Content uploaded by Ming-Fai Liu
Author content
All content in this area was uploaded by Ming-Fai Liu on Sep 25, 2024
Content may be subject to copyright.
More than fruity scents: floral biology, scent and spectral reflectance of 1
twelve Annonaceae species 2
Ming-Fai Liu1,2, Junhao Chen3,4, Chun-Chiu Pang5, Tanya Scharaschkin6, Richard M. K. 3
Saunders1 4
5
1: Flora Conservation Department, Kadoorie Farm & Botanic Garden, Lam Kam 6
Road, Lam Tsuen, Hong Kong. 7
2: Area of Ecology & Biodiversity, School of Biological Sciences, The University 8
of Hong Kong, Pokfulam Road, Hong Kong. 9
3: Singapore Botanic Gardens, National Parks Board, 1 Cluny Road, Singapore 10
259569, Singapore. 11
4: Department of Biological Sciences, National University of Singapore, 16 12
Science Drive 4, Singapore 117558, Singapore 13
5: Coalition for Research on Ecology and Wildlife Limited, Room F, 25/F, 18 14
Farm Road, Tokwawan, Kowloon, Hong Kong. 15
6: Botanical Research, Art and Training, 54 Mill Road, Collinsvale, TAS, 7012, 16
Australia. 17
18
Correspondence 19
1: Ming-Fai Liu, Flora Conservation Department, Kadoorie Farm & Botanic 20
Garden, Lam Kam Road, Lam Tsuen, Hong Kong; 21
Email: nedlmf0@gmail.com 22
23
2: Richard M.K. Saunders, Area of Ecology & Biodiversity, School of Biological 24
Sciences, The University of Hong Kong, Pokfulam Road, Hong Kong. 25
Email: saunders@hku.hk 26
27
ORCIDs 28
Ming-Fai Liu: https://orcid.org/0000-0002-4732-2512 29
Junhao Chen: https://orcid.org/0000-0001-8822-169X 30
Chun-Chiu Pang: https://orcid.org/0000-0002-5286-1238 31
Tanya Scharaschkin: https://orcid.org/0000-0003-3081-0066 32
Richard M. K. Saunders: https://orcid.org/0000-0002-8104-7761 33
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprintthis version posted September 20, 2024. ; https://doi.org/10.1101/2024.09.16.613362doi: bioRxiv preprint
Abstract 34
The pantropical family Annonaceae possesses a broad array of floral phenotypes and 35
pollination specialisations. Being the most species-rich lineage in the magnoliids, 36
Annonaceae are a major flowering plant component of tropical rainforests, and are important 37
in the plant-pollinator network of this biome. Insects such as beetles, flies, bees, cockroaches 38
and thrips have been reported as their pollinators. Although there has been considerable effort 39
to assess their interactions with pollinators, attempts to characterise the visual and olfactory 40
communication channels between them are scarce. Here, we report the floral phenology and 41
characterise the spectral reflectance and odour of 12 Annonaceae species from five genera, 42
viz. Meiogyne, Monoon, Polyalthia, Pseuduvaria, and Uvaria. Pseuduvaria species are either 43
monoecious or dioecious while the four genera produce hermaphroditic flowers. The 44
Meiogyne species are likely pollinated by small beetles; the Polyalthia and Pseuduvaria 45
species are likely pollinated by both beetles and flies; and the Uvaria species is likely 46
pollinated by both beetles and bees. All species produce flowers that are not reflective in the 47
300–350 nm UV spectrum, and have various spectral reflectance profile across the remaining 48
UV and visible spectra. We detected diverse floral volatiles across all the species assessed. 49
The flowers of the small beetle-pollinated species, including Meiogyne species and 50
Polyalthia xanthocarpa, emitted mostly branched-chain esters, including isobutyl acetate and 51
isoamyl acetate. The Pseuduvaria species emitted scent reminiscent of rotten fruit, largely 52
consisting of 2,3-butanediol, a common by-product in yeast fermentation. The flowers of 53
Uvaria concava, which were visited by both bees and beetles, released mainly straight-chain 54
esters, including methyl hexanoate and methyl (Z)-4-octenoate. A corrugated inner petal 55
outgrowth was observed in Meiogyne and Pseuduvaria; this functions as a food body reward 56
in Meiogyne, but as a nectary gland for Pseuduvaria. This study provides insights into how 57
Annonaceae flowers attract different guilds of pollinators. 58
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprintthis version posted September 20, 2024. ; https://doi.org/10.1101/2024.09.16.613362doi: bioRxiv preprint
Introduction 59
The pantropical family Annonaceae is an early-divergent angiosperm lineage of trees and 60
lianas, comprising ca. 2500 species from 110 genera (Couvreur et al., 2019). Annonaceae 61
species are one of the major components of tropical lowland to lower montane rainforests 62
(Sosef et al., 2017). Almost all Annonaceae species possess a basic floral architecture of one 63
whorl of three sepals, two whorls of three petals, numerous spirally arranged stamens and 64
often free carpels (van Heusden, 1992). These flowers are largely pollinated by small beetle 65
pollinators from Curculionidae, Nitidulidae, Staphylinidae and Chrysomelidae (Gottsberger, 66
2012; Saunders, 2012). Because beetle pollinators spend considerable time in the flowers and 67
can often inflict damage to floral tissues, Annonaceae flowers have evolved protective 68
structures like fleshy corolla and shield-like stamen connectives (Gottsberger, 1999). These 69
adaptations are particularly pronounced in Neotropical species that are pollinated by large 70
scarab beetles from Cetoniinae, Dynastinae, Rutelinae and Trichiinae (Gottsberger, 1999). 71
While small beetles pollinate the majority of Annonaceae species, some are reported to be 72
pollinated by thrips (Gottsberger, 1999) or small flies (Silberbauer-Gottsberger et al., 2003), 73
and more rarely, flowers can be pollinated by bees (Teichert et al., 2009) or cockroaches 74
(Nagamitsu et al., 1997). Pollinators are often rewarded with pollen, stigmatic exudates, 75
floral nectar and thickened nutritious floral tissues (Gottsberger and Webber, 2018; Saunders, 76
2020). The floral chamber created by the corolla often additionally serves as tryst sites, and 77
more rarely as oviposition sites (Saunders, 2020). Highly specialised pollination systems 78
have been previously identified in Annonaceae, such as floral mimicry of fruits (Goodrich 79
and Jürgens, 2018, and references therein), fermentation substrates (Goodrich et al., 2006), 80
mushrooms (Teichert et al., 2012) and aerial litter (Liu et al., 2024). Other derived pollination 81
strategies, including private channel communication with beetles (Maia et al., 2012) and 82
euglossine bees (Teichert et al., 2009) have also been reported. Among these floral 83
specialisations, mimicry of fruits was proposed to be the ancestral condition for the family 84
(Johnson and Schiestl, 2016), and was likely pervasive, as around half of the recorded species 85
emit fruity floral aromas (Goodrich, 2012). 86
87
How flowers achieve specialised pollination is a research interest for pollination ecologists. 88
In particular, visual and olfactory cues have been recognised for their roles in mediating these 89
interactions (Schaefer and Ruxton, 2011). Annonaceae flowers display diverse combinations 90
of colour and odour, which might have enabled the evolution of specialisation in pollinators. 91
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprintthis version posted September 20, 2024. ; https://doi.org/10.1101/2024.09.16.613362doi: bioRxiv preprint
Strong floral fragrance is often noticeable in Annonaceae flowers. Based on a review by 92
Goodrich (2012), there appears to be a myriad of floral odour reported in the family based on 93
textual description. Although delineation of the chemical components of floral aromas is rare 94
for Annonaceae, volatiles from over 12 chemical classes have been reported so far (Goodrich, 95
2012). The type of pollination adaptation seems to be associated with molecules of specific 96
classes. For instance, proposed fruit mimics appear to be associated with aliphatic esters 97
(Jürgens, 2000; Johnson and Schiestl, 2016); flowers mimicking fermenting substates 98
reportedly emit fermentation by-products, including short-chain alcohols and ketones 99
(Goodrich et al., 2006); and monoterpenoids are associated with aerial litter mimics (Liu et 100
al., 2024) and flowers pollinated by Euglossines bees (Teichert et al., 2009). 101
102
Floral coloration is also diverse within the family, and is more frequently documented in the 103
literature. Floral colours, including yellow, red, pink, green, orange, white and dark maroon 104
or indigo flowers have been recorded in Annonaceae (van Heusden, 1992). It has been 105
reported that colour can shape the choice decision of one of the major pollinating beetle 106
family Nitidulidae (Döring et al., 2012; Vuts et al., 2022), highlighting the importance of 107
floral visual cues. Surprisingly, virtually no attempt has been made to characterise 108
Annonaceae floral colour based on spectral reflectance with only one exception (Liu et al., 109
2024). Given the diversity of pollination strategies, floral colour is an overlooked area which 110
can potentially help elucidate how Annonaceae flowers interact with their pollinators. 111
112
Previous studies have revealed that Annonaceae species in Australia display a variety of 113
floral adaptations to small beetle, fly or bee pollinators (Silberbauer-Gottsberger et al., 2003). 114
For instance, Meiogyne and Polyalthia (as Haplostichanthus), both generally with yellowish 115
corolla, are reportedly pollinated by small beetles from the family Nitidulidae and 116
Curculionidae, respectively. Pseuduvaria is reportedly pollinated by flies, and Uvaria is 117
reportedly pollinated by stingless bees. The aim of the current study is to characterise the 118
