Cardenolide fingerprint of monarch butterflies reared on common milkweed,Asclepias syriaca L.

Monarch butterfly,Danaus plexippus (L.), larvae were collected during August 1983 from the common milkweed,Asclepias syriaca L., across its extensive North American range from North Dakota, east to Vermont, and south to Virginia. This confirms that the late summer distribution of breeding monarchs in eastern North America coincides with the range of this extremely abundant milkweed resource. Plant cardenolide concentrations, assayed by spectrophotometry in 158 samples from 27 collection sites, were biased towards plants with low cardenolide, and ranged from 4 to 229 g/ 0.1 g dry weight, with a mean of 50 g/0.1 g. Monarch larvae reared on these plants stored cardenolides logarithmically, and produced 158 adults with a normally distributed concentration range from 0 to 792 g/0. l g dry butterfly, with a mean of 234 g/0.1 g. Thus butterflies increased the mean plant cardenolide concentration by 4.7. The eastern plants and their resultant butterflies had higher cardenolide concentrations than those from the west, and in some areas monarchs sequestered more cardenolide from equivalent plants. Plants growing in small patches had higher cardenolide concentrations than those in larger patches, but this did not influence butterfly concentration. However, younger plants and those at habitat edges had higher cardenolide concentrations than either older, shaded, or open habitat plants, and this did influence butterfly storage. There were no apparent topographical differences reflected in the cardenolides of plants and butterflies. Twenty-eight cardenolides were recognized by thin-layer chromatography, with 27 in plants and 21 in butterflies. Butterflies stored cardenolides within the more polar 46% of the plantR _d range, these being sequestered in higher relative concentrations than they occurred in the plants. By comparison with published TLC cardenolide mobilities, spots 3, 4, 9, 16, 24 or 25, 26, and 27, may be the cardenolides syrioside, uzarin, syriobioside, syriogenin, uzarigenin, labriformidin, and labriformin, respectively. Cochromatography with cardenolide standards indicated that desglucosyrioside did not occur in the plants but did occur in 70% of the butterflies, and aspecioside was in 99% of the plants and 100% of the butterflies. The polar aspecioside was the single most concentrated and diagnostic cardenolide in both plants and butterflies. ButterflyR _d values were dependent on those of the plant, and both showed remarkable uniformity over the range of areas sampled. Thus contrary to previous reports,A. syriaca has a biogeographically consistent cardenolide fingerprint pattern. The ecological implications of this for understanding the monarch's annual migration cycle are significant.     

Cardenolide content and thin-layer chromatography profiles of monarch butterflies,Danaus plexippus L., and their larval host-plant milkweed,Asclepias asperula subsp.Capricornu (woods.) woods., in north central Texas

This paper is the second in a series on cardenolide fingerprinting of monarch butterflies and their host-plant milkweeds in the eastern United States. Spectrophotometric determinations of the gross cardenolide content ofAsclepias asperula plants in north central Texas indicated wide variation ranging from 341 to 1616 g/0.1 g dry weight. The mean plant cardenolide concentration (886 g/0.1 g) is the highest for any milkweed species on which monarch cardenolide profiles have been produced. Forty-one butterflies reared individually on these plants contained a skewed distribution of cardenolide concentrations ranging from 231 to 515 g/0. 1 g dry weight with a mean of 363g/0.1 g. The uptake of cardenolide by the butterflies was independent of plant concentration, suggesting that saturation occurs in cardenolide sequestration by monarchs when feeding on cardenolide-rich host-plants. Female monarchs contained significantly greater mean cardenolide concentrations (339 g/0.1 g) than did males (320 g/0.1 g). The mean dry weight of the male butterflies (0.211 g) was significantly greater than the female mean (0.191) so that the mean total cardenolide contents of males (675 fig) and females (754 g) were not significantly different. Butterfly size was not significantly correlated to butterfly cardenolide concentration when differences due to sex and individual host-plant concentration were removed. Thin-layer chrornatograms of 24 individual plant-butterfly pairs developed in two solvent systems resolved 22 individual spots in the plants and 15 in the butterflies.A. asperula plants appear to contain several relatively nonpolar cardenolides of the calotropagenin series which are metabolized to more polar derivatives in the butterflies. Quantitative evaluation of theR _f values, spot intensities, and probabilities of occurrence in the chloroform-methanol-formamide TLC system produced a cardenolide fingerprint clearly distinct from those previously established for monarchs reared on otherAsclepias species. Our data support the use of fingerprints to make ecological predictions concerning larval host-plant utilization.A. asperula subsp.capricornu andA. viridis Walt, are the predominant early spring milkweeds throughout most of the south central United States. Cardenolide-rich monarchs reared on these two species may be instrumental in establishing and reinforcing visual avoidance of adults by naive predators throughout their spring and summer breeding cycle in eastern North America.Lepidoptera: Danaidae.Apocynales: Asclepiadaceae.