1a

Rs

M

CuA

CuP

Pcu

A1

A2

Chou et al. (1985 )

C

Sc

R + M

R

M

Cu

Clavus suture

A

A

Margin

Dworakowska (1988 )

CA

Pc + CP

ScP + R + M

ScP + R + MA

ScP + RA

RA

RP + MA

MP

CuA

CuP

AA

AP’

AP”

Ding (2006 )

C

Sc + R + M

Sc + R

Sc

Sc2

R1

M (‘Rs, M1, M2, M3′ = M1, M2, M3, M4)

Cu1 (‘Cu1a’ = CuA1)

Cu2

IA

IIA

Margin

Interpretation and recommended terminology

CA

Pc + CP

ScP + R + M + CuA

ScP + R

ScP + RA

RA

RP (+MA)

MP

CuA (forks in CuA1 and CuA2)

CuP

Pcu

A1

A2

To facilitate the annotation of innate immunity genes in insects, we initially created an Insect Immunity Database (IIID) composed of the published immune repertoires of four insect models spanning several different orders: Drosophila melanogaster, Diptera [18] (link), [19] (link), Anopheles gambiae, Diptera [16] (link), [20] (link), Apis mellifera, Hymenoptera [17] (link), [21] (link), and Acrythosiphon pisum, Hemiptera [22] (link). Our criteria for inclusion were that the species have a complete, publicly-available genome sequence, that the innate immune genes have been previously identified in computational or molecular studies, and that each species has an extensive review of its global immune pathways available as a resource. Sequence information was obtained through NCBI for the 105 immunity genes described for Acrythosiphon pisum[22] (link), 317 genes for Anopheles gambiae[20] (link), [23] (link), 379 genes for Drosophila melanogaster[18] (link), [19] (link), and 174 genes for Apis mellifera[17] (link), [21] (link). In total, 975 genes were included in the dataset used to analyze the Nasonia genomes. Each gene was categorized into its primary, secondary and tertiary pathways of putative function (i.e. Toll pathway, IMD pathway, humoral response, JAK/STAT, and cell cycle regulation) and into finite classes of function based upon its putative role in an immune response. Such classes include recognition (identifying potential pathogens and stressors), signaling (communicating between recognition and response), and response (molecules that interact with the pathogen or stressor).

According to the seven-superfamily system of Nepomorpha proposed by Hebsgaard et al. (2004) [5 (link)], the representatives of each superfamily were selected (Table 2). Representatives of the infraorders Gerromorpha, Leptopodomorpha, Cimicomorpha and Pentatomomorpha were also included (Table 2). A representative of the suborder Archaeorrhyncha (Insecta: Hemiptera), Lycorma delicatula (White), was chosen to root the trees.
A single individual of each species was preserved in 95% ethanol at -20°C and total genomic DNA was extracted using the method based on CTAB [40 (link)]. PCRs were performed with TaKaRa LA PCR Kit Ver.2.1 following the manufacturer's recommendations. The primers are listed in additional file 5. PCR products were electrophoresed in 0.7% agarose gel, purified, and then both strands were sequenced with primer walking by Beijing Sunbiotech Co. Ltd.
The complete sequences of each gene were used for phylogenetic analysis (excluding stop codons of the PCGs). All PCGs were aligned based on amino acid sequence alignments in MEGA version 4.0 [41 (link)]. The rRNAs and the tRNAs were aligned with CLUSTAL X version 1.83 [42 (link)] under the default settings. Ambiguously aligned regions of PCGs and rRNA genes were carefully adjusted by hand. Transfer RNA alignments were corrected according to secondary structure. The aligned sequences were concatenated as four matrices used in phylogenetic analyses: 1) The PCG123RT matrix, including all three codon positions of PCGs, rRNA genes, and tRNA genes; 2) the PCG12RT matrix, including the first and the second codon positions of PCGs, rRNA genes, and tRNA genes; 3) the PCG123 matrix, including all the three codon positions of PCGs; 4) the PCG12 matrix, including the first and the second codon positions of PCGs.
MrBayes Version 3.1.1 [43 (link)] and a PHYML online web server [44 (link)] were employed to reconstruct the phylogenetic trees under the GTR model. In Bayesian inference, two simultaneous runs of 3,000,000 generations were conducted for each matrix. Trees inferred prior to stationarity were discarded as burn-in, and the remaining trees were used to construct a 50% majority-rule consensus tree. In ML analysis, the parameters were estimated during analysis and the node support values were assessed by bootstrap resampling (BP) [45 (link)] calculated using 100 replicates.

