Orb-weaving Spider Nephila Clavipes Research Articles

Custom-designed, mass silk production in genetically engineered silkworms.

Genetically engineered silkworms have been widely used to obtain silk with modified characteristics especially by introducing spider silk genes. However, these attempts are still challenging due to limitations in transformation strategies and difficulties in integration of the large DNA fragments. Here, we describe three different transformation strategies in genetically engineered silkworms, including transcription-activator-like effector nuclease (TALEN)-mediated fibroin light chain (FibL) fusion (BmFibL-F), TALEN-mediated FibH replacement (BmFibH-R), and transposon-mediated genetic transformation with the silk gland-specific fibroin heavy chain (FibH) promoter (BmFibH-T). As the result, the yields of exogenous silk proteins, a 160 kDa major ampullate spidroin 2 (MaSp2) from the orb-weaving spider Nephila clavipes and a 226 kDa fibroin heavy chain protein (EvFibH) from the bagworm Eumeta variegate, reach 51.02 and 64.13% in BmFibH-R transformed cocoon shells, respectively. Moreover, the presence of MaSp2 or EvFibH significantly enhances the toughness of genetically engineered silk fibers by ∼86% in BmFibH-T and ∼80% in BmFibH-R silkworms, respectively. Structural analysis reveals a substantial ∼40% increase in fiber crystallinity, primarily attributed to the presence of unique polyalanines in the repetitive sequences of MaSp2 or EvFibH. In addition, RNA-seq analysis reveals that BmFibH-R system only causes minor impact on the expression of endogenous genes. Our study thus provides insights into developing custom-designed silk production using the genetically engineered silkworm as the bioreactor.

Effect of Loading Rate on Mechanical Properties and Fracture Morphology of Spider Silk

Spider silks have been shown to have impressive mechanical properties. In order to assess the effect of extension rate, both quasi-static and high-rate tensile properties were determined for single fibers of major (MA) and minor (MI) ampullate single silk from the orb weaving spider Nephila clavipes . Low rate tests have been performed using a DMA Q800 at 10(-3) s(-1), while high rate analysis was done at 1700 s(-1) utilizing a miniature Kolsky bar apparatus. Rate effects exhibited by both respective silk types are addressed, and direct comparison of the tensile response between the two fibers is made. The fibers showed major increases in toughness at the high extension rate. Mechanical properties of these organic silks are contrasted to currently employed ballistic fibers and examination of fiber fracture mechanisms are probed via scanning electron microscope, revealing a globular rupture surface topography for both rate extremums.

Developmental response to low diets by giant Nephila clavipes females (Araneae: Nephilidae)

Abstract Female-biased sexual size dimorphism is common in arthropods, apparently driven by fecundity selection in females. Selective pressures that limit growth are less often considered. One factor that researchers have rarely considered is the possible role of energetic limits on growth. The orb weaving spider Nephila clavipes (Linnaeus 1767) is extremely sexually size dimorphic. Males are “normal” sized spiders and females are up to ten times longer, having passed through several additional juvenile instars. This extreme size dimorphism presents the opportunity to test for intrinsic energetic costs of gigantism. Prior studies have shown that males successfully reach maturity on a range of diets, while female dietary requirements increase rapidly with increasing size. We here examine the effects of variation in food availability on juvenile female development by randomly assigning spiderlings from six different families (from six distinct populations) to quantitatively varying but qualitatively identic...

Effects of male size and female dispersion on male mate-locating success in Nephila clavipes

The reproductive success of sexually reproducing organisms depends on their capacity to locate potential partners. Research on the mating systems of web-building spiders, like other terrestrial invertebrates, has focused around male courtship and the signalling behaviours that facilitate copulation. In contrast, both mate searching and mate location behaviours are largely ignored and rarely quantified in field studies. We conducted a series of field removal experiments to explore the effects of male body size and female dispersion on mate locating behaviour of the golden orb-weaving spider (Nephila clavipes). The number of males that arrived on females’ webs was influenced by travel distance, male body size, male body weight and female dispersal. More males returned to webs that were closer to the release point; smaller males experienced a greater weight loss; and larger numbers of males arrived at webs where males had been previously present. Female dispersion influenced the size but not the number of arriving males: males were no more likely to arrive at solitary than aggregations of webs, but males arriving at solitary webs were typically larger than those arriving at aggregations. We discuss the implications of various male traits that could act as selective pressures on male mate location and mate searching behaviours.

