Pollination biology of the endangered orchid Cypripedium japonicum in a fragmented forest of Japan

Pollination biology studies of the endangered orchid Cypripedium japonicum were conducted in its natural habitat using pollinator observation and hand‐pollination experiments. The observed fruit set was as follows: artificial outcross‐pollinated, 100%; artificial self‐pollinated, 100%; pollinator‐excluded, 0%; and emasculated flowers, 0%. These results show that this species, although self‐compatible, is neither autogamous nor agamospermous. The fruit set for open‐pollinated flowers was 14.9%, which suggests that the study population was subject to pollinator limitation. The nectarless flowers of C. japonicum were exclusively visited and pollinated by the queens of two bumblebee species (Bombus ardens and B. diversus diversus). It is probable that the nectarless flowers of C. japonicum attract pollinators through a generalized food deceptive system.     

Autonomous self‐pollination in the nectarless orchid Pogonia minor

The pollination biology of the nectarless orchid Pogonia minor was investigated in central Japan. The investigation revealed that the solitary flowers failed to attract pollinators, while high rates of fruit set were observed in the natural population. Comparable levels of fruit set were obtained in bagged, artificial self‐pollinated and artificial cross‐pollinated plants, indicating that the species is not pollinator‐limited for fruit set under natural conditions. Autonomous self‐pollination in P. minor resulted from a reduced rostellum, which allowed contact between the pollinia and the stigma. Self‐pollination is thought to be an adaptive response that provides reproductive assurance under conditions of pollinator limitation. Since pollen limitation is generally known to be frequent among deceptive orchids, strong pollen limitation is probably a driving force in the autonomous self‐pollination mechanism in the nectarless orchid P. minor.     

Pollination ecology of two nectariferous Cimicifuga sp. (Ranunculaceae) and the evolution of andromonoecy

The pollination ecology of Cimicifuga americana Michx. and C. laciniata Wats., two nectariferous self-incompatible species, was studied. Both species are pollinated by bumblebees, the former by workers and the latter by males. All bees imbibe nectar and workers also collect pollen, meanwhile performing pollination. Large syrphids act as co-pollinators. C. americana is andromonoecious, and it is suggested that male-flower production at the end of the flowering period contributes a disproportionate fitness component because of dichogamy-induced female-biased functional sex-ratio in the hermaphroditic flowers at that time.     

Effects of natural rates of geitonogamy on fruit set in Asclepias speciosa (Apocynaceae): evidence favoring the plant's dilemma

The role of geitonogamy in the evolution of inflorescence design is not well understood. The plant's dilemma hypothesis proposes that evolution of larger inflorescences is driven by selection for greater pollinator attraction, but constrained by higher rates of geitonogamy experienced by larger inflorescences. Here we investigate the role of geitonogamy on fruit set in natural populations of Asclepias speciosa. We compared fruit set from three pollination treatments: (1) inflorescences bagged before and after receiving 6 hand outcross pollinia (Bag), (2) inflorescences unbagged and receiving 6 hand outcross pollinia (Open), and (3) naturally pollinated inflorescences (Control). The Bag and Open treatments initiated significantly more fruits than the Control. Bag aborted significantly fewer fruits than Open or Control. Fruit set was significantly higher in Bag than Open, and Open had significantly higher fruit set than Control. From these results, we conclude that (1) high rates of geitonogamy significantly increase fruit abortion and reduce fruit set in natural populations of A. speciosa and (2) natural populations are compatible pollen limited. Both findings are consistent with the plant's dilemma hypothesis.     

Pollination biology of the deceptive orchid Changnienia amoena

The pollination biology of Changnienia amoena , an endangered terrestrial orchid, was investigated at two sites in the Shennongjia Mountains, Hubei, central China. The results show that the orchid is exclusively pollinated by bumblebees. In Longmenhe, Bombus ( Diversobombus ) trifasciatus is the primary effective pollinator, whereas B.  ( Tricornibombus ) imitator is the only pollinator in Guanmenshan. These two bumblebees can be treated as a functional group because they carry pollinaria on the same position on their bodies and have similar pollinating behaviours. The morphological traits of the flower adapt precisely to the pollinators. Visitation by bumblebees is mainly in the first half of the flowering period of C. amoena . Pollinarium removal and pollinia deposition take place when bumblebees withdraw from the flower. Crossing experiments show that C. amoena is a self-compatible and outcrossing species. Fruit set in this species is low and may result from limited pollinators because 87.5% of individuals set fruits under hand-pollination, but only 6–12% of individuals set fruits under natural conditions. These results imply that it is important to conserve the pollinator community together with the plants when conservation management for this endangered species is undertaken. Artificial pollination would also be a useful measure to facilitate restoration of the small populations.  © 2006 The Linnean Society of London, Botanical Journal of the Linnean Society , 2006, 150 , 165–175.     

