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Ultraviolet reflectance mediates pollinator visitation

in Mimulus guttatus

Article in Plant Species Biology · September 2013

Impact Factor: 1.27 · DOI: 10.1111/j.1442-1984.2012.00375.x

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Jason Rae Jana C Vamosi

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Plant Species Biology (2012) ••, ••–•• doi: 10.1111/j.1442-1984.2012.00375.x

1 Ultraviolet reflectance mediates pollinator visitation in

2 Mimulus guttatus psbi_375 1..9

3
4 JASON M. RAE and JANA C. VAMOSI
5 Department of Biological Sciences, University of Calgary, Calgary, Alberta, Canada
6
7 Abstract

8 Floral colors are widely believed to be an adaptation to attract pollinators. Recently, our
9 understanding of floral reflectance has broadened to include colors that are beyond the
10 spectrum that human eyes can perceive (such as ultraviolet (UV) reflectance), yet we still
11 know relatively little about which plant species reflect UV light or its effectiveness in
12 attracting pollinators. We investigated the effect of UV reflectance in Mimulus guttatus in
13 a number of different populations in British Columbia, Canada. We found that M. gut-
14 tatus had distinct regions of the corolla where UV light was reflected and absorbed. When
15 we manipulated the degree of contrast between the reflection and absorption area, we
16 found that pollinator visitation was severely disrupted, in terms of frequency and forag-
17 ing patterns observed. Despite the bright yellow (bee-green) coloration and visible nectar
18 guides in M. guttatus, we conclude that UV reflectance is critical in pollinator attraction.
19 Keywords: floral constancy, geographical variation, Mimulus guttatus, pollinator attraction, UV
20 reflectance.
21 Received 7 June 2001; revision received 15 December 2001; accepted 8 January 2012

22

23 Introduction The metaphor of a busy marketplace with vendors (the 46

plants) and customers (the pollinators) operating transac- 47
24 The vast majority of flowering plants (~80–90%) employ
tions for goods and services has been applied to plant– 48
25 insects, birds, and bats to transport and deposit pollen on
pollinator relationships (Chittka & Schurkens 2001). From 49
26 the stigma of conspecifics (Ollerton et al. 2011). Floral
the pollinator’s perspective, optimal foraging is achieved 50
27 color represents one of the key avenues by which plants
if they are able to distinguish between species of flowers 51
28 advertise their rewards to pollinators, affecting the attrac-
that they have previously learned contain suitable nectar 52
29 tion of different functional groups of pollinators (Fenster
(Leonard et al. 2011), and floral constancy can result from 53
30 et al. 2004) as well as influencing the floral constancy of
a pollinator’s focus to obtain the “best bargain” for their 54
31 visitation (Gumbert et al. 1999; Gegear 2005; Gegear &
effort investment (Chittka & Schurkens 2001). From the 55
32 Burns 2007). Investigating the importance of floral color to
plants’ perspective, it is advantageous to avoid cross- 56
33 the reproductive ecology of a particular plant species or
pollination with incompatible species of flowering plants 57
34 the community ecology of coflowering species (McEwen
by facilitating conspecific visitation patterns of the polli- 58
35 & Vamosi 2010) is complicated, however, by the fact that
nators (Flanagan et al. 2009). This goal of generating “cus- 59
36 many plant species reflect in the ultraviolet (UV) part of
tomer loyalty,” or floral constancy, can be accomplished in 60
37 the spectrum, which many pollinators can perceive but
many ways, most notably by visual differences between 61
38 humans cannot. Investigations of floral color that incor-
different species of flower as well as morphological dif- 62
39 porate the detection of UV light reflectance of petals,
ferences and complexity of nectar access (Wilson & Stine 63
40 sepals, and tepals is thus necessary to refine our under-
1996). Because insect pollinators use these visual senses to 64
41 standing of how selection has operated on these advertis-
identify and locate the flowers they forage on, these 65
42 ing structures in different flowering plant communities.
flowers must be easily distinguishable from the surround- 66

