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Review
. 2019 Apr;94(2):555-574.
doi: 10.1111/brv.12467. Epub 2018 Sep 26.

Functional principles of steerable multi-element probes in insects

Affiliations
Review

Functional principles of steerable multi-element probes in insects

Uroš Cerkvenik et al. Biol Rev Camb Philos Soc. 2019 Apr.

Abstract

Hemipterans, mosquitoes, and parasitic wasps probe in a variety of substrates to find hosts for their larvae or food sources. Probes capable of sensing and precise steering enable insects to navigate through solid substrates without visual information and to reach _targets that are hidden deep inside the substrate. The probes belong to non-related taxa and originate from abdominal structures (wasps) or mouthparts (hemipterans and mosquitoes), but nevertheless share several morphological characteristics. Although the transport function clearly differs (egg laying and acquisition of liquid food), the functional demands on the mechanical behaviour of the probe within the substrate tend to be similar. The probe needs to be thin to limit substrate deformation, and long, in order to attain substantial path lengths or depths. We linked the morphology across taxa to the different functional requirements, to provide insights into the biology of probing insects and the evolution of their probes. Current knowledge of insect probes is spread over many taxa, which offers the possibility to derive general characteristics of insect probing. Buckling during initial puncturing is limited by external support mechanisms. The probe itself consist of multiple (3-6) parts capable of sliding along one another. This multi-part construction presumably enables advancement and precise three-dimensional steering of the probe through the substrate with very low net external pushing forces, preventing buckling during substrate penetration. From a mechanical viewpoint, a minimum of three elements is required for 3D steering and volumetric exploration, as realised in the ovipositors of wasps. More elements, such as in six-element probes of mosquitoes, may enhance friction in soft substrates. Alternatively, additional elements can have functions other than 'drilling', such as saliva injection in mosquitoes. Despite the gross similarities, probes show differences in their cross sections, tip morphologies, relative lengths of their elements, and the shape of their interconnections. The hypothesis is that the probe morphology is influenced by the substrate properties, which are mostly unknown. Correlating the observed diversity to substrate-specific functional demands is therefore currently impossible. We conclude that a multipart probe with sliding elements is highly effective for volumetric substrate probing. Shared functional demands have led to an evolutionary convergence of slender multi-element probes in disparate insect taxa. To fully understand 3D probing, it is necessary to study the sensory and material properties, as well as the detailed kinematics and dynamics of the various probes in relation to the nature of the selective pressure originating from the species-specific substrates. Such knowledge will deepen our understanding of probing mechanisms and may support the development of slender, bio-inspired probes.

Keywords: buckling avoidance; hemipterans; mosquitoes; multi-element probes; parasitic wasps; spatial probing; steering.

