Comparative morphology of sensilla on antenna,maxillary palp and labial palp of larvae of Eucryptorrhynchus scrobiculatus (Olivier) and E. brandti (Harold) (Coleoptera: Curculionidae)

Eucryptorrhynchus scrobiculatus (Olivier) and E. brandti (Harold) are two wood boring pests of Ailanthus altissima (Mill.) Swingle (tree of heaven) and the variety Ailanthus altissima var. Qiantouchun. These beetles attack healthy trees and bore into roots and trunks during the larval stage. We studied the typology, distribution and morphostructure of the sensilla on the antennae, maxillary palps and labial palps of E. scrobiculatus and E. brandti larvae using scanning and transmission electron microscopy. The results showed the following: (i) the antennae of the two weevil larvae had two types of sensilla, sensilla basiconica (S.b.1 and S.b.2) and sensilla twig basiconica (S.tb.1‐S.tb.3), with S.tb.4 observed only on the antennae of E. brandti larvae; (ii) the maxillary palps had three types of sensilla, S.b.2, S.tb. (S.tb.2, S.tb.3 and S.tb.5) and digitiform sensilla; (iii) the labial palps had two types of sensilla, S.b.2 and S.tb. (S.tb.2, S.tb.3 and S.tb.5); (iv) the quantity and distribution of sensilla on the antennae, maxillary palps and labial palps remained constant between E. scrobiculatus and E. brandti larvae; and (v) sensilla basiconica had distinct sidewall pores, an apical pore was observed on sensilla twig basiconica, and digitiform sensilla were oval in shape, with a distinct apical pore. Based on the microstructure of the cuticle wall and dendrite, we hypothesized that these sensilla functioned as olfactory, gustatory and hygro‐/thermo‐receptors, respectively. We discuss the relationships among types of sensilla and the types of damage caused by the larvae inside the host tree to understand olfactory and gustatory receptor mechanisms. The results of this study will provide a firm basis for future electrophysiological studies.     

Gustatory organs of Drosophila melanogaster: fine structure and expression of the putative odorant-binding protein PBPRP2

In Drosophila, as in most insects, gustation is mediated by sensory hairs located on the external and internal parts of the proboscis and on the legs and wings. We describe in detail the organization and ultrastructure of the gustatory sensilla on the labellum and legs and the distribution of PBPRP2, a putative odorant-binding protein, in the gustatory organs of Drosophila. The labellum carries two kinds of sensilla: taste bristles and taste pegs. The former have the typical morphology of gustatory sensilla and can be further subdivided into three morphological subtypes, each with a stereotyped distribution and innervation. Taste pegs have a unique morphology and are innervated by two receptor cells: one mechanoreceptor and the other a putative chemoreceptor cell. PBPRP2 is abundantly expressed in all adult gustatory organs on labellum, legs, and wings and in the internal taste organs on the proboscis. In contrast to olfactory organs, where PBPRP2 is expressed in the epidermis, this protein is absent from the epidermis of labial palps and legs. In the taste bristles of the labellum and legs, PBPRP2 is localized in the crescent-shaped lumen of the sensilla, and not in the lumen where the dendrites of the gustatory neurons are found, making a function in stimulus transport unlikely in these sensilla. In contrast, PBPRP2 in peg sensilla is expressed in the inner sensillum-lymph cavity and is in contact with the dendrites. Thus, PBPRP2 could be involved as a carrier for hydrophobic ligands, e.g., bitter tastants, in these sensilla.     

Restricted diet breadth of the larvae of Antheraea assamensis and the role of the labrum‐epipharynx and galeal sensilla

The silkworm, Antheraea assamensis Helfer (Lepidoptera: Saturniidae), grows primarily on Persea bombycina and Litsea polyantha. To understand if the restricted diet breadth is due to the specific role of gustatory sensilla of the larvae of A. assamensis, the same fifth instar larvae retaining only labrum‐epipharynx or galeal sensilla were subjected to food choice tests. The foods used were leaves of two host‐plant and two non‐host‐plant species. Mean per cent consumption and per cent of choosing larvae were used as parameters for drawing conclusions. The finding indicated involvement of the labrum‐epipharynx for acceptance and galeal sensilla for rejection of a non‐host‐plant species. Scanning electron microscope studies revealed the presence of two sensilla on the galea, one lateral and one medial sensilla styloconicum and two gustatory sensilla in the epipharynx of A. assamensis. The study revealed the key role of galeal sensilla in the restrictive diet‐breadth of A. assamensis.     

