See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/346577790

Floral traits and chemical cues associated with rock bee (Apis dorsata
Fabricius) for the host selection in West Bengal, India

Article in Grana · December 2020

DOI: 10.1080/00173134.2020.1823466

CITATIONS READS

0 259

6 authors, including:

Ujjwal Layek Sourabh Bisui

Rampurhat college Vidyasagar University
19 PUBLICATIONS 80 CITATIONS 6 PUBLICATIONS 12 CITATIONS

SEE PROFILE SEE PROFILE

Rajib Mondal Nandita Das

Vidyasagar University Vidyasagar University
2 PUBLICATIONS 4 CITATIONS 1 PUBLICATION 0 CITATIONS

SEE PROFILE SEE PROFILE

Some of the authors of this publication are also working on these related projects:

UGC Rajiv Gandhi National Fellowship View project

All content following this page was uploaded by Ujjwal Layek on 03 December 2020.

The user has requested enhancement of the downloaded file.

 Grana

ISSN: (Print) (Online) Journal homepage: https://www.tandfonline.com/loi/sgra20

Floral traits and chemical cues associated with

rock bee (Apis dorsata Fabricius) for the host
selection in West Bengal, India

Ujjwal Layek , Sourabh Bisui , Rajib Mondal , Nandita Das , Subrata Kumar
De & Prakash Karmakar

To cite this article: Ujjwal Layek , Sourabh Bisui , Rajib Mondal , Nandita Das , Subrata
Kumar De & Prakash Karmakar (2020): Floral traits and chemical cues associated with
rock bee (Apis�dorsata Fabricius) for the host selection in West Bengal, India, Grana, DOI:
10.1080/00173134.2020.1823466

To link to this article: https://doi.org/10.1080/00173134.2020.1823466

View supplementary material

Published online: 02 Dec 2020.

Submit your article to this journal

View related articles

View Crossmark data

Full Terms & Conditions of access and use can be found at

https://www.tandfonline.com/action/journalInformation?journalCode=sgra20
Grana, 2020
https://doi.org/10.1080/00173134.2020.1823466

Floral traits and chemical cues associated with rock bee (Apis dorsata
Fabricius) for the host selection in West Bengal, India

UJJWAL LAYEK 1, SOURABH BISUI2, RAJIB MONDAL2, NANDITA DAS3,

SUBRATA KUMAR DE4 & PRAKASH KARMAKAR2
1
Department of Botany, Rampurhat College, Birbhum, India, 2Department of Botany & Forestry, Vidyasagar University,
Midnapore, India, 3Centre for Life Sciences, Vidyasagar University, Midnapore, India,, 4Department of Zoology,
Vidyasagar University, Midnapore, India

Abstract
In the present work, bee foraging plants were determined by palynological analysis of colony stored samples (honey and pollen)
of Apis dorsata collected from southern West Bengal, India. Then flowering phenology and pollen morphologies were described
to evaluate the forage preference of the bee species. In addition, chemical cues were analysed using gas chromatography-mass
spectrometry (GC-MS). Foraging strategy of the bee species was profitable in that almost all the bee-visited plants were utilised
for both nectar and pollen sources. The bee species became generalist visitor of several plants having diverse life form, flower
characters and pollen morphologies. However, most of the bee-visited plants were of trees with synchronous type of flowering.
Most plants had small-sized, yellow, cream and white coloured flowers and pollen was openly presented. We did not find any
patterns in the morphological traits of bee collected pollen, though the pollen types were predominantly of medium sized with
trizonocolporate apertural pattern and reticulate surface ornamentation. Floral volatile organic compounds (VOCs) of bee
visited plants were also diversified. Compounds frequently found in the floral VOC bouquets were amphetamine-3-methyl;
butane, 2-cyclopropyl; 2,3-butanediol; cyclohexan, 1-methyl-5-(1-methylethyl); D-limonene; methyl (2E)-2-methoxy-
2-butenoate; phenol, 4-[2-(methylamino)ethyl]; phthalic acid, di(2-propylpentyl)ester; propanamide, N-(aminocarbonyl)
and pyrrolo[1,2-a]pyrazine-1,4-dione, hexahydro-3-(2-methylpropyl). In conclusion, floral traits and chemical cues of plants
influence the host selection specificity (to collect floral rewards) of the rock bees.

Keywords: flowering phenology, palynological analysis, pollen morphology, volatile organic compound

In tropical ecosystems, wild bees play an important
role in maintaining animal and plant communities.
The giant honey bee, Apis dorsata Fabricius is found
throughout southern Asia (Hepburn & Radloff
2011). The bee species is an important pollinator
(Corlett 2011; Partap 2011) and honey producer
(Crane 1999; Robinson 2012). However, Apis dor­
sata colonies cannot be domesticated for bee pro­
ducts and pollination purposes. A management
system which utilises anthropogenic nesting sub­
strates for rock bee is known as rafter beekeeping,
which was reported from Cambodia, Indonesia and
Vietnam (Crane et al. 1993; Tan et al. 1997; Jump &
Waring 2004; Gratzer et al. 2019). In the Melaleuca
forest of Vietnam, beekeepers use rafters which have
been raised before the Melaleuca trees start to
bloom. By using this technique, they harvest honey
two to three times from the same colony per season
(Tan et al. 1997). In addition to honey, the wax and
bee pollen are also sold at a significant price.
To harvest more bee products, it is essential to
know the vital floral sources to the bee species. The
palynological analysis of honey and pollen loads
provides reliable data on the nectar and pollen
sources used by honey bees (Jones & Bryant
1996). Some palynological works from West Bengal
were done by Pal and Karmakar (2013), Layek
et al. (2016), Layek and Karmakar (2016).
Research work on plant preferences of rock bees
is poorly documented. Therefore, more work is

Correspondence: Prakash Karmakar, Department of Botany & Forestry, Vidyasagar University, Midnapore 721102, India. E-mail: karmakar_p@yahoo.co.in

(Received 21 January 2020; accepted 27 August 2020)