phenotypic adaptations for various types of pollination systems in Annonaceae. We selected 119
12 species from the genera Meiogyne, Monoon, Polyalthia, Pseudvaria and Uvaria in 120
Australia. These systems offer arenas for studying specialised bee pollination and potential 121
floral mimicry of fruit and fermenting substrates. Their floral biology, floral spectral 122
reflectance and floral odour are characterised here, laying a foundation for the future study of 123
floral traits and pollination ecology of specialised pollination systems. Furthermore, 124
magnoliids including Annonaceae preserve many architectural elements of ancestral 125
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprintthis version posted September 20, 2024. ; https://doi.org/10.1101/2024.09.16.613362doi: bioRxiv preprint
angiosperm flowers (Sauquet et al., 2017). Characterisation of the pollination strategies 126
employed by early-divergent angiosperms like magnoliids may therefore offer insights into 127
the pollination system of the ancestral flower. 128
129
Materials and methods 130
Study species and study sites 131
We examined three Meiogyne Miq. species (Meiogyne cylindrocarpa (Burck) Heusden, 132
Meiogyne trichocarpa (Jessup) D.C.Thomas & R.M.K. Saunders, Meiogyne stenopetala 133
(F.Muell.) Heusden), one Monoon Miq. species (Monoon patinatum (Jessup) B.Xue & 134
R.M.K.Saunders), two Polyalthia Blume species, (Polyalthia hispida B.Xue & 135
R.M.K.Saunders, Polyalthia xanthocarpa B.Xue & R.M.K.Saunders), four Pseuduvaria Miq. 136
species (Pseuduvaria hylandii Jessup, Pseuduvaria glabrescens (Jessup) Y.C.F.Su & 137
R.M.K.Saunders, Pseuduvaria mulgraveana Jessup, Pseuduvaria villosa Jessup) and two 138
Uvaria species (Uvaria concava Teijsm. & Binn. and Uvaria rufa Blume) (See Table 1 for 139
detailed information on the studied plants. Observations were conducted in the wild for 140
individuals of Me. stenopetala in Mount Tamborine in 2013 (permit number: 141
WITK08297010). All other observations were made on cultivated individuals at various sites: 142
Gardens by the Bay, Singapore, in 2018; Garry and Nada Sankowsky Arboretum, Tolga, 143
Australia, in 2019 and 2023; Cairns Botanic Gardens, Australia, in 2023; and Graham Wood 144
Arboretum, Wondecla, Australia, in 2023 (see Table 1 for details). 145
146
Phenology 147
To assess floral ontology, 10 flowers for each species were tagged and observed for seven 148
consecutive days. Observations were made daily to assess the duration of the pistillate and 149
staminate phase. Stigmatic peroxidase activity has been widely used as a gauge for stigma 150
receptivity (Dafni and Maués, 1998). However, the stigmas of some species, especially 151
Meiogyne spp., displayed peroxidase activity even during bud stage based on our preliminary 152
assessment with Peroxtesmo KO paper (MacheryNagel, Düren, Germany). The onset of 153
stigmatic receptivity was therefore defined by stigmatic exudate secretion, while the end of 154
stigmatic receptivity was determined by the abscission of stigmas. Staminate activity was 155
similarly determined by anther dehiscence and the abscission of petals. A pistillate phase is 156
defined here as the period that the flower has only the pistillate function, and likewise a 157
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprintthis version posted September 20, 2024. ; https://doi.org/10.1101/2024.09.16.613362doi: bioRxiv preprint
staminate phase is defined as the period that the flower only exhibits the staminate function. 158
An overlap phase describes the period that the flower has both the pistillate and staminate 159
functions, while an interim phase refers to the non-receptive period between the pistillate and 160
staminate phase. Floral thermogenesis was assessed using a handheld digital thermometer 161
every two hours throughout the day in the floral chamber, with additional readings ca. 10 cm 162
away from the flower to measure ambient temperature. 163
164
Floral visitors 165
Floral visitors were recorded during the entire observation period. A representative number of 166
floral visitors were collected and either preserved in 50% isopropanol for identification or 167
frozen at -20°C and silica-dried for inspection of pollen deposition under a dissecting 168
microscope. Probable pollinators were assessed by these criteria: 1) visitation to flowers of 169
both sexes or sexual phases; 2) contact with floral sexual organs; and 3) presence of pollen 170
grains on visitors in pistillate-phase flowers. Since the pistillate phase precedes the staminate 171
phase, the presence of these pollen-laden insects provide evidence for inter-floral transfer of 172
pollen grains. 173
174
Spectral reflectance of the petals 175
Pollinators often access Annonaceae floral reproductive organs via apical and basal apertures 176
created by the inner petals (Saunders, 2012). For genera with similar colour and morphology 177
between the inner and outer petals, namely Meiogyne, Monoon, Polyalthia and Uvaria, petal 178
spectral reflectance was represented by that of the inner petals. The abaxial surface of the 179
inner petals was measured for genera with drooping inner petals, viz. Meiogyne, Monoon and 180
Polyalthia, whereas the adaxial surface of the inner petals were measured for Uvaria, which 181
has spreading inner petals. Pseuduvaria has varying coloration among different parts of the 182
petals, and therefore three perianth areas were measured: outer petal adaxial surface, inner 183
petal abaxial surface, and inner petal gland. Differences between sexes and sexual phases 184
were not assessed due to logistic constraints. Spectral reflectance of the flowers at 300–700 185
nm was measured using an Ocean Optics FLAME-S-UV-VIS-ES spectrometer (Dunedin, FL, 186
USA), a DH-mini UV-VIS-NIR light source and an R200-7-UV-VIS probe positioned at 45°. 187
For each species, individual flowers (n=1–8) selected randomly from one to five biologically 188
distinct individuals were used as replicates. 189
190
Floral odour 191
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprintthis version posted September 20, 2024. ; https://doi.org/10.1101/2024.09.16.613362doi: bioRxiv preprint
To sample the floral volatiles, up to 10 intact flowers were enclosed in a “Toppits” oven bag 192
for each sample. To control for ambient contaminants, an empty bag was likewise sampled. 193
Two slits were made in the headspace bag, with one end connecting to a charcoal cartridge 194
(ORBO 32, Supelco, USA), supplying filtered air, and the other connected to a PAS 500 air 195
pump (Spectrex, USA). Floral volatiles were collected in a Porapak-Q cartridge (ORBO 196
1103, 50/80, Supelco, USA) at a flow rate of 100 m/min for 4 h. The porapak-Q cartridge 197
was then sealed and transported to the laboratory at The University of Hong Kong. The floral 198
volatiles then were eluted in 1 ml hexane (HPLC-grade, Sigma-Aldrich, USA). The eluents 199
were then concentrated to 200 µl by nitrogen blowdown. For each species, technical 200
replicates were made. The internal standard toluene was added to each sample at 34.68 ng/µl. 201
202
Gas Chromatography-Mass Spectrometry was performed on an Agilent 6890N/5973 gas 203
chromatograph-mass selective detector to characterise floral headspace. For each sample, 1 204
µl eluent was injected into the inlet at 230 °C for 1 min, and then injected into a DB-WAX 205
column (30 m × 0.25 mm, 0.25
μ
m thick film; J&W) in splitless mode. Helium was supplied 206
at a flow rate of 1 ml/min as the carrier gas. Oven temperatures were held at 60 °C for 3 min, 207
followed by a ramp up to 250 °C at 10 °C/min, and finally held for 7 min. Peaks were 208
manually integrated and identified tentatively using the Wiley and National Institute of 209
Standards and Technology mass spectral libraries (NIST, USA; 2020), with an 85% quality 210
threshold. Compound identities were verified by published standardised retention index 211
values or by co-injecting with commercially available standards. Molecules were quantified 212
with the internal standard toluene (see also Kantsa et al., 2018). The relative abundance of 213
each molecule was calculated. Volatiles recovered in the ambient control sample were 214
excluded in other samples. 215
216
Results 217
The floral biology of the studied species is summarised in Table 1. Most species assessed 218
displayed strong levels of protogyny and none were thermogenic. All Meiogyne, Monoon, 219
Polyalthia and Uvaria species assessed here bear hermaphroditic flowers, whereas the 220
Pseuduvaria species bear unisexual flowers (Ps. hylandii and Ps. glabrescens) or a 221
combination of staminate and hermaphroditic flowers (Ps. mulgraveana). 222
223
Floral biology and pollinator activities—Meiogyne species 224
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprintthis version posted September 20, 2024. ; https://doi.org/10.1101/2024.09.16.613362doi: bioRxiv preprint
The three Meiogyne species (Fig. 1a–c) produced yellow flowers with longitudinally grooved 225
basal outgrowths on the adaxial surface of the inner petals (Fig. 1c, Fig. 2a, c) and emitted a 226
banana-like scent. Their pistillate phase lasted two days, while the duration of staminate 227
phase differs among species (1–3 days) (Table 1). They exhibited a short overlap between the 228
pistillate and staminate phases that lasts ca. 0–5 hours. Small beetles were frequently 229
observed visiting the flowers of Me. cylindrocarpa (Curculionidae and Nitidulidae), Me. 230
trichocarpa (Curculionidae and Nitidulidae), and Me. stenopetala (Nitidulidae and 231
Staphylinidae) in both staminate and pistillate phase, during which they contacted the 232
