Pollen on-twine for food provisioning and oviposition of predatory mites in protected crops

The establishment of predatory mites on protected crops is affected by the availability of shelter and alternative food. We suggest that a pollen covered twine, coined here “pollen on-twine”, may provide these necessities when attached to the plant. We evaluated the effect of twine types and two pollen species on the establishment of Euseius scutalis (Athias-Henriot) and Amblyseius swirskii Athias-Henriot on pre-flowering pepper plants in a growth chamber. For both phytoseiids, rayon jute twine was found more beneficial while corn and oak pollen did not differ in their effect. Though populations of both predators were best promoted when twine and pollen were applied separately, E. scutalis population increased by more than tenfold and A. swirskii doubled when plants were applied with pollen on-twine. We propose that after further refining, pollen on-twine can serve as a feasible solution for predatory mite establishment on protected crops.     

Variability in bumblebee pollination buzzes affects the quantity of pollen released from flowers

Buzz-pollination is a plant strategy that promotes gamete transfer by requiring a pollinator, typically bees (Hymenoptera: Apoidea), to vibrate a flower’s anthers in order to extract pollen. Although buzz-pollination is widespread in angiosperms with over 20,000 species using it, little is known about the functional connection between natural variation in buzzing vibrations and the amount of pollen that can be extracted from anthers. We characterized variability in the vibrations produced by Bombus terrestris bumblebees while collecting pollen from Solanum rostratum (Solanaceae), a buzz-pollinated plant. We found substantial variation in several buzzing properties both within and among workers from a single colony. As expected, some of this variation was predicted by the physical attributes of individual bumblebees: heavier workers produced buzzes of greater amplitude. We then constructed artificial “pollination buzzes” that varied in three parameters (peak frequency, peak amplitude, and duration), and stimulated S. rostratum flowers with these synthetic buzzes to quantify the relationship between buzz properties and pollen removal. We found that greater amplitude and longer duration buzzes ejected substantially more pollen, while frequency had no directional effect and only a weak quadratic effect on the amount of pollen removed. These findings suggest that foraging bumblebees may improve pollen collection by increasing the duration or amplitude of their buzzes. Moreover, given that amplitude is positively correlated with mass, preferential foraging by heavier workers is likely to result in the largest pollen yields per bee, and this could have significant consequences for the success of a colony foraging on buzz-pollinated flowers.     

Consumption of Bt Rice Pollen Containing Cry1C or Cry2A Protein Poses a Low to Negligible Risk to the Silkworm Bombyx mori (Lepidoptera: Bombyxidae)

By consuming mulberry leaves covered with pollen from nearby genetically engineered, insect-resistant rice lines producing Cry proteins derived from Bacillus thuringiensis (Bt), larvae of the domestic silkworm, Bombyx mori (Linnaeus) (Lepidoptera: Bombyxidae), could be exposed to insecticidal proteins. Laboratory experiments were conducted to assess the potential effects of Cry1C- or Cry2A-producing transgenic rice (T1C-19, T2A-1) pollen on B. mori fitness. In a short-term assay, B. mori larvae were fed mulberry leaves covered with different densities of pollen from Bt rice lines or their corresponding near isoline (control) for the first 3 d and then were fed mulberry leaves without pollen. No effect was detected on any life table parameter, even at 1800 pollen grains/cm² leaf, which is much higher than the mean natural density of rice pollen on leaves of mulberry trees near paddy fields. In a long-term assay, the larvae were fed Bt and control pollen in the same way but for their entire larval stage (approximately 27 d). Bt pollen densities ≥150 grains/cm² leaf reduced 14-d larval weight, increased larval development time, and reduced adult eclosion rate. ELISA analyses showed that 72.6% of the Cry protein was still detected in the pollen grains excreted with the feces. The low exposure of silkworm larvae to Cry proteins when feeding Bt rice pollen may be the explanation for the relatively low toxicity detected in the current study. Although the results demonstrate that B. mori larvae are sensitive to Cry1C and Cry2A proteins, the exposure levels that harmed the larvae in the current study are far greater than natural exposure levels. We therefore conclude that consumption of Bt rice pollen will pose a low to negligible risk to B. mori.     

Influence of Pollen Nutrition on Honey Bee Health: Do Pollen Quality and Diversity Matter?

