Quantitative trait locus mapping and functional genomics of an organophosphate resistance trait in the western corn rootworm, Diabrotica virgifera virgifera.

The western corn rootworm, Diabrotica virgifera virgifera, is an insect pest of corn and population suppression with chemical insecticides is an important management tool. Traits conferring organophosphate insecticide resistance have increased in frequency amongst D. v. virgifera populations, resulting in the reduced efficacy in many corn-growing regions of the USA. We used comparative functional genomic and quantitative trait locus (QTL) mapping approaches to investigate the genetic basis of D. v. virgifera resistance to the organophosphate methyl-parathion. RNA from adult methyl-parathion resistant and susceptible adults was hybridized to 8331 microarray probes. The results predicted that 11 transcripts were significantly up-regulated in resistant phenotypes, with the most significant (fold increases ≥ 2.43) being an α-esterase-like transcript. Differential expression was validated only for the α-esterase (ST020027A20C03), with 11- to 13-fold greater expression in methyl-parathion resistant adults (P < 0.05). Progeny with a segregating methyl-parathion resistance trait were obtained from a reciprocal backcross design. QTL analyses of high-throughput single nucleotide polymorphism genotype data predicted involvement of a single genome interval. These data suggest that a specific carboyxesterase may function in field-evolved corn rootworm resistance to organophosphates, even though direct linkage between the QTL and this locus could not be established.

Role of a γ-aminobutryic acid (GABA) receptor mutation in the evolution and spread of Diabrotica virgifera virgifera resistance to cyclodiene insecticides.

The western corn rootworm, Diabrotica virgifera virgifera, is a damaging pest of cultivated corn that was controlled by applications of cyclodiene insecticides from the late 1940s until resistance evolved ∼10 years later. Range expansion from the western plains into eastern USA coincides with resistance development. An alanine to serine amino acid substitution within the Rdl subunit of the gamma-aminobutyric acid (GABA) receptor confers resistance to cyclodiene insecticides in many species. We found that the non-synonymous single nucleotide polymorphism (SNP) G/T at the GABA receptor cDNA position 838 (G/T(838)) of D. v. virgifera resulted in the alanine to serine change, and the codominant SNP allele T(838) was genetically linked to survival of beetles in aldrin bioassays. A phenotypic gradient of decreasing susceptibility from west to east was correlated with higher frequencies of the resistance-conferring T(838) allele in the eastern-most populations. This pattern exists in opposition to perceived selective pressures since the more eastern and most resistant populations probably experienced reduced exposure. The reasons for the observed distribution are uncertain, but historical records of the range expansion combined with the distribution of susceptible and resistant phenotypes and genotypes provide an opportunity to better understand factors affecting the species' range expansion.

Effects of photoperiod on boll weevil (Coleoptera: Curculionidae) development, survival, and reproduction.

Effects of photoperiod on development, survival, feeding, and oviposition of boll weevils, Anthonomus grandis grandis Boheman, were assessed under five different photophases (24, 14, 12, 10, and 0 h) at a constant 27 degrees C temperature and 65% RH in the laboratory. Analyses of our results detected positive relationships between photoperiod and puncturing (mean numbers of oviposition and feeding punctures per day), and oviposition (oviposition punctures/oviposition+feeding punctures) activities, and the proportion of squares attacked by boll weevil females. When boll weevil females developed in light:darkness cycles, they produced a significantly higher percentage of eggs developing to adulthood than those developed in 24-h light or dark conditions. In long photoperiod (24:0 and 14:10 h), the number of female progeny was significantly higher and their development time was significantly shorter than those developed in short photoperiod (0:24 and 10:14 h). Lifetime oviposition was significantly highest at 12- and 14-h photophase, lowest at 0- and 10-h photophase, and intermediate at 24 h of light. Life table calculations indicated that boll weevil populations developed in a photoperiod of 14:10 and 12:12 (L:D) h will increase an average of two-fold each generation (Ro) compared with boll weevils developed in 24:0- and 10:14-h photoperiods and 15-fold compared with those at 0:24 h. Knowledge of the photoperiod-dependent population growth potential is critical for understanding population dynamics to better develop sampling protocols and timing insecticide applications.

Reproductive potential of overwintering, F1, and F2 female boll weevils (Coleoptera: Curculionidae) in the Lower Rio Grande Valley of Texas.