stamens and stigmas. Pollen-laden curculionid beetles from the tribe Ochyromerini and 233
pollen-laden nitidulid beetles from the subfamily nitidulinae observed in pistillate-phase 234
flowers were likely the pollinators of both Me. cylindrocarpa and Me. trichocarpa. The 235
staphylinid beetles from the subfamily Omaliinae and the nitidulid beetles from the subfamily 236
Epuraeinae were likely the pollinators of Me. stenopetala. These floral pollinators consumed 237
pollen grains on the flowers where the beetles also sought shelter and copulated. Stigmas of 238
all three Meiogyne species produced a thin film of stigmatic exudate, which became more 239
copious towards the end of the pistillate phase. Consumption of stigmatic exudate was 240
invariably observed in curculionid, nitidulid and staphylinid beetle visitors. Consumption of 241
floral tissues, specifically petals and stigmas, were observed, primarily by curculionid 242
beetles, and less frequently by nitidulid and staphylinid beetles, leaving gnaw marks on the 243
floral tissues of the three Meiogyne species. However, consumption of the inner petal 244
corrugation was particularly pronounced compared to the surrounding floral tissues (Fig. 2). 245
Nectar secretion was not observed on the inner petal corrugation of Me. cylindrocarpa, Me. 246
trichocarpa and Me. stenopetala, thus they likely functioned as a food body reward for the 247
floral visitors. 248
249
Floral biology and pollinator activities—Polyalthia species 250
Polyalthia xanthocarpa (Fig. 1f) exhibited a 2–3-day pistillate phase followed by a 1-day 251
staminate phase (Table 1). The transition between pistillate and staminate phase was not 252
observed directly and could have occurred at night: it is therefore unclear whether an interim 253
or overlap phase was present. As with Meiogyne, Po. xanthocarpa bore yellow flowers with a
254
banana-like odour (Fig. 1f) and were visited by curculionid beetles that accessed the floral 255
sexual organs in both the staminate and pistillate phases (Table 1). Pollen grains were not 256
observed on the limited number of pistillate-phase visiting beetles, however. The curculionid 257
beetles were observed to consume pollen and utilise the flower for shelter. Secretion of 258
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprintthis version posted September 20, 2024. ; https://doi.org/10.1101/2024.09.16.613362doi: bioRxiv preprint
stigmatic exudate in Po. xanthocarpa was weak and the consumption of stigmatic exudate 259
was not observed. In contrast, Polyalthia hispida bore yellow flowers without detectable 260
odour (Fig. 1e). Its anthesis comprised a three-day pistillate phase followed by a two-day 261
staminate phase. No floral visitors were observed. 262
263
Floral biology and pollinator activities—Uvaria species 264
Uvaria concava bore flowers with red petals and emitted a fruity odour (Fig. 1k). It had a 2–265
3-day pistillate phase, followed by a prolonged overlap phase (1–2 days), in which stigmas 266
remained attached to the ovaries as the anthers dehisce. The drying and abscission of stigmas 267
usually occurred one day after the overlap phase and were sometimes synchronised with petal 268
abscission. In most flower, petal abscission occurred two days after the staminate function 269
initiated. Because of the variation in the timing of stigma drying, the duration of staminate 270
phase varied between 0–1 day. Nitidulid beetles (subfamily Nitidulinae) were the primary 271
floral visitors observed: they accessed the floral sexual organs and were present throughout 272
anthesis. They were probable pollinators since pollen-laden individuals were found in 273
pistillate-phase flowers. We observed the visitation of a single honeybee (Apis sp.) during the 274
overlap phase; although the bee contacted the floral sexual organs while collecting pollen, we 275
failed to capture it and were unable to assess pollen deposition. 276
277
Uvaria rufa flowers bore orange petals with tints of red and green (Fig. 1l). The flower 278
emitted a fruity odour and had a one-day pistillate phase, followed by a one-day interim 279
phase and a four-day staminate phase. Only one unidentified fly was observed, which visited 280
during the interim phase. It is unclear whether pollen grains were deposited onto this fly 281
visitor. 282
283
Floral biology and pollinator activities—Pseuduvaria 284
Pseuduvaria species (Fig. 1g–j) displayed an array of sexual systems (Table 1). Pseuduvaria 285
hylandii was assessed to be dioecious, while Ps. mulgraveana and Ps. glabrescens were 286
assessed to be structurally andromonoecious, in which the same individuals bear staminate 287
flowers that lack carpels, as well as bisexual flowers with a few stamens. Pseuduvaria 288
hylandii had prolonged anthesis for both pistillate and staminate flowers (> 6 days), but we 289
were unable to assess the anthesis duration (and floral phenology) for Ps. glabrescens due to 290
logistic limitations. Anthesis in staminate flowers of Ps. mulgraveana lasts two days, whereas 291
in hermaphroditic flowers the pistillate phase lasts for two days, followed immediately by 292
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprintthis version posted September 20, 2024. ; https://doi.org/10.1101/2024.09.16.613362doi: bioRxiv preprint
petal abscission, with anthers dehiscing two days after petal abscission. We were only able to 293
assess one staminate individual of Ps. villosa and were therefore unable to determine its 294
sexual system. 295
296
The flowers of all four Pseuduvaria species possessed white outer petals (Fig. 1g–j). They 297
also possessed white to reddish inner petals with dark maroon inner petal glands on their 298
adaxial surface (fig. 2d). These flowers emitted a strong fermentation odour that resembled 299
rotten fruits. They had a mitriform corolla, with large basal apertures that allowed larger 300
insects to access the floral chamber. The inner petals were apically connivent and the three 301
pairs of glands on the inner petals form a platform in the floral chamber (Fig. 1h, Fig. 2d–f). 302
Nectar secretion was observed on the inner petal glands and surrounding dark red petal 303
tissues of the four Pseuduvaria species (Fig. 2e). Drosophilid flies and nitidulid beetles were 304
observed to be the primary floral visitors of Ps. glabrescens, Ps. hylandii and Ps. 305
mulgraveana (Table 1). These insects were observed accessing the floral chamber and 306
consuming nectar, often positioning themselves underneath the sexual organs to access the 307
nectary. Pollen grains were observed to be deposited either onto the back of the insects (Fig. 308
2f), or onto the inner petal glands where the pollen grains were then picked up secondarily by 309
the flies and beetles on their legs. The drosophilid flies and nitidulid beetles were observed 310
moving across the stigmas and stamens with their legs touching the sexual organs, which may 311
assist pollen transfer. Some drosophilid flies are large enough to brush the back of their 312
thorax or abdomen against the sexual organs, which may additionally deposit pollen grains 313
onto the stigmas. Both drosophilid flies and nitidulid beetles were observed to access the 314
sexual organs of pistillate and staminate flowers for Ps. glabrescens, Ps. hylandii and Ps. 315
mulgraveana. While pollen grains were observed on the drosophilid flies and nitidulid beetles 316
found in pistillate-phase flowers of Ps. hylandii, pollen deposition was observed only on 317
nitidulid beetles from pistillate-phase flowers of Ps. glabrescens and Ps. mulgraveana. Other 318
dipteran visitors, namely lauxaniid and neriid flies, were also observed in Ps. hylandii, but 319
they are too large to access the floral chamber and thus unlikely to be pollinators. 320
Unfortunately, we failed to observe any floral visitors for Ps. villosa, though it has similar 321
floral colour and odour to the other Pseuduvaria species (Fig. 1j). 322
323
Floral biology and pollinator activities—Monoon 324
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprintthis version posted September 20, 2024. ; https://doi.org/10.1101/2024.09.16.613362doi: bioRxiv preprint
Monoon patinatum produced flowers with greenish petals (Fig. 1d). Only two staphylinid 325
beetle visitors from the subfamily Tachyporinae were observed during the pistillate phase 326
(Table 1). Pollen grains were not observed on their bodies. 327
328
Floral Colour 329
Various types of floral coloration were observed in the studied species, including red (Uvaria 330
concava), orange (U. rufa), yellow (Meiogyne cylindrocarpa, Me. trichocarpa, Me. 331
stenopetala, Polyalthia hispida and Po. xanthocarpa), green (Monoon patinatum), and a 332
mixtue of white and maroon (Pseuduvaria hylandii, Ps. glabrescens, Ps. mulgraveana and 333
Ps. villosa). The spectral reflectance of the petals of 12 Annonaceae species is shown in Fig. 334
3. The petals of all species are not UV-reflective from 300–350 nm; most of the reflectance 335
was observed in the visible spectrum (400–700 nm). Note that while yellow-flowered species 336
possess similar coloration to human perception, their spectral reflectance can be quite varied, 337
highlighting the importance of quantification of reflective spectra. 338
339
Floral Odour 340
The floral headspace composition of 11 Annonaceae species is summarised in Table 2. We 341
detected a total of 97 volatiles from seven classes of compounds. We did not detect any major 342
volatiles in the headspace of Polyalthia hispida against the ambient air control, and therefore 343
did not present the headspace data here. 344
345