© 2020 Collegium Palynologicum Scandinavicum

2 U. Layek et al.

needed regarding the forage preferences of the bee Palynological analysis of honey and pollen samples
species.
Palynological processing of honey samples followed
The purpose of this study was to identify the plant
the method of Louveaux et al. (1978) with modifi­
species visited by Apis dorsata in southern West
cations suggested by Jones and Bryant (2004). First,
Bengal, India. Then, we describe the life form, flow­
10 g of honey was dissolved in 10 ml of distilled
ering phenology, floral scent components and pollen
water and stirred to dissolve the honey. Then, 50 ml
morphologies of the bee-visited plants to determine
of 95% ethyl alcohol (ETOH) was added and the
the forage preferences of the rock bees. We think
resulting honey–ETOH solution was stirred. All the
that the plant host selection specificity for a honey
ETOH-diluted sub-samples were centrifuged at
bee species depends on several factors including
4000 rpm (1658 g) for 5 min. The resulting sedi­
flower availability index and pollen availability
ments were subjected to acetolysis (Erdtman 1960)
index (Layek et al. 2020a). Here, we hypothesised
with a 9:1 ratio (v/v) of acetic anhydride to concen­
that floral traits and chemical cues of plants also act
trated sulphuric acid (purity: 98%; manufacturer:
as decision making drivers to select a floral host by
Merck Specialities Private Limited). After thor­
the rock bees.
oughly mixing, the mixture containing tubes were
placed into a water bath (at 100 °C) for 3 min. Then
the samples were centrifuged at 4000 rpm (1658 g)
Materials and methods for 5 min. After decanting the supernatants, sedi­
Collection of honey and pollen samples ments were rinsed with distilled water, centrifuged
and decanted, and then again rinsed with 95%
Ten honey samples were collected from ten wild ETOH. After centrifuging and decanting, the pollen
hives of Apis dorsata during January to April, 2017. sediment was taken on a small piece of glycerine
We selected the hives from seven districts (Bankura, jelly and transferred to the centre of a glass slide.
Birbhum, Hooghly, Paschim Burdwan, Paschim Subsequently warmed gently to melt the jelly con­
Medinipur, Purba Burdwan, Purba medinipur) in taining the pollen sediment and then covered with
southern West Bengal (Table I). From each hive, a cover glass. The cover glass was sealed with par­
approximately 50 g of honey was collected from affin wax. Using reference slides prepared from the
honey cells using a micropipette or dropper and local flora, pollen grain identification was per­
constitute the one pipetted honey sample. For the formed. Microphotographs were taken using
collection of pollen samples, pollen cells were rup­ a Nikon Eclipse LV100 POL polarising microscope.
tured and ≥ ten pollen cells content of each hive Pollen was classified according to the pollen type
were taken together as a single pollen sample. For­ system (Joosten & De Klerk 2002; De Klerk &
maldehyde–acetic acid–alcohol (FAA) solution was Joosten 2007), where pollen grains from one or
added to each pollen sample (Pal & Karmakar 2013) more species is grouped together according to
and then stirred well to homogenise the samples. shared morphological features. For each pipetted
honey sample, we counted approximately 100 pollen
Table I. Collection of hive products (honey and pollen) during grains (Layek et al. 2020b). Then, frequency classes
different seasons in West Bengal, India. were established based on the percentage of each
pollen type in a given sample (Louveaux et al.
Hive
no. Date of collection Season Location
1978): predominant pollen type (> 45%), secondary
pollen type (16–45%), important minor pollen type
1 13 January 2017 Winter Hargram, Purba (3–15%) and minor pollen type (< 3%). The sample
Burdwan comprising predominant pollen type was called uni­
2 15 January 2017 Winter Nandanpur, Hooghly
3 16 January 2017 Winter Ramnagar, Paschim
floral honey and absence of predominant pollen type
Burdwan was called multifloral honey (Louveaux et al. 1978;
4 10 February 2017 Winter Goshpur, Hooghly Von der Ohe et al. 2004; White 2005).
5 19 February 2017 Spring Chakdah, Purba For the analysis of pollen samples, five to ten
Medinipur drops of homogenised pollen solution were sus­
6 13 March 2017 Spring Pingla, Paschim
Medinipur
pended in 50 ml of 95% ethanol and thoroughly
7 28 March 2017 Spring Haridaspur, Purba mixed (Jones & Bryant 2004). This solution was
Medinipur then centrifuged at 4000 rpm (1658 g) for 5 min.
8 15 April 2017 Summer Moyna, Purba After decanting the supernatant, the sediment was
Medinipur processed using acetolysis method as recommended
9 16 April 2017 Summer Suri, Birbhum
10 29 April 2017 Summer Jenadihi, Bankura
by Erdtman (1960). Glycerine jelly was used to
prepare the samples for microscopic analysis as

a mounting medium. After identification and count­
ing of the pollen grains (approximately 300 pollen
grains per sample), sample wise percentages of the
pollen types were determined. Then we classified
the obtained pollen types into four groups: very
frequent (> 45%), frequent (> 15–45%), less fre­
quent (3–15%) and rare (< 3%).
To describe morphology (size, shape, amb, sur­
face ornamentation and aperture types) of pollen
types we used standard terminologies (Erdtman
1952; Faegri & Iversen 1975; Punt et al. 2007). In
some pollen types (especially for per-prolate or per-
oblate), one axis is much larger than the other. We
think multiplication value in between polar axis
(PA) and equatorial diameter (ED) will gives an
idea of its size more accurately. Therefore, based
on the multiplication values, we classified the pollen
grains into three size groups viz. small (PA × ED <
625 µm2), medium (PA × ED 625–2500 µm2) and
large (PA × ED > 2500 µm2).
For both honey and pollen samples, we determine
pollen niche width using the diversity index (H′) of
Shannon and Weaver (1949) as follows:
X
n
H0 ¼ ðpi: ln piÞ
i selection by the honey bee Apis dorsata, we selected
where H′ is the index of diversity, pi is the propor­
tion of each pollen type found in the sample. This
proportion is given by ni
N , where ni is the number of
grains counted for i pollen type and N is the total
number of pollen grains counted in the sample, ln is
the natural logarithm.
Equitability index (J′) of Pielou (1977) was used
to assess the homogeneity of the honey and pollen
samples using the following equation:
J 0 ¼ H 0 =H 0max
where H′ is the index of diversity and H′max is the
natural logarithm for the total number of pollen
types present in the sample. The J′ index ranges
between 0 to 1, i.e. heterogeneous use to homoge­
neous use of the resources.
Collection of data about flowering phenology
We surveyed about 1.5 km around the bee colonies
during the study period. We noted all flowering
plants to roughly estimate whether a plant species
is preferred or not by the bee species. This is not
comprehensive but an important observation as it
supports preferentially selection by the bees of
some species over others. Some phenological data
including flowering type, size, shape, colour and
longevity of flower and pollen presentation of the
Plant host selection by Apis dorsata 3
bee-visited plants (determined by palynological ana­
lyses, as well as field observations) were collected.
Flowering types (viz. synchronous, continuous, and
extended) were based on the classification of Opler
et al. (1980). To determine the longevity of an
individual flower, we marked the matured flower
buds (in the case of large sized flowers) by tagging
them individually with number tags. In the case of
small sized flowers, we tagged inflorescences and
flowers were identified by coding them (in bud con­
dition) with small black dots of ink on their petiole
or calyx. For each flower we recorded the date of
opening and date of senescence. Senescence was
defined when the corolla lost its lustrous look and
had fallen off. The shapes of the flowers were
described with slight modification of Dafni (1992).
According to the length of floral parts, we classified
the flowers into three size-based groups (Layek &
Karmakar 2018a; Layek et al. 2020a) viz. small (<
1 cm), medium (1–3 cm), and large (> 3 cm). The
colours of flowers were described by colour names
as they appeared to the human observer. Pollen
presentation of the bee plants was determined by
close and careful notice of the opened flowers.
To determine the role of flower fragrances on host
seven rock bee preferred (bee-visited) plants viz.
Borassus flabellifer L., Cocos nucifera L., Foeniculum
vulgare Mill., Millettia pinnata (L.) Panigrahi,
Momordica charantia L., Sesamum indicum L. and
Tamarindus indica L. and three non-preferred (non-
visited or rarely visited) plants viz. Abelmoschus escu­
lentus (L.) Moench, Argemone maxicana L. and
Hibiscus vitifolius L.). For floral scent component’s
analysis, freshly opened flowers were enclosed
within a 20 ml Thomson Fisher Scintific glass vial.
After 6–12 h of incubation at room temperature, the
vial was placed into TriPlus RSH Autosampler to
inject the sample into a Trace 1300 Gas Chromato­
graph by a splitless injector. The gas chromatograph
was connected to an ISQ QD single quadrupole
mass spectrometer. Volatile compounds were sepa­
rated in the gas chromatograph using an EC-WAX
column and helium as carrier gas. We used an
empty glass vial as a control in the analysis. Volatile
organic compounds (VOCs) were identified by com­
paring the SI, RSI values and mass spectra with
a computerised mass spectrometry (MS)-database
using NIST 2017 library.
Statistical analysis
Statistical analyses of the honey and pollen samples
were conducted to obtain the arithmetic mean and
standard deviation. To estimate the relationship
4 U. Layek et al.