43
ing environment (Spaethe et al. 2001). 67
Most hymenopterans have trichromatic vision that rec- 68
44 Correspondence: Jana C. Vamosi ognizes ultraviolet (UV) light with peak sensitivity at 69
45 Email: jvamosi@ucalgary.ca 340 nm, blue light at 430 nm and yellow light at 535 nm 70

© 2012 The Authors

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2 J . M . R A E A N D J . C . VA M O S I

1 (Peitsch et al. 1992). In order to absorb UV light and create (Streisfeld & Kohn 2005; Cooley et al. 2008). Our investi- 55
2 patterns visible to bees, plants commonly use pigments gations focused on choice tests that (i) examine the pres- 56
3 such as flavonoids and dearmatized isoprenylated phlo- ence and variation in UV reflectance of M. guttatus within 57
4 roglucinols (Gronquist et al. 2001). A common floral the region and (ii) manipulate the UV signals to examine 58
5 pattern includes areas of low UV reflectance (high absor- its functional role in pollinator attraction and guidance. 59
6 bance) in the center of a floral display surrounded by Mimulus guttatus is being rapidly developed as a model 60
7 areas of high reflectance, or a “bulls-eye pattern” (Lunau species for genetic mapping (Hall et al. 2010), and a thor- 61
8 1992). The surrounding environment (e.g., sand), ough knowledge of how floral traits vary and operate 62
9 however, can also reflect UV radiation (Frolich 1976). It is along its geographical range will help guide future inves- 63
10 believed that the reflectance of the background interferes tigations of variation in the presence and expression of 64
11 more with blue and yellow hues, thereby decreasing the alleles. 65
12 magnitude of color contrast in the eye of a bee, while 66
13 flowers with UV are much more vibrant to the bee eye
Methods 67
14 (Chittka et al. 1994). While research indicates that UV light
15 signals are interpreted by hymenopterans in much the Populations of Mimulus guttatus were located on Hornby 68
16 same way as the rest of the visible light colors in nature Island, Gabriola Island, Quadra Island, and Elk Falls in 69
17 (Kevan et al. 2001), research in other taxa have indicated British Columbia, Canada. To receive a visual model rep- 70
18 that UV reflectance may have a different function than resentative of bee vision, flowers of M. guttatus were 71
19 other wavelengths (Borgia 2008). Some studies have found photographed using an ultraviolet lens attached to a 72
20 that when UV light is blocked from the environment, bees Nikon D50 digital camera. Although digital cameras are 73
21 are relatively quick to adjust using only visible light to not commonly used in studies of UV reflectance, recent 74
22 differentiate between flowers, allowing them to maintain studies have demonstrated the sensitivity of digital SLR 75
23 equivalent foraging efficiency as when the full spectrum cameras with CCD and CMOS sensors to UV light 76
24 of light was available (Dyer & Chittka 2004). However, (Tetley & Young 2008). In fact, these silicon based digital 77
25 when UV reflectance from flowers is manipulated in the sensors are able to record wavelengths between 200 and 78
26 field, a drastic reduction in pollinator visits can be 1100 nm, much wider than the visible spectrum (Tetley & 79
