Inventory of leaf and flower odorants in plants associated with the life cycle of Japanese Papilio butterflies

Abstract The odorants of eight Japanese mainland native species (Citrus x deliciosa, Zanthoxylum ailanthoides, Z. schinifolium, Z. piperitum, Phellodendron amurense, Orixa japonica, Skimmia japonica, and Boenninghausenia albiflora), one tropical species (Euodia meliifolia), and one invasive species (Ruta graveolens) of the Rutaceae family and three Japanese mainland native species (Angelica keiske, Heracleum lanatum, Anthriscus sylvestris, and one invasive species (Foeniculum vulgare) of the Apiaceae family were analyzed using gas chromatography–mass spectrometry with dynamic–headspace and thermal–desorption methods. These plants are hostplants to Japanese Papilio butterflies. Herein, these 14 plants were classified into six major groups based on the odorant volatiles, which did not correspond to the current phylogenetic classification. Similarly, floral odorant analysis of the six plant species (Clerodendrum trichotomum, Cayratia japonica, Robinia pseudoacacia, Lonicera japonica, C. deliciosa, Z. ailanthoides) visited by Papilio butterflies for nectaring, revealed the presence of linalool in all the flowers. Floral volatiles in C. deliciosa and Z. ailanthoides exhibited moderate resemblance to their respective leaf volatiles. Interestingly, our results in C. trichotomum was not in complete agreement with previous reports, emphasizing the need for newer methods of extraction and analysis.</jats:p

Inventory of leaf and flower odorants in plants associated with the life cycle of Japanese Papilio butterflies

Abstract The odorants of eight Japanese mainland native species (Citrus x deliciosa, Zanthoxylum ailanthoides Siebold &amp; Zucc, Z. schinifolium Sieb. et Zucc., Z. piperitum L., Phellodendron amurense Ruprecht, Orixa japonica Thunb., Skimmia japonica Thunb., and Boenninghausenia albiflora (Hook.) Rchb. ex Meisn.), one tropical species (Euodia meliifolia (Hance) Benth.), and one invasive species (Ruta graveolens) of the Rutaceae family and three Japanese mainland native species (Angelica keiskei (Miq.) Koidz., Heracleum lanatum W. Bartram, Anthriscus sylvestris (Blume) DC), and one invasive species (Foeniculum vulgare Mill.) of the Apiaceae family were analyzed using gas chromatography–mass spectrometry with dynamic–headspace and thermal–desorption methods. These plants are hostplants to Japanese Papilio butterflies. Herein, these 14 plants were classified into six major groups based on the odorant volatiles, which did not correspond to the current phylogenetic classification. Similarly, floral odorant analysis of the six plant species (Clerodendrum trichotomum Thunb., Cayratia japonica (Thunb.) Gagnep., Robinia pseudoacacia L., Lonicera japonica Thunb., C. deliciosa, Z. ailanthoides) visited by Papilio butterflies for nectaring, revealed the presence of linalool in all the flowers. Floral volatiles in C. deliciosa and Z. ailanthoides exhibited moderate resemblance to their respective leaf volatiles. Interestingly, our results in C. trichotomum was not in complete agreement with previous reports, emphasizing the need for newer methods of extraction and analysis.</jats:p

Sasakia Charonda (Apaturinae: Nymphalidae) and its Three Related Butterfly Larvae Use Halitosis to Repel Predators

Abstract Accidentally, we discovered that Sasakia charonda (Nymphalidae: Apaturinae) larvae disturbed by ants or humans released volatile compounds from their mouth; thus, we tried to identify these halitosis. We collected halitosis directly from the mouths of S. charonda larvae into volatile-collecting tubes. Trapped halitosis were subjected to gas chromatography-mass spectrometry (GC–MS). We confirmed the identity of eleven substances by comparison to GC of known standards, and inferred them to be mainly alcohols and aldehydes/ketones, with main chains of 4–5 carbons. Three of the chemicals in these halitosis, 2-butanol, 1-penten-3-ol and 3-pentanone, affected the behavior of Pristomyrmex punctatus and Formica japonica ants that co-inhabited the S. charonda rearing cage. We concluded that the substances we identified in this study were used as defensive halitosis, analogous to osmeterium emissions specific to Papilionidae butterflies. Based on smell, Holikawa found that Hestina assimilis and H. persimilis larvae have closely related halitosis. Thus, we also analyzed the halitosis of these two species as well as Apatura metis, another Apaturinae, using the same methods. We found that these species also release halitosis. The composition of the substances of H. assimilis and H. persimilis were somewhat similar to that of S. charonda, whereas that of A. metis differed. Some of the substances also induced defensive behavior in these species of Apaturinae larvae.</jats:p

K+ excretion: the other purpose for puddling behavior in Japanese Papilio butterflies.

To elucidate the purpose of butterfly puddling, we measured the amounts of Na+, K+, Ca2+, and Mg2+ that were absorbed or excreted during puddling by male Japanese Papilio butterflies through a urine test. All of the butterflies that sipped water with a Na+ concentration of 13 mM absorbed Na+ and excreted K+, although certain butterflies that sipped solutions with high concentrations of Na+ excreted Na+. According to the Na+ concentrations observed in naturally occurring water sources, water with a Na+ concentration of up to 10 mM appears to be optimal for the health of male Japanese Papilio butterflies. The molar ratio of K+ to Na+ observed in leaves was 43.94 and that observed in flower nectars was 10.93. The Na+ amount in 100 g of host plant leaves ranged from 2.11 to 16.40 mg, and the amount in 100 g of flower nectar ranged from 1.24 to 108.21 mg. Differences in host plants did not explain the differences in the frequency of puddling observed for different Japanese Papilio species. The amounts of Na+, K+, Ca2+, and Mg2+ in the meconium of both male and female butterflies were also measured, and both males and females excreted more K+ than the other three ions. Thus, the fluid that was excreted by butterflies at emergence also had a role in the excretion of the excessive K+ in their bodies. The quantities of Na+ and K+ observed in butterfly eggs were approximately 0.50 μg and 4.15 μg, respectively; thus, female butterflies required more K+ than male butterflies. Therefore, female butterflies did not puddle to excrete K+. In conclusion, the purpose of puddling for male Papilio butterflies is not only to absorb Na+ to correct deficiencies but also to excrete excessive K+

Amounts of the four metal ions in the eggs of five Japanese <i>Papilio</i> species.

Amounts of the four metal ions in the eggs of five Japanese Papilio species.</p

Amounts of Na⁺, K⁺, Mg²⁺, and Ca²⁺ in 100 g flower nectar used by Japanese Papilioninae species.

<p>Amounts of Na⁺, K⁺, Mg²⁺, and Ca²⁺ in 100 g flower nectar used by Japanese Papilioninae species.</p

Amounts of the four metal ions in the meconium of Japanese female <i>Papilio</i> butterflies.

<p>Amounts of the four metal ions in the meconium of Japanese female <i>Papilio</i> butterflies.</p

Absorption and excretion of Na⁺, K⁺, Mg²⁺, and Ca²⁺ in <i>Papilio</i> butterflies (YS01).

<p>Absorption and excretion of Na⁺, K⁺, Mg²⁺, and Ca²⁺ in <i>Papilio</i> butterflies (YS01).</p