The feeding and oviposition activity of overwintering boll weevils, Anthonomus grandis grandis (Boheman), and seasonal fluctuations in development, survival, and reproduction of progeny of overwintering and first- and second-generation boll weevil females were determined in the laboratory at 27 degrees C, 65% RH, and a photoperiod of 12:12 (L:D) h. During the cotton-free period in the Lower Rio Grande Valley, female boll weevils without access to cotton resorb their unlaid eggs and enter reproductive diapause. However, when they were provided daily with greenhouse-grown cotton squares, commencement of oviposition began after 7, 15, or 20 d, depending on when they were captured. Females captured later in the winter fed longer before laying eggs than those captured in the early fall, suggesting that it may take females longer to terminate diapause the longer they have been dormant. The rate of feeding by females was significantly less during the winter months, and this may have affected the rate of diet-mediated termination of dormancy. Females of the first and second generations after the overwintering generation produced a significantly higher percentage of progeny surviving to adulthood and a higher proportion of these progeny were females. Offspring development time from overwintering female parents was significantly longer than that from first and second generations under the same laboratory conditions. The total number of lifetime eggs produced by females of the second generation during the cotton-growing season were approximately 9.9-fold higher than for overwintering females and 1.5-fold higher than for first-generation females. Life table calculations indicated that the population of second-generation boll weevils increased an average of 1.5-fold higher each generation than for females of the first generation and 22.6-fold higher than for overwintering females. Our data showed variation in boll weevil survival, development, and reproductive potential among the overwintering and first- and second-generation females, suggesting inherent seasonal fluctuations in these parameters.

Size-dependent feeding and reproduction by boll weevil (Coleoptera: Curculionidae).

The considerable variation in adult size of the boll weevil, Anthonomus grandis grandis Boheman, has been well documented, but the influences of adult size on reproductive rate are not known. We examined the relationship between the size of boll weevils and their feeding and oviposition. Weevils weighed to the nearest milligram were grouped into five categories based on pupal weight: < or =5, 6-10, 11-15, 16-20, and >20 mg. Numbers of lifetime punctures produced in flower buds (squares) of cotton, Gossypium hirsutum L., by both sexes of adults tended to increase with pupal weight. Boll weevil females with pupal weights >10 mg produced progeny with significantly higher survival to adulthood and also produced a higher percentage of female progeny than those with pupal weights < or =10 mg. The population growth indices for females having pupal weights >10 mg averaged 1.8-fold higher than those of females weighing < or =10 mg. Survivorship of adults of both sexes also tended to increase with pupal weight. The percentage of females laying eggs on any given day averaged 2.1 times higher when their pupal weights were >10 mg than when their pupal weights were < or =10 mg. Although small size negatively affected female reproductive potential, even extremely small females produced some viable offspring. However, the penalties of small adult size, in terms of longevity and reproductive potential, suggest that cultural practices that result in the production of small adults may be used to impact weevil populations.

Molecular characteristics of insect vitellogenins and vitellogenin receptors.