Honey bee colonies are highly dependent upon the availability of floral resources from which they get the nutrients (notably pollen) necessary to their development and survival. However, foraging areas are currently affected by the intensification of agriculture and landscape alteration. Bees are therefore confronted to disparities in time and space of floral resource abundance, type and diversity, which might provide inadequate nutrition and endanger colonies. The beneficial influence of pollen availability on bee health is well-established but whether quality and diversity of pollen diets can modify bee health remains largely unknown. We therefore tested the influence of pollen diet quality (different monofloral pollens) and diversity (polyfloral pollen diet) on the physiology of young nurse bees, which have a distinct nutritional physiology (e.g. hypopharyngeal gland development and vitellogenin level), and on the tolerance to the microsporidian parasite Nosema ceranae by measuring bee survival and the activity of different enzymes potentially involved in bee health and defense response (glutathione-S-transferase (detoxification), phenoloxidase (immunity) and alkaline phosphatase (metabolism)). We found that both nurse bee physiology and the tolerance to the parasite were affected by pollen quality. Pollen diet diversity had no effect on the nurse bee physiology and the survival of healthy bees. However, when parasitized, bees fed with the polyfloral blend lived longer than bees fed with monofloral pollens, excepted for the protein-richest monofloral pollen. Furthermore, the survival was positively correlated to alkaline phosphatase activity in healthy bees and to phenoloxydase activities in infected bees. Our results support the idea that both the quality and diversity (in a specific context) of pollen can shape bee physiology and might help to better understand the influence of agriculture and land-use intensification on bee nutrition and health.     

The Role of a Pollen-Expressed Cullin1 Protein in Gametophytic Self-Incompatibility in Solanum

We previously isolated a pollen factor, ui6.1, which encodes a Cullin1 protein (CUL1) that functions in unilateral interspecific incompatibility (UI) in Solanum. Here we show that CUL1 is also required for pollen function in self-incompatibility (SI). We used RNA interference (RNAi) to reduce CUL1 expression in pollen of Solanum arcanum, a wild SI tomato relative. Hemizygous T₀ plants showed little or no transmission of the transfer DNA (T-DNA) through pollen when crossed onto nontransgenic SI plants, indicating that CUL1-deficient pollen are selectively eliminated. When crossed onto a related self-compatible (SC) accession lacking active S-RNase, pollen transmission of the T-DNA followed Mendelian ratios. These results provide further evidence for functional overlap between SI and UI on the pollen side and suggest that CUL1 mutations will reinforce SI-to-SC transitions in natural populations only if preceded by loss of pistil S-RNase expression.     

Use of maize pollen by adult Chrysoperla carnea (Neuroptera: Chrysopidae) and fate of Cry proteins in Bt-transgenic varieties

We investigated the use of maize pollen as food by adult Chrysoperla carnea under laboratory and field conditions. Exposure of the insects to insecticidal Cry proteins from Bacillus thuringiensis (Bt) contained in pollen of transgenic maize was also assessed. Female C. carnea were most abundant in a maize field when the majority of plants were flowering and fresh pollen was abundant. Field-collected females contained an average of approximately 5000 maize pollen grains in their gut at the peak of pollen shedding. Comparable numbers were found in females fed ad libitum maize pollen in the laboratory. Maize pollen is readily used by C. carnea adults. When provided with a carbohydrate source, it allowed the insects to reach their full reproductive potential. Maize pollen was digested mainly in the insect's mid- and hindgut. When Bt maize pollen passed though the gut of C. carnea, 61% of Cry1Ab (event Bt176) and 79% of Cry3Bb1 (event MON 88017) was digested. The results demonstrate that maize pollen is a suitable food source for C. carnea. Even though the pollen grains are not fully digested, the insects are exposed to transgenic insecticidal proteins that are contained in the pollen.     

Pollen subsidies promote whitefly control through the numerical response of predatory mites

Several studies have shown that biological control of pests can be improved by supplying extra food to natural enemies. This increases population levels of the enemies, resulting in decreases in pest densities. In theory, however, supplying food can also have negative effects on biological control. We specifically tested for such negative effects, using a predator–prey system consisting of the whitefly Bemisia tabaci (Gennadius) and a predatory mite Amblyseius swirskii (Athias-Henriot). This predator attacks eggs and young instars of the whitefly, but also feeds on pollen. We added pollen to populations of predators and whiteflies on isolated cucumber plants. Although the set-up of our experiments would favour the occurrence of a negative effect of the addition of pollen on biological control, we found increased control throughout the experiment. This shows that the control of whiteflies by A. swirskii can be improved by supplementing the predators with pollen.     

Better safe than sorry? A Fabaceae species exhibits unfavourable pollen properties for developing bee larvae despite its hidden anthers

Empirical evidence suggests that pollen chemistry plays an important role in shaping the pollen host spectra of many bee species. Although the underlying mechanisms are poorly understood, pollen diets of several plant taxa have experimentally been found to impede larval development of unspecialized bees. The pollen of all plant taxa, for which such a detrimental effect on bee larval development has been observed so far, is freely accessible in the flowers and thus easily harvestable for flower visitors, suggesting that this pollen might be chemically protected in order to reduce its loss to pollen-feeding animals. In the present study, we compared larval performance of five solitary bee species on pollen diets of the two Fabaceae species Onobrychis viciifolia and Lotus corniculatus, which have their anthers concealed inside the flowers, with that on control diets composed of host pollen provisions. As the complex flower morphology of the two Fabaceae species already considerably narrows the spectrum of pollen harvesting bee taxa, which might supersede costly chemical protection of the pollen, we expected bees that usually do not exploit Fabaceae to develop well on Onobrychis and Lotus pollen diets. Larval survival on the Onobrychis pollen diet was successful for all five bee species tested. In contrast, larval survival on the Lotus pollen diet was reduced in three species despite the fact that Lotus flowers are more difficult to exploit for pollen than Onobrychis flowers. We conclude that there is no trade-off between pollen concealment and pollen defence in Lotus and that pollen of morphologically complex flowers with a restricted visitor spectrum is not necessarily an easy-to-use nutritional source.     

The Impact of the Invasive Alien Plant,Impatiens glandulifera,on Pollen Transfer Networks

Biological invasions are a threat to the maintenance of ecological processes, including pollination. Plant-flower visitor networks are traditionally used as a surrogated for pollination at the community level, despite they do not represent the pollination process, which takes place at the stigma of plants where pollen grains are deposited. Here we investigated whether the invasion of the alien plant Impatiens glandulifera (Balsaminaceae) affects pollen transfer at the community level. We asked whether more alien pollen is deposited on the stigmas of plants on invaded sites, whether deposition is affected by stigma type (dry, semidry and wet) and whether the invasion of I. glandulifera changes the structure of the resulting pollen transfer networks. We sampled stigmas of plants on 10 sites invaded by I. glandulifera (hereafter, balsam) and 10 non-invaded control sites. All 20 networks had interactions with balsam pollen, although significantly more balsam pollen was found on plants with dry stigmas in invaded areas. Balsam pollen deposition was restricted to a small subset of plant species, which is surprising because pollinators are known to carry high loads of balsam pollen. Balsam invasion did not affect the loading of native pollen, nor did it affect pollen transfer network properties; networks were modular and poorly nested, both of which are likely to be related to the specificity of pollen transfer interactions. Our results indicate that pollination networks become more specialized when moving from the flower visitation to the level of pollen transfer networks. Therefore, caution is needed when inferring pollination from patterns of insect visitation or insect pollen loads as the relationship between these and pollen deposition is not straightforward.     

Some pollinators are more equal than others: Factors influencing pollen loads and seed set capacity of two actively and passively pollinating fig wasps

The nursery pollination system of fig trees (Ficus) results in the plants providing resources for pollinator fig wasp larvae as part of their male reproductive investment, with selection determining relative investment into pollinating wasps and the pollen they carry. The small size of Ficus pollen suggests that the quantities of pollen transported by individual wasps often limits male reproductive success. We assessed variation in fig wasp pollen loads and its influence on seed production in actively pollinated (Ficus montana) and passively pollinated (Ficus carica) dioecious fig trees.The ratios of number of male flowers on number of female flowers in a glasshouse-maintained F. montana population were highly variable. When fig wasps were introduced into receptive female figs, the resulting seed numbers were strongly linked to the numbers of pollinators that had been seeking access to pollen, relative to the number of anthers in their natal figs. In F. carica estimates of the amounts of