between different variables, we followed Karl Pear­
son’s correlation coefficient method. Analysis of
similarity (agglomerative hierarchical clustering)
among the hive’s pollen types was performed using
the Jaccard similarity coefficient.
Results
From palynological evaluation of ten honey samples,
34 pollen types were recorded (Table II). The num­
ber of pollen types per sample varied from four to
ten with a mean value of 7.2 ± 1.93. Among the ten
honey samples, six were unifloral. Predominant pol­
len types were Borassus flabellifer (sample nos 7
and 8), Brassica type (sample nos 1 and 3), Holopte­
lea integrifolia (sample no. 6) and Sesamum indicum
(sample no. 10). Besides, other important secondary
pollen types were Alangium salviifolium, Azadirachta
indica, Cocos nucifera, Coriandrum sativum, Millettia
pinnata, Phoenix sylvestris, Raphanus sativus, Schlei­
chera oleosa, Solanum lycopersicum and Terminalia
arjuna. Twenty pollen types represented as only
‘important minor’ and/or ‘minor’ pollen type.
From ten pollen samples 33 pollen types were
obtained (Table III). The number of pollen types
per sample varied from 4 to 11 with a mean value of
7.1 ± 2.13. Very frequent pollen types were Borassus
flabellifer (sample nos 7 and 8) and Brassica type
(sample nos 1 and 3). Other frequent pollen types
were Alangium salviifolium, Ceiba pentandra, Citrus
type, Cocos nucifera, Millettia pinnata, Phoenix sylves­
tris, Raphanus sativus, Sesamum indicum and Termi­

Table II. Pollen types obtained from honey samples of Apis dorsata in West Bengal, India.

Honey samples

Plant family Pollen type 1 2 3 4 5 6 7 8 9 10

Acanthaceae Hemigraphis hirta M

Hygrophila auriculata M M M M
Amaranthaceae Celosia argentea M
Anacardiaceae Anacardium occidentale M
Mangifera indica M M
Apiaceae Coriandrum sativum S M IM IM
Foeniculum vulgare M
Arecaceae Borassus flabellifer IM P P IM S
Cocos nucifera M S
Phoenix sylvestris M S M IM IM M
Asteraceae Mikania scandens M
Tridax procumbens IM IM
Xanthium strumarium IM
Brassicaceae Brassica type P S P
Raphanus sativus IM S IM M M
Combretaceae Terminalia arjuna IM S
Cornaceae Alangium salviifolium S
Cucurbitaceae Momordica charantia M
Dipterocarpaceae Shorea robusta IM
Fabaceae Leucaena leucocephala M IM
Millettia pinnata S
Tamarindus indica IM
Lecythidaceae Barringtonia acutangula M
Malvaceae Ceiba pentandra M IM IM
Sida cordifolia M
Meliaceae Azadirachta indica S
Myrtaceae Eucalyptus type M M M
Syzygium cumini IM
Pedaliaceae Sesamum indicum M P
Rutaceae Aegle marmelos M IM
Citrus type M IM S IM
Sapindaceae Schleichera oleosa S M
Solanaceae Solanum lycopersicum M S S
Ulmaceae Holoptelea integrifolia P
Total pollen types 5 6 4 8 9 10 9 6 8 7
Diversity H′ 0.49 1.26 0.40 1.70 1.60 1.13 1.49 0.78 1.64 1.52
Equitability J′ 0.30 0.70 0.29 0.82 0.73 0.49 0.68 0.44 0.79 0.78

P, predominant pollen; S, secondary pollen; IM, important minor pollen; M, minor pollen.
 Plant host selection by Apis dorsata 5

Table III. Pollen types obtained from pollen samples of Apis dorsata in West Bengal, India.

Pollen samples

Plant family Pollen type 1 2 3 4 5 6 7 8 9 10

Acanthaceae Hemigraphis hirta R

Hygrophila auriculata LF R R R LF
Apiaceae Coriandrum sativum F LF LF LF LF
Foeniculum vulgare R
Arecaceae Borassus flabellifer F VF VF LF F
Cocos nucifera R F
Phoenix sylvestris LF F LF F LF
Asteraceae Mikania scandens R
Tridax procumbens LF LF
Xanthium strumarium LF LF
Brassicaceae Brassica type VF F VF
Raphanus sativus F F R R LF
Combretaceae Terminalia arjuna F F
Cornaceae Alangium salviifolium F
Cucurbitaceae Momordica charantia R
Dipterocarpaceae Shorea robusta LF
Euphorbiaceae Croton bonplandianum LF
Fabaceae Leucaena leucocephala LF R
Millettia pinnata F
Tamarindus indica
Lecythidaceae Barringtonia acutangula R
Malvaceae Ceiba pentandra LF LF F
Sida cordifolia LF
Meliaceae Azadirachta indica LF
Myrtaceae Eucalyptus type LF LF LF
Syzygium cumini LF LF
Pedaliaceae Sesamum indicum LF F
Poaceae Poa type LF
Rutaceae Aegle marmelos LF R
Citrus type LF F F LF LF
Sapindaceae Schleichera oleosa LF LF
Solanaceae Solanum melongena LF
Ulmaceae Holoptelea integrifolia F
Total pollen types 5 5 4 7 8 11 9 6 8 8
Diversity H′ 1.22 1.24 0.80 1.80 1.96 2.24 1.43 1.22 1.98 1.72
Equitability J′ 0.76 0.77 0.58 0.92 0.94 0.93 0.65 0.68 0.95 0.83

VF, very frequent pollen; F, frequent pollen; LF, less frequent pollen; R, rare pollen.

