Table of contents

 

 

Antenna morphology and pheromone-based mate location in Polyphemus moths.

 

Fellow: Erin Heydenreich

Mentor:  Dr. Douglas Emlen

Department of Biology

University of Montana, Missoula

 

 

ABSTRACT

Mate location in moths is accomplished by long distance detection of low concentrations of pheromones emitted by females upon emergence.  Pheromone based mating systems have been suggested to be under strong sexual selection (Phelan, 1997).  There are several aspects of moth biology that suggest that larger body size and larger antennae will be selected for in males.  In this study I predicted that polyphemus (Antheraea plyphemus: Family Saturniidae) males able to successfully locate females would have proportionately larger antennae than males sampled from the population at random. Captive, reared males represented the base population and males caught with captive females represented the “successful”, or sexually selected, population.  Wild males were captured using female-baited traps in Mclay Flats of the Blue Mountain Recreation Area, the Rattlesnake Recreation Area, and Rock Creek, in Missoula, Montana. Neither absolute nor relative antenna size differed between “successful” (i.e. baited) and control males, suggesting weak or absent sexual selection for antenna size in this population.  Study of moth pheromones is a useful tool in monitoring populations and has important implications for agriculture and conservation.

 

 

Key phrases: sexual selection, antenna morphology, sex pheromones, moth mating systems, and saturniidae.

 

Antenna morphology and pheromone-based mate location in Polyphemus moths.

 

Fellow: Erin Heydenreich

Mentor:  Dr. Douglas Emlen

Department of Biology

University of Montana, Missoula

 

 

ABSTRACT

Mate location in moths is accomplished by long distance detection of low concentrations of pheromones emitted by females upon emergence.  Pheromone based mating systems have been suggested to be under strong sexual selection (Phelan, 1997).  There are several aspects of moth biology that suggest that larger body size and larger antennae will be selected for in males.  In this study I predicted that polyphemus (Antheraea plyphemus: Family Saturniidae) males able to successfully locate females would have proportionately larger antennae than males sampled from the population at random. Captive, reared males represented the base population and males caught with captive females represented the “successful”, or sexually selected, population.  Wild males were captured using female-baited traps in Mclay Flats of the Blue Mountain Recreation Area, the Rattlesnake Recreation Area, and Rock Creek, in Missoula, Montana. Neither absolute nor relative antenna size differed between “successful” (i.e. baited) and control males, suggesting weak or absent sexual selection for antenna size in this population.  Study of moth pheromones is a useful tool in monitoring populations and has important implications for agriculture and conservation.

 

 

Key phrases: sexual selection, antenna morphology, sex pheromones, moth mating systems, and saturniidae.

 

 

 

INTRODUCTION

Most moth species use pheromone based mating systems.  Females remain stationary upon emergence and attract males by the use of chemical pheromones (Phelan, 1997).  Males are equipped with large plumed antennae that are highly sensitive to very low levels of pheromone (females generally emit picogram to nanogram concentrations of pheromones) and have been known to fly long distances to locate females (Hogue, 1993; Phelan,1997).

Most moths are sexually dimorphic for antennae size and morphology: males generally have longer and wider antennae then females, and many more chemoreceptors.  The intensity of selection on male ability to find mates is reflected by these strong sexual dimorphisms in the number and sensitivity of chemoreceptors. For example, in one species of saturniids, male antennae contain over 64,000 sensory hairs (females have less than half that), 80 percent of these are specific for sex pheromone detection (Evans, 1984).  It has been suggested that low release of female pheromone is a form of sexual selection in which females select for strong searchers (Phelan, 1997).

There are several features of moth biology that suggest that pheromone-based mate location will select preferentially for the largest males. First, sensory receptors are located on the sensory hairs of the antennae. Large antennae may contain more hairs with more sensory receptors, and therefore have a more acute sensitivity to female pheromone. Second, antennae size scales allometricaly with body size in most saturniids (Heydenreich, unpublished data) as it does in other insects (Hogue, 1993), resulting in males with disproportionately large antenna. Third, both flight capacity and flight speed have been related to body size in several moth species (Gu, and Danthanarayana, 1992; Kuenen and Carde, 1993).  This suggests that larger individuals may not only be more sensitive to low pheromone concentrations, but they may be able to fly longer distances, and perhaps under a wider range of weather conditions.

Based on these features of moth biology, I predicted that male saturniids able to successfully locate females, will either have proportionately larger antenna for a given body size, or will be larger over all in both body and antennae size than the average male. In this study I will compare two populations of males to test this prediction.  The relationship between antennae size and body size will be compared between wild males caught with female bated traps and a base population of males raised in captivity.

Although sexual selection has often been suggested as an important force during the evolution of moth pheromones, relatively few moth species have been studied with respect to pheromone use and behaviors associated with mate attraction (Phelan, 1997). Studies of evolution of mate-signaling systems and speciation in insect pheromones are rare in general, and therefore can pose interesting and unanswered questions (Phelan, 1997).  Besides scientific curiosity, use of moth pheromones is an important tool in detecting and controlling pest populations in agriculture (Barnes et al., 1992).  Although few saturiniids are pests, monitoring populations is still important for conservation reasons.  A number moth species, especially saturniids, are becoming endangered in the United States (Bellows et al., 1984).