27 observed (Johnson & Andersson 2002). Thus, the role of Young 2008; Virginia Semiconductors 2011). Modern 80
28 UV reflectance in pollination requires further study in a cameras and their lenses usually have UV and IR block- 81
29 variety of environments and species. ing coatings or components, however, to ensure only the 82
30 While most attention is paid to the role of reflectance in visible spectrum of light is able to pass through to the 83
31 attracting pollinators, less is known about the evolution- camera sensor (Tetley & Young 2008). The Nikon camera 84
32 ary significance of floral color in manipulating aspects of model used in this experiment contained relatively 85
33 pollinator behavior, such as floral constancy (Temeles & minimal internal UV blocking coatings by factory design 86
34 Rankin 2000). The UV absorbing areas in the center of the and was very similar in specifications to those demon- 87
35 flower may serve such a function by providing a nectar strated by Tetley and Young (2008) to have a strong 88
36 guide near the anthers and stigma (Jones & Buchmann ability to record UV radiation. 89
37 1974). This area of UV absorbing pigment has even been The commercial lens, however, did pose a significant 90
38 hypothesized to increase the rates at which a bee becomes barrier to UV photography necessitating replacement 91
39 properly oriented with respect to the anthers (and, from with a lens composed of a glass photocopier lens, a Schott 92
40 the pollinators perspective, the nectar). Selection for such BG38 filter to block infrared (IR), and a Hoya U340 filter to 93
41 signals may be generated to prevent floral damage, block visible light. Spectrometer analysis of the entire lens 94
42 increase anther/stigma contact time, or increase floral assembly revealed transmittance between the range of 330 95
43 constancy (or all three). to 390 nm, a range that corresponds relatively well to the 96
44 To date, few studies have investigated the degree to hymenopteran eye, which peaks in sensitivity to UV 97
45 which UV reflectance determines function (Johnson & around 340 nm (Peitsch et al. 1992). It is important to note 98
46 Andersson 2002) and some studies have found surpris- that the lens setup used does not transmit the wavelengths 99
47 ingly little effect (Borgia 2008). Whether UV patterns are in the lowest 10–15 nm of the bee range of vision very 100
48 generally favored through selection thus requires infor- well, nevertheless it does still provide a useful rough esti- 101
49 mation from a large array of species. Here, we examine mate of the UV patterns that a bee would perceive. Any IR 102
50 the effect of UV reflectance in a new species, Mimulus contamination was removed from the photographs by iso- 103
51 guttatus in several sites in southwestern British Columbia. lating the color channel UV was recorded in and convert- 104
52 Mimulus guttatus has yellow, zygomorphic flowers and ing the photograph to grayscale for more accurate 105
53 belongs to a genus where there appears to be much varia- interpretation of the UV patterns in a manner similar to 106
54 tion in floral color, including the ability to reflect UV Tetley and Young (2008). 107