nalia arjuna. Twenty-one pollen types were repre­
sented as ‘less frequent’ and/or ‘rare’ pollen type.
Considering both honey and pollen samples, a total
of 37 pollen types assigned to 23 families were recorded
from southern West Bengal during winter to summer
seasons (Figure 1). Among the identified pollen types,
29 were common in both honey and pollen samples.
Five pollen types’ viz. Anacardium occidentale, Celosia
argentea, Mangifera indica, Solanum lycopersicum and
Tamarindus indica were found only in honey samples.
Three pollen types viz. Croton bonplandianum, Poa type
and Solanum melongena were found only within pollen
samples. The number of pollen types between the
honey samples and pollen samples were positively cor­
related (r = 0.92, n = 10). According to Jaccard’s simi­
larity index among the hive’s pollen types, hive nos 4
and 5 (collected during February from Hooghly and
Purba Medinipur districts, respectively) were very simi­
lar in pollen composition (Figure 2). Hive nos 6 and 7
(collected during March from Paschim Medinipur and
Purba Medinipur districts, respectively) also showed
higher similarity index. The pollen composition of the
hives collected during January (hive nos 1, 2 and 3)
completely differs from hives collected during April
(hive nos 8, 9 and 10).
Within honey samples, the value of diversity
indices (H′ values) varied between 0.40 and 1.70,
with an average of 1.20 ± 0.49 (Table II). For pollen
samples, the value ranged from 0.80 to 2.24 with
a mean of 1.56 ± 0.45 (Table III). Higher values
obtained in the case of sample nos 4 (Goshpur,
Hooghly), 5 (Chakdah, Purba Medinipur), 6 (Pingla
of Paschim Medinipur) and 9 (Suri, Birbhum). The
lowest value was recorded in sample no. 3 (H′ = 0.40
6 U. Layek et al.

Figure 1. Photomicrographs of the some pollen types found in honey and pollen samples of Apis dorsata. A. Alangium salviifolium. B, C.
Anacardium occidentale. D, E. Azadirachta indica. F. Borassus flabellifer. G, H. Brassica type. I. Ceiba pentandra. J, K. Citrus type. L. Cocos
nucifera. M. Coriandrum sativum. N. Holoptelea integrifolia. O, P. Millettia pinnata. Q. Phoenix sylvestris. R. Sesamum indicum. S,
T. Terminalia arjuna. Scale bars ‒ 10 µm.

for honey, 0.80 for pollen) which was collected from (mean = 0.60 ± 0.20, n = 10) for honey samples
Ramnagar of Burdwan district. The value of equit­ and between 0.58 and 0.95 (mean = 0.80 ± 0.13,
ability index (J′) ranged between 0.29 and 0.82 n = 10) for pollen samples. This indicated that the

Figure 2. Dendrogram of similarity (using the Jaccard index) for the hive’s pollen types obtained from West Bengal, India.
foragers were more uniform in their pollen collections
rather than nectar collections.
The plants visited by the bee species were much
diversified with respect to plant habit, flowering
phenology and pollen morphology. According to
plant life forms of the 37 foraged plant taxa most
were trees (20 taxa), followed by herbs (14 taxa),
climbers (two taxa) and shrubs (one taxon). Most of
them show synchronous type of flowering with
2–3 days longevity of individual flowers. The bee
foraged plant showed much variation in flower
shapes. However, most common were brush and
dish shaped flowers. Among the foraged plants, 20
taxa were with small sized flowers and 17 plant taxa
were with medium sized flowers. Considering flower
colours as perceived by humans, yellow, cream and
white flowers were dominant within the bee flora.
Pollen presentation of the bee-visited plants were of
the mostly exposed type (28 taxa), followed by well-
hidden type (5 taxa) and partially-hidden type (4
taxa).
Considering the morphological features of pollen
grains of bee-visited plants, all pollen types were
monad and isopolar. Pollen types belonging to
dicot plant families showed radial symmetry,
whereas all monocot pollen types (except Poa type,
Poaceae) were bilateral symmetry. Most pollen types
were medium sized, followed by small sized and
large sized. The shape of pollen grains ranged from
oblate to perprolate with dominance of prolate
spheroidal (n = 8), spheroidal (n = 6) and oblate
spheroidal (n = 5). Most of the pollen grains were
Plant host selection by Apis dorsata 7
with circular amb, followed by triangular, oval and
trilobate outline. The most preponderant type of
apertural pattern had been identified as trizonocol­
porate. Members of Acanthaceae (Hemigraphis hirta
and Hygrophila auriculata) and Combretaceae (Ter­
minalia arjuna) showed heterocolpate aperture type.
Among the monocot plant families, bilaterally sym­
metrical pollen grains have sulcate type of aperture
and radially symmetrical pollen grains (Poa type,
Poaceae) have ulcus type of aperture. Only a single
pollen type (Croton bonplandianum) shows inapertu­
rate type of pollen grains. Exine ornamentations of
the observed pollen types were also greatly varied.
Though, most common sculpturing types were reti­
culate, echinate and striate.
Through gas chromatography (GC)-MS analysis of
floral volatiles, a total of 119 compounds were identi­
fied (Table IV). The number of obtained VOCs per
plant varied from 9 to 44. In general, bee plants
emitted higher number of VOCs as floral scent
(18.57 ± 11.70) compared to non or less visited plants
(11.33 ± 2.08). Among the identified compounds
from bee visited plants, most common are ampheta­
mine-3-methyl; phenol, 4-[2-(methylamino)ethyl];
propanamide, N-(aminocarbonyl); butane, 2-cyclo­
propyl; 2,3-butanediol and methyl (2E)-2-methoxy-
2-butenoate. The former three compounds are also
recorded from non or less visited plants. Within stu­
died bee-visited plants, the most abundant com­
pounds were butane, 2-cyclopropyl; 2,3-butanediol;
cyclohexan, 1-methyl-5-(1-methylethyl); D-limonene;
methyl (2E)-2-methoxy-2-butenoate; phthalic acid, di
 8
Table IV. Floral volatile organic compounds of some honey bee preferred (bee-visited) and non-preferred (non-visited or rarely visited) plants.

Preferred plants Non-preferred plants

Borassus Cocos Foeniculum Millettia Momordica Sesamum Tamarindus Abelmoschus Argemone Hibiscus
Volatile organic compounds flabellifer nucjfera vulgare pinnata charantia indicum indica esculentus mexicana vitifolius

Acetaldehyde - - - - + - - - - -
Acetic formic anhydride - - - - + - - - + -
U. Layek et al.