 

MATERIALS AND METHODS

Back-round and natural history of saturniid moths 

Polyphemus moths (Antheraea polyphemus) of the family Saturniidae occur throughout the North America. They generally occur in riparian habitats for both larval and adult stages.  As larva they feed on oak, willow, and birch. They are ready to mate upon emergence and only live for a few weeks. Adults emerge and fly from mid May to late July, coinciding with the time of this study.

Polyphemus moths are large and easy to handle, therefore they can be measured alive released. All saturniids are convenient to study because they require no feeding (in adults the proboscus is reduced and they are unable to feed). Also caged virgin female moths have been found to attract and trap mates for many moth families, including Saturniids (Martin et al., 1992).

Polyphemus moths exhibit sexual dimorphism (Figure 1).  The males have larger body sizes and proportionately larger antennae.  Specifically, antennae length scales positively with body size in males (b=0.626; r2=0.177; p=0.0001; N=78), but not in females (b=0.291; r2=0.02; p=0.4711; N=27).  Therefore, large males have much larger antennae than either females or smaller males (Figure 1).

Experimental Design

Seventy-five Polyphemus cocoons were raised in an enclosed outdoor tent consisting of a rainproof roof and mosquito-netted walls.  The cocoons were placed in trays of peat moss and sprayed with water daily until hatching.  Upon emergence, males were measured using millimeter scale metal calipers.  Measurements were taken of the wing length from the base of the wing to the tip; body length from between the antennae to the tip of the thorax; width of the thorax measured between the wings in the under side; antennae length and width.  The males remained in the tent until death and were separated from the females by a mosquito-net curtain.  These males will be termed “base males” and represent the general (i.e. unselected) population.

            To measure sexual selection in these moths, I placed un-mated females in the field and recorded the phenotypes of the subset of males that were successful in locating them.  By comparing these males to the base population, I could test whether the antenna morphology of the “successful” males differed from that of the males in the base population.  In other words, to measure whether variation in antenna size contributed to successful mate location.

Female Polyphemus were measured in the same manner as the males with the exception of antennae width which is negligible.  Females were then placed in traps, to prevent escape, and set one of the selected field sites.  Traps consisted of a cylindrical wire frame one-foot in diameter and about a foot and a half tall.  The entrance to the trap was an inverted cone three-inches in diameter, which prevented the moths from escaping. The frame was covered in a mesh cloth and hung from a tree branch with fishing line.

Traps were placed in Mclay flats in the Blue Mountain recreational area, the lower Rattlesnake Recreational forest, and Rock Creek forest.  All three sites shared the common characteristics of an open meadow surrounded by forest with a stream or river within fifty yards.  Female-baited traps were placed about 100 yards apart, three to five feet from the ground.  Between one and five traps were set per night, depending on the number of females that had emerged that day (i.e. the total number available at the time).  Females were left out all night and collected and brought back to the tent during the day.  Each female was used from one to three nights in a row; also depending on the number emerged. A total of thirteen females were used in traps.  Traps were set from May 24th to June 16th.  Males were collected from the traps and transported back to the tent where all measurements were made.  These will be termed “baited males”.  Males were marked using fluorescent non-toxic paint and released the same day in the general area where they were caught.

To supplement the sample of base population males (and attempt to include wild males in the sample representing the base population), I used black-lights hung in front of a white sheet between trees were used to attract males, and the same collection and measurement procedure was employed.  Black lights were used in the same three study sights on nights when no female bated traps were out.  Light traps were set on June 9th, 15th, 20th, and July 5th.

Analysis

Scaling relationships between antennae length and width and antennae length and body size for males and females were determined using regression analysis.  Analyses of covariance were used to test for differences in the slope and intercept of the regressions for antennae length and body size between the base males and the baited males.

 

RESULTS

Ten out of the thirteen females used in traps attracted males.  The average number of males caught per trap was 7 (median 5).  The most males collected in one night were 15.  Males were rarely found inside the trap.  Instead, they tended to be piled on the outside of the trap on top of one another, and directly above the female.  Only four males were successfully caught using the light trap at Rock Creek, and these were included in the base population sample.

Sexually selected males (i.e. males successful in locating a female) were not detectably different from the base population.  The slope of the scaling relationship of baited males did not differ from that of the control males (Figure 2; 1factor ANCOVA with body width as covariate and antenna width as the dependent variable, f1, 74= 2.568; p=0.1133).  The intercepts of these relationships also were not significantly different (1 factor ANCOVA with body width as covariate and antenna width as the dependent variable, f1, 74=2.690; p=0.1052).