The recent cloning and sequencing of several insect vitellogenins (Vg), the major yolk protein precursor of most oviparous animals, and the mosquito Vg receptor (VgR) has brought the study of insect vitellogenesis to a new plane. Insect Vgs are homologous to nematode and vertebrate Vgs. All but one of the insect Vgs for which we know the primary structure are cleaved into two subunits at a site [(R/K)X(R/K)R or RXXR with an adjacent beta-turn] recognized by subtilisin-like proprotein convertases. In four of the Vgs, the cleavage site is near the N-terminus, but in one insect species, it is near the C-terminus of the Vg precursor. Multiple alignments of these Vg sequences indicate that the variation in cleavage location has not arisen through exon shuffling, but through local modifications of the amino acid sequences. A wasp Vg precursor is not cleaved, apparently because the sequence at the presumed ancestral cleavage site has been mutated from RXRR to LYRR and is no longer recognized by convertases. Some insect Vgs contain polyserine domains which are reminiscent of, but not homologous to, the phosvitin domain in vertebrate Vgs. The sequence of the mosquito VgR revealed that it is a member of the low-density lipoprotein receptor (LDLR) family. Though resembling chicken and frog VgRs, which are also members of the LDLR family, it is twice as big, carrying two clusters of cysteine-rich complement-type (Class A) repeats (implicated in ligand-binding) instead of one like vertebrate VgRs and LDLRs. It is very similar in sequence and domain arrangement to the Drosophila yolk protein receptor (YPR), despite a non-vitellogenin ligand for the latter. Though vertebrate VgRs, insect VgR/YPRs, and LDLR-related proteins/megalins all accommodate one cluster of eight Class A repeats, fingerprint analysis of the repeats in these clusters indicate they are not directly homologous with one another, but have undergone differing histories of duplications, deletions, and exon shuffling so that their apparent similarity is superficial. The so-called epidermal growth factor precursor region contains two types of motifs (cysteine-rich Class B repeats and YWXD repeats) which occur independently of one another in diverse proteins, and are often involved in protein-protein interactions, suggesting that they potentially are involved in dimerization of VgRs and other LDLR-family proteins. Like the LDLR, but unlike vertebrate VgRs and the Drosophila YPR, the mosquito VgR contains a putative O-linked sugar region on the extra-cellular side of the transmembrane domain. Its function is unclear, but may protect the receptor from membrane-bound proteases. The cytoplasmic tail of insect VgR/YPRs contains a di-leucine (or leucine-isoleucine) internalization signal, unlike the tight-turn tyrosine motif of other LDLR-family proteins. The importance of understanding the details of yolk protein uptake by oocytes lies in its potential for exploitation in novel insect control strategies, and the molecular characterization of the proteins involved has made the development of such strategies a realistic possibility.

Mosquito vitellogenin receptor: purification, developmental and biochemical characterization.

Vitellogenin receptors (VgRs) play a critical role in egg development of oviparous animals by mediating endocytosis of the major yolk protein precursor, vitellogenin. A modification of the method for extracting the mosquito (Aedes aegypti) VgR from ovary membranes resulted in an 11-fold higher yield and 56-fold increase in relative purity of the VgR, in turn permitting purification, antibody production, and microsequencing. A Kd of 15 nM was estimated from binding assays for the enriched VgR, indicating a very high affinity for its ligand. Immunoprecipitation of [14C]VgR using anti-VgR polyclonal antibodies followed by SDS-PAGE under reducing conditions and fluorography demonstrated that the 205 kDa VgR does not consist of subunits held together with disulfide bonds. However, an immunoblot of the native VgR suggests that it exists as an approximately 390 kDa noncovalent homodimer in its native state. Immunoblot assays confirmed that the VgR is present only in ovarian tissue. A quantitative immunoassay of VgR extracts showed that VgR was present in previtellogenic ovaries on the day of emergence, increasing from 2 ng to more than 10 ng per ovary by day 5. After initiation of vitellogenesis and onset of Vg uptake, VgR quantity increased rapidly between 8 and 24 h after a blood meal, then began to decline between 24 and 36 h. Immunocytochemistry confirmed the presence of substantial amounts of the VgR in 4-day-old previtellogenic oocytes. In both previtellogenic and vitellogenic ovaries, the VgR was present only in the oocyte, primarily in the cortex.

Molecular characterization of the VLDL receptor homolog mediating binding of lipophorin in oocyte of the mosquito Aedes aegypti.

Lipophorin (Lp) functions as a yolk protein precursor in the mosquito Aedes aegypti and it is internalized via receptor-mediated endocytosis (Insect Biochem. Mol. Biol., 30 (2000) 1161). We cloned and molecularly characterized a putative mosquito ovarian lipophorin receptor (AaLpRov) cDNA. The cDNA has a length of 3468 bp coding for a 1156-residue protein with a predicted molecular mass of 128.9 kDa. The deduced amino acid sequence of the cDNA revealed that it encodes a protein homolog of the LDL receptor superfamily, and that it harbors eight cysteine-rich ligand binding repeats at the N-terminus like vertebrate VLDL receptors. The deduced amino acid sequence of this mosquito ovarian receptor is most similar to that of the locust lipophorin receptor (LmLpR) (64.3%), and is only distantly related to the mosquito vitellogenin receptor (VgR) (18.3%), another ovarian LDLR homolog with a different ligand. The AaLpRov cDNA was expressed in a TnT Coupled Reticulocyte Lysate system, and co-immunoprecipitation experiments confirmed that the receptor protein specifically binds Lp. Developmental expression profiles clearly showed that AaLpRov transcripts are present in the vitellogenic ovary, with peak expression at 24-36 h post blood meal. In situ hybridization indicated that AaLpRov transcripts are present only in female germ line cells. Distance-based phylogenetic analyses suggest that the insect LpR and vertebrate LDL/VLDL receptor lineages separated after divergence from the insect VgR lineage.