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Figures

Figure 1
Figure 1
General principles of puncturing, insertion, and bending of the probe in two dimensions. (A) Puncturing. The probe with Young's modulus E, second moment of area I, and length L (real probes are longer than depicted here) is positioned at a suitable location on the substrate and pressed against it (F push and surface reaction force F sub). The probe outside the substrate is stabilised along its length (black vertical lines). (B) Advancement within the substrate. Further insertion is achieved using a ‘push‐pull’ mechanism (Vincent & King, 1995). F ext, pushing or pulling force on the valve; F sub, friction force of the substrate along the probe shaft; F ngb, inner inter‐element friction force; F tip, force on the probe tip region. (C) Hypothesised bending mechanisms of insect probes. In all cases, the tip motion depends on the force generated between probe and substrate. The amount of bending (ii–iv) can be controlled by adjusting the amplitude of pro‐/retraction of individual elements. Steering (ii–iv) can be achieved by the interplay of at least three elements (for clarity, only two are depicted). (i) An asymmetrical geometry of the probe tip region (bevel) leads to asymmetrical forces acting on it (F tip) during insertion, causing the tip to bend from a straight path. Steering can be achieved via rotation of the probe about the longitudinal axis (Webster et al., 2006). (ii) ‘Preloaded’ elements (Pollard, 1969) curve towards each other when a low enough opposing force is present. This results in asymmetric forces as in (i). (iii) Longitudinally restricted movement of the elements by ‘preapical stops’ (Quicke, Fitton & Harris, 1995). The connections cause the build‐up of tension and compression within the elements, thus generating bending moments within the ovipositor. (iv) Differential sclerotisation of elements causes the probe to bend when stiff (arches) and flexible regions (nodes) are aligned with each other (Quicke, 1991).
Figure 2
Figure 2
Examples of insect probes. (A, B) Ovipositor of the parasitoid wasp Megarhyssa atrata (Ichneumonidae) Nénon, Kacem & Lannic (1997). (A) Scanning electron microscopy (SEM) image of the serrated dorsal (bottom, Dv) and ventral (top, Vv) valves. PO, pores; CF, cuticular formations. (B) Cross section through the distal end of the ovipositor. One of the ventral valves is missing. CE, cuticular epithelium; CU, cuticle; LC, longitudinal intravascular canal; SC, intracuticle canal; SE, secretion; v1, ventral valve; v2, dorsal valve. (D, E) Mouthparts of Philagra albinotata (Uhler) (Wang et al., 2015). (D) SEM image of the mouthparts outside the protective labium, with flared out mandibles and maxillae (middle) that are usually kept together. (E) Cross section through the stylets fascicle (location not given in the original article). The mandibles envelop the maxillae that form the food canal (Fc) and the salivary canal (Sc). Asterisks denote the dendritic canals. (G, H) Mosquito mouthparts (proboscis) of Anopheles stephensi (Liston) (Krenn & Aspöck, 2012). Maxillae (Mx) and mandibles (Md) enveloping the labrum (Lr; not indicated in G) and hypopharynx (Hy; not distinguishable in G). La, labium; Fc, food canal. (C, F, and I) Generalised schematics of probe cross sections for Hymenoptera (C), Hemiptera (F) and mosquitoes (I). Ec, egg canal; L, lumen; Nc, neural canal; Mc, membranous connection. All scale bars in µm.
Figure 3
Figure 3
Probing capabilities of hymenopterans and hemipterans. (A, B) Probing of parasitic wasp Diachasmimorpha longicaudata in artificial media (Cerkvenik et al., 2017). (A) A three‐dimensional example of a probing session during which the ovipositor was partially retracted and reinserted in a different direction (blue lines). The endpoint of an individual insertion can be described by its horizontal distance to the start point (r), and the depth in the substrate (d). From these parameters the insertion angle (α) and the position vector of the insertion trajectory endpoint (R) can be calculated. (B) Top view of the insertion endpoints of many different wasps showing no directional preference and a range difference between the stiff (red) and soft (blue) substrates. (C) Parasitic wasp Idarnes flavicollis (Mayr) inserting its ovipositor into a fig fruit (Elias et al., 2012). The ovipositor takes a sinuous path between inflorescences indicating active steering during insertion. Scale not given in original publication (D) Hemipteran Homalodisca coagulata (Say) stylet insertions into sunflower stem (light micrograph) (Leopold et al., 2003). The salivary sheaths (Ss) show a clear branched pattern when probing for xylem vessels (Xy). Pp, pith parenchyma. (E, F) Hemipteran Aulacaspis tubercularis (Newstead) probing a mango leaf (Juárez‐Hernández et al., 2014). (E) Compilation of multiple high‐resolution micrographs of stylet insertion (marked with a black line) through a mango leaf cleared with sodium hypochlorite Scale not given in original publication. A, insertion site; B, last field showing evidence of the stylets. (F) A micrograph showing part of the stylet bundle (arrow).
Figure 4
Figure 4
Variation in element size and inter‐element connections in probes of different taxa. (A–D) Cross sections through ovipositors of various parasitic wasps showing different kinds of inter‐element connections. Note that a basic rail‐like shape is present in all. The differences lie in the size of the connections, their orientation [diverging (A) or converging (D)], and the distance between them. A, B and C are from Quicke et al. (1994): 142, 103, and 93, respectively. Scale bars A–D: 10 µm. (E–H) Inter‐element connections between mouthparts of hemipterans. Note the complex shape of the connection between the maxillae (E, F). The ‘rail type’ can also be present, for example, between the mandibles and maxillae (G). In H, the mandibles (rm) are not connected to the maxillae (rmx) that also contain the salivary canal (sc); Figures from Cobben (1978): 138C (indicated magnification 1400×) and 143C (indicated magnification 4680×). (I–L) Cross‐section of the mosquito proboscis. The stylets are weakly connected and are held together by the labium on the outside of the substrate, but can flare out when inside the tissue: a, labrum; b, maxilla; c, hypopharynx; d, mandible; e, labium. Details of the maxilla–labrum connection are shown in J (detail of I) and K (detail of L). Indicated magnification of I: 495×. Scale bar in L: 10 µm. I and J are from Hudson (1970), fig. 1.1. Species: (A) Coleocentrus sp. (Acaenitinae); (B) Lycorina sp. (Lycorininae); (C) Oedemopsis sp. (Tryphoninae); (D) Diachasmimorpha longicaudata (Braconidae) (E–G) species of Thaumastocoridae; (H) Hebrus ruficeps Thomson (Hebridae), section at the base of the fourth labial segment; (I, J) Aedes atropalpus (Coquillet); (K, L) Culex pipiens (Linnaeus).
Figure 5
Figure 5
Tip variations within insect orders. (A–D) Large variability is observed in the ovipositor tips of phytophagous and parasitoid wasps ranging from smooth valves, to small serrations, to strong serrations (Ghara et al., 2011). The serrations can be present on either the upper valve (uv), the lower valves (lv), or all three valves. (A) Ceratosolen fusciceps (Mayr) (pollinating wasp), (B) Apocryptophagus fusca (Girault) (galler wasp), (C) Apocryptophagus agraensis (Joseph) (parasitoid), (D) Apocrypta westwoodi (Grandi) (parasitoid). (E, F) Different types of serrations on the outer wall of the mandibles of two species of hemipterans: (E) Oncopeltus Fasciatus (Dallas) (Angelini & Kaufman, 2004; contrast enhanced) and (F) Sogatella furcifera (Horváth) (Dai et al., 2014) (G–I) Variation in shape of mosquito maxillae. Species differ in the number and size of stylet serrations. (G) Aedes albopictus (Skuse) (Kong & Wu, 2009), (H) Aedes Atropalpus (Coquillett) (Hudson, 1970), (I) Anopheles farauti (Laveran) (Lee & Craig, 1983). Scale bars: 10 µm. Scale for E and H was not given in the original papers.

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