Aliphatic ester was the most abundant class of volatiles in Meiogyne cylindrocarpa, Me. 346
trichocarpa, Me. stenopetala, Polyalthia xanthocarpa, Uvaria concava and U. rufa. The 347
headspaces of all Meiogyne species and Po. xanthocarpa contained a varying degree of 348
esters, including isobutyl acetate (Me. cylindrocarpa: 7.87 ± 2.16%; Me. trichocarpa: 11.86 349
± 4.37%; Me. stenopetala: 56.71 ± 1.5%) and isoamyl acetate (Me. cylindrocarpa: 79.58 ± 350
7.65%; Me. trichocarpa: 84.07 ± 4.01%; Me. stenopetala: 28.17 ± 4.13%; Po. xanthocarpa: 351
99.28 ± 0.02%). For U. concava, the floral headspaces were dominated by two esters, namely 352
methyl-(Z)-4-octenoate (25.36 ± 0.32%), and methyl hexanoate (54.69 ± 0.32%), while for U. 353
rufa, the most abundant volatiles were hexyl acetate (21.27 ± 0.25%), butyl acetate (20.65 ± 354
0.21%), and ethyl octanoate (20.04 ± 0.22%). In Meiogyne and Polyalthia, branched-chain 355
aliphatic esters dominated the floral odour, while in Uvaria, the floral odours primarily 356
consisted of straight-chain the aliphatic esters. For Monoon patinatum, the headspace mainly 357
comprised the monoterpenes
β
-ocimene (45.50 ± 0.61%) and allo-ocimene (11.89 ± 0.59%), 358
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprintthis version posted September 20, 2024. ; https://doi.org/10.1101/2024.09.16.613362doi: bioRxiv preprint
and 2-methyl-butanoic acid (16.48 ± 0.53%). The oxygenated aliphatic compound was the 359
most abundant volatile class for Pseuduvaria hylandii, Ps. glabrescens, Ps. mulgraveana and 360
Ps. villosa, consisting of the molecule 2,3-butanediol (84.83–96.98% of the headspaces). 361
362
Discussion 363
Floral phenology 364
Most species assessed in this study exhibited a 3–5 day anthesis period and none exhibited 365
circadian trapping, a time-dependent trapping mechanism in which pollinators are trapped 366
and released in accordance to the insects’ peak circadian activities (Lau et al., 2017). The 367
duration of anthesis for most species in the current study is similar to the typical non-trapping 368
Annonaceae species (
≥
2 days) (Pang and Saunders, 2014; Lau et al., 2017). As the family is 369
self-compatible (Pang and Saunders, 2014), temporal separation of sexual phases is an 370
important adaptation to reduce autogamy. An interim phase was observed in Uvaria rufa, 371
during which the stigmas are no longer receptive and anther dehiscence has not begun, 372
potentially offering additional assurance to prevent autogamy (Pang and Saunders, 2014). 373
However, in some of the other species, including Meiogyne spp. and U. concava, an overlap 374
phase was observed, during which carpels and stamens are simultaneously functional within a 375
flower. This overlap phase could offer reproductive assurance by autogamy when pollinator 376
availability is scarce. It was previously reported that U. concava was pollinated by stingless 377
bees (Meliponinae) in its natural population (50 km from our site of study) (Silberbauer-378
Gottsber et al., 2003). Because bee visitors to Annonaceae often collect pollen only as a food 379
reward, and seldom visit pistillate-phase flowers, the overlap phase has been suggested to be 380
essential for bee pollination (Gottsberger, 2014). It was, however, reported that meliponine 381
visitors also collect stigmatic exudate as a reward (Silberbauer-Gottsberger et al., 2013), 382
suggesting the flower offers reward to the meliponine visitors even during pistillate phase. 383
The role of the prolonged overlap phase for U. concava is therefore contentious. 384
385
Floral phenology also played a role in the sexual function of the flower, especially for 386
Pseuduvaria species. Previous studies have shown that Ps. mulgraveana is structurally 387
andromonoecious (Pang et al., 2013), meaning that staminate flowers and bisexual flowers 388
are borne on the same plant. The stamens of bisexual flowers, however, release functional 389
pollen grains only after petal abscission (Pang et al., 2013), rendering the plant functionally 390
monoecious, which is corroborated by our observations. 391
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprintthis version posted September 20, 2024. ; https://doi.org/10.1101/2024.09.16.613362doi: bioRxiv preprint
392
Inner petal corrugation and glands 393
Inner petal outgrowth was observed in Meiogyne spp. and Pseuduvaria spp. in the current 394
study. This outgrowth has independently evolved in around 20 genera, including Annona L., 395
Alphonsea Hook.f. & Thomson, Asimina Adans., Asteranthe Engl. & Diels, Duguetia A.St.-396
Hil., Maasia Mols, Kessler & Rogstad, Meiogyne Miq., Miliusa Lesch. ex A.DC., Orophea 397
Blume, Porcelia Ruiz & Pav., Pseuduvaria Miq., Sapranthus Seem., Stenanona Standl., 398
Tetrameranthus R.E.Fr., Tridimeris Baill., Uvaria L., Wangia X.Guo & R.M.K.Saunders, 399
and Xylopia L. (van Heusden, 1992; Schatz and Maas, 2010; Xue et al., 2017). In 400
Pseuduvaria, the inner petal outgrowth has been reported to be a nectary gland (Silberbauer-401
Gottsberger et al., 2003). In the current study, the four Pseuduvaria species also possessed 402
this floral tissue, which invariably functioned as nectary, corroborating the previous study. 403
Likewise, all three Meiogyne species assessed in our study possessed weakly folded to 404
longitudinally grooved basal growths on the adaxial surface of the inner petals. Although this 405
structure has previously been interpreted as a gland (van Heusden, 1992, 1994), histological 406
studies have failed to identify secretory ducts or hairs (Xue et al., 2021). The inner petal 407
corrugation may potentially offer tactile cues for the floral mimicry of aerial litter in 408
Meiogyne heteropetala (Liu et al., 2024), but the role of this tissue remains enigmatic for 409
most Meiogyne species. Our observations indicate that pollinators preferentially consume the 410
inner petals over other floral structures (Fig. 2), suggesting that the corrugation likely 411
functions as a food body in Me. cylindrocarpa, Me. trichocarpa and Me. stenopetala. In 412
Annonaceae, specialised nutritious petal tissues are common in species that are pollinated by 413
scarab beetles in the neotropics, including Annona, Cymbopetalum, Duguetia, and Malmea 414
species (Gottsberger and Webber, 2018). Among these lineages with specialised food body, 415
there is potential convergence in tissue structure. The inner petal corrugation of these species 416
is anatomically similar to those found in Meiogyne, lacking secretory openings, ducts or 417
trichromes, and comprising a mixture of cells full of starch granules and cells enriched with 418
tannins (Gottsberger and Webber, 2018; Xue et al., 2021). 419
420
Exploitation of pollinators associated with fruits 421
The fruity floral aromas and beetle pollination of Annonaceae have led some authors to 422
propose some Annonaceae flowers mimic fruits (Johnson and Schiestl, 2016; Goodrich and 423
Jürgens, 2018). In particular, fruit mimicry has been postulated in Anaxagorea, Annona, 424
Artabotrys, Duguetia and Xylopia and is likely to be prevalent in the Annonaceae (Johnson 425
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprintthis version posted September 20, 2024. ; https://doi.org/10.1101/2024.09.16.613362doi: bioRxiv preprint
and Schiestl, 2016; Goodrich and Jürgens, 2018; Chen et al., 2020). These flowers are 426
pollinated by small beetles such as Curculionidae, Nitidulidae and Staphylinidae that are 427
known to use fruits as a food source or oviposition site (Phelan and Lin, 1991; Racette et al., 428
1992; Frank and Thomas, 1999; Mutinelli et al., 2015). Previous studies have recorded small-429
beetle visitors in Polyalthia xanthocarpa flowers (as “Haplostichanthus spec. (“Cape 430
tribulation”)”; Morawetz, 1988), and curculionid beetles have been observed visiting the 431
flowers in Cape Tribulation, Australia (pers. comm., Dr David Tng). For Meiogyne, nitidulid 432
and curculionid beetles have been reported to be the pollinators of Me. cylindrocarpa in 433
tropical rainforests in West Malesia (Momose, 2005; Roubik et al, 2005). Beetles from these 434
families were also observed in the current study. 435
436
In this study, floral aromas resembling ripe fruits were recorded in Me. cylindrocarpa, Me. 437
trichocarpa, Me. stenopetala, Po. xanthocarpa. Small beetle pollinators (families Nitidulidae, 438
Curculionidae and Staphylinidae) were observed as floral visitor of Me. cylindrocarpa, Me. 439
trichocarpa, Me. stenopetala and Po. xanthocarpa (Table 1). Their floral odour resembled 440
strong banana-like scent to human perception, and is largely composed of either isoamyl 441
acetate or isobutyl acetate (Table 2). These two molecules have been reported in various 442
fruits, including banana (Jordán et al., 2001), papaya (Katague and Kirch, 1965), pear (Zhang 443
et al., 2023) and wild soursop (Pino et al., 2002), and is implicated as the semiochemicals of 444
Curculionidae and Nitidulidae (Rochat et al., 2000; Torto et al., 2007). The spectral 445
reflectance of Meiogyne species, especially Me. cylindrocarpa and Me. trichocarpa, 446
exhibited low UV reflectance, with stronger reflectance from 500–700 nm (Fig. 3). The UV-447
absorbing yellow spectral profile is a characteristic for pollen (Dötterl et al., 2014) and was 448
also reported in fruits such as bananas (Xie et al., 2018). Some nitidulid and curculionid 449
beetles are known to associate with fruits (Phelan and Lin, 1991; Racette et al., 1992; 450
Mutinelli et al., 2015), and nitidulidae has previously been demonstrated to be attracted to 451
various shade of yellow (Döring et al., 2012; Vuts et al., 2022). The current evidence 452