Sensilla on maxillary and labial palps in a helicophagous ground beetle larva (Coleoptera,Carabidae)

Sensilla on the labial and maxillary palp of Carabus lefebvrei Dejean, 1826 larvae were investigated using scanning and transmission electron microscopy. Two identical sets of sensilla were present on the tips of both palp pairs, and four morphological types of sensilla were identified: sensilla basiconica types 1 and 2, sensilla coeloconica and sensilla digitiformia. Ultrastructure indicates that the sensilla basiconica type 1 and coeloconica have a chemical role as gustatory and olfactory receptors, respectively, while sensilla basiconica type 2 are mechanoreceptors and the sensilla digitiformia are probably thermo‐ and hygroreceptors. Their function is discussed in relation to specialized prey detection and habitat adaptations.     

Aspects on the Ultrastructure of the Cephalic Region in Two Species of Acrobelinae (Nematoda,Cephalobidae)

The lip structures termed labial probolae, characteristic for the subfamily Acrobelinae, and the anterior sense organs have been studied by electron microscopy. The labial probolae consist of an amorphous tissue with denser strengthenings. They do not contain any sensory structures or musculature. The anterior sense organs consists of: (1) six inner labial sensilla, each with one receptor ending in a pore on a papilla; (2) six outer labial sensilla, each with one receptor ending in the cuticle; (3) four cephalic sensilla, each with two receptors, one ending in a pore on a papilla and the other in the cuticle; (4) two amphids, each with 12 sensory neurons basally; (5) accessory sensilla ventro-laterally. The combination of several chemoreceptive anterior sensilla and the morphological diversity of the labial probolae within the subfamily may aid in the utilization of different soil microhabitats.     

Multimodal sensory organs in larvae of some insect species

The morphological diversity of antennal sensory organs in larvae of Galeruca sp. (Coleoptera, Chrysomelidae), Limnephilus centralis (Trichoptera, Limnephilidae), and Eristalis tenax (Diptera, Syrphidae) was studied. A sensory cone on the antenna was found in all the species studied. The cuticular part of the antennal cone is pierced with pores. No significant differences in the shape of the antennal cone were revealed. The innervation patterns of the antennal cone varied from single cells in the leaf beetle larvae to groups of cells with a common scolopoid sheath in the fly larvae. The cone combines several types of sensilla that perform mechanoreceptor, olfactory, and gustatory functions. The morphology and ultrastructural organization of the antennal cone in relation to the heterogeneity of the larval habitats are described.     

Sensing the Underground - Ultrastructure and Function of Sensory Organs in Root-Feeding Melolontha melolontha (Coleoptera: Scarabaeinae) Larvae

Introduction

Below ground orientation in insects relies mainly on olfaction and taste. The economic impact of plant root feeding scarab beetle larvae gave rise to numerous phylogenetic and ecological studies. Detailed knowledge of the sensory capacities of these larvae is nevertheless lacking. Here, we present an atlas of the sensory organs on larval head appendages of Melolontha melolontha. Our ultrastructural and electrophysiological investigations allow annotation of functions to various sensory structures.

Results

Three out of 17 ascertained sensillum types have olfactory, and 7 gustatory function. These sensillum types are unevenly distributed between antennae and palps. The most prominent chemosensory organs are antennal pore plates that in total are innervated by approximately one thousand olfactory sensory neurons grouped into functional units of three-to-four. In contrast, only two olfactory sensory neurons innervate one sensillum basiconicum on each of the palps. Gustatory sensilla chaetica dominate the apices of all head appendages, while only the palps bear thermo-/hygroreceptors. Electrophysiological responses to CO₂, an attractant for many root feeders, are exclusively observed in the antennae. Out of 54 relevant volatile compounds, various alcohols, acids, amines, esters, aldehydes, ketones and monoterpenes elicit responses in antennae and palps. All head appendages are characterized by distinct olfactory response profiles that are even enantiomer specific for some compounds.