Effects of burial and soil condition on postharvest mortality of boll Weevils (Coleoptera: Curculionidae) in fallen cotton fruit.

Effects of soil condition and burial on boll weevil, Anthonomus grandis grandis Boheman, mortality in fallen cotton, Gossypium hirsutum L., fruit were assessed in this study. During hot weather immediately after summer harvest operations in the Lower Rio Grande Valley of Texas, burial of infested fruit in conventionally tilled field plots permitted significantly greater survival of weevils than in no-tillage plots. Burial of infested squares protected developing weevils from heat and desiccation that cause high mortality on the soil surface during and after harvest in midsummer and late summer. A laboratory assay showed that burial of infested squares resulted in significantly greater weevil mortality in wet than in dry sandy or clay soils. Significantly fewer weevils rose to the soil surface after burial of infested bolls during winter compared with bolls set on the soil surface, a likely result of wetting by winter rainfall. A combination of leaving infested fruit exposed to heat before the onset of cooler winter temperatures and burial by tillage when temperatures begin to cool might be an important tactic for reducing populations of boll weevils that overwinter in cotton fields.

Ligand-binding domains in vitellogenin receptors and other LDL-receptor family members share a common ancestral ordering of cysteine-rich repeats.

Insect vitellogenin and yolk protein receptors (VgR/YPR) are newly discovered members of the low-density lipoprotein receptor (LDLR) family, which is characterized by a highly conserved arrangement of repetitive modular elements homologous to functionally unrelated proteins. The insect VgR/YPRs are unique in having two clusters of complement-type cysteine-rich (class A) repeats or modules, with five modules in the first cluster and seven in the second cluster, unlike classical LDLRs which have a single seven-module cluster, vertebrate VgRs and very low density lipoprotein receptors (VLDLR) which have a single eight-module cluster, and LDLR-related proteins (LRPs) and megalins which have four clusters of 2-7, 8, 10, and 11 modules. Alignment of clusters across subfamilies by conventional alignment programs is problematic because of the repetitive nature of the component modules which may have undergone rearrangements, duplications, and deletions during evolution. To circumvent this problem, we "fingerprinted" each class A module in the different clusters by identifying those amino acids that are both relatively conserved and relatively unique within the cluster. Intercluster reciprocal comparisons of fingerprints and aligned sequences allowed us to distinguish four cohorts of modules reflecting shared recent ancestry. All but two of the 57 modules examined could be assigned to one of these four cohorts designated A, B, C, and D. Alignment of clusters based on modular cohorts revealed that all clusters are derived from a single primordial cluster of at least seven modules with a consensus arrangement of CDCADBC. All extant clusters examined are consistent with this consensus, though none matches it perfectly. This analysis also revealed that the eight-module clusters in vertebrate VgRs, insect VgR/YPRs, and LRP/megalins are not directly homologous with one another. Assignment of modules to cohorts permitted us to properly align 32 class A clusters from all four LDLR subfamilies for phylogenetic analysis. The results revealed that smaller one-cluster and two-cluster members of the family did not originate from the breakup of a large two-cluster or four-cluster receptor. Similarly, the LRP/megalins did not arise from the duplication of a two-cluster insect VgR/YPR-like progenitor. Rather, it appears that the multicluster receptors were independently constructed from the same single-cluster ancestor.

Genetic sources of pheromone variation inColias eurytheme butterflies.

The courtship pheromone ofColias eurytheme butterflies varies greatly among males in both the quantities and relative proportions of its three chemical components [n-heptacosane (C27), 13-methylheptacosane (13MH),n-nonacosane (C29)]. Narrow-sense heritabilities were high for the blend of 13MH and C27 and for the component quantities in one population (Kansas) but were low for the other population tested (Arizona). Genetic correlations between the three components were high in both populations, indicating a substantial degree of additive genetic influence on the component blends. High variability among populations in phenotypic correlations suggests that much of the male-to-male variation in the courtship pheromone may be attributable to environmental or developmental sources. Pheromone phenotypes do not seem to be associated with alba genotype.

Developmental and environmental sources of pheromone variation inColias eurytheme butterflies.