Acetic acid - + - - + - - - + -
5-Acetooxymethyl-2,6,10-trimethyl - - - + - - - - - -
-2,9-undecadien-6-ol
Acetophenone - - - + - - - - - -
6-Acetyl-8a-methyloctahydro-1(2H)- - - - - - - - - +
naphthalenone
(4E,6E)-Allocimene - - + - - - - - - -
2-Amino-1-(4-methylphenyl)propane - - - + - - - - - -
Amphetamine-3-methyl + - + + + + + + + -
Anhydro-D-mannosan ++ - - - - - - - - -
1,4-Anhydro-1-threibl - - - - + - - - - -
(2-Aziridinyl ethyl)amine - - - - - - + - -
Benzaldehyde, 4-hydroxy - - - - - + - - - -
Benzene, 1-methoxy-4-(1-propenyl) - - + - - - - - - -
1,2-Benzisothiazole - - - + - - - - - -
1,1ʹ-Biphenyl, 4-methyl - - - + - - - - - -
Butane, 2-cyclopropyl - - ++ - - +++ +++ - - -
2,3-Butanediol ++ + - - +++ - - - - -
Butanoic acid, 2-(amino oxy) - + - - - - - - - -
Cetene - - - + - - - - - -
3-(1-Chloroethenyl)-3-methyl-1,2,4-trioxolane - - - - + - - - - -
5α-Cholestan-3α-ol, 2-methylene - - - - - - - - - +
Cyclohexan, 1-methyl-5-(1-methylethyl) - - +++ - - - - - - -
3,4-Diamino-1,2,4 (4H)-triazole - - - + - - - - - -
1,4:3,6-Dianhydro-α-D-glucopyranose + - - - - - - + + -
Dibutyl phthalate ++ - - - - - - - - -
3,6-Diisopropylpiperazin-2,5-dione - - + - - - - - -
2,3-Dimethoxypropan-1-ol - - - - - - + - - -
2,3-Dimethyldecane - - - + - - - - - -
3,5-Dimethyl-2,6-dioxa-3,5-diazaheptane - - - - + - - - - -
2,2-Dimethyl-3-methoxy-cyclopropane - - - + - - - - - -
-1-carboxylic acid
1,7-Dimethylnaphthalene - - - + - - - - - -
2,2-Dimethylpropyl 2,2-dimethyl propanesulfinyl - - - ++ - - - - - -
sulphone
Dimethyl sulphoxide - - - - - - - - + -
Dodecanal - - - + - - - - - -

(Continued )
Table IV. (Continued ).

Preferred plants Non-preferred plants

Borassus Cocos Foeniculum Millettia Momordica Sesamum Tamarindus Abelmoschus Argemone Hibiscus
Volatile organic compounds flabellifer nucjfera vulgare pinnata charantia indicum indica esculentus mexicana vitifolius

1-Dodecanone, 2-(imidazol-1-yl)- + - - - - - - - - -
1-(4-methoxyphenyl)
3-Ethoxy-4-methoxyphenol - - - ++ - - - - - -
Ethoxy(oxo)acetic acid - - - - - - + - - -
2-Ethyl-1-hexanol - - - + - - - - - -
Ethyl hexopyranoside - - - - - - - +++ - -
3-Ethyl-7-hydroxyphthalide + - - - - - - - - -
Ethyl-2-hydroxybenzylsulfone - + - - - - - - - -
α-Farnesene - - + - - - - - - -
(E,Z)-α-Farnesene - - - + - - - - - -
Fenchone - - + - - - - - - -
Fenchyl acetate - - + - - - - - - -
Formamide - - - - + - - - + -
4-Formylphenyl 3-chloropropanoate - - - - - + - - - -
2-Heptadecenal - - - - - + - - - -
Heptane, 3,3-dimethyl - - - + - - - - - -
4-Heptenal, (E) - + - - - - - - - -
9-Hexacosene - - - - - - - - ++ ++
Hexadecane - - - + - - - - - -
n-Hexadecanoic acid - - - - - - - - +++ -
1-Hexadecanol - - - + - - - - - -
2,3-Hexanediol - + - - - - - - - -
1-Hexyl-2-nitrocyclohexane - - - - - + - - - -
4-Hydroxy-α-bromoethylphenone - - - - - + - - - -
4-Hydroxy tranylcypromine - - - - - - - - - +
1H-Indane, 1-methylene - - - + - - - - - -
5-Isopropyl-6-methyl-5-hapten-3-yn-2-one + - - - - - - - - -
3-Isonitrosobenzoylacetone - - - + - - - - - -
Labda-8(17),12-diene-15,16-dial - - - - - - - - - +
D-Limonene - - +++ - - - - - - -
3-Methoxy-N,N-diethylbenzylamine + - - - - - - - - -
(2-Methoxy-2-propenyl)benzene - - - - - - - - - +
Methyl glyoxylate - - - - - - + - - -
1-Methyl-4-isopropyl-cyclohexyl - - - - - - - - - +
2-hydroperfluorobutanoate
2-Methyl-3-(3-methyl-but-2-enyl)-2-(4-methyl- - - + - - - - - - -
pent-3-enyl)-oxetane
2-Methyl-1-undecanol - - - + - - - - - -
4-Methyl-2,3-pentanediol + - - - - - + - - -
Plant host selection by Apis dorsata

5-Methyl-1-phenylhexa-1,3,4-triene - - - + - - - - - -
5-Methyl-2-pyrimidone - + - - - - - - - -
9

(Continued )
Table IV. (Continued ).
10

Preferred plants Non-preferred plants

Borassus Cocos Foeniculum Millettia Momordica Sesamum Tamarindus Abelmoschus Argemone Hibiscus
Volatile organic compounds flabellifer nucjfera vulgare pinnata charantia indicum indica esculentus mexicana vitifolius

8-Methyl-5,6,7,8-tetrahydro- - - - + - - - - - -
2,4-quinazolinedione
S-Methyl-3-methylbutanethioate + - - - - - - - - -
U. Layek et al.

Methyl (2E)-2-methoxy-2-butenoate +++ - + ++ - + - - + -

1-Methylene-2b-hydroxymethyl-3,3-dimethyl- - - - - - - - - - +
4b-(3-methylbut-2-enyl)-cyclohexane
α-Methylnaphthalene - - - + - - - - - -
1-(2-Methoxyphenyl)-1-propanol - - - - - - - - - +
Methyl silane - - - - - - - ++ - -
Methylthiomethyl acetyl sulphide ++ - - - - - - - - -
4-Nitrophenyl 2-methoxybenzoate ++ - - - - - - - - -
Nonacos-1-ene + - - - - - - - - -
Nonadecane, 2-methyl - - - + - - - - - -
1-Nonanamine - + - - - - - - - -
4-Nonene, 2-methyl, (Z) - + - - - - - - - -
Nonylcyclopentane - - - + - - - - - -
2-Norpinene, 2,6-dimethyl-6-(4-methyl- - - + - - - - - - -
3-pentenyl)
α-Ocimene - - ++ + - - - - - -
1,2,3,5,6,7,8,8a-Octahydro 4-azulenylacetate - - - - - + - - - -
(3E,5E)-1,3,5-Octatriene - - - + - - - - - -
Oxiranemethanol, (R) + - - - - - - - + -
Pentadecane, 3-methyl - - - + - - - - - -
α-Pinene - - + - - - - - - -
Phenol, 4-[2-(methylamino)ethyl] + + + + + + + + + -
Phenoxyacetone - - - + - - - - - -
1-Phenyl-1-decanol - ++ - - - - - - - -
3-Phenyl-4-hydroxyacetophenone - - - + - - - - - -
Photocitral B - - - - - - - - - ++
Phthalic acid, di(2-propylpentyl)ester - - +++ - +++ - ++ - -
Propanamide, N-(aminocarbonyl) + + + + + + + + + -
Propane - - - - - - - + - -
Propanoic acid, 2-(amino oxy) - - - - - - - - + -
Propylamine, 3-chloro-2,2,3-trifluoro-N, - + - - - - - - - -
N,1,1-tetramethyl
Propylene carbonate - - - + - - - - - -
Propylglycolate - - - - + - - - - -
Pyrrolidine, 2-(methylaminomethyl) - - - - - + - - - -
Pyrrolo[1,2-a]pyrazine-1,4-dione, hexahydro- - - + - - +++ +++ - - -
3-(2-methylpropyl)
Tetradecane - - - + - - - - - -

(Continued )
 Plant host selection by Apis dorsata 11

(2-propylpentyl)ester and pyrrolo[1,2-a]pyrazine-

vitifolius
Hibiscus

+++
1,4-dione, hexahydro-3-(2-methylpropyl).