suggests that Me. cylindrocarpa, Me. trichocarpa, Me. stenopetala and Po. xanthocarpa 453
likely attract their pollinators at least by exploiting visual and olfactory cues that indicate 454
fruits. 455
456
Exploitation of pollinators associated with fermented substrates 457
Records of floral visitors to Pseuduvaria are more comprehensive than for most other 458
Annonaceae genera in Australia. In a previous pollination study on another Australian 459
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprintthis version posted September 20, 2024. ; https://doi.org/10.1101/2024.09.16.613362doi: bioRxiv preprint
Pseuduvaria species, Ps. froggattii, it was shown that the flowers are primarily pollinated by 460
drosophilid flies (Silberbauer-Gottsberger et al., 2003). Similarly, Ps. glabrescens was 461
previously reported to be visited primarily by small flies (as Pseuduvaria spec. (“Davies 462
Creek”); Morawetz, 1988). Pseuduvaria mulgraveana, on the other hand, was reported to be 463
pollinated by the nitidulid beetle, Aethina australis, and was only infrequently visited by 464
flies. Our observations are largely congruent with these studies: Ps. hylandii, Ps. glabrescens 465
and Ps. mulgraveana were observed to be visited by both nitidulid beetles and drosophilid 466
flies. Pollen deposition on the nitidulid beetles was observed in the pistillate-phase flowers of 467
all three species, while drosophilid flies appeared to be the pollinator only for Ps. hylandii. It 468
was previously postulated that flies could pollinate Ps. villosa, but floral visitors were not 469
observed in our study. 470
471
Floral coloration in the Pseuduvaria species varies across different areas of petal tissues 472
(Figs. 1 & 3). The abaxial surface of the inner petals in most species demonstrate largely 473
similar reflectance spectra with the adaxial surface of the outer petals, except for Ps. hylandii. 474
In contrast to these areas, the inner petal glands of all four Pseuduvaria species studied are 475
burgundy red with generally lower reflectance across all wavelengths. Additionally, all four 476
Pseuduvaria species emit an odour reminiscent of rotten fruits. Rotting fruits have been 477
reported to display a reduction in overall reflectance across the visible spectrum under fungal 478
infection (Liu et al., 2020). The contrast between the dark burgundy inner petal gland and the 479
remaining bright floral tissues may have evolved to imitate rotten patches on fruits. 480
Interestingly, in Pseuduvaria, nectar secretion is localised in the darkened area (Fig. 2), 481
suggesting there is co-localisation of gustatory cue and visual cue, which may help position 482
the pollinators underneath the reproductive organs and facilitate pollen transfer. The floral 483
odours of all four Pseuduvaria species are largely composed of 2,3-butanediol (Table 2), 484
which is a common volatile emitted by yeast and fermenting substrates (Goodrich et al., 485
2006, 2023). This molecule is one of the major volatiles of Asimina triloba (Goodrich et al., 486
2006), a maroon-petalled floral mimic of fermentation substrates (Goodrich et al., 2023). As 487
with Ps. glabrescens, Ps. hylandii and Ps. mulgraveana, Asimina triloba is pollinated by 488
small flies and small beetles (Martin, 2021; Goodrich et al., 2023). This suggests 489
convergence in floral colour and odour between Asimina and Pseuduvaria for attracting the 490
same pollinator guilds. The four Pseuduvaria species may therefore be potential candidates 491
for floral mimics of fermentation substrates. 492
493
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprintthis version posted September 20, 2024. ; https://doi.org/10.1101/2024.09.16.613362doi: bioRxiv preprint
Exploitation of bee and beetle pollinators 494
Uvaria concava was reported to be pollinated by stingless bees in the wild (Silberbauer-495
Gottsberger et al., 2003) but is here inferred to be pollinated by nitidulid beetles in cultivation 496
(Table 1). This species possesses bright-red petals, reflecting the visible spectrum mostly at 497
600–700 nm (Fig. 3). However, neither the tribe Meliponini (stingless bees) nor the family 498
Apidae have photoreceptors within this range (van der Kooi et al., 2021). Though the 499
photosensitivity of Nitidulidae is unknown, most coleopteran families with known visual 500
systems are unable to detect this part of the visible spectrum (van der Kooi et al., 2021). It is 501
therefore unclear how floral visual cues assist pollinator attraction in U. concava. The floral 502
odour of U. concava is largely composed of methyl hexanoate and methyl (Z)-4-octenoate. 503
While there is little documentation how meliponine bees and nitidulid beetles respond to 504
these molecules, methyl hexanoate is reported to be a pheromone for male bumblebees 505
(Valterová et al., 2001). Among Annonaceae species that emit esters, branched-chain 506
aliphatic esters are typically produced (Table 2; Jürgens et al, 2000), but Uvaria spp. 507
primarily produce straight-chain aliphatic esters instead. Interestingly, the branched-chain 508
aliphatic ester isoamyl acetate (the dominant floral scent component of Meiogyne spp. and 509
Polyalthia xanthocarpa) is a bee alarm pheromone that reduces foraging in the honeybee 510
species Apis mellifera and Apis cerana (Gong et al., 2017). Nonetheless, whether this 511
structural change in floral esters contributes to the shift to bee pollination remains to be 512
investigated. Future studies comparing the contrast between spectral reflectance of the petals 513
and reproductive organs as well as colour-based and scent-based bioassays could help assess 514
the role played by visual and olfactory cues in bee-pollinated Annonaceae species. 515
516
Conclusions 517
The basic floral biology, petal spectral reflectance and floral scent of selected species in five 518
Annonaceae genera are characterised here in detail. Our study has identified multiple species 519
that have floral traits resembling fruits (Meiogyne spp. and Po. xanthocarpa) and fermented 520
substrates (Meiogyne spp. and Po. xanthocarpa). The specialised floral cues and adaptations 521
identified in this study provide preliminary evidence for potential floral mimicry of fruits and 522
fermented substrates, and offer insights into how floral cues may attract floral visitors. These 523
findings serve as a prelude to future studies on specialised pollination systems, and provide 524
baseline information on floral traits in the pantropical family Annonaceae. Future studies 525
involving the characterisation of colour and odour of co-ocurring fruits and fermenting 526
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprintthis version posted September 20, 2024. ; https://doi.org/10.1101/2024.09.16.613362doi: bioRxiv preprint
substrates would be important to assess whether these species exploit their pollinators 527
through floral mimicry. 528
529
Acknowledgements 530
We would like to express our gratitude to Garry and Nada Sankowsky, Graham Wood, 531
Charles Clarke (Cairns Botanic Gardens), and Janelle Jung (Gardens by the Bay) for allowing 532
us to work on the living collections of Annonaceae species in their arboreta or botanic 533
gardens. We are grateful for Frank Zich from Australian Tropical Herbarium, and staff from 534
Mount Tamborine National Park and the Queensland Department of Environment, Science 535
and Innovation for their assistance, and Jessie Lai and Laura Wong for their technical 536
support. This research is funded by the Hong Kong Research Grants Council 537
(HKU17112616), awarded to R.M.K.S. 538
539
References 540
Chen, J. Liu, M.-F., and Saunders, R M. K. (2020). Contrasting floral biology of Artabotrys 541
species (Annonaceae): Implications for the evolution of pollinator trapping. Plant Species 542
Biology, 35, 210–223. 543
544
Couvreur, T. L. P., Helmstetter, A. J., Koenen, E. J. M., Bethune, K., Brandão, R. D., Little, 545
S. A., Sauquet, H. and Erkens, R. H. J. (2019). Phylogenomics of the major tropical plant 546
family Annonaceae using targeted enrichment of nuclear genes. Frontiers in Plant Science, 9, 547
431757. 548
549
Dafni, A., and Maués, M. M. (1998). A rapid and simple procedure to determine stigma 550
receptivity. Sexual Plant Reproduction, 11(3), 177–180. 551
552
Döring, T. F., Skellern, M., Watts, N., & Cook, S. M. (2012). Colour choice behaviour in the 553
pollen beetle Meligethes aeneus (Coleoptera: Nitidulidae). Physiological Entomology, 37(4), 554
360–378. 555
556
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprintthis version posted September 20, 2024. ; https://doi.org/10.1101/2024.09.16.613362doi: bioRxiv preprint
Dötterl, S., Glück, U., Jürgens, A., Woodring, J., and Aas, G. (2014). Floral reward, 557
advertisement and attractiveness to honey bees in dioecious Salix caprea. PloS One, 9(3), 558
e93421. 559
560
Frank, J. H., and Thomas, M. C. (1999). Rove Beetles of Florida, Staphylinidae (Insecta: 561
Coleoptera: Staphylinidae). DPI Entomology Circulars, 343, 1–11. 562
563
Gong, Z., Wang, C., Dong, S., Zhang, X., Wang, Y., Hu, Z., and Tan, K. (2017). High 564
concentrations of the alarm pheromone component, isopentyl acetate, reduces foraging and 565
dancing in Apis mellifera Ligustica and Apis cerana Cerana. Journal of Insect Behavior, 30, 566
188–198. 567
568
Goodrich, K. R., and Jürgens, A. (2018). Pollination systems involving floral mimicry of 569
fruit: aspects of their ecology and evolution. New Phytologist, 217(1), 74–81. 570
571
Goodrich, K. R., Zjhra, M. L., Ley, C. A., and Raguso, R. A. (2006). When flowers smell 572