© 2012 The Authors Plant Species Biology ••, ••–••

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V I S I TAT I O N I N M I M U L U S G U T T A T U S 3

1 Table 1 General description of the geographical variation in M. guttatus populations and pollinator abundances in southwestern British
2 Columbia
3
4 Mean population Mean observation Mean floral Mean number of
5 Number of size of M. guttatus period per site density per pollinators approaching
6 Location quadrats (number of individuals) (min) quadrat quadrats

7 Gabriola 4 67 105 27.3 10.3

8 Hornby 5 65 72 18.8 3.6
9 Quadra 2 30 120 14.0 6.5
10 Elk Falls 5 125 60 32.0 1.0
11

12

13 To obtain a more quantitative assessment of color reflec- were recorded. The data were collected during daylight 55
14 tance of M. guttatus, reflectance spectra of flower petals hours between 9:00 am and 5:00 pm from May 20, 2010 56
15 were obtained with an Ocean Optics Jaz 2000-Series spec- to June 12, 2010. The data recorded included the number 57
16 trometer, with a UV-Vis light source and a fiber optic of individual pollinators that approached and landed on 58
17 reflection probe for every 0.22 nm between 300 nm to a particular floral type within the quadrat, how many 59
18 700 nm wavelengths. Reflectance spectra of unaltered and flowers of each type (flower- or stem-treated) they 60
19 manipulated flowers (both top and bottom petals) can visited within the quadrat and whether the pollinator 61
20 then be used to calculate the amount by which the three correctly oriented on each visited flower. Within each 62
21 types of photoreceptors of bee eyes (UV, blue, and green) quadrat, the differences in pollinator visitation between 63
22 are excited by light input, thereby determining how the flower- and stem-treated plants were analyzed in a 64
23 petals are perceived in bee color space (methods for cal- paired design using the statistical program R 2.0.9.0 (R 65
24 culating loci on the bee color hexagon are outlined in Development Core Team 2010). Finally, we examined the 66
25 Chittka 1992; Chittka et al. 1994). function of UV contrast in orienting pollinators by exam- 67
26 Mimulus guttatus exhibits a patchy distribution within ining the proportion of first visits where the pollinators 68
27 southwestern British Columbia, with patches surrounded landed “correctly” in control and treated flowers using a 69
28 by moist rocky surfaces and mosses. We located ten sepa- contingency test. Here, a “correct” orientation was 70
29 rate populations, with population sizes ranging from ~20– landing on the bottom petal with the pollinator’s head 71
30 225 individuals. Within each population, 0.5 m ¥ 0.5 m pointed towards the nectar, while an “incorrect” orien- 72
31 quadrats were placed within randomly selected patches of tation was landing on the wrong petal or with the head 73
32 M. guttatus (see Table 1 for the number of quadrats per pointed away from the nectar sources. Finally, we exam- 74
33 population). Quadrats within the same site (but in differ- ined the relationship between the success of their first 75
34 ent patches) were located 3–10 m apart to ensure that no landing and how many visits they subsequently per- 76
35 single individual was repeatedly sampled. Within each formed within the quadrat. 77
36 quadrat, the total number of flowers was recorded and Both stem- and flower-treated flowers often began to 78
37 half the flowers were randomly selected and covered with wilt prior to 180 min of observation of the quadrats. 79
1
38 1 a light spray coating of “Kinesys for kids” 30 SPF Though the floral density varied in the different sites 80
39 (KINeSYS Pharmaceutical Inc., Richmond, BC, Canada) (Table 1), the number of flowers in each treatment group 81
40 sunscreen, which had one the highest toxin-free scores of was equivalent per quadrat (with the exception of one 82
41 any sunscreens on the market (and is also fragrance- and quadrat where two flowers in the flower-treatment group 83
42 alcohol-free). The flowers were coated with one or two were accidentally damaged during observation). Differ- 84
43 pumps from a squirt bottle held at a distance of about ences in total number of visits and the number of inde- 85
44 50 cm. In the remaining flowers within the patch, we con- pendent pollinator foraging bouts between the floral 86
45 trolled for any residual scent in the sunscreen by instead types (stem vs. flower treated) were not normally distrib- 87
46 coating a section of stem directly below the flower with uted and this was not adequately corrected with standard 88
47 the same sunscreen. We ensured that this sunscreen transformations; thus nonparametric paired Wilcoxon 89
48 coating treatment was sufficient to decrease the UV reflec- tests were conducted. In cases where we thought there 90
49 tance (Fig. 2) and change the degree of color contrast may be important covariates, we analyzed multivariate 91
50 between top and bottom petals (Fig. 3). Flowers that were generalized linear models with the “glm” procedure in R 22 92
51 already wilted and falling off were omitted from the study (R Development Core Team 2010) software. We assumed a 93
52 and not recorded with either the control or altered groups. Gaussian error distribution when examining continuous 94
53 Each quadrat was then observed for up to 2 h, or until response variables and a Poisson error distribution when 95
54 visible signs of wilting, and all pollinator visitations examining count data (Crawley 2007). 96

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4 J . M . R A E A N D J . C . VA M O S I

(a)
(a)
Colour

(b) (b)

1 3 Fig. 1 Visible light (top) and ultraviolet reflectance (bottom) pat-

2 terns of Mimulus guttatus.