开花时间与伴生种对鹤首马先蒿传粉和生殖成功的影响

开花物候是物种间相互作用的重要生活史特征和适合度因子,在全球气候变化的背景下而备受关注.为探讨开花时间如何存种内和种间水平上影响植物的传粉和生殖成功,我们连续3年(2003-2005)对不同花期和伴生种存在情况下的鹤首马先蒿(Pedicularis gruina)的传粉者访花忠实度、受粉率、坐果率、单果种子产量和果实被啃食频率进行了比较研究.结果表明鹤首马先蒿的坐果率主要受其传粉环境的影响:在没有伴生种时,不同时期鹤首马先蒿的坐果率没有显著差异,34-38％的花可以坐果;在有伴生种存在时既可以显著提高其坐果率,也可显著降低其坐果率,这取决于传粉者类型以及伴生种密穗马先蒿(P.densispica)花期的差异.密穗马先蒿具有花蜜和花粉双重报酬,在群落中可以作为主体物种吸引传粉者并间接促进与其伴生的鹤首马先蒿的传粉和生殖成功.同样无蜜的管花马先蒿(P.siphonantha)和鹤首马先蒿伴生,则是通过提高群落水平对传粉者的吸引力进而直接促进鹤首马先蒿的传粉和生殖成功.此外,研究结果也表明开花时间对坐果率没有显著影响,但是显著影响单果种子产量和果实被啃食的频率.在相同的传粉条件下早花期植株单果种子产量显著高于晚花期的种子产量,同时早花期的果实被啃食的频率显著增加.     

Three pollination morphs in Cimicifuga simplex; incipient speciation due to inferiority in competition

Cimicifuga simplex, a Japanese herb, has three different pollinator guilds. At high altitudes it is pollinated by bumblebees. In lowlands, Impatiens textori, a much superior nectar-producer, outcompetes it for bumblebees. In sunny lowland habitats, flowers of a second morph produce a two-component fragrance which facilitates pollination by fritillaries. In experiments, the two components separately had limited effect on alighting rate, while application of both together elevated the attractiveness of non-fragrant flowers to the same as for naturally fragrant flowers. In heavily shaded lowland habitats, a third morph with nonfragrant flowers does not blom until after I. textori, and is pollinated by occasional bumblebees. The data strongly suggest that inferiority in competition has induced incipient speciation in this plant species.     

Does community‐level floral abundance affect the pollination success of a rewardless orchid,Calanthe reflexa Maxim.?

Decreases in pollinator abundance may particularly constrain plants that lack floral rewards, since they are poor competitors for pollinators in the plant community. Here, we documented the pollination ecology of a rewardless orchid, Calanthe reflexa Maxim., and examined effects of forest understory degradation by deer browsing on pollination success of the species in the light of a change in the abundance of neighboring flowering plants in 2010 and 2011. Bombus species were the only pollinators at each site and the flowering phenology of C. reflexa did not overlap with that of other rewarding plants. Pollinator visit rates (assessed by time‐lapse photography), and pollinia removal rate were higher in the undegraded understory site than the degraded site in both years, while the fruit set ratio did not differ between the sites in 2011. Coverage by neighboring flowering plants was extremely low in the degraded site. Our results suggest that, although its flowering phenology and consequently lower interspecific competition of C. reflexa with rewarding plants for attracting bumblebees, neighboring flowering plants may play an important role for maintaining the visitation frequency of bumblebees of C. reflexa and contribute to its pollination success.     