Body size, age, ambient temperature, wing wear, and flight activity were investigated as possible sources of variation in the quantities and relative proportions of the three chemical components [n-heptacosane (C27), 13-methylheptacosane (13MH), andn-nonacosane (C29)] of the male courtship pheromone ofColias eurytheme butterflies. Size of the male has very little influence on the amount of any of the pheromone components present on the wings. Most of the deposition of all three components onto the surface of the hindwing occurs between 3 and 9 hr after emergence from the pupa. 13MH is deposited more rapidly than C27 and C29, and C27 more rapidly than C29. After the first 12 hr posteclosion, the pheromone phenotype of an individual male remains relatively constant through at least 96 hr of age. Experiments showed that none of the three chemicals volatilize to any appreciable extent at temperatures likely to be experienced in the field. The pheromones of actively searching and courting males did not differ from those of less-active feeding and resting males, suggesting that volatilization induced by flight activity is not an important source of pheromone variation. Loss of scales with age does seem to affect pheromone phenotype, but not in a readily interpretable way. Although the quantity of 13MH was lower in worn males than in fresh, C27 was higher.

Genetic structuring of boll weevil populations in the US based on RAPD markers.

Abstract Randomly amplified polymorphic DNA (RAPD) analysis was performed to infer the magnitude and pattern of genetic differentiation among boll weevil populations from eighteen locations across eight US states and north-east Mexico. Sixty-seven reproducible bands from six random primers were analysed for genetic variation within and between weevil populations. Genetic and geographical distances among all populations were positively correlated, reflecting a pattern of isolation by distance within a larger metapopulation. Gene flow between south-central, western and eastern regions is limited, but migration between locations within regions appears to be relatively frequent up to distances of approximately 300-400 km. However, estimates of effective migration were much lower than those estimated from mtDNA-RFLP data reported previously.

Disruptive sexual selection in Colias eurytheme butterflies.

Sexual selection on male pheromone composition in Colias eurytheme (Pieridae) butterflies has the remarkable effect of increasing the variability of this trait. Sexual selection on important traits is generally thought to have a strong stabilizing effect on intraspecific variation of those characters. In this species, however, the male courtship pheromone is highly variable in the relative proportions of its three chemical constituents. Stabilization and/or canalization of this polygenic character in a population is impeded by the disproportionate mating success of males in one portion of the character distribution with alba (white morph) females, and of those in the opposite portion with orange (colored morph) females.

Internalization and recycling of vitellogenin receptor in the mosquito oocyte.

The major yolk protein precursor in mosquito oocytes, vitellogenin (Vg), is internalized by a 205-kDa membrane-bound receptor (VgR). Recently, VgR has been isolated permitting the production of polyclonal anti-VgR antibodies. To elucidate the pathway of VgR internalization and recycling in mosquito oocytes during Vg uptake, we carried out an immunogold electron-microscopic study, labeling both Vg and VgR in ultrathin frozen sections of ovarian tissue. VgR immunolabeling demonstrated that the oocyte plasma membrane was subdivided into microdomains, with VgR being located between and at the lower portions of the oocyte microvilli. During the early stages of internalization, Vg and VgR were observed together in coated pits, coated vesicles, and early endosomes. Fusion of early endosomes created transitional yolk bodies (TYB) in which Vg and VgR became segregated. VgR label was present in the numerous tubular compartments that protruded from the TYBs. These tubular organelles extended to and fused with the plasma membrane, suggesting that they represented the vehicle for VgR recycling. Vg label was not observed in the tubular compartments. Instead, Vg accumulated in the core of the TYB, a region free of VgR label. Mature yolk bodies (MYB) were heavily labeled for Vg, but completely lacked any VgR label, indicating that MYB are storage compartments that do not participate in receptor recycling. Thus, our immunocytochemical data clearly visualize the steps in Vg/VgR internalization, dissociation, sorting, and recycling of the receptor to the plasma membrane.

Extensive sequence conservation among insect, nematode, and vertebrate vitellogenins reveals ancient common ancestry.