Conclusions

Chemosensory capacities in M. melolontha larvae are as highly developed as in many adult insects. We interpret the functional sensory units underneath the antennal pore plates as cryptic sensilla placodea and suggest that these perceive a broad range of secondary plant metabolites together with CO₂. Responses to olfactory stimulation of the labial and maxillary palps indicate that typical contact chemo-sensilla have a dual gustatory and olfactory function.     

Comparative morphology of sensilla on antenna,maxillary palp and labial palp of larvae of white‐spotted and yellow‐spotted Asian long‐horned beetle,Anoplophora glabripennis Motschulsky (Coleoptera: Cerambycidae)

The Asian long‐horned beetle (ALB) is one of the most important wood‐boring insects worldwide that damages broad‐leaved trees, primarily poplar, willow, elm and maple. Based on the color of the spots on the elytra, the beetles are separated into white‐spotted (ALB‐W) and yellow‐spotted (ALB‐Y) Asian long‐horned beetles. In order to clarify the morphology of sensilla on antenna, maxillary palp and labial palp of ALB‐W and ALB‐Y larvae, we studied the typology, morphology, number and distribution of the sensilla by scanning electron microscopy. The results showed that: (i) the antennae of two biotypes had five types of sensilla, including three types of sensilla basiconica (b.) and two types of sensilla twig basiconica (s.tb); numbers of b.1, b.2, b.3 and s.tb1 on antenna were not significantly different (P > 0.05) between two biotypes, and the numbers of s.tb2 were significantly different (P < 0.05); (ii) the maxillary palp of two biotypes had four types of sensilla, including sensilla styloconica (st.), two types of s.tb and digitiform sensilla (ds.); the numbers of st. and ds. on the maxillary palp were not significantly different (P > 0.05) between two biotypes, and the numbers of s.tb1 and s.tb2 were significantly different (P < 0.05); (iii) the labial palp of two biotypes had four types of sensilla, including b., st. and two types of s.tb, and the numbers of b.3, st., s.tb1 and s.tb2 on the labial palp were not significantly different (P > 0.05) between two biotypes. We discuss the relationships between sensilla and damage caused by the larvae inside the host trees.     

Role of proneural genes in the formation of the larval olfactory organ of <Emphasis Type="Italic">Drosophila</Emphasis>

In this paper, we address the role of proneural genes in the formation of the dorsal organ in the Drosophila larva. This organ is an intricate compound comprising the multineuronal dome—the exclusive larval olfactory organ—and a number of mostly gustatory sensilla. We first determine the numbers of neurons and of the different types of accessory cells in the dorsal organ. From these data, we conclude that the dorsal organ derives from 14 sensory organ precursor cells. Seven of them appear to give rise to the dome, which therefore may be composed of seven fused sensilla, whereas the other precursors produce the remaining sensilla of the dorsal organ. By a loss-of-function approach, we then analyze the role of atonal, amos, and the achaete-scute complex (AS-C), which in the adult are the exclusive proneural genes required for chemosensory organ specification. We show that atonal and amos are necessary and sufficient in a complementary way for four and three of the sensory organ precursors of the dome, respectively. AS-C, on the other hand, is implicated in specifying the non-olfactory sensilla, partially in cooperation with atonal and/or amos. Similar links for these proneural genes with olfactory and gustatory function have been established in the adult fly. However, such conserved gene function is not trivial, given that adult and larval chemosensory organs are anatomically very different and that the development of adult olfactory sensilla involves cell recruitment, which is unlikely to play a role in dome formation. N. Grillenzoni and V. de Vaux contributed equally to this work.     