fermented: the chemistry and ontogeny of yeasty floral scent in pawpaw (Asimina triloba: 573
Annonaceae). International Journal of Plant Sciences, 167(1), 33–46. 574
575
Goodrich, K. R., Ellis, I., DeHaas, A., Senski, R., and Savage, J. (2023). False advertising 576
with fermented scents: floral mimicry in pawpaw (Asimina triloba: Annonaceae) 577
pollination. International Journal of Plant Sciences, 184(6), 485–497. 578
579
Gottsberger, G. (1999). Pollination and evolution in neotropical Annonaceae. Plant Species 580
Biology, 14(2), 143–152. 581
582
Gottsberger, G. (2012). How diverse are Annonaceae with regard to pollination? Botanical 583
Journal of the Linnean Society, 169(1), 245–261. 584
585
Gottsberger, G. (2014). Evolutionary steps in the reproductive biology of 586
Annonaceae. Revista Brasileira de Fruticultura, 36, 32–43. 587
588
Gottsberger, G., and Webber, A. C. (2018). Nutritious tissue in petals of Annonaceae and its 589
function in pollination by scarab beetles. Acta Botanica Brasilica, 32, 279–286. 590
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprintthis version posted September 20, 2024. ; https://doi.org/10.1101/2024.09.16.613362doi: bioRxiv preprint
591
Johnson, S. D., and Schiestl, F. P. (2016). Floral mimicry. Oxford University Press. 592
593
Jordán, M. J., Tandon, K., Shaw, P. E., and Goodner, K. L. (2001). Aromatic profile of 594
aqueous banana essence and banana fruit by gas chromatography-mass spectrometry (GC-595
MS) and gas chromatography-olfactometry (GC-O). Journal of Agricultural and Food 596
Chemistry, 49(10), 4813–4817. 597
598
Jürgens, A., Webber, A. C., and Gottsberger, G. (2000). Floral scent compounds of 599
Amazonian Annonaceae species pollinated by small beetles and 600
thrips. Phytochemistry, 55(6), 551–558. 601
602
Kantsa, A., Raguso, R. A., Dyer, A. G., Olesen, J. M., Tscheulin, T., and Petanidou, T. 603
(2018). Disentangling the role of floral sensory stimuli in pollination networks. Nature 604
Communications, 9(1), 1041. 605
606
Katague, D.B. and Kirch, E.R. (1965), Chromatographic analysis of the volatile components 607
of papaya fruit. Journal of Pharmaceutical Sciences, 54(6), 891–894. 608
609
Lau, J. Y., Guo, X., Pang, C. C., Tang, C. C., Thomas, D. C., and Saunders, R. M. K. (2017). 610
Time-dependent trapping of pollinators driven by the alignment of floral phenology with 611
insect circadian rhythms. Frontiers in Plant Science, 8, 1119. 612
613
Liu, Q., Zhou, D., Tu, S., Xiao, H., Zhang, B., Sun, Y., Pan., L., and Tu, K. (2020). 614
Quantitative visualization of fungal contamination in peach fruit using hyperspectral 615
imaging. Food Analytical Methods, 13, 1262–1270. 616
617
618
Liu, M. F., Chen, J., Goodrich, K. R., Chiu, S. K., Pang, C. C., Scharaschkin, T., and 619
Saunders, R. M. K. (2024). Aerial litter mimicry: a novel form of floral deception mediated 620
by a monoterpene synthase. bioRxiv, 2024.06.12.596753; doi: 621
https://doi.org/10.1101/2024.06.12.596753 622
623
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprintthis version posted September 20, 2024. ; https://doi.org/10.1101/2024.09.16.613362doi: bioRxiv preprint
Maia, A. C. D., Dötterl, S., Kaiser, R., Silberbauer-Gottsberger, I., Teichert, H., Gibernau, 624
M., do Amaral Ferraz Navarro, D. M., Schlindwein, C., and Gottsberger, G. (2012). The key 625
role of 4-methyl-5-vinylthiazole in the attraction of scarab beetle pollinators: a unique 626
olfactory floral signal shared by Annonaceae and Araceae. Journal of Chemical Ecology, 38, 627
1072–1080. 628
629
Martin, K. R. (2021). When flowers play Dead: microbes as architects of a ‘deceptive’ floral 630
phenotype (Doctoral dissertation, Cornell University). 631
632
Momose, K. 2005. Beetle pollination in tropical rain forests. In Roubik, D.W., Sakai, S. and 633
Karim, A.A.H. (Eds.), Pollination ecology and the rain forest (pp. 104–110). New York: 634
Springer. 635
636
Morawetz, W. (1988). Karyosystematics and evolution of Australian Annonaceae as 637
compared with Eupomatiaceae, Himantandraceae, and Austrobaileyaceae. Plant Systematics 638
and Evolution, 159, 49–79. 639
640
Mutinelli, F., Federico, G., Carlin, S., Montarsi, F., and Audisio, P. (2015). Preliminary 641
investigation on other Nitidulidae beetles species occurring on rotten fruit in Reggio Calabria 642
province (south-western Italy) infested with small hive beetle (Aethina tumida). Journal of 643
Apicultural Research, 54(3), 233–235. 644
645
Nagamitsu, T., and Inoue, T. (1997). Cockroach pollination and breeding system of Uvaria 646
elmeri (Annonaceae) in a lowland mixed
dipterocarp forest in Sarawak. American Journal 647
of Botany, 84(2), 208–213. 648
649
Pang, C. C., Scharaschkin, T., Su, Y. C., and Saunders, R. M. K. (2013). Functional monoecy 650
due to delayed anther dehiscence: a novel mechanism in Pseuduvaria mulgraveana 651
(Annonaceae). PLoS One, 8(3), e59951. 652
653
Pang, C. C., and Saunders, R. M. K. (2014). The evolution of alternative mechanisms that 654
promote outcrossing in Annonaceae, a self-compatible family of early-divergent 655
angiosperms. Botanical Journal of the Linnean Society, 174(1), 93–109. 656
657
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprintthis version posted September 20, 2024. ; https://doi.org/10.1101/2024.09.16.613362doi: bioRxiv preprint
Phelan, P. L., and Lin, H. (1991). Chemical characterization of fruit and fungal volatiles 658
attractive to dried-fruit beetle, Carpophilus hemipterus (L.)(Coleoptera: Nitidulidae). Journal 659
of Chemical Ecology, 17, 1253–1272. 660
661
Pino, J. A., Marbot, R., and Agüero, J. (2002). Volatile components of wild soursop (Annona 662
montana Macf.) fruit. Journal of Essential Oil Research, 14(4), 257–258. 663
664
Racette, G., Chouinard, G., Vincent, C., and Hill, S. B. (1992). Ecology and management of 665
plum curculio, Conotrachelus nenuphar [Coleoptera: Curculionidae], in apple 666
orchards. Phytoprotection, 73(3), 85–100. 667
668
Rochat, D., Meillour, P. N. L., Esteban-Duran, J. R., Malosse, C., Perthuis, B., Morin, J. P., 669
and Descoins, C. (2000). Identification of pheromone synergists in American palm weevil, 670
Rhynchophorus palmarum, and attraction of related Dynamis borassi. Journal of Chemical 671
Ecology, 26, 155–187. 672
673
Roubik, D. W., Sakai, S. and Karim, A. A. H. (2005). Appendix A. In Roubik, D. W., Sakai, 674
S. and Karim, A. A. H. (Eds.), Pollination ecology and the rain forest: Sarawak studies (pp. 675
223–245). New York: Springer. 676
677
Saunders, R. M. K. (2012). The diversity and evolution of pollination systems in 678
Annonaceae. Botanical Journal of the Linnean Society, 169(1), 222–244. 679
680
Saunders, R. M. K. (2020). The evolution of key functional floral traits in the early divergent 681
angiosperm family Annonaceae. Journal of Systematics and Evolution, 58(4), 369–392. 682
683
Sauquet, H., Von Balthazar, M., Magallón, S., Doyle, J. A., Endress, P. K., Bailes, E. J., de 684
Morais, E. B. et al. (2017). The ancestral flower of angiosperms and its early 685
diversification. Nature Communications, 8(1), 16047. 686
687
Schaefer, H. M., and Ruxton, G. D. (2011). Plant-animal communication. OUP Oxford. 688
689
Schatz, G. E., and Maas, P. J. M. (2010). Synoptic revision of Stenanona 690
(Annonaceae). Blumea, 55(3), 205–223. 691
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprintthis version posted September 20, 2024. ; https://doi.org/10.1101/2024.09.16.613362doi: bioRxiv preprint
692
Silberbauer-Gottsberger, I., Gottsberger, G., and Webber, A. C. (2003). Morphological and 693
functional flower characteristics of new and old world Annonaceae with respect to their mode 694
of pollination. Taxon, 52(4), 701–718. 695
696
Sletvold, N., Trunschke, J., Smit, M., Verbeek, J., and Ågren, J. (2016). Strong pollinator-697
mediated selection for increased flower brightness and contrast in a deceptive 698
orchid. Evolution, 70(3), 716–724. 699
700
Sosef, M. S., Dauby, G., Blach-Overgaard, A., van Der Burgt, X., Catarino, L., Damen, T., 701
Deblauwe, V., Dessein, S., Dransfield, J., Droissart, V., Duarte, M. C., Engledow, H., Fadeur, 702
G., Figueira, R., Gereau, R. E., Hardy, O. J., Harris, D. J., de Heij, J., Janssens, S., Klomberg, 703
Y., Ley, A. C., Mackinder, B. A., Meerts, P., van de Poel, J. L., Sonké, B., Stévart, T., 704
Stoffelen, P., Svenning, J.-C., Sepulchre, P., Zaiss, R., Wieringa, J. J. and Couvreur, T. L. 705
(2017). Exploring the floristic diversity of tropical Africa. BMC Biology, 15, 1–23. 706
707
Teichert, H., Dötterl, S., Zimma, B., Ayasse, M., and Gottsberger, G. (2009). 708
Perfume
collecting male euglossine bees as pollinators of a basal angiosperm: the case of 709
Unonopsis stipitata (Annonaceae). Plant Biology, 11(1), 29–37. 710
711
Teichert, H., Dötterl, S., Frame, D., Kirejtshuk, A., and Gottsberger, G. (2012). A novel 712
pollination mode, saprocantharophily, in Duguetia cadaverica (Annonaceae): a stinkhorn 713
(Phallales) flower mimic. Flora-Morphology, Distribution, Functional Ecology of 714
Plants, 207(7), 522–529. 715
716
Torto, B., Arbogast, R. T., Alborn, H., Suazo, A., Van Engelsdorp, D., Boucias, D., 717
Tumlinson, J. H., and Teal, P. E. (2007). Composition of volatiles from fermenting pollen 718
dough and attractiveness to the small hive beetle Aethina tumida, a parasite of the honeybee 719
Apis mellifera. Apidologie, 38(4), 380–389. 720
721
Valterová, I., Urbanová, K., Hovorka, O., and Kindl, J. (2001). Composition of the labial 722
gland secretion of the bumblebee males Bombus pomorum. Zeitschrift für Naturforschung 723
C, 56(5–6), 430–436. 724
725
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprintthis version posted September 20, 2024. ; https://doi.org/10.1101/2024.09.16.613362doi: bioRxiv preprint