Diversity of Molecular Transformations Involved in the Formation of Spider Silks

Spiders that spin orb webs secrete seven types of silk. Although the spinning process of the dragline thread is beginning to be understood, the molecular events that occur in spiders' opisthosomal glands, which produce the other fibers, are unknown due to a lack of data regarding their initial and final structures. Taking advantage of the efficiency of Raman spectromicroscopy in investigating micrometer-sized biological samples, we have determined the secondary structure of proteins in the complete set of glands of the orb-weaving spider Nephila clavipes. The major and minor ampullate silks in the sac of their glands have identical secondary structures typical of natively unfolded proteins. Spidroins are converted into fibers containing highly oriented β-sheets. The capture spiral represents a distinct structural singleton. The proteins are highly disordered prior to spinning and undergo no molecular change or alignment upon spinning. The cylindrical, aciniform, and piriform proteins are folded in their initial state with a predominance of α-helices, but whereas the cylindrical gland forms a fiber similar to the major ampullate thread, the aciniform and piriform glands produce fibers dominated by moderately oriented β-sheets and α-helices. The conformation of the proteins before spinning is related to intrinsic characteristics of their primary structure. Proteins that are unfolded in the gland have repeat sequences composed of submotifs and display no sequence regions with aggregation propensity. By contrast, the folded proteins have neither submotifs nor aggregation-prone sequence regions. Taken together, the Raman data show a remarkable diversity of molecular transformations occurring upon spinning.

Spider Web Glue: Two Proteins Expressed from Opposite Strands of the Same DNA Sequence

The various silks that make up the web of the orb web spiders have been studied extensively. However, success in prey capture depends as much on the web glue as on the fibers. Spider silk glue, which is considered one of the strongest and most effective biological glues, is an aqueous solution secreted from the orb weaving spider's aggregate glands and coats the spiral prey capturing threads of their webs. Studies identified the major component of the glue as microscopic nodules made of a glycoprotein. This study describes two newly discovered proteins that form the glue-glycoprotein of the golden orb weaving spider Nephila clavipes . Our results demonstrate that both proteins contain unique 110 amino acid repetitive domains that are encoded by opposite strands of the same DNA sequence. Thus, the genome of the spider encodes two distinct yet functionally related genes by using both strands of an identical DNA sequence. Moreover, the closest match for the nonrepetitive region of one of the proteins is chitin binding proteins. The web glue appears to have evolved a substantial level of sophistication matching that of the spider silk fibers.

Environmental conditions impinge on dragline silk protein composition

The silk formed in the major ampullate (MA) gland of the orb weaving spider Nephila clavipes is composed of two silk fibroins, which are called major ampullate spidroins 1 (MaSp1) and 2 (MaSp2). Analysis of proteolytic peptides and reactivity to spidroin type specific antibodies indicated that MaSp2 constituted only a minor part in the spinning dope as well as in the spun filaments. Upon starvation, a change in the silk's characteristic features was observed that was concomitant of a decrease in the contribution of MaSp2. The silk became less elastic and stiffer, which will better tailor its usability for the safety line, albeit at the expense of its employment as the web frame threads. In addition, since MaSp2 production requires greater ATP consumption, such a shift in the protein ratio cuts down on the energy costs to produce the silk. From this change in protein composition the spider might therefore benefit twice, by synthesizing 'cheaper' silk that into the bargain has properties that potentially can better support foraging in times of food shortage.

The adaptive advantage of prolonged mating: a test of alternative hypotheses

When a female can mate with more than one male, males are thought to reduce intrasexual competition by prolonging mating, that is, mating longer than necessary for sperm release. Two principal hypotheses have been proposed to explain the advantage of prolonged mating. The extended mate guarding hypothesis suggests that with longer mating, the female is less likely to be inseminated by another male. The ejaculate transfer hypothesis suggests that with longer mating, more of the male's sperm are stored. To our knowledge, this is the first study to test these alternatives simultaneously by manipulating duration of mating and examining effects on sperm release, storage and female polyandry. We found support for the mate guarding hypothesis in the golden orb-weaving spider Nephila clavipes; the longer the mating with the first male, the lower the likelihood of the female copulating with another male. This was not due to influence on female receptivity but on the ability of subsequent males to couple. Inconsistent with the ejaculate transfer hypothesis was the observation that duration of mating with the first male did not linearly correlate with release or storage of its sperm. We suggest that prolonged mating does not result in sexual conflict in this species.

QUANTITATIVE SHIFTS IN ORB-WEB INVESTMENT DURING DEVELOPMENT IN NEPHILA CLAVIPES (ARANEAE, NEPHILIDAE)

Abstract When there are direct conflicts in resource allocation to foraging effort versus growth and development, the relative allocation to foraging may change in a predictable manner with development. Orb-webs provide a physical record of resource allocation to foraging, and their synthesis requires the investment of physiologically important resources. Spiders in strongly seasonal habitats must complete development prior to the end of the season, and may be expected to alter foraging effort to maximize the probability of successful reproduction. Comparison of populations of the orb-weaving spider Nephila clavipes (Araneae, Nephilidae) in very seasonal versus less seasonal habitats allows testing for changes in allocation of resources to foraging effort during development. Orb-web size increases with increasing spider size, with little variation in slope among populations. However, in univoltine populations inhabiting strongly seasonal habitats, the size of the orb web is not a simple function of spider...