Sensory structures involved in prey detection on the labial palp of the ant‐hunting beetle Siagona europaea Dejean 1826 (Coleoptera,Carabidae)

Giglio, A., Ferrero E.A., Perrotta, E., Talarico, F.F. and Zetto Brandmayr, T. 2010. Sensory structures involved in prey detection on the labial palp of the ant‐hunting beetle Siagona europaea Dejean 1826 (Coleoptera, Carabidae). —Acta Zoologica (Stockholm) 91 : 328–334 The ultrastructure and distribution of sensilla on the labial palps of a myrmecophagous carabid beetle, Siagona europaea, were investigated using scanning and transmission electron microscopy techniques. Five types of sensilla were identified: three types of sensilla basiconica on the apical sensory area and two types, one sensillum trichodeum and one coeloconicum, on the external palp surface. On morphological grounds, the s. basiconica type 1 were considered as olfactory, the type 2 as gustatory, the type 3 and the s. trichodeum as mechanoreceptive, and the s. coeloconicum as a thermo/hygroreceptor. Their function is discussed in relation to prey detection and habitat adaptations.     

Sensory organs of the antennae and mouthparts of beetle larvae (Coleoptera)

The sensilla located on the antennae and maxillary and labial palps of the larvae of 64 beetle species from 22 families were studied using electron microscopy. The larvae of beetles living in different habitats and having different trophic specializations possess a uniform structure of the sensory organs. They are composed of two groups of sensilla on the apical and subapical segments of the antennae, one apical group of sensilla on both maxillary and labial palps, and one or several digitiform sensilla on the lateral surface of the maxillary and, occasionally, labial palp. The external morphology of the sensory organs is adaptive and represents modifications of the initial type. Band-shaped sensilla or placoid sensilla, clearly different from the initial sensory organs, appear in some taxa as rare exceptions, while other groups display either partial reduction of the receptor organs (Gyrinidae) or reduction of the cuticular parts of the sensilla (Cantharidae).     

An accessory olfactory pathway in Lepidoptera: the labial pit organ and its central projections in Manduca sexta and certain other sphinx moths and silk moths

Summary In the hawkmoth, Manduca sexta, the third segment of each labial palp contains a pit, which houses a densely packed array of sensilla. We have named this structure the labial pit organ (LPO). The sensilla within the pit are typical of olfactory receptors, characterized by a grooved surface, wall pores, and pore tubules. Axons arising from receptor cells that innervate these sensilla project bilaterally to a single glomerulus in each antennal lobe. We have compared this central projection with that in three other species of Manduca (M. quinquemaculata, M. dilucida, and M. lanuginosa) and in the silkmoths Antheraea polyphemus and Bombyx mori. A bilateral projection to a single glomerulus in each antennal lobe is present in all cases. We suggest that the LPO serves as an accessory olfactory organ in adult Lepidoptera.     

Sexual dimorphism of antennal,tarsal and ovipositor chemosensilla in the African stemborer,Busseola fusca (Fuller) (Lepidoptera: Noctuidae)

The number and distribution of chemosensilla located on different organs of Busseola fusca (Fuller) (Lepidoptera: Noctuidae) males and females are described based on observations using scanning electron microscopy, selective staining with silver nitrate, and gustatory electrophysiological recording. The antennae and the fifth tarsomere of the prothoracic legs of both sexes bear chemosensilla: uniporous chaetica and multiporous trichoidea sensilla. However, there is a sexual dimorphism in the number and size of sensilla on these organs. The distal part of the ovipositor has uniporous gustatory chemosensilla of the chaetica type. The involvement of these sensilla in oviposition site selection by B. fusca is discussed.     