Van Heusden, E. C. H. (1992). Flowers of Annonaceae: morphology, classification, and 726
evolution. Blumea. Supplement, 7(1), 1–218. 727
728
Van Heusden, E. C. H. (1994). Revision of Meiogyne (Annonaceae). Blumea: Biodiversity, 729
Evolution and Biogeography of Plants, 38(2), 487–511. 730
731
Van Der Kooi, C. J., Stavenga, D. G., Arikawa, K., Beluši
č
, G., and Kelber, A. (2021). 732
Evolution of insect color vision: from spectral sensitivity to visual ecology. Annual Review of 733
Entomology, 66, 43–5461. 734
735
Vuts, J., Szarukán, I., Marczali, Z., Csonka, É. B., Szilágyi, A., Imrei, Z., Nagy, A. and Tóth, 736
M. (2022). Differences in colour preference among pollen beetle species (Coleoptera: 737
Nitidulidae). Journal of Applied Entomology, 146(3), 301–309. 738
739
Xue, B., Shao, Y. Y., Saunders, R. M., and Tan, Y. H. (2017). Alphonsea glandulosa 740
(Annonaceae), a new species from Yunnan, China. PLoS One, 12(2), e0170107. 741
742
Xie, C., Chu, B., and He, Y. (2018). Prediction of banana color and firmness using a novel 743
wavelengths selection method of hyperspectral imaging. Food Chemistry, 245, 132–140. 744
745
Xue, B., Shao, Y. Y., Xiao, C. F., Liu, M. F., Li, Y., and Tan, Y. H. (2021). Meiogyne 746
oligocarpa (Annonaceae), a new species from Yunnan, China. PeerJ, 9, e10999. 747
748
Zhang, W., Yan, M., Zheng, X., Chen, Z., Li, H., Mao, J., Qin, H., Zhu, C., Du, H., and Abd 749
El-Aty, A. M. (2023). Exploring the aroma fingerprint of various Chinese pear cultivars 750
through qualitative and quantitative analysis of volatile compounds using HS-SPME and 751
GC× GC-TOFMS. Molecules, 28(12), 4794. 752
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprintthis version posted September 20, 2024. ; https://doi.org/10.1101/2024.09.16.613362doi: bioRxiv preprint
753
Fig. 1. Flowers of Annonaceae species studied. (a) Meiogyne cylindrocarpa. (b) Meiogyne 754
trichocarpa with curculionid and nitidulid visitors. (c) Meiogyne stenopetala with a 755
staphylinid visitor. (d) Monoon patinatum. (e) Polyalthia hispida. (f) Polyalthia xanthocarpa. 756
(g) Pseuduvaria glabrescens with nitidulid visitors. (h) Pseuduvaria hylandii with 757
drosophilid visitors. (i) Pseuduvaria mulgraveana. (j) Pseuduvaria villosa. (k) Uvaria 758
concava. (l) Uvaria rufa at interim phase with a fly visitor. Photo credit: Photo credits: (a, b, 759
d, e, g–l): Ming-Fai Liu; (c): Chun-Chiu Pang; (f): Garry & Nada Sankowsky. 760
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprintthis version posted September 20, 2024. ; https://doi.org/10.1101/2024.09.16.613362doi: bioRxiv preprint
761
Fig. 2. Inner petal outgrowths. (a) Adaxial surface of inner petal of Meiogyne cylindrocarpa. 762
(b) Gnaw marks (magenta arrows) created by curculionid beetles on the inner petals of 763
Meiogyne trichocarpa. (c) Gnaw marks (magenta arrows) created by staphylinid beetles on 764
the inner petals of Meiogyne stenopetala: longitudinal ridges of the basal corrugation have 765
been gnawed away. (d) Pair of nectary glands on the adaxial surface of the inner petal of 766
Pseuduvaria hylandii. (e) Nectar secreted by Pseuduvaria mulgraveana (yellow arrows). (f) 767
Drosophilid fly consuming nectar in a Pseuduvaria hylandii flower. Photo credit: (a–f): 768
Ming-Fai Liu. 769
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprintthis version posted September 20, 2024. ; https://doi.org/10.1101/2024.09.16.613362doi: bioRxiv preprint
770
Fig. 3. Spectral reflectance of 12 Annonaceae species. (a) Meiogyne cylindrocarpa. (b) 771
Meiogyne trichocarpa. (c) Meiogyne stenopetala. (d) Monoon patinatum. (e) Polyalthia 772
hispida. (f) Polyalthia xanthocarpa. (g) Pseuduvaria glabrescens. (h) Pseuduvaria hylandii. 773
(i) Pseuduvaria mulgraveana. (j) Pseuduvaria villosa. (k) Uvaria concava. (l) Uvaria rufa. 774
For species with drooping inner petals and no colour differentiation between outer and inner 775
petal whorls, the inner petal abaxial surface was measured (a–f; i.e. Meiogyne, Monoon and 776
Polyalthia). For species with spreading inner petals and no colour differentiation between 777
outer and inner petal whorls, the inner petal adaxial surface (k–l; i.e. Uvaria) was measured. 778
For species with dissimilar petal whorls (g–j; i.e. Pseuduvaria), three perianth areas were 779
measured (solid line: inner petal abaxial surface; dotted line: outer petal adaxial surface; 780
dashed line: inner petal gland); the sample size (n) for these species refer to the sample size 781
for these three perianth areas in the aforementioned order. Photo credits: (a, b, d, e, g–l): 782
Ming-Fai Liu; (c): Chun-Chiu Pang; (f): Garry & Nada Sankowsky. 783
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprintthis version posted September 20, 2024. ; https://doi.org/10.1101/2024.09.16.613362doi: bioRxiv preprint
Table 1. Floral biology of Meiogyne, Monoon, Polyalthia, Pseuduvaria and Uvaria species. Abbreviations: CBG: Cairns Botanic Gardens, NE 784
Queensland, Australia; GBB: Gardens by the Bay, Singapore; MT: Mt. Tamborine, SE Queensland, Australia; SA: Sankowsky Arboretum, 785
Tolga, NE Queensland, Australia; WA: Graham Wood Arboretum, Wondecla, NE Queensland, Australia; +: present, -: absent, † : presence on 786
both pistillate-phase and staminate-phase flowers, * : contact with reproductive organs, ‡: pistillate-phase visitors were pollen-laden. 787
Meiogyne
cylindrocarpa Meiogyne
trichoc arpa Meiogyne
stenopetala Monoon
patinatum Polyalthia
hispida Polyalthia
xanthocarpa Pseuduvaria
hylandii Pseuduvaria
glabrescens Pseuduvaria
mulgraveana Pseuduvaria
villosa Uvaria concava Uvaria rufa
Site of st udy
(n= individual
stud ied)
SA (n=8), GBB
(n=3), WA (n=4) SA (n=8) SA (n=1),
WA (n=1),
MT (n=20)
SA (n=1) SA (n=2) S A (n=1) SA (n=5)
SA (n=1),
CBG (n=1) SA (n=3) SA (n=1) SA (n=2) SA (n=2)
H
abit
treelet/shrub
T
reelet
treelet
tree
treelet
treelet
treelet
treelet
treelet
treelet
climber
climber
Flower position axillary/apical axillary/apical axillary/apical ramiflorous axillary/apical axillary/apical/
ramiflorous cauliflorous axillary/apical axillary/apical ramiflorous axillary/apical axillary/apical/
ramifloro us
Sexual system bisexual flower bisexual flower bisexual flower bisexual flower bisexual flower bisexual flower unisexual flower,
dioeci ous andromonoecious andromonoecious ? (only seen
staminate treelet) bisexual flower bisexual flower
Pistillate phase
durati on 2d 2d 2d not assessed 3d 2–3d >6d not assessed 2d not assessed 2–3d 1d
Staminate phase
durati on 2d 1d 3d not assessed 2d 1d >6d not assessed 2d not assessed 0–1d 4d
Overla p phase
duration
ca. 0–3h ca. 5h ca. 1–3h not assessed ? ? not applicable not applicable - not assessed 1–2d -
Interim phase
duration
- - - not assessed ? ? not applica ble not applicable 2d not assessed - 1d
Protogyny + + + not assessed + + not applicable not applicable + not assessed + +
Pollinator trapping - - - - - - - - - - - -
Thermoge nesis
-
-
-
-
-
-
-
-
-
-
-
-
Floral od our banana-like bana na-like banana-like od orous odourless banana-like rott en fruit rotten fruit rotten fruit rotten fruit fruity fruity
Inner pet al
growth/gland +, smooth–weakly
longitud inally
grooved
+, smooth–weakly
longitud inally
grooved
+, longitudinall y
grooved - - - +,
smooth & raised +,
smooth & raised +,
smooth & raised +,
smooth & raised - -
Inner pet al
corrugation
function
food body food body food body corrugation
absent corrugation
absent cor rugation
absent nectary nectary nectary nectary corrugation
absent corrugation
absent
Floral visitors
(n=total nu mber of
floral vi sitors
observed )
n=42 (sites:
SA+WA)
Coleoptera:
Curculi onidae:
Ochyrome rini
(14.3%)†, * , ‡
Curculi onidae:
Carpophilus sp.
(4.8%)†, *
Nitidulidae:
Nitid ulinae
(81%)†, *, ‡
n=29
Coleoptera:
Curculi onidae:
Ochyromerini
(17.2%)†, *, ‡
Nitidulidae:
Nitidulinae
(82.8%)†, *, ‡
n=14 (site:
MT+WA)
Coleoptera:
Staphylinidae:
Omaliinae
(64.3%)†, *, ‡
Nitidulidae:
Epuraeinae
(35.7%)†, *, ‡
n=2
Coleoptera:
Staphyli nidae:
Tachyporinae
(100%)*
not obser ved n=8
Coleoptera:
Curculionidae:
Ochyrome rini
(100%)†,*
n=58
Diptera:
Drosophilidae:
Drosophilinae
(60.3%)†, *, ‡
Lauxani idae
(3.4%)†
Neriidae (3.4%)†
Coleoptera:
Nitidulidae:
Nitidulinae
(31.0%) †, *, ‡
Hymenopte ra:
Formicidae
(1.7%)
n=19 (site: CBG)
Diptera:
Drosophilidae:
Drosophilinae
(21%)†, *
Coleopter a:
Nitidulidae:
Nitidulinae
(79%)†, *, ‡
n=33
Dipt era:
Drosophilidae:
Drosophilinae
(21.2%)†, *
Coleoptera:
†,*Nitidulidae:
Nitidulinae
(72.7%)†, *, ‡
Hymenoptera:
Formicidae (6.1%)
not observed n=13
Coleopter a:
Nitidulidae:
Nitidulinae
(92.3%)†, *, ‡
Hymenoptera:
Apis sp.
(7.7%)†, *
n=1
Diptera:
unknown sp.
(100%)
788
789
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprintthis version posted September 20, 2024. ; https://doi.org/10.1101/2024.09.16.613362doi: bioRxiv preprint
Table 2. Floral scent composition of Meiogyne, Monoon, Polyalthia, Pseuduvaria and Uvaria species. The average percentage of the relative 790
peak area for each volatile and its standard error are presented below. Italicised molecules were verified by co-injecting with commercially 791
available standards. Unknown molecules were denoted by the top ten most abundant ion fragments, followed by their relative intensity in 792
parentheses. n refers to the number of technical replicates performed, followed by the number of individuals the odour was collected from. 793
standard
retention
index
(DB-WAX)
Relat ive content (%)
M
eiogyn
e
cylindrocarpa
(n=15;15)
M
eiogyn
e
trichocarpa
(n=8;8)
M
eiogyn
e
stenopetala
(n=5;2)
M
onoo
n
patinatum
(n=3;1)
P
olyalthi
a
xanthocarpa
(n=5;1)
P
seuduvari
a
hylandii.