Experimental analysis of mating patterns in a clonal plant reveals contrasting modes of self‐pollination

Hermaphrodite plants commonly practice self‐fertilization (selfing), but the mechanisms responsible vary depending on the mode of self‐pollination, pollinator behavior, and degree of clonality. Whether selfing occurs within (autogamy) or between flowers (geitonogamy) is of evolutionary significance because their fitness consequences differ. We used floral manipulations and genetic markers to determine the relative contribution of autogamy and within‐ versus between‐ramet geitonogamy to the selfing rate of the bumblebee‐pollinated, clonal herb Aconitum kusnezoffii. Data on flowering phenology and bumblebee foraging were also collected to determine opportunities for different modes of self‐pollination. Autogamy accounted for only 12% of the selfing rate with the remainder resulting from geitonogamy. Whole‐ramet emasculation of clones with multiple ramets reduced selfing by 78%, indicating that within‐ramet geitonogamy contributed significantly (68%) to total selfing. Selfing of single‐ramet plants was 44% less than multiple‐ramet plants, indicating that the contribution of between‐ramet geitonogamy was substantially less (20%) than within‐ramet geitonogamy, probably because of bumblebee foraging behavior. Our results demonstrate for the first time in a clonal plant that within‐ramet geitonogamy is substantially greater than between‐ramet geitonogamy and highlight the importance of considering the influence of clonal architecture and pollinator foraging on modes of self‐pollination.     

Community-level flowering phenology and fruit set: Comparative study of 25 woody species in a secondary forest in Japan

We investigated the flowering phenology, pollinator visitation, and fruit set of 25 animal-pollinated woody species in a warm temperate secondary forest in Japan. Various species flowered sequentially from February to October. The principal pollinators were bumblebees, honey-bees, flies and/or beetles and birds; bumblebees and flies/beetles pollinated most trees. The duration of flowering was shorter for species that bloomed in the middle of the season than it was for species that bloomed earlier or later in the season. The timing of flowering was more synchronous within species that had a shorter flowering duration; this was also detected when phylogenetically independent contrasts were calculated. This could be important for the effective pollination of species with a short flowering duration because such species bloom sequentially over a short period of less than 1month around May. Fruit set was related not to pollinator type, sex expression, flowering sequence (in order of the date of peak flowering) or flowering duration, but to the relative abundance of the species in the forest. This correlation was detected for fly- and beetle-pollinated species but not for bumblebee-pollinated species. Thus, relatively rare plant species with opportunistic pollinators might experience limited fruit set because of insufficient pollinator services. Bagging experiments conducted on eight hermaphrodite species revealed that the fruit set of bagged flowers was nearly zero, lower than that of control flowers. These results indicate the importance of pollinators for successful reproduction and thus for the coexistence of plants in this secondary forest.     

Flower herbivory and seed predation in Silene vulgaris (Caryophyllaceae): effects of pollination and phenology

Pollination and seed predation were studied in Silene vulgaris populations during two seasons, one with much lower pollinator abundance than the other. Among the pollinators, noctuid moths of the genus Hadena also acted as seed predators. Nectar-foraging female moths oviposited in flowers, and their larvae consumed flowers and seed capsules.
Despite a lower percentage of pollinated flowers in the year of low pollinator abundance, similar numbers of flowers set fruit in both years, because fewer flower buds and flowers were eaten by Hadena larvae during the year of low pollinator visitation. The number of seed capsules preyed upon was also lower in the year with low pollinator abundance, resulting in a higher seed set. The positive correlation between the percentage of pollinated flowers and the percentage of seed capsules destroyed was also observed when comparing flowers opening in different parts of the season.
Early flowering plant individuals had the same pollination success but suffered higher seed predation than late flowering ones. Selection for maximized pollination success through synchronous flowering, is probably the main reason for the compressed flowering period in 5. vulgaris , but the high level of predation early in the season may further increase the reproductive success of synchronous flowering individuals.     

FLORAL BIOLOGY,HUMMINGBIRD POLLINATION AND FRUIT PRODUCTION OF TRUMPET CREEPER (CAMPSIS RADICANS,BIGNONIACEAE)

Trumpet creeper is self-incompatible and bears long, tubular, orange flowers from June to September. Flowering peaks rapidly, then declines and continues at low levels for several weeks. The initial burst of flowering may attract pollinators that return even during subsequent reduced flowering. Most flowers open before noon and nectar production totals 110 μl of 26% sucrose equivalents per flower, an exceptionally high production for a temperate zone plant. Production ceases within 20–30 hr of flower opening, but corollas persist for several days and may serve to attract pollinators. Effective pollination reduces the period of stigma receptivity and speeds closing of stigma lobes. Only 1–9% of flowers produced mature fruits at four sites in Illinois and Missouri. Roughly 400 pollen grains had to be deposited on a receptive stigma to cause fruit development beyond an initial period of high abortion. At two sites, 17% and 89% of stigmas received over 400 pollen grains. Assuming 50% of deposited grains were from the same plant, fruit production at one site was clearly pollinator limited, that at the second site may have been. Ruby-throated Hummingbirds (Archilochus colubris) deposited ten times as much pollen per stigma per visit as honeybees (Apis mellifera) and bumblebees (Bombus spp.). Fruit set was highest where rubythroat visitation was most frequent. Trumpet creeper appears primarily adapted for hummingbird pollination, but can also be adequately pollinated by honeybees and bumblebees. This is one of the first attempts to relate pollen-depositing capabilities of pollinators of any plant to pollen requirements for fruit production. Several characteristics suggest that trumpet creeper may be adapted to pollination at low densities (often called traplining) in its presumed original, woodland, habitat.     