3 Fig. 2 (a) Reflectance spectra of top and bottom petals in control 28

and treated flowers and (b) the degree of color separation as seen 29
4 Results by bees. In (b) the circles indicate top petals and squares indicate 30
bottom petals (control and treatment flowers shown in black and 31
5 Mimulus guttatus flowers have zygomorphic flowers and grey, respectively). 32
6 thus do not have the common “bulls-eye” or “target-type”
33
7 pattern to their reflectance (Penny 1983). Instead, the side
8 and top petals exhibit strong UV reflectance while the were of the genus Bombus, representing > 90% of all visits. 34
9 lower petal has a very low UV reflectance, providing a One site on Gabriola Island, however, had a small propor- 35
10 great deal of contrast for pollinators that can see UV light tion of visitations (four individuals) from Apis mellifera. 36
11 (Fig. 1 and 2). Also, the nectar guides present in the visible Syrphid flies were also rarely observed at two of the Elk 37
12 spectrum were not evident under UV photography. The Falls provincial park sites. In general, M. guttatus received 38
13 pattern of UV reflection and absorption was uniform a low frequency of visitation from pollinators compared to 39
14 throughout the sites observed in this study. other early blooming species in the area (e.g., Plectritis 40
15 Manipulation of M. guttatus flowers with sunscreen congesta, unpublished results), with control patches only 41
16 was very effective at decreasing the area of UV reflectance receiving a total of 68 visits in 1320 min of observation, or 42
17 (Fig. 2 and 3) and should decrease the ability of pollina- an average of about three visits per hour. Pollinators were 43
18 tors to detect these flowers and/or confuse their ability to observed to visit 1–11 flowers in a quadrat before flying 44
19 properly orient themselves to find the nectaries. In total, off beyond our line of vision (Table 1). 45
20 pollinator visitations were observed within 16 quadrats in The total number of times that control flowers were 46
21 the 10 populations. Four of these sites were on Gabriola visited per patch (mean = 4.74 visits/h) was significantly 47
22 Island, five on Hornby Island, two on Quadra Island and greater than the total number of times treated flowers 48
23 five on Vancouver Island at Elk Falls Provincial Park. (mean = 1.02 visits/h) in the same patch were visited 49
24 Quadrats within these patches had between 10 and 80 (V1,15 = 75.5, P = 0.005). Pollinator abundance varied from 50
25 flowers within the quadrat and these were always divided site to site (Table 1), yet even when we control for the total 51
26 equally into stem- versus flower-treated groups. By far, number of pollinators foraging within the quadrats, the 52
27 the most common pollinators observed at all four sites mean number of visits-per-bee to control flowers 53

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V I S I TAT I O N I N M I M U L U S G U T T A T U S 5

(a)
Colour

(b)

Fig. 4 Pairwise differences in the mean number of visits per bee 29

between control and sunscreen treated flowers in the 16 quadrats 30
used in this study. 31