The eggs of most oviparous animals are provisioned with a class of protein called vitellogenin (Vg) which is stored as the major component of yolk. Until recently, deduced amino acid sequences were available only from vertebrate and nematode Vgs, which proved to be homologous. The sequences of several insect Vgs are now known, but early attempts at pairwise alignments with vertebrate and nematode Vgs have been problematic, leading to conflicting conclusions about how closely insect Vgs are related to the others. In this paper we demonstrate that insect Vg sequences can be confidently aligned with one another along their entire lengths and with multiple vertebrate and nematode Vg sequences along most of their spans. Although divergence is high, conservation among insect, vertebrate, and nematode Vg sequences is widespread with a preponderance of glycine, proline, and cysteine residues among strictly conserved amino acids, establishing conclusively that Vgs from the three phyla are homologous. Areas of least-certain alignment are primarily in and around insect and vertebrate polyserine domains which are not homologous. Phylogenetic reconstructions of Vgs based on sequence identities indicate that the insect lineage is the most diverged and that the mammalian serum protein, apolipoprotein B-100, arose from a Vg ancestor after the nematode/vertebrate divergence.

A hexamerin protein, AgSP-1, is associated with diapause in the boll weevil(1).

The objective of this research was to identify a reliable biochemical indicator for diapause (dormancy) in the boll weevil, Anthonomus grandis. Hemolymph polypeptides from reproductive and diapausing weevils were compared using denaturing sodium dodecyl sulpfate (SDS)-polyacrylamide gel electrophoresis (PAGE). A 77-kDa protein, which proved to be a hexamerin (AgSP-1), strongly correlated with morphological diapause characters in both male and female adult weevils. N-terminal sequence analysis identified the first 25 amino acids of the mature protein and was used to develop an antibody to AgSP-1. Anti-AgSP-1 reacted only with hemolymph from diapausing weevils of both sexes but not with hemolymph from reproductive weevils. Also, the yolk protein, vitellogenin (VG), inversely correlated with AgSP-1. When hemolymph VG was high, AgSP-1 was absent or barely perceptible.Juvenile hormone regulates VG synthesis in most insect species. Juvenile hormone is reported to stimulate reproductive maturation in the boll weevil (Physiological Entomology 22 (1997) 261) and to be absent during diapause (Physiological Entomology 22 (1997a) 269). Therefore, the juvenile hormone (JH) mimic, methoprene, was used to assess the role of JH activity in the boll weevil for terminating diapause, stimulating reproductive maturation and possibly influencing AgSP-1 titers. Application of methoprene was not effective in activating reproductive development. Hemolymph from methoprene-treated, females contained VG and AgSP-1 titers that were similar to acetone-treated and untreated control weevils.Using a genomic DNA library and 3' RACE, two clones were isolated that yielded the complete sequence of AgSP-1 as well as a portion of the 5' untranslated region. Northern blot analysis confirmed the presence of a 2.5 kB transcript for AgSP-1 in the fat body of diapausing weevils. AgSP-1 was also present in the fat body of reproductive weevils, but to a lesser extent. No sex-related differences in gene expression were observed; diapausing weevils of both sexes showed similar levels of AgSP-1 expression. An inverse correlation was observed between the VG transcript and AgSP-1 mRNA. VG was highly expressed in the fat body of reproductive females and only slightly expressed in tissue from diapausing females. Our data suggests that AgSP-1 is a diapause-specific protein in adult weevils and that JH, alone, is not effective in terminating diapause.

Molecular characterization of the mosquito vitellogenin receptor reveals unexpected high homology to the Drosophila yolk protein receptor.

The mosquito (Aedes aegypti) vitellogenin receptor (AaVgR) is a large membrane-bound protein (214 kDa when linearized) that mediates internalization of vitellogenin, the major yolk-protein precursor, by oocytes during egg development. We have cloned and sequenced two cDNA fragments encompassing the entire coding region of AaVgR mRNA, to our knowledge the first insect VgR sequence to be reported. The 7.3-kb AaVgR mRNA is present only in female germ-line cells and is abundant in previtellogenic oocytes, suggesting that the AaVgR gene is expressed early in oocyte differentiation. The deduced amino acid sequence predicts a 202.7-kDa protein before posttranslational processing. The AaVgR is a member of the low density lipoprotein receptor superfamily, sharing significant homology with the chicken (Gallus gallus) VgR and particularly the Drosophila melanogaster yolk protein receptor, in spite of a very different ligand for the latter. Distance-based phylogenetic analyses suggest that the insect VgR/yolk protein receptor lineage and the vertebrate VgR/low density lipoprotein receptor lineage diverged before the bifurcation of nematode and deuterostome lines.