(n=3;3)
P
seuduvari
a
glabrescens
(n=4;2)
P
seuduvari
a
mulgraveana
(n=4;3)
P
seuduvari
a
villosa
(n=4;1)
U
vari
a
concava
(n=3;2)
Uvaria rufa
(n=4;2)
Aliphatic hydrocarbon
1
-
T
etradece
ne
1348 — 0.19 ± 0.19 — — — — — — — — —
T
etradeca
ne
1401 — 0.33 ± 0.22 — — — — — — — — —
Oxygenated Aliphatic Compound
2,3-Butanediol 1589 — — — — — 95.97 ± 0.91 84.83 ± 0.80 96.98 ± 0.21 95.32 ± 0.15 — —
2-Methyl-butanoic acid 1678 — — — 16.48 ± 0.53 — — — — — — —
2-Methyl-(E)-7-hexadecene 1725 — — — — — — — — — —
0.05 ± 0.00
Ester
Isobutyl acetate 1014 7.87 ± 2.16 11.86 ± 4.37 5 6.71 ± 1.50 — — — — — — — —
Ethyl butanoate 1039 — — — — — — — — — — 7.35 ± 0.38
Butyl acetate 1073 0.22 ± 0.24 — — — — — — — — — 20.65 ± 0.21
Isoamyl acetate 1109 79.58 ± 7.65 84.07 ± 4.01 28.17 ± 4.13 — 99.28 ± 0.02 — — — — — —
Methyl hexanoate 1191 — — — — — — — — — 54.69 ± 0.32 4.70 ± 0.08
Butyl butanoate 1223 — — — — — — — — — — 1.94 ± 0.07
2-methyl-(2
E
)-butenyl acetate 1225 0.82 ± 0.70 — — — — — — — — — —
Ethyl hexanoate 1238 — — — — — — — — — — 12.58 ± 0.29
2-methyl-(2
Z
)-butenyl acetate 1254 1.16 ± 0.70 — — — — — — — — — —
Hexyl acetate 1277 0.07 ± 0.87 — 0.87 ± 0.12 — — — — — — — 21.27 ± 0.25
Isoamyl isovalerate 1316 1.09 ± 0.85 — — — — — — — — — —
Methyl octanoate 1396 — — — — — — — — — 4.30 ± 0.29 0.56 ± 0.02
Hexyl butanoate 1423 — — — — — — — — — — 6.38 ± 0.10
Methyl (Z)-4-octenoate 1438 — — — — — — — — — 25.36 ± 0.32 —
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprintthis version posted September 20, 2024. ; https://doi.org/10.1101/2024.09.16.613362doi: bioRxiv preprint
Ethyl octanoate 1442 — — — — — — — — — — 20.04 ± 0.22
Hexyl 2-methylbutanoate 1452 — — 0.44 ± 0.07 — — — — — — — —
Octyl acetate
1482
—
—
—
—
—
—
—
—
—
—
1.48 ± 0.06
trans-2-Hexenyl isovalerate 1508 — — 2.34 ± 0.12 — — — — — — — —
2,3-Butanediyl diacetate 1530 1.26 ± 1.37 — 0.09 ± 0.05 — — — — — — — —
linalyl anthranilate 1565 — 0.01 ± 0.01 — — — — — — — — —
Hexyl hexanoate 1618 — — — — — — — — — — 0.86 ± 0.02
Octyl butanoate
1625
—
—
—
—
—
—
—
—
—
—
0.66 ± 0.03
Hexyl tiglate 1632 — — — — — — — — — — 0.16 ± 0.02
Ethyl decanoate 1646 — — — — — — — — — — 0.23 ± 0.01
Ethyl trans-4-decenoate 1675 — — — — — — — — — — 0.09 ± 0.00
6-Me thyl-4 -hepten yl
pentanoate 1678 — — — — — — — — — — 0.20 ± 0.00
Ethyl benzoate
1685
—
—
—
—
—
—
—
—
—
—
0.08 ± 0.01
(
E
)-Methylgeranate 1706 — — — — — — — — — — 0.30 ± 0.00
Geranyl 2-methylbutanoate 1735 — 0.03 ± 0.01 — — — — — — — — —
Geranyl acetate 1766 1.56 ± 1.97 1.24 ± 0.59 — — — — — — — — —
2-Phenylethyl acetate 1835 — — 0.52 ± 0.01 — — — — — — — —
Phenylethyl isovalerate 2011 — — 0.54 ± 0.27 — — — — — — — —
hexyl benzoate 2098 — — — — — — — — — — 0.05 ± 0.01
Phenylpropa noid
Methyl eugenol 2026 — — — — — —
1.06 ± 0.09 — — — —
Monoterpene
β
-
M
yrce
ne
1170 — 0.15 ± 0.11 — — — — — — — — —
L
imone
ne
1205 — 1.24 ± 0.61 — — — — — — — — —
1,8-cineole 1210 — 0.01 ± 0.01 — — — 2.42 ± 1.22 11.78 ± 0.38 — — — —
t
ra
ns
-
β
-
O
cime
ne
1241 — 0.06 ± 0.04 — — — — — — — — —
cis-
β
-Ocimene 1259 — 0.14 ± 0.10 — 45.50 ± 0.61 — — — — — — —
terpinolene
1283
—
0.05 ± 0.03
—
—
—
—
—
—
—
—
—
allo-Ocimene 1384 — 0.18 ± 0.13 — 11.89 ± 0.59 — — — — — — —
Menthone
1456
—
0.03 ± 0.02
—
—
—
—
—
—
—
1.04 ± 0.07
—
α
-
T
erpine
ol
1711 — — — — — — — — — — —
camphor 1532 — 0.02 ± 0.02 — — — — — — — — —
menthacamphor 1654 — 0.06 ± 0.04 — — — — — — — — —
geranylacetone 186 8 — — 0.18 ± 0.04 3.01 ± 0.12 — — — — — — —
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprintthis version posted September 20, 2024. ; https://doi.org/10.1101/2024.09.16.613362doi: bioRxiv preprint
Sesquiterpene
α
-Cubebene 1471 1.11 ± 1.31 — 0.06 ± 0.06 — — — — — — — —
δ
-
elemene
1485
1.14 ± 1.36
—
—
—
—
—
—
—
—
—
—
Copaene 1511 — — 0.83 ± 0.35 — 0.32 ± 0.01 0.26 ± 0.26 1.12 ± 0.05 0.65 ± 0.05 1.05 ± 0.10 — —
β
-bourbonene 1518 1.25 ± 1.45 — — — — — — — — — —
β
-Cubebene 1554 1.25 ± 1.50 — — — 0.04 ± 0.00 — — — — — —
Cyperene 1549 — — — — — 0.42 ± 0.42 — — — — —
β
-
ylangene
1578
1.32 ± 1.57
—
—
—
—
—
—
—
—
—
—
Calarene 1602 1.48 ± 1.60 — — — — — — — — — —
(
E)
-
β
-
c
aryophylle
ne
1608 2.87 ± 1.51 — — 6.08 ± 0.16 0.14 ± 0.01 — — — — 1.86 ± 0.05 —
γ
-elemene 1647 1.42 ± 1.72 — — — — — — — — — —
cis-
β
-Farnesene 1672 — 0.02 ± 0.01 — — — — — — — 3.39 ± 0.00 —
humulene
1681
2.26 ± 1.70
—
—
2.83 ± 0.27
0.04 ± 0.00
—
—
—
—
—
—
γ
-Muurolene 1709 — — — — 0.07 ± 0.00 — — — — — —
δ
-Selinene 1724 — — 0.08 ± 0.03 — — — — — — — —
Germacrene D 1732 1.86 ± 1.83 — — 3.87 ± 0.14 0.05 ± 0.00 — — — — 0.31 ± 0.16 —
Copaene/
γ
-Muurolene 1732 — — 0.10 ± 0.03 — — — — — — — —
α
-Muurolene 1744 — — 0.10 ± 0.03 — — — — — — 0.79 ± 0.02 —
α
-Farnesene 1756 — — — 7.40 ± 0.07 — — — — — — 0.03 ± 0.00
Bicylogermacrene 1758 1.59 ± 1.91 — 0.10 ± 0.04 — — — — — — — —
δ
-Cadinene 1777 1.65 ± 1.96 — 0.37 ± 0.10 1.83 ± 0.18 0.06 ± 0.00 — — — — — 0.04 ± 0.01
Cada-1,4-diene 1805 — — 0.08 ± 0.02 — — — — — — — —
Miscellaneous Compound
Naphthalene 1768 — — 0.06 ± 0.02 1.11 ± 0.09 — — — — — 0.21 ± 0.11 0.02 ± 0.00
β
-Ionene 1872 — — — — — — — — — 0.49 ± 0.03 —
Unknown
43(24), 57(20), 41(14), 56(11),
42(8), 70(7), 55(6), 86(4),
61(3), 69(3)
1176 0.56 ± 0.51 — 0.09 ± 0.06 — — — — — — — —
57(26), 43(19), 41(12), 56(11),
70(7), 42(7), 71(6), 55(5),
85(4), 86(3)
1201 0.16 ± 0.72 — 1.95 ± 0.54 — — — — — — — —
57(23), 43(22), 41(13), 56(12),
71(8), 42(6), 85(5), 69(4),
55(4), 68(4)
1244 — — 1.55 ± 0.33 — — — — — — — —
57(20), 70(15), 43(14), 85(12), 1299 — — 1.17 ± 0.34 — — — — — — — —
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprintthis version posted September 20, 2024. ; https://doi.org/10.1101/2024.09.16.613362doi: bioRxiv preprint
41(10), 71(9), 56(7), 55(6),
42(5), 86(3)
73(29), 57(17), 41(11), 43(11),
56(9), 147(7), 42(5), 32(5),
86(4), 207(3)
1316 — — — — — — 0.61 ± 0.21 — — — —
43(24), 67(16), 57(15), 82(12),
41(11), 56(7), 29(4), 42(4),
55(3), 39(3)
1319 — — 0.37 ± 0.14 — — — — — — — —
57(19), 43(18), 41(15), 56(11),
105.1(8), 42(7), 119(7), 29(6),
32(6), 86(4)
1344 — — — — — 0.47 ± 0.24 — 1.09 ± 0.08 1.90 ± 0.10 — —
43(21), 57(18), 56(15), 41(13),
70(7), 42(6), 55(6), 32(5),
69(4), 86(4)
1381 — — 0.01 ± 0.01 — — — — — — — —
43(52), 87(11), 57(9), 70(6),
41(5), 56(5), 42(3), 55(3),
86(3), 45(3)
1387 — — 2.11 ± 0.46 — — 0.12 ± 0.06 — — — — —
57(28), 43(15), 41(13), 56(12),
85(6), 71(6), 42(6), 55(6),
32(4), 86(3)
1405 0.19 ± 0.85 — 0.61 ± 0.15 — — 0.19 ± 0.10 0.59 ± 0.20 1.12 ± 0.05 1.74 ± 0.07 — —
57(20), 82(15), 67(14), 41(11),
43(10), 85(9), 56(7), 55(5),
32(4), 42(4)
1475 1.66 ± 1.25 — 0.08 ± 0.08 — — 0.09 ± 0.05 — 0.17 ± 0.10 — 1.09 ± 0.19 0.02 ± 0.02
57(20), 43(15), 41(15), 56(10),
80(9), 32(8), 29(6), 79.1(6),
42(6), 71(5)
1493 — — — — — — — — — 0.48 ± 0.11 —
57(21),43(17),56(12),85(11),4
1(11),103(9),84(6),42(5),55(5),
32(5)
1499 1.81 ± 1.36 — 0.27 ± 0.05 — — — — — — 0.44 ± 0.10 —
57(22), 43(16), 41(16), 56(12),
32(8), 55(7), 42(6), 71(5),
119(5), 69(5)
1510 — — — — — — — — — 1.34 ± 0.11 —
57(23), 85(14), 43(14), 41(11),
70(10), 56(8), 71(5), 112(5),
42(5), 55(5)
1555 — — 0.15 ± 0.08 — — 0.06 ± 0.06 — — — — —
111(26), 81(13), 154(12), 1607 0.14 ± 1.73 — — — — — — — — 3.59 ± 0.07 —
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprintthis version posted September 20, 2024. ; https://doi.org/10.1101/2024.09.16.613362doi: bioRxiv preprint
95(10), 112(8 ), 123(7), 79(7),
41(6), 94(6), 53(5)
84(15), 57(13), 123(13),
41(12), 43(11), 42(8), 56(8),
81(8), 95(6), 32(6)
1614 — — — — — — — — — — 0.05 ± 0.00
79(16), 108(13), 57(13),
41(12), 43(11), 150(8), 67(8),
56(7), 32(7), 29(6)
1736 — — — — — — — — — 0.39 ± 0.02 0.06 ± 0.00
57(19), 43(16), 41(12), 32(11),
56(11), 108(8 ), 42(6), 45(6),
91(6), 44(5)
1745 — — — — — — — — — 0.21 ± 0.12 0.01 ± 0.00
43(16), 57.1(13), 56(11),
41(11), 117.1 (10), 84.1(10),
71(8), 55(8), 145.1(7), 69(7)
1821 — — — — — — — — — — 0.15 ± 0.01
794
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprintthis version posted September 20, 2024. ; https://doi.org/10.1101/2024.09.16.613362doi: bioRxiv preprint