Two Bee-Pollinated Plant Species Show Higher Seed Production when Grown in Gardens Compared to Arable Farmland

Background

Insect pollinator abundance, in particular that of bees, has been shown to be high where there is a super-abundance of floral resources; for example in association with mass-flowering crops and also in gardens where flowering plants are often densely planted. Since land management affects pollinator numbers, it is also likely to affect the resultant pollination of plants growing in these habitats. We hypothesised that the seed or fruit set of two plant species, typically pollinated by bumblebees and/or honeybees might respond in one of two ways: 1) pollination success could be reduced when growing in a floriferous environment, via competition for pollinators, or 2) pollination success could be enhanced because of increased pollinator abundance in the vicinity.

Methodology/Principal Findings

We compared the pollination success of experimental plants of Glechoma hederacea L. and Lotus corniculatus L. growing in gardens and arable farmland. On the farms, the plants were placed either next to a mass-flowering crop (oilseed rape, Brassica napus L. or field beans, Vicia faba L.) or next to a cereal crop (wheat, Triticum spp.). Seed set of G. hederacea and fruit set of L. corniculatus were significantly higher in gardens compared to arable farmland. There was no significant difference in pollination success of G. hederacea when grown next to different crops, but for L. corniculatus, fruit set was higher in the plants growing next to oilseed rape when the crop was in flower.

Conclusions/Significance

The results show that pollination services can limit fruit set of wild plants in arable farmland, but there is some evidence that the presence of a flowering crop can facilitate their pollination (depending on species and season). We have also demonstrated that gardens are not only beneficial to pollinators, but also to the process of pollination.     

Pollinator‐mediated interactions between cultivated papaya and co‐flowering plant species

Many modern crop varieties rely on animal pollination to set fruit and seeds. Intensive crop plantations usually do not provide suitable habitats for pollinators so crop yield may depend on the surrounding vegetation to maintain pollination services. However, little is known about the effect of pollinator‐mediated interactions among co‐flowering plants on crop yield or the underlying mechanisms. Plant reproductive success is complex, involving several pre‐ and post‐pollination events; however, the current literature has mainly focused on pre‐pollination events in natural plant communities. We assessed pollinator sharing and the contribution to pollinator diet in a community of wild and cultivated plants that co‐flower with a focal papaya plantation. In addition, we assessed heterospecific pollen transfer to the stigmatic loads of papaya and its effect on fruit and seed production. We found that papaya shared at least one pollinator species with the majority of the co‐flowering plants. Despite this, heterospecific pollen transfer in cultivated papaya was low in open‐pollinated flowers. Hand‐pollination experiments suggest that heterospecific pollen transfer has no negative effect on fruit production or weight, but does reduce seed production. These results suggest that co‐flowering plants offer valuable floral resources to pollinators that are shared with cultivated papaya with little or no cost in terms of heterospecific pollen transfer. Although HP reduced seed production, a reduced number of seeds per se are not negative, given that from an agronomic perspective the number of seeds does not affect the monetary value of the papaya fruit.     

Successful pollination of the Neotropical crop Solanum quitoense by Bombus terrestris: behaviour,efficiency and yield