Female morphology, web design, and the potential for multiple mating in Nephila clavipes: do fat-bottomed girls make the spider world go round?

In animal species where females mate with multiple males, female mating success might be expected to covary with aspects of female morphology, such as size or shape. Spiders are especially interesting in this regard, as the females of several spider groups weave intricate webs that often accommodate multiple male spiders, all of whom are potential mates. Because web design is likely to be dependent on female size/shape, we use multivariate methods to assess the relationships among female morphology, web design, and reproductive ecology over a range of body sizes in the orb-weaving spider Nephila clavipes . Of the measured variables, only abdomen size explained a significant amount of the variation in number of males on a web, and this relationship holds even after statistically accounting for body size. Because abdomen size is an indicator of body condition in spiders, we suggest that condition is likely to be an important factor relating to potential mating success in female spiders. We found no evidence for an association between web design and number of males on a web, although our data indicate that larger females build webs that are both larger and further from the ground than those of smaller females. © 2006 The Linnean Society of London, Biological Journal of the Linnean Society, 2006, 87 , 95 − 102.

Structure-activity relationships of pyrrolizidine alkaloids in insect chemical defense against the orb-weaving spider Nephila clavipes.

Pyrrolizidine alkaloids (PAs) are known to protect Arctiidae moths and Danainae and Ithomiinae butterflies against the orb-weaving spider Nephila clavipes (Araneae, Araneidae, Tetragnathinae), which liberates adults of these insects unharmed from its web. We tested against this spider the role of eight PAs and one derived structure [an 89:11 mixture of the 12-membered macrocyclic diester free base integerrimine and senecionine and the respective N-oxide; two hydrolysis products from this mixture (the necine base retronecine, its respective N-oxide, and a mixture of integerriminic and senecionic necic acids); the 12-membered macrocyclic senkirkine; the 9-O-monoester free base senecioylretronecine and its respective N-oxide; and the 9-O-monoester free base callimorphine (a PA biosynthesized only by insects from retronecine)]. The mixture integerrimine-senecionine N-oxide seems to be more active than the respective free base [LibD50 (liberation dose 50) = 0.042 and 0.153 microg/dry weight of prey, respectively], but the difference in activity between the N-oxide and free base of the 9-O-monoester senecioylretronecine was slight (LibD50 = 0.167 and 0.104, respectively). Senkirkine, an otonecine base PA that does not form N-oxide and is not found in insects, was the less active, showing the highest LibD50 (0.354). The difference in antipredator activity between N-oxides and free bases from macrocyclic diesters and monoesters may be correlated with physicochemical properties of these molecules in interaction with the Nephila receptors. For the active structures, there was a significant correlation between dosage and antipredator activity. Both forms of retronecine and a necic acid mixture were inactive, supporting the hypothesis that PAs biosynthesized by insects from retronecine were originally produced and stored in order to optimize chemical defense. Comparison of dose/activity data with reported amounts of PAs in butterflies suggested that, in general, PA-specialist insects are protected against predation by Nephila.

Mathematical Simulation of Thermoregulatory Behavior in an Orb-Weaving Spider

1. In order to better document the thermoregulatory effectiveness of observed behaviours compared to other possible behaviours, we develop a model of interception of solar radiation. We demonstrate its use with data from a population of the orbweaving spider Nephila clavipes inhabiting a mid-altitude Mexican desert. 2. Nephila clavipes exhibits three behaviours that appear related to thermoregulation in this extreme climate: microhabitat selection, orb-web orientation and postural adjustments. 3. Large females more often built in open spaces without shade and small juveniles built within shrubs in shaded web sites. Webs built by large females tended to have a north-south orientation, whereas among small juveniles there was no correlation in web orientation. 4. Only individuals in unshaded sites exhibit postural thermoregulation, in which the spider's abdomen tracks the elevation of the sun. The simulation revealed that interception of solar radiation by large females was reduced by solar tracking, but that the north-south orientation of the orb did not reduce the interception of solar radiation relative to an east-west orientation.