We assembled a taxon set of 771 species across 94 out of 109 recognized extant families (sensu Huber¹⁸ , with modifications by Chen et al.^{79 (link)}, Pilgrim et al.^{80 (link)}, and Sann et al.^{81 (link)}), belonging to all 22 recognized superfamilies within the Hymenoptera^{18 ,80 (link)}, and six non-hymenopteran outgroups. Our taxon sampling aimed for the representation of major lineages within families while sampling across the respective root nodes on the family level, covering between 0.06–50% (=1–150 representatives) of the described species diversity. While we generated UCE sequence data de novo for most taxa, some sequences have already been published in other studies by some of us: 126 aculeate wasps^{38 (link),82 (link),83} , 25 chalcidoids^{84 –86 (link)}, 76 cynipoids^{87 (link)}, 26 Ichneumonidae^{88 (link)–90 (link)} and 142 Braconidae^{91 (link)}. We further included six representatives of other insect orders as outgroups by mining UCEs in silico from published genomes: Coleoptera (Agrilus planipennis), Diptera (Aedes albopictus), Lepidoptera (Papilio glaucus), Hemiptera (Homalodisca vitripennis), Psocodea (Pediculus humanus corporis), and Blattodea (Blattella germanica). Supplementary Data 1 list voucher information and NCBI accession numbers for all sequences, while more detailed specimen data is provided for sequences newly released in this article. All specimens were collected with the required permits and in accordance with local regulations at the time of their collection, and vouchers have been deposited in major collections.

In 2015 and 2016, spider amount was assessed by a drop cloth method (in 2015 in both Vineyards A and B: 11, 22, 29 June, and 6 July; in 2016 in Vineyard A: 6, 20, 28 June, and 6 July; in 2016 in Vineyard C: 7, 21 June, and 1, 8 July). Only in 2015, both spiders and generalist predatory insects were monitored using yellow sticky traps that were replaced weekly (Vineyards A and B, from 4 June to 18 August). Yellow sticky traps were used in 2015 because they are an effective sampling method for leafhoppers which were considered only during this year [11 (link)]. For both sampling methods, the first sampling was carried out before the first kaolin application. The drop cloth method consisted of manually shaking a grapevine canopy five times, grabbing the apical part of the trunk, and collecting fallen spiders on a pale cloth sheet (65 × 45 cm). At each sampling date, the spiders collected on 10 grapevines in the central part of each subplot were separately preserved in vials with 70% aqueous ethanol. Yellow sticky traps (20 × 10 cm) were smeared with glue (Temo-O-Cid^®, Kollant Srl, Vigonovo, VE, Italy) and hung on the horizontal wires of the grapevine trellis at about 1.5 m from the ground level to be inside the canopy, but not covered by leaves. At each sampling date, one trap per subplot was installed.
Spiders and generalist predatory insects were observed in the laboratory under a dissecting microscope. Spiders collected with the drop cloth method were identified to different taxonomic levels, i.e., adults were assigned to genus or species and juvenile individuals to family or genus. In addition, they were grouped on the basis of hunting strategy (i.e., web-builders and hunters, with the latter further divided into ambushers and active hunters), and body size (i.e., ≤4 mm, small; 4.1–7 mm, medium; 7.1–10 mm, large). The distance from the front of the carapace, without the chelicerae, to the end of the abdomen was measured. Among predatory insects captured by yellow sticky traps, the following taxa were identified: Aeolothrips sp. (Thysanoptera: Aeolothripidae), Scymninae (Coleoptera: Coccinellidae), and Coccinellidae non-Scymninae, Chrysoperla carnea s.l. (Stephens) (Neuroptera: Chrysopidae), and Orius sp. (Hemiptera: Anthocoridae). Among spiders captured by yellow sticky traps, web-builders were distinguished from hunters, which were separately counted at the family level.

We compiled all available karyotypes from online databases [25 (link),26 (link),27 (link),28 (link),29 (link)]. To allow for comparisons across clades, we used the haploid chromosome count of the homogametic sex as a surrogate for the karyotype. For the remainder of the paper, we will refer to this as the chromosome number. For the rate estimates described below, we used the most recent large time-calibrated phylogenies available for each clade: Coleoptera: [30 (link)], Lepidoptera: [31 (link)], Odonata: [32 (link)], Hymenoptera: [13 (link)], Hemiptera: [33 (link)], Blattodea: [33 (link)], Orthoptera: [26 (link)], and Diptera: [34 (link)].