The South American lulo (Solanum quitoense Lam.) is a crop plant of the Andes of Ecuador and Colombia, pollinated by South American bumblebees, such as, Bombus atratus Franklin. The cultivation of lulo outside of its native range, for example in European glasshouses, requires the presence of efficient pollinators to enable high fruit set and yield. Until now, the suitability of Bombus terrestris L., native to Europe and commonly used in agriculture, has been untested for this purpose. In this study, the pollen‐collecting behaviour of B. terrestris when visiting lulo flowers was investigated. It was shown that B. terrestris adopted the lulo as a pollen source, and on average visited three flowers per minute, had five buzzing events per stay and foraged for 15 s on a single flower, independently of the previous number of visits and level of bruising to the anthers. The pollination efficiency of five different treatments was evaluated: (i) exclusion of bees, (ii) single and (iii) multiple visits of B. terrestris, (iv) self‐ and (v) cross‐pollination by hand. The results clearly demonstrated that, for fruit set, pollination is crucial. It was also found that lulo flowers can be successfully self‐pollinated, but give 25% fewer fruit set compared with pollination via multiple bumblebee visits, or cross‐pollination by hand. Fruit set, seed set and fruit size were as high with pollination by B. terrestris as with cross‐pollination by hand, indicating that this bumblebee is an appropriate pollinator for lulo. However, B. terrestris was conspicuously less effective when a flower was visited only once. Therefore, when growing lulos commercially, multiple bumblebee visits should be encouraged, but it is likely that the behaviour of B. terrestris would ensure this anyway. Our results indicate that B. terrestris is a suitable and efficient pollinator for the production of lulo fruits.     

Specialized pollination by carpenter bees in Calanthe striata (Orchidaceae), with a review of carpenter bee pollination in orchids

Calanthe striata has nectarless flowers that are self‐compatible, but pollinator dependent. Field observations showed that the flowers were pollinated exclusively by the carpenter bee Xylocopa appendiculata circumvolans, although the bees occasionally wasted pollen by delivering to the stigmatic surface pollinaria that retained their anther caps. Fruit set ratios at the population level varied spatiotemporally, but were generally low (8.3–17.3%). Calanthe striata blooms in spring when post‐overwintering carpenter bees have not yet started foraging for brood production. It can therefore exploit an abundance of opportunistic/naïve foragers. This timing may also increase the possibility of pollinator visits, because no rewarding co‐flowering plants are available in the orchid habitats. A literature review of Orchidaceae pollinated by carpenter bees revealed that at least 14 species of Orchidoideae and Epidendroideae have evolved flowers specialized for carpenter bee pollination. They typically have shallow pink/magenta flowers with a foothold for pollinators; pollinaria are attached to the head, ventral thorax or base of the middle legs of carpenter bees when they insert their heads and/or proboscises into flowers; pollination success is generally low, a probable consequence of the deceptive pollination systems. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013 , 171 , 730–743.     

濒危植物独花兰的传粉生物学初步观察

独花兰(Changnienia amoena Chien)为我国特有的单种属植物,近年因生境破碎化和过度采挖,其野生资源日渐减少。迄今对独花兰极为有限的研究表明,其结实率很低或根本不结实,其传粉媒介也一直未被发现。2002年3～4月,我们对神农架2个移植居群和5个天然居群进行了传粉生物学的定点观察,发现雌性三条熊蜂(Bombus (Diversobombus)trifasciatus Smith)、仿熊蜂(Bombus (Tricornibombus) imitator Pittion)和蜜蜂均访问独花兰,但只有二条熊蜂身体粘有花粉块,是独花兰的有效传粉者。三条熊蜂的访问频率很低,在113h的观察中只有9次访问,但在一个天然居群(population 3)中曾观察到一天4次的最高访问频率;访问主要在12：00～15：00出现,但在花上停留时间很短,不超过10s。在末被授粉的情况下,独花兰花朵大约3周后自然枯萎,但受粉后3、4d内即出现～系列形态和颜色的变化,包括花梗逐渐伸长,子房在花梗逐渐停止伸长后开始膨大等,表明花梗伸长可作为结实(授粉成功)的指标。人工授粉实验表明,自花、异花受粉后花梗均伸长,而套袋隔离花的则花梗不伸长,说明独花兰是自交亲和的异交种,需要昆虫传粉。根据传粉者的访问频率、居群中果实的分布,尤其是花距内无花蜜等特征,我们认为独花兰是一种欺骗性传粉的兰花。相对于其他欺骗性传粉的兰花,独花兰的自然结实率并不很低(26．98％),这与居群规模小会提高欺骗性传粉兰花结实率这一观点吻合。花粉块的输出数远高于结实数,这说明独花兰存在一定的花粉浪费。     

