Floral fragrances in two closely related fruit fly orchids, Bulbophyllum hortorum and B. macranthoides (Orchidaceae): assortments of phenylbutanoids to attract tephritid fruit fly males

  • Tomoko Katte
  • Keng Hong Tan
  • Zhi-Hui Su
  • Hajime Ono
  • Ritsuo NishidaEmail author
Original Research Paper


Floral chemical components are important cues used by plants to attract pollinators. One outstanding case is “fruit fly orchids” in the genus of Bulbophyllum to attract their pollinators by releasing characteristic fragrances. Dacini fruit flies are main or exclusive pollinators which are strongly attracted to certain natural chemicals, either methyl eugenol (ME: a phenylpropanoid) or raspberry ketone (RK: a phenylbutanoid). Furthermore, zingerone (ZN: a phenylbutanoid) has been characterized as the attractant for both ME- and RK-sensitive fruit fly species. In the present study, we examined chemical profiles of two closely related Bulbophyllum orchids—B. hortorum, and B. macranthoides subsp. tollenoniferum—distributed in Papua New Guinea and the Southeast Asian countries, respectively. We first observed that RK-sensitive flies were attracted to these orchids by ex situ cultivation in Penang, Malaysia. These Bulbophyllum orchids contained RK and/or ZN as their main floral components. Other than these attractants, multiple phenylbutanoids including potential attractants for RK-sensitive species were identified from these orchids. Therefore, we examined attractiveness of potential phenylbutanoid attractants to an RK-sensitive melon fly, Zeugodacus cucurbitae, using laboratory-reared flies. Furthermore, we analyzed molecular phylogenetic relationships among phenylpropanoid- or phenylbutanoid-producing orchids to see a relation between chemical profiles and phylogenetic classification in the related species.


Bulbophyllum Floral volatiles Phenylbutanoids Pollination Dacini fruit flies 



We thank Jaap J. Vermeulen for confirmation of the species and subspecies of the Bulbophyllum species; and Jane Royer for the Dacus species identification. We thank Atsushi Honma and Yasutsune Sadoyama of Okinawa Prefectural Plant Protection Center for providing Z. cucurbitae. We also thank Ayako Sasaki for technical assistance. R. Nishida was partly supported by the Grant-in-Aid for Scientific Research from JSPS (Nos. 19310142 and 23380035) from the Ministry of Education, Culture, Sports, Science and Technology of Japan. H. Ono was partly supported by JSPS KAKENHI Grant Number 26450466.

Supplementary material

13355_2019_653_MOESM1_ESM.tif (4.2 mb)
Supplementary material 1: Fig. S1 Laboratory bioassay to evaluate the attractiveness of the phenylbutanoids. a The plastic container (75 mm × 75 mm × 45 mm) with a lid having a hole (35 mm diameter) bored and covered with a plastic mesh was used for the laboratory bioassays. b Flies were supplied with water filled in a 2.0-ml microtube and food. To test each compound, a filter paper (21 mm diameter) treated with 100 µg of a test compound was placed on the plastic mesh of the lid of the container. c Several males were attracted to the plastic mesh just at bottom of the treated filter paper (arrow), and voraciously fed on an active compound applied to the filter paper. (TIFF 4351 kb)
13355_2019_653_MOESM2_ESM.tif (100 kb)
Supplementary material 2: Fig. S2 A trace amount of cue-lure was detected from two floral specimens (BH2, BH8 and BH10) of the fruit fly orchids, Bulbophyllum hortorum. ZN: zingerone; CL: cue-lure. (TIFF 100 kb)
13355_2019_653_MOESM3_ESM.tif (613 kb)
Supplementary material 3: Fig. S3 Amounts of the major phenylbutanoids—AA, RK, ZN and SN—in the whole flowers of the fruit fly orchids, Bulbophyllum hortorum (a) and B. macranthoides subsp. tollenoniferum (b). AA: anisyl acetone; RK: raspberry ketone; ZN: zingerone; SN: syringerone. (TIFF 612 kb)
13355_2019_653_MOESM4_ESM.tif (517 kb)
Supplementary material 4: Fig. S4 Average numbers of attracted flies at 5 min intervals to each attractant—calculated from data as shown in Fig. 4. Different alphabets indicate significant differences between compounds (p < 0.05 by Tukey’s HSD test). (TIFF 516 kb)
13355_2019_653_MOESM5_ESM.pdf (26 kb)
Supplementary material 5 (PDF 25 kb)
13355_2019_653_MOESM6_ESM.pdf (6 kb)
Supplementary material 6 (PDF 6 kb)


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Copyright information

© The Japanese Society of Applied Entomology and Zoology 2019

Authors and Affiliations

  1. 1.Laboratory of Chemical Ecology, Graduate School of AgricultureKyoto UniversityKyotoJapan
  2. 2.Mobula Research Sdn Bhd.PenangMalaysia
  3. 3.JT Biohistory Research HallOsakaJapan

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