12
+
-
-
-

-
-
-
-
-
Non-preferred plants

Argemone
mexicana
Discussion

13
-
-
-

-
-
-
-
-
From the present studies, 37 pollen types were
recorded as bee-visited plants to Apis dorsata during
winter to summer. Most of them were already
Abelmoschus
esculentus

recorded from West Bengal by Pal and Karmakar

09
-
-
-

-
-
-
-
-
(2013), Layek and Karmakar (2016, 2018b), Layek
et al. (2016), Bisui et al. (2019). Only a few viz.
Anacardium occidentale, Scheichera oleosa and Sola­
num lycopersicum were newly added taxa from West
Tamarindus
indica

Bengal. The majority of the plant taxa were foraged

09
-
-
-

-
-
-
-
-
by the bee species to collect both nectar and pollen
grains, which indicate the profitable foraging strat­
egy of the bee species. For this reason, the number
Sesamum
indicum

of pollen types was almost the same in honey sample

16
+

+
-

-
-
-

and pollen sample collected from the same hive.

Most of the honey samples were unifloral in pollen
composition with predominance of Borassus flabelli­
Momordica
charantia

fer, Brassica sp., Holoptelea integrifolia and Sesamum

12

indicum. Unifloral honey of Borassus flabellifer type

-
-
-

-
-
-
-
-

and Sesamum indicum type were well established

Preferred plants

from West Bengal (Layek & Karmakar 2016) as

well as outside of Bengal (Ramanujam et al. 1992).
Millettia
pinnata

++

++

44

However, palynologically unifloral honey of Holop­

+

+
+
-

-
-

telea integrifolia type may be the first time reported

from West Bengal. Furthermore, dominance of uni­
Foeniculum

floral honeys indicates the higher honey yielding

vulgare

17

potential of the study areas during winter to

-
-
-

-
-
-
-
-

spring–summer. Regarding the diversity of pollen

type, hive samples (honey and pollen) collected dur­
ing the same season showed greater similarity index
nucjfera
Cocos

13

and the hives of different seasons showed little or no

-
-
-

-
-
-
-
-

similarities. That means different sampling sites of

seven different districts in southern West Bengal do
flabellifer
Borassus

not significantly differ floristically. The sampling

19
+
-
-
-

-
-

-
-

Relative abundance: high (+++), medium (++), low (+).

areas characterised by almost similar altitude, cli­

mate conditions and soil pattern which favour simi­
lar type of floral vegetation. However, season wise
2,4,4-Trimethyl-3-hydroxymethyl-5a-(3-methyl-

blooming plant species were varied due to variation

(2E,6E)-3,7,11-Trimethyl-9-(phenylsulfonyl)-

of climatic conditions. During winter (January–mid

2,2,3,3-Tetramethylhexahydro[1,4]dioxino

Trifluoro[(2E)-2-methyl-2-butenyl]silane

February), temperature and humidity level was very

Volatile organic compounds

low. Summer (start from mid April) is the hottest

2,2,3,4-Tetramethyl cyclobutanone

season. Spring (mid February–mid April) is

a transitional season between winter and summer.
Total number of compounds
but-2-enyl)-cyclohexene

The amplitude of the temporal pollen niche was

2,6,10-dodecatrien-1-ol
Table IV. (Continued ).

lower in honey samples than the pollen samples. The

[2,3-b][1,4]dioxine

obtained values for honey samples (ranged 0.40–1.70)

1,3,5-Trioxepane

were lower than the values reported by Novais et al.

4-Tetradecanol

3-Tridecanone

3-Undecanone
1-Undecanol

(2013) and Sosa-Nájera et al. (1994) for stingless

bees. We also hope that the sample wise pollen diver­
sity will be lower than month wise diversity, used by
the earlier-mentioned authors. Niche diversity
12 U. Layek et al.
depends on the availability of floral resources and
length of flowering period of utilised taxa.
Evenness values (J′) followed the same trend as the
diversity index. The values of J′ were also lower in
honey samples than the pollen samples. Our obtained
values (for honey samples, ranged 0.30–0.82) were
similar to the values reported by Rezende et al. (2019)
for stingless bees. However, the values were much
higher than the value found by Ferreira and Absy
(2018), who studied pollen niche of Melipona interrupta
in central Amazon. The high evenness values indicated
that the honeybee Apis dorsata used nectar and pollen
sources more uniformly. Furthermore, higher mean
value of J′ in pollen samples than the value recorded
in honey samples allude that the bee species more
homogeneously collect pollen than nectar.
Among the foraged plants, the majority were trees
and flowers of those tree species providing ample
amount of resources to the bee species. Most of
the bee-visited plants exhibit synchronous type of
flowering with greater longevity of flowers. The syn­
chronised flowering of different species could
increase the resource density and local pollinator
attraction (Schemske 1981; Sakai et al. 1999).
The majority of the foraged plants had small sized
flowers. However, small flowers are usually orga­
nised in dense inflorescences to intensify advertise­
ment by the plants (Faegri & Van der Pijl 1979).
The shape of the flowers were diverse, i.e. the bee
species does not show specific preference on any
shape type. Gould (1985) also reported that the
honeybees are very poor in distinguishing shapes.
Perhaps they have an innate and persistent prefer­
ence for figural intensity with longer outlines (Hertz
1935). However, brush and dish shaped flowers
were more common. Brush shaped flowers have
abundant exposed stamens, which seem to allow
for an easy access of the bee species to collect abun­
dant pollen. Dish flowers are generally polyphilic
and attract wide array of visitors. Colour of the bee
visited flowers was much diversified, though most
common were yellow, cream and white colours.
Yellow colour is also documented as the most pre­
ferred flower traits of honey bees (Balamurali et al.
2018) and stingless bees (Vossler 2012; Layek &
Karmakar 2018a). The bee plants also showed sig­
nificant variation in pollen morphological characters
with predominance of monad, isopolar, radially
symmetrical, medium sized, trizonocolporate aper­
ture and reticulate surface ornamentation. When we
focused on pollen morphological characters of vig­
orously foraged plants, there was no specific choice
for the bee species on pollen morphological traits
like pollen size, shape, surface ornamentation or
apertural pattern.
Both bee visited plants and non or less visited
plants emit large number of volatile substances
belong to diverse groups like acid, alkene, alde­
hyde, ester, ketone and terpene. Some of them
(viz. alloocimine, limonene, ocimene, α-pinene, α-
farnesene) were already reported by different work­
ers (Qin et al. 2014; Li et al. 2016; Berhal et al.
2017). Some volatile compounds (amphetamine-
3-methyl; phenol, 4-[2-(methylamino)ethyl]; pro­
panamide, N-(aminocarbonyl); butane, 2-cyclo­
propyl) are well represented by both bee visited
plants and non or less visited plants. Other few
compounds (butane, 2-cyclopropyl; 2,3-butanediol
and methyl (2E)-2-methoxy-2-butenoate) domi­
nated within volatile spectra of bee visited plants.
The honey bee species may be attracted by the
earlier-mentioned three compounds in addition to
other volatile compounds. Only a few floral vola­
tiles are responsible to attract a particular insect
group (Eltz et al. 1999; Schiestl et al. 2003).
Others are common biogenic VOCs that become
attractive for a large array of generalist visitors (Li
et al. 2008; Johnson & Hobbhahn 2010). The
volatile compounds act as chemical cues that facil­
itate floral location by creating concentration gra­
dients that visitors perceive with their sensory
receptors (Chittka & Raine 2006).
Conclusion
Winter and spring–summer in southern West Bengal
are the most productive seasons for honey bees to
produce honey from a variety of flowering plants.
During this time important bee foraging plants were
Alangium salviifolium, Borassus flabellifer, Brassica
sp., Ceiba pentandra, Citrus sp., Cocos nucifera, Cor­
iandrum sativum, Holoptelea integrifolia, Millettia pin­
nata, Phoenix sylvestris, Raphanus sativus, Sesamum
indicum, Solanum lycopersicum and Terminalia arjuna.
Almost all bee-visited plants were utilised by the bee
species as a source of nectar and pollen grains. Due
to diversified visitation pattern showed by the bee
species together with wide plant habits, varied flow­
ering shapes and diverse VOCs, it may be concluded
that Apis dorsata is an obligatory generalist flower
visitor.
Acknowledgements
The authors are thankful to authorities of Vidyasagar
University (VU) for providing necessary laboratory
facilities. The authors are indeed thankful to USIC
section of VU as well as to Mr Dipankar Mandal for
analysis of scent chemistry.