濒危植物独花兰的传粉生物学初步观察

独花兰(Changnienia amoena Chien)为我国特有的单种属植物,近年因生境破碎化和过度采挖,其野生资源日渐减少.迄今对独花兰极为有限的研究表明,其结实率很低或根本不结实,其传粉媒介也一直未被发现.2002年3～4月,我们对神农架2个移植居群和5个天然居群进行了传粉生物学的定点观察,发现雌性三条熊蜂(Bombus(Diversobombus)trifasciatus Smith)、仿熊蜂(Bombus(Tricornibombus)imitator Pittion)和蜜蜂均访问独花兰,但只有三条熊蜂身体粘有花粉块,是独花兰的有效传粉者.三条熊蜂的访问频率很低,在113 h的观察中只有9次访问,但在一个天然居群(population 3)中曾观察到一天4次的最高访问频率;访问主要在12:00～15:00出现,但在花上停留时间很短,不超过10 s.在未被授粉的情况下,独花兰花朵大约3周后自然枯萎,但受粉后3、4 d内即出现一系列形态和颜色的变化,包括花梗逐渐伸长,子房在花梗逐渐停止伸长后开始膨大等,表明花梗伸长可作为结实(授粉成功)的指标.人工授粉实验表明,自花、异花受粉后花梗均伸长,而套袋隔离花的则花梗不伸长,说明独花?兰是自交亲和的异交种,需要昆虫传粉.根据传粉者的访问频率、居群中果实的分布,尤其是花距内无花蜜等特征,我们认为独花兰是种欺骗性传粉的兰花.相对于其他欺骗性传粉的兰花,独花兰的自然结实率并不很低(26.98%),这与居群规模小会提高欺骗性传粉兰花结实率这一观点吻合.花粉块的输出数远高于结实数,这说明独花兰存在一定的花粉浪费.     

No evidence for simultaneous pollen and resource limitation in Aciphylla squarrosa: A long‐lived,masting herb

Abstract For successful reproduction animal pollinated plants must provide resources for both pollinator attraction and offspring production, and theory suggests that resources and pollen delivery limit reproduction simultaneously. We conducted a series of experiments involving supplemental pollination, flower removal, fertilizer addition and foliage removal to investigate the interaction of resources and pollen on fruit‐set of Aciphylla squarrosa, a long‐lived, dioecious, masting herb in Wellington, New Zealand. Reducing floral display decreased open‐pollinated fruit‐set, suggesting that display size is a reflection of an optimal investment between attraction and fecundity. In combination with supplemental pollination, resource reduction and fertilization addition did not alter fruit‐set, suggesting that changes in resource availability did not limit reproduction in the current year. In addition, supplemental pollination of non‐manipulated treatments did not increase fruit‐set, demonstrating that plants were not naturally pollen limited. While we found that simultaneous pollen and resource limitation did not occur within a season, this is possibly mitigated by life history patterns including mast flowering and a storage taproot. Multiple year studies are required to further examine simultaneous resource and pollen limitation.     

The effects of landscape on bumblebees to ensure crop pollination in the highland agricultural ecosystems in China

Honey bees and wild bees provide critical pollination services to agricultural ecosystems; however, the relative contributions of different bee taxa are not well understood. The natural habitats surrounding farmland support food and nesting resources for wild bees and therefore play an important role in the maintenance of crop pollination. In this study, we selected Cucurbita pepo L. (squash) as a model crop to investigate the relative importance of honey bees and bumblebees in pollinating the crop. Thirteen fields, which were surrounded by a gradient of natural habitat, were investigated on the Yunnan‐Guizhou Plateau in China. We measured the visit densities of honey bees and bumblebees, the number of pollen grains deposited in a single visit by the two bee taxa, as well as the overall pollen grains deposited on stigmas during a flowering day, and then used Bayesian inference to decouple the pollen grains deposited by either the honey bees or the bumblebees. Compared with honey bees, bumblebees deposited a higher number of pollen grains on stigmas in a single visit, but had a lower visit density than honey bees. Meanwhile, the bumblebee visit density increased along the proportion of natural habitat, while the honey bee visit density was not affected by the surrounding natural habitat. Data simulations using Bayesian inference showed that on a flowering day, the number of pollen grains deposited by bumblebees increased with the proportion of natural habitat in the surrounding landscape, but the number of pollen grains deposited by honey bees did not. Moreover, the total numbers of pollen grains deposited by honey bees or bumblebees alone were all below 2000 (the critical level to satisfy the pollination requirement of this crop). Pollen calculations demonstrated that the number of pollen grains deposited by the two bee taxa was greater than 2000 in fields surrounded by more than 13% natural habitat (grasslands and forests). The results revealed that bumblebees ensured C. pepo pollination in combination with honey bees in the highland agricultural ecosystems.