32

landing on the side petals or the back of the flower. In 33

addition, the bee’s initial orientation appeared to deter- 34
mine bee foraging behavior to some extent; those bees 35
that failed to orient themselves properly on their first 36
visit, visited fewer flowers within the quadrat before 37
leaving than those that oriented themselves correctly on 38
1 Fig. 3 A comparison of the visible light (top) and ultraviolet
2 (bottom) reflectance patterns of two control Mimulus guttatus their first visit (mean successive visits = 1.04 in the 39
3 flowers (centre, red arrows) and two treatment flowers (lower control group versus 0.14 in the treatment group; 40
4 right, blue arrows). W = 5623.5, P < 0.0001). While we might be cautious in 41
interpreting this particular result because the sunscreen 42
5
treatment may alter taste and deter pollinators from 43
6 (mean = 0.96 visits/bees/h) was significantly greater than repeat visits, initial orientation of the pollinator had a 44
7 the mean number of visits-per-bee to treatment flowers significant effect on their number of repeat visits, even 45
8 (mean = 0.42 visits/bees/h) in the same patch (V1,15 = 59, when their reflectance level (treatment or control 46
9 P = 0.023; Fig. 4). Even if only the first visits by pollinators flowers) was accounted for (glm: Z = -6.33, P < 0.0001). In 47
10 are considered, giving an idea of the general attractive- other words, bees that stumbled to find the nectar on 48
11 ness of the flowers, the number of bees approaching and their first try usually left the M. guttatus patch, regardless 49
12 beginning their foraging bouts on control flowers of whether their first visit was upon a control or 50
13 (mean = 2.28 bees/h) was significantly greater than that of sunscreen-treated flower. 51
14 the treatment flowers (mean = 0.90 bees/h) in the same 52
15 patch (V1,15 = 63, P = 0.008). We did not find any effect of
Discussion 53
16 the floral density of the patch on either the differences
17 observed in the number of bees visiting (glm: t1,15 = 0.572, Mimulus guttatus is being developed as a model genetic 54
18 P = 0.576) or the mean visitation per bee (glm: t1,15 = 0.385, system for the study of ecological and evolutionary genet- 55
19 P = 0.706). ics in nature with its relatively small genome, ecological 56
20 Excluding data from two pollinators that were hard to diversity, and ease of propagation (Wu et al. 2008). We find 57
21 assess, we found that the individuals visiting treatment that M. guttatus has an added dimension of intriguing 58
22 flowers oriented themselves improperly significantly complexity with the observation that the petals have 59
23 more frequently than those visiting control flowers (like- defined regions of UV reflectance and absorption not 60
24 lihood ratio test: c1,94 = 29.24, P < 0.0001). Pollinators visible to the human eye and these regions appear to be 61
25 landing on control flowers oriented correctly 98% (67/68 integral to pollination success. Surprisingly, the UV pat- 62
26 pollinators) of the time, while those landing on treatment terns did not overlay or complement the nectar guide 63
27 flowers oriented correctly 50% (14/28 pollinators) of the markings that were visible to the human eye. While the 64
28 time, with the remaining visits consisting of pollinators visitor assemblages in this study were heavily dominated 65

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6 J . M . R A E A N D J . C . VA M O S I