Pyrrolizidine alkaloids in the arctiid mothHyalurga syma

The arctiid mothHyalurga syma (subfamily Pericopinae) sequesters pyrrolizidine alkaloids (PAs) from its larval food plantHeliotropium transalpinum (Boraginaceae). Colorimetric quantification of total PAs in the larvae, pupae, and adults ofHyalurga revealed mean values of about 286-445μg per individual (1.4-2.6% of dry weight). The PA mixtures found in the moth and its larval food plant were evaluated by GC-MS. Food-piant leaves were found to contain the diastereoisomeric retronecine esters indicine (IIIa), intermedine (IIIb), and lycopsamine (IIIc), and the heliotridine ester rinderine (IIId) only as minor constituents, whereas 3'-acetylrinderine (IVc) (68% of total PAs) and the respective 3'-acetyl esters of indicine (IVa) and intermedine (IVb) (both 17%) were the major alkaloids. Supinine (IIa) is detectable in traces only. The PA mixtures in eggs, larvae, pupae, and imagines ofHyalurga were identical: indicine, intermedine, and lycopsamine accompanied by considerable amounts of supinine and amabiline or coromandalinine (IIb/IIc) were the major components. Only larvae were found to store small quantities of a 3'-acetyl derivative. Rinderine and its 3'-acetyl ester were never found in the insects. Low concentrations of the arctiidspecific PA callimorphine (I) were present in larvae, pupae, and imagines. The differences in the PA patterns of the insects and their larval food plant suggest thatHyalurga is capable of modifying plant-derived PAs by inversion of the 7-OH configuration (conversion of the necine base heliotridine into retronecine), and perhaps the inversion of the 3'-OH [conversion of (+)-trachelanthic acid into (-)-viridifloric acid], although the possibility of a selective sequestration of the respective retronecine esters cannot be excluded. Some trials with the orb-weaving spiderNephila clavipes, a common neotropical predator, showed that both freshly emerged and field-caught adults ofHyalurga syma are liberated unharmed by the spider. The liberation could be related to the presence of PAs in the moths.

Chemical attraction of kleptoparasitic flies to heteropteran insects caught by orb-weaving spiders.

Insects of the heteropteran families Pentatomidae (stink bugs) and Coreidae (squash bugs), when being eaten by the orb-weaving spider Nephila clavipes, attract flies of the family Milichiidae. The flies aggregate on the bugs and, as kleptoparasites, share in the spider's meal. Stink bugs and squash bugs typically eject defensive sprays when attacked; they do so when caught by Nephila, but the spray only minimally affects the spider. Evidence is presented indicating that it is the spray of the bugs that attracts milichiids to the spider's catch.

Web Orientation by Nephila clavipes in Southeastern Texas

-We undertook a longitudinal study of marked free-living individuals of the orb-weaving spider Nephila clavipes (Linnaeus), investigating the importance of individual web-site structure in determining the orientation of the web. A new method of analyzing pairwise circular data allowed comparisons of successive webs at a given site or by a given spider. Successive webs by the same spider at the same site were oriented in the same direction and successive, nonoverlapping occupants of the same site oriented their webs in the same direction. Additionally, spiders that built in aggregations of webs tended to orient their webs in the same direction. However, as an individual changed web sites, the orientation of webs at successive sites were not correlated. The analysis of orientation within each microhabitat revealed slightly significant correlations of web orientations, but there was no correlation of web orientations over the entire population. We conclude that the conditions at individual web sites were the dominant factor in determining the orientation of webs at this location.

Variation in foraging investment during the intermolt interval and before egg-laying in the spiderNephila clavipes (Araneae: Araneidae)

Repeated observations were made of marked, freeliving females in three disjunct populations of the orb-weaving spider Nephila clavipes,recording orb size and structure and barrier web complexity. These longitudinal studies revealed regular patterns of variation in orb size associated with the molting cycle and cycle of egg maturation. Juveniles maintained in an insectary under a controlled feeding regime exhibited similar variation in orb size. All spiders ceased orb renewal prior to molting and the majority replaced the viscid orb with a nonviscid platform prior to molting. Differences among the populations in molting web structure are discussed in light of differences in the physical and biological environments. Cessation of orb renewal corresponded temporally with reported physiological changes associated with the molt cycle and possibly reflect changes in resource partitioning between growth and foraging during the molt cycle.

Egg-laying of the golden silk spider, Nephila clavipes L. (Araneae, Araneidae): Functional analysis of the egg sac

The egg-laying sequence of the orb-weaving spider Nephila clavipes was observed in the field setting. The functions of certain modal characteristics of the egg sac were assessed through the collection of normative field data on overwintering egg sacs and through the effects of egg sac manipulations on spiderling production. Components of the egg sac were manipulated before and after egg hatching to determine if they promoted offspring survival at different times in development. Naturally occurring and induced changes in the position and structure of the egg sac resulted in similar decreases in productivity. Certain components of the egg sac were important primarily before hatching, and others throughout the entire overwintering period. These results and the limitations of functional analyses of behaviours are discussed.