Larval head anatomy of Heterogynis penella (Zygaenoidea, Heterogynidae), and a general discussion of caterpillar head structure (Insecta, Lepidoptera)

The internal head anatomy (and the peculiar integumental structure of the epicranial notch region) of Heterogynis penella larvae are described; special attention is paid to the skeleto‐muscular and nervous systems and to the cephalic glands. Transverse ligaments connect the apodemes of the mandibular adductor muscles of both sides and the anterior maxillo‐labial articulations of both sides. The two ligaments are linked to each other by a thin, apparently acellular membrane. An accessory, trilobed mandibular gland is present. A putative stretch receptor, connecting the oblique dorsal cibarial dilators of both sides, is described for the first time in a lepidopterous larva and its importance in assessing the homology of these muscles is discussed. The presence of cibarial sensilla, previously predicted in other caterpillars on the basis of behavioural experiments and observations of the nerve pattern, is confirmed. The structural diversity of larval head anatomy in ditrysian Lepidoptera is discussed, with particular emphasis on the innervation of the corpora cardiaca and corpora allata and of the sensilla of the head capsule.     

Sexual dimorphism of adult labial palps of the peach fruit moth Carposina sasakii Matsumura (Lepidoptera: Carposinidae) with notes on their sensilla

The labial palps and their sensilla of the peach fruit moth Carposina sasakii Matsumura, a serious pest of pome fruits in eastern Asia, were investigated using light microscopy and scanning and transmission electron microscopy. The labial palps are three‐segmented and exhibit distinct sexual dimorphism in length: much longer in the female than in the male. Four types of sensilla were found on the labial palps: campaniform, squamiform, chaetic and flattened sensilla. The campaniform sensilla are present on the first segment alone. The squamiform sensilla are located on all the three segments. The chaetic sensilla are mainly present on the third segment. The flattened sensilla are grooved with wall pores and situated in a labial palp‐pit organ, which is located at the apex of the distal segment of the labial palp. The sexual dimorphism of the labial palp and the labial palp‐pit organ was briefly discussed.     

The accessory organ,a scolopidial sensory organ,in the cave cricket Troglophilus neglectus (Orthoptera: Ensifera: Rhaphidophoridae)

Mechanoreceptor organs occur in great diversity in insect legs. This study investigates sensory organs in the leg of atympanate cave crickets (Troglophilus neglectus KRAUSS, 1879) by neuronal tracing. Previously, the subgenual and the intermediate organs were recognised in the subgenual organ complex, lacking the tympanal membranes present for example in the tibial hearing organs of Gryllidae and Tettigoniidae. We document the presence of the accessory organ in T. neglectus. This scolopidial organ is located in the posterior tibia close to the subgenual organ and can be identified by position, innervation and orientation of the dendrites of sensory neurons. The main motor nerve in the leg innervates a part of the subgenual organ and the accessory organ. The dendrites of sensory neurons in the accessory organ are characteristically bent in proximo‐dorsal direction, while the subgenual organ dendrites run distally along the longitudinal axis of the leg. The accessory organ contains 6–10 scolopidial sensilla, and no differences in neuroanatomy occur between the three thoracic leg pairs. Hence, the subgenual organ complex in cave crickets is more complex than previously known. The wider taxonomic distribution of the accessory scolopidial organ among orthopteroid insects is inconsistent, indicating its repeated losses or convergent evolution.     

Two antagonistic gustatory receptor neurons responding to sweet‐salty and bitter taste in Drosophila

In Drosophila, gustatory receptor neurons (GRNs) occur within hair‐like structures called sensilla. Most taste sensilla house four GRNs, which have been named according to their preferred sensitivity to basic stimuli: water (W cell), sugars (S cell), salt at low concentration (L1 cell), and salt at high concentration (L2 cell). Labellar taste sensilla are classified into three types, l‐, s‐, and i‐type, according to their length and location. Of these, l‐ and s‐type labellar sensilla possess these four cells, but most i‐type sensilla house only two GRNs. In i‐type sensilla, we demonstrate here that the first GRN responds to sugar and to low concentrations of salt (10–50 mM NaCl). The second GRN detects a range of bitter compounds, among which strychnine is the most potent; and also to salt at high concentrations (over 400 mM NaCl). Neither type of GRN responds to water. The detection of feeding stimulants in i‐type sensilla appears to be performed by one GRN with the combined properties of S + L1 cells, while the other GRN detects feeding inhibitors in a similar manner to bitter‐sensitive L2 cells on the legs. These sensilla thus house two GRNs having an antagonistic effect on behavior, suggesting that the expression of taste receptors is segregated across them accordingly. © 2004 Wiley Periodicals, Inc. J Neurobiol, 2004     