Disclosure statement
No potential conflict of interest was reported by the
authors.
Supplemental Material
Supplemental data for this article can be accessed
here
References
Balamurali GS, Nicholls E, Somanathan H, De Ibarra NH. 2018.
A comparative analysis of colour preferences in temperate and
tropical social bees. The Science of Nature 105(1–2): 8.
doi:10.1007/s00114-017-1531-z.
Berhal C, De Clerk C, Fauconnier ML, Levicek C, Boullis A,
Kaddes A, Jijakli HM, Verheggen F, Massart S. 2017. First
characterisation of volatile organic compounds emitted by
banana plants. Scientific Reports 7(1): 46400. doi:10.1038/
srep46400.
Bisui S, Layek U, Karmakar P. 2019. Comparing the pollen
forage pattern of stingless bee (Trigona iridipennis Smith)
between rural and semi-urban areas of West Bengal, India.
Journal of Asia-Pacific Entomology 22(3): 714–722.
doi:10.1016/j.aspen.2019.05.008.
Chittka L, Raine NE. 2006. Recognition of flowers by
pollinators. Current Opinion in Plant Biology 9(4):
428–435. doi:10.1016/j.pbi.2006.05.002.
Corlett RT. 2011. Honeybees in natural ecosystems. In:
Hepburn R, Radloff SE, eds. Honeybees of Asia, 215–226.
Berlin: Springer.
Crane E. 1999. The world history of beekeeping and honey
hunting. New York: Routledge.
Crane E, Van Luyen V, Mulder V, Ta TC. 1993. Traditional
management system for Apis dorsata in submerged forests in
Southern Vietnam and Central Kalimantan. Bee World 74(1):
27–40. doi:10.1080/0005772X.1993.11099151.
Dafni A. 1992. Pollination ecology – a practical approach.
Oxford: Oxford University Press.
De Klerk P, Joosten H. 2007. The difference between pollen
types and plant taxa: A plea for clarity and scientific
freedom. Eiszeitalter und Gegenwart Quaternary Science
Journal 56: 162–171.
Eltz T, Whitten WM, Roubik DW, Lisenmair KE. 1999. Fra­
grance collection, storage, and accumulation by individual
male orchid bees. Journal of Chemical Ecology 25(1):
157–176. doi:10.1023/A:1020897302355.
Erdtman G. 1952. Pollen morphology and plant taxonomy -
Angiosperms (An introduction to palynology I). Stockholm:
Almqvist & Wiksell.
Erdtman G. 1960. The acetolysis method. A Revised
Description. Svensk Botanisk Tidskrift 54: 561–564.
Faegri K, Iversen J. 1975. Textbook of pollen analysis. Copenha­
gen: Munksgaard.
Faegri K, Van der Pijl L. 1979. Principles of pollination ecology
(3rd edition). New York: Pergamon Press.
Ferreira MG, Absy MA. 2018. Pollen niche of Melipona (Meli­
kerria) interrupta (Apidae: Meliponini) bred in a meliponary in
a terra-firme forest in the central Amazon. Palynology 42(2):
199–209. doi:10.1080/01916122.2017.1332694.
Gould JL. 1985. How bees remember flower shapes. Science 227
(4693): 1492–1494. doi:10.1126/science.227.4693.1492.
Plant host selection by Apis dorsata 13
Gratzer K, Susilo F, Purnomo D, Fielder S, Brodschneider R.
2019. Challenges for beekeeping in Indonesia with autochtho­
nous and introduced bees. Bee World 96(2): 40–44.
doi:10.1080/0005772X.2019.1571211.
Hepburn HR, Radloff SE. 2011. Biogeography. In: Hepburn R,
Radloff SE, eds. Honeybees of Asia, 51–68. Berlin: Springer.
Hertz M. 1935. Die Untersuchungen über den Formensinn der
Honigbiene. Naturwissenschaften 36(36): 618–624.
doi:10.1007/BF01493245.
Johnson SD, Hobbhahn N. 2010. Generalized pollination, floral
scent chemistry, and a possible case of hybridization in the
African orchid Disa fragrans. South African Journal of Botany
76(4): 739–748. doi:10.1016/j.sajb.2010.07.008.
Jones G, Bryant V. 1996. Melissopalynology. In: Jansonius J,
McGregor D, eds. Palynology: Principles and applications,
933–938. Houston, TX: American Association of Strati­
graphic Palynologists Foundation 3.
Jones GD, Bryant VM Jr. 2004. The use of ETOH for the
dilution of honey. Grana 43(3): 174–182. doi:10.1080/
00173130410019497.
Joosten H, De Klerk P. 2002. What’s in a name? Some thoughts
on pollen classification, identification, and nomenclature in
quaternary palynology. Review of Palaeobotany and Palynol­
ogy 122(1–2): 29–45. doi:10.1016/S0034-6667(02)00090-8.
Jump D, Waring C. 2004. Rafter beekeeping in Combodia. Bee
Craft 86: 4–5.
Layek U, Karmakar P. 2016. Bee plants used as nectar sources by
Apis florea Fabricius in Bankura and Paschim Medinipur dis­
tricts, West Bengal. Geophytology 46: 1–14.
Layek U, Karmakar P. 2018a. Nesting characteristics, floral
resources, and foraging activity of Trigona iridipennis Smith
in Bankura district of West Bengal, India. Insectes sociaux
65(1): 117–132. doi:10.1007/s00040-017-0593-4.
Layek U, Karmakar P. 2018b. Pollen analysis of Apis dorsata
Fabricius honeys in Bankura and Paschim Medinipur dis­
tricts, West Bengal. Grana 57(4): 298–310. doi:10.1080/
00173134.2017.1390604.
Layek U, Manna SS, Karmakar P. 2020a. Pollen foraging beha­
viour of honey bee (Apis mellifera L.) in southern West Bengal,
India. Palynology 44(1): 114–126. doi:10.1080/
01916122.2018.1533898.