 

DISCUSSION

There was no significant difference in antenna size for males that find females in the wild and males representing the general population in Polyphemus moths.  This result has two major implications.  First, that there may be little or no selective pressure for larger antennae size in males.  Second, some factor other than antennae size may influence female-location success in males.

            The first implication suggests that all males, regardless of size would do equally well in locating a female.  One reason for this may be that females are emitting such a large concentration of pheromone that all males in the area are readily capable of detecting it. A study of pheromone sensitivity in blueberry leaftier moths (Tortricidae) showed that relatively low concentrations of pheromone result in antennal saturation (Ponder and Seabrook, 1991).  This suggests that males are affected by very small amounts of pheromone, and therefore all males may be able to detect females regardless of antennae size. 

The same study tested female antenna sensitivity to her own pheromone and found that there was no response.  This suggests that there is sexual dimorphism for chemoreceptor cells on the antennae. Large antennae do indicate a greater number of receptors; however, it may be the sensitivity of receptors, rather than the number, that is important in mate location.

Sensitivity of receptor cells may be the important key factor in mate location in polyphemus moths and may have nothing to do with antennae size in males.  Tests similar to those done by Ponder and Seabrook would have to be done with polyphemus to determine the role that sensors may play in differential mate location.  However, no studies have been done to test sensitivity of antennae to pheromone with any saturniid.  One reason may be that production of synthetic pheromones is very costly.  Another reason may be that satuiniids are generally of little economical interest because very few are agricultural or forest pests.

A final possibility is that male ability to locate a female may not be the primary trait under selective pressure. An alternative involves male ability to discriminate the correct species of female. It has been proposed that pheromone mating systems are under strong stabilizing selection (Phelen, 1997).    This means that males should be optimally sensitive to the most common blend of pheromone in the population.  Therefore males that are tuned to the right blend for their species will be the ones selected for.  In this case, sensitivity to a particular blend of pheromone may have no correlation to antennae size, since a qualitative mixture would be more important than a qualitative amount.

Given the large p-values for the analysis of covariance for base verses baited males, it is unlikely that the results are due to errors in experimental design.  However it is possible that the base males used were not representative of the general population for unseen reasons.  Because the light traps were unsuccessful, it was impossible to get a good sample of the base population in Missoula.  However, the four that were caught fell on the same line as the rest of the base males, suggesting that the reared males were not different from the wild males.  Another possible flaw may be that the traps were too close together to detect a significant treatment effect.  In other words, the pheromone concentration in the trapping site may have been stronger than normal because of the number of females placed together in one area.

In conclusion, antenna size does not appear to be a factor that is selected for in male Polyphemus moths.  The results of this study suggest that some other aspect of phenotype (i.e. sensitivity of chemoreceptors) may be more important in mate location than antenna size and therefore number of receptors.  A study manipulating concentrations of pheromones and a better sample of the base population, would be required to determine anything further.

 

LITERATURE CITED

 

 

Barnes, Martin M., Jocelyn Miller, Philipp Kirsch, and David Hawks.  1992.  Codling 

Moth (Lepidoptera: Torticidae)  controll by dessemination of synthetic female sex pheromone.  Journal of Economic Entomology 85: 1274-1277.

 

 

Bellows Jr., Tom S., John Cowens and Ellis Huddeleston.  1984.  Flight activity and

 dispersal of Range caterpillar.  Canadian Entomologist 116: 246-252.

 

Evans, Howard.  1984.  Insect Biology: A Textbook of Entomology.  Colorado State

 University,  Addison Wesley Publishing Co., CO.

 

Gu, H. and W. Danthanarayana. 1992.  Influence of larval rearing conditions on the body

size and flight capacity of Epiphyas postuittana moths.  Australian Journal of Zoology 40: 573-580.

 

Heydenrecih, Erin. 1999.  Allometry and symmetry in body size and antennae in

Saturiniiad moths. Unpublished data.  Monteverde, Costa Rica.

 

Hogue, Charles L.  1993.  Latin American Insects and Entomology.  University of

California Press, CA.

 

Kuenen, L.P.S., and R.T.  Carde.  1993.  Effects of moth size on velocity and steering

during upwind flight toward a sex pheromone source by Lymantria dispar (Lepidoptera:  Lymantriideae).  Journal of Insect Behavior 6: 177-193.

 

Phelen, P.L. 1997.  “Evolution of mate-signaling in moths.” In: Choe, J and B.J. Crespi

 eds., Mating Systems in Insects and Arachnids.  Cambridge University Press,

Cambridge, MA.

 

Ponder, B.M. and W.D. Seabrook.  1991.  Sensitivity of blueberry leafeatier moths

(Lepidoptera : Tortricidea) to their own sex pheromone: mating bioessay,

 electroantennogram, and trap attractancy studies.  Canadian Entomologist 123:

 231-238.

 

 

 

Acknowledgements:

This work was funded by an IBS-CORE Undergraduate Research Fellowship to Erin Heydenreich through a grant from the Howard Hughes Medical Institute to The University of Montana.