1 by Bombus (as in other studies, Wise et al. 2011), other frequently than those visiting the control flowers. When 55
2 studies have found M. guttatus pollinated by higher pro- the pollinator is unable to determine where to land to 56
3 portions of Apis mellifera (Martin 2004) and the degree to access the nectar (as in the treatment flowers) they must 57
4 which pollinators respond to UV versus visible light may spend a greater amount of time at each flower searching 58
5 vary. Similarly, different species of Bombus may react dif- for nectar, which will decrease pollination efficiency and 59
6 ferently to floral color variation but this was not some- the likelihood of invoking floral constancy. This result is 60
7 thing we detected in our observations during this study. consistent with the idea that the two contrasting colors 61
8 Overall, our results are consistent with the hypothesis between the upper and lower petals of M. guttatus serve 62
9 that UV reflectance provides contrast with background to signal pollinators where to land to access the nectar. 63
10 vegetation to attract pollinators in M. guttatus. We also Interestingly, previous work in M. luteus found that the 64
11 speculate that contrast with the lower petal functions as a visible nectar guides were more important in a similar 65
12 nectar guide. Because the control acts to help ensure the way in increasing bird visitation (Medel et al. 2007). We 66
13 scent of the sunscreen is not deterring the pollinator visi- find that the UV nectar guides in M. guttatus may be as 67
14 tation, decreased visitation is likely due to the altered UV important in bee attraction because pollinators reduced 68
15 reflectance. Decreased floral constancy is also possible as the number of flower visitations when individuals failed 69
16 pollinators that visited altered flowers had a reduced to properly orient themselves to the first flower. 70
17 capacity to land in the correct orientation and tended to It is acknowledged that a species of flowering plants 71
18 leave the patch altogether. Thus, the contrast in UV reflec- can receive a greater number of visitations by increasing 72
19 tance and absorption appears to attract the attention of its visibility to potential pollinators (Sih & Baltus 1987; 73
20 pollinators, as well as guide them to the nectar source Chittka et al. 2001). This relationship creates selection 74
21 (Penny 1983), two measures that are likely to increase the pressure for flowers with increasingly prominent visual 75
22 efficiency by which pollen is delivered in M. guttatus. patterns. The costs and benefits of UV reflectance and 76
23 Detecting whether the contrast with the background veg- absorption (as opposed to other colors) are not well 77
24 etation or with the lower petal is most important to attrac- understood, however. Because UV sensitivity is an ances- 78
25 tion or floral constancy will require more careful tral trait in arthropods (Kevan et al. 2001), it may represent 79
26 experimentation, which will be difficult considering that an important signal between plants and insects, albeit one 80
27 sunscreen treatments are more effective at changing that is invisible to mammals. 81
28 the reflectance spectra of UV-reflecting regions than Ultraviolet signals possibly serve an adaptive function 82
29 UV-absorbing regions (Fig. 2). of allowing insect pollinators to spot the flowers easier, 83
30 While it has been demonstrated previously that patch but at the same time preventing mammal herbivores from 84
31 size is positively correlated with the visitation rates of exploiting this UV signal to increase their herbivory on 85
32 Bombus (Sih & Baltus 1987), our paired experimental the flowers. Considering that previous studies have indi- 86
33 design kept the relative number of control and treatment cated that vertebrate florivory can be responsible for 87
34 flowers constant in each quadrat, controlling for variation reducing the floral abundance by up to 90% (Wang & 88
35 in floral density of unaltered M. guttatus flowers sur- Mopper 2008), the effects of reducing conspicuousness to 89
36 rounding our focal quadrat varied from site to site. One mammals remains a very understudied avenue of 90
37 factor we did not address, however, was the community research. However, the subsequent selection for UV 91
38 context in terms of coflowering species. Other studies signals that increase flower conspicuousness may be con- 92
39 have found that that the degree of pollinator sharing is strained by the increased attraction of insect herbivores 93
40 influenced by color overlap (Peter & Johnson 2008; (Strauss et al. 2004). 94
41 Campbell et al. 2010) and the preference for unaltered Ultraviolet pigments could serve a dual purpose, 95
42 M. guttatus flowers may have been influenced by the pro- however, as they can act also as chemical deterrents to 96
43 portion of similarly colored coflowerers in the neighbor- some herbivores (Gronquist et al. 2001). Plants with these 97
44 hood. While an intriguing avenue of future study, we pigments in their flowers may experience multiple ben- 98
45 observed the M. guttatus has specific habitat require- efits, chemical defense from predators, attraction of polli- 99
46 ments and often the same complement of surrounding nators and increased floral constancy. This study provides 100
47 species across the different sites. Thus, we do not antici- a greater understanding of the effects of UV reflectance 101
48 pate that coflowering variation had an undue influence across the wide variation of patterns of UV reflectance in a 102
49 on our results. species where it may become possible to locate and 103
50 The mean number of treated flowers each pollinating manipulate the genes for UV reflectance. Future studies 104
51 bee visited while inside the patch was also significantly might use genetic knock-outs or clever crosses with 105
52 lower from the mean number of control flowers visited. Mimulus species that do not reflect UV (Cooley et al. 2008) 106
53 Bees that visited sunscreen-treated flowers oriented them- will be useful in characterizing the genes and ecological 107
54 selves incorrectly to access the nectar significantly more role of UV reflectance. 108

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1 Acknowledgments Johnson S. D. & Andersson S. (2002) A simple field method for 59

manipulationg ultraviolet reflectance of flowers. Canadian 60
2 We thank L. Adderley for help in the field. This study was Journal of Botany 80: 1325–1328. 61
3 funded by NSERC-CANPOLIN grant to JCV (this repre- Jones C. E. & Buchmann S. L. (1974) Ultraviolet floral patterns as 62
4 sents publication #29 of NSERC-CANPOLIN). functional orientation cues in hymenopterous pollination 63
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