Layek U, Mondal R, Karmakar P. 2020b. Honey sample collec­
tion methods influence pollen composition in determining
true nectar-foraging bee plants. Acta Botanica Brasilica, 34
(3): 478–486. doi:10.1590/0102-33062020abb0086.
Layek U, Nandi T, Karmakar P. 2016. Pollen forage and storage
pattern of Apis dorsata Fabricius in Bankura and Paschim Medi­
nipur districts, West Bengal. International Journal of Pure and
Applied Bioscience 4: 59–71. doi:10.18782/2320-7051.2384.
Li P, Luo Y, Bernhardt P, Kou Y, Perner H. 2008. Pollination of
Cypripedium plectrochilum (Orchidaceae) by Lasioglossum spp.
(Halictidae): The roles of generalist attractants versus restric­
tive floral architecture. Plant Biology 10(2): 220–230.
doi:10.1111/j.1438-8677.2007.00020.x.
Li Y, Ma H, Wan Y, Li T, Liu X, Sun Z, Li Z. 2016. Volatile
organic compounds emissions from Luculia pinceana flower
and its changes at different stages of flower development.
Molecules 21(4): 531. doi:10.3390/molecules21040531.
Louveaux J, Maurizio A, Vorwohl G. 1978. Methods of
Melissopalynology. Bee World 59(4): 139–157. doi:10.1080/
0005772X.1978.11097714.
Novais JS, Absy MA, Santos FAR. 2013. Pollen grains in honeys
produced by Tetragonisca angustala (Latreille, 1811) (Hyme­
noptera: Apidae) in tropical semi-arid areas of north-eastern
Brazil. Arthropod-plant Interactions 7(6): 619–632.
doi:10.1007/s11829-013-9276-x.
 14 U. Layek et al.
Opler PA, Frankie GW, Baker HG. 1980. Comparative phono­
logical studies of treelet and shrub species in tropical wet and
dry forests in the lowlands of Costa Rica. Journal of Ecology
68(1): 1167–1188. doi:10.2307/2259250.
Pal PK, Karmakar P. 2013. Pollen analysis in understanding the
foraging behaviour of Apis mellifera in Gangetic West Bengal.
Geophytology 42: 93–114.
Partap V. 2011. The pollination role of honeybees. In:
Hepburn R, Radloff SE, eds. Honeybees of Asia, 227–256.
Berlin: Springer.
Pielou EC. 1977. Mathematical ecology (2nd edition).
New York: Willey-Interscience Publishing, John Wiley &
Sons.
Punt W, Hoen PP, Blackmore S, Nilsson S, Le Thomas A. 2007.
Glossary of pollen and spore terminology. Review of Palaeo­
botany and Palynology 143(1–2): 1–81. doi:10.1016/j.
revpalbo.2006.06.008.
Qin XW, Hao CY, He SZ, Wu G, Tan LH, Xu F, Hu RS. 2014.
Volatile organic compound emissions from different stages of
Cananga odorata flower development. Molecules 19(7):
8965–8980. doi:10.3390/molecules19078965.
Ramanujam CGK, Reddy PR, Kalpana TP. 1992. Pollen analysis
of apiary honeys from East Godavari District, AP. Journal of
the Indian Institute of Science 72: 289–299.
Rezende ACC, Absy MA, Ferreira MG, Marinho HA,
Santos OA. 2019. Pollen of honey from Melipona seminigra
merrillae Cockerell, 1919, Scaptotrigona nigrohirta Moure,
1968 and Scaptotrigona sp. Moure, 1942 (Apidae: Melipo­
nini) reared in Sataré Mawé indigenous communities, Ama­
zon, Brazil. Palynology 43(2): 255–267. doi:10.1080/
01916122.2018.1458664.
Robinson WS. 2012. Migrating giant honey bees (Apis dorsata)
congregate annually at stop over site in Thailand. PloS One 7
(9): e44976. doi:10.1371/journal.pone.0044976.
View publication stats
Sakai S, Momose K, Yumoto T, Nagamitsu T, Nagamasu H,
Hamid AA, Nakashi T, Inoue T. 1999. Plant reproductive
phenology over four years including an episode of general
flowering in a low land dipterocarp forest, Sarawak,
Malaysia. American Journal of Botany 86(10): 1414–1436.
doi:10.2307/2656924.
Schemske DW. 1981. Floral convergence and pollinator sharing
in two bee pollinated tropical herbs. Ecology 62(4): 946–954.
doi:10.2307/1936993.
Schiestl FP, Peakall R, Mant JG, Ibarra F, Schulz C, Franke S,
Franke W. 2003. The chemistry of sexual deception in an
orchid-wasp pollination system. Science 302(5644):
437–438. doi:10.1126/science.1087835.
Shannon CE, Weaver W. 1949. The mathematical theory of
communication. Urbana, IL: University of Illinois Press.
Sosa-Nájera MS, Martínez-Henández E, Lozano-García MS,
Cuadriello-Aguiar JI. 1994. Nectaropolliniferous sources
used by Trigona (Tetragonisca) Angustula in Chiapas, South­
ern México. Grana 33(4–5): 225–230. doi:10.1080/
00173139409429003.
Tan NQ, Chinh PH, Thai PH, Mulder V. 1997. Rafter beekeep­
ing with Apis dorsata: Some factors affecting the occupation of
rafters by bees. Journal of Apicultural Research 36(1): 49–54.
doi:10.1080/00218839.1997.11100930.
Von der Ohe W, Persano Oddo L, Piana ML, Morlot M,
Martin P. 2004. Harmonized methods of melissopalynology.
Apidologie 35(Suppl. 1): S18–S25. doi:10.1051/
apido:2004050.
Vossler FG. 2012. Flower visits, nesting and nest defence behaviour
of stingless bees (Apidae: Meliponini): Suitability of the bee
species for meliponiculture in the Argentinean Chaco region.
Apidologie 43(2): 139–161. doi:10.1007/s13592-011-0097-6.
White JW. 2005. Honey. In: Graham JM, ed. The hive and the
honey bee, 869–927. Hamilton, IL: Dadant & Sons.