The contribution of gustatory input to larval acceptance and female oviposition choice of potential host plants in Papilio hospiton (Géné)

The Lepidopteran Papilio hospiton uses only plants belonging to the Apiaceae and the Rutaceae families as hosts. Both adult females and larvae are equipped with gustatory receptor neurons (GRNs) capable of detecting sugars, bitters and salts, thus providing information for evaluating the chemical composition of the plant. Since the activation of these neurons may affect insect behavior, the aim of this study were: (a) to study the gustatory sensitivity of both females and larvae to the sap of two Apiaceae, Foeniculum vulgare (fennel) and Daucus carota (carrot), that are not used as host plants; (b) to cross‐compare the spike activity evoked from these two plants with that evoked by Ferula communis (ferula), the host plant preferred by ovipositing females of P. hospiton and where the larvae perform best; (c) finally, to confirm that the gustatory system can provide the central nervous system with the necessary information to evaluate differences between plant saps. The results show that: (a) fennel and carrot both evoke a higher neural activity from the bitter‐sensitive neurons and lower from the sugar‐sensitive neurons with respect to ferula, in both adult females and larvae; (b) on the basis of the different patterns of neural activity generated in tarsal, lateral and medial sensilla by fennel and carrot versus ferula, both adult and larvae possess enough information to discriminate among these plants; (c) adult females of P. hospiton lay eggs where the larvae have the greatest growth success and this confirms the importance of taste sensitivity in host plants selection.     

Transgenic maize lines with cell‐type specific expression of fluorescent proteins in plastids

Plastid number and morphology vary dramatically between cell types and at different developmental stages. Furthermore, in C4 plants such as maize, chloroplast ultrastructure and biochemical functions are specialized in mesophyll and bundle sheath cells, which differentiate acropetally from the proplastid form in the leaf base. To develop visible markers for maize plastids, we have created a series of stable transgenics expressing fluorescent proteins fused to either the maize ubiquitin promoter, the mesophyll‐specific phosphoenolpyruvate carboxylase (PepC) promoter, or the bundle sheath‐specific Rubisco small subunit 1 (RbcS) promoter. Multiple independent events were examined and revealed that maize codon‐optimized versions of YFP and GFP were particularly well expressed, and that expression was stably inherited. Plants carrying PepC promoter constructs exhibit YFP expression in mesophyll plastids and the RbcS promoter mediated expression in bundle sheath plastids. The PepC and RbcS promoter fusions also proved useful for identifying plastids in organs such as epidermis, silks, roots and trichomes. These tools will inform future plastid‐related studies of wild‐type and mutant maize plants and provide material from which different plastid types may be isolated.     

Transient GFP expression in Nicotiana plumbaginifolia suspension cells: the role of gene silencing,cell death and T-DNA loss

The transient nature of T-DNA expression was studied with a gfp reporter gene transferred to Nicotiana plumbaginifolia suspension cells fromAgrobacterium tumefaciens. Individual GFP-expressing protoplasts were isolated after 4 days' co-cultivation. The protoplasts were cultured without selection and 4 weeks later the surviving proto-calluses were again screened for GFP expression. Of the proto-calluses initially expressing GFP, 50% had lost detectable GFP activity during the first 4 weeks of culture. Multiple T-DNA copies of the gfp gene were detected in 10 of 17 proto-calluses lacking visible GFP activity. The remaining 7 cell lines contained no gfp sequences. Our results confirm that transiently expressed T-DNAs can be lost during growth of somatic cells and demonstrate that transiently expressing cells frequently integrate multiple T-DNAs that become silenced. In cells competent for DNA uptake, cell death and gene silencing were more important barriers to the recovery of stably expressing transformants than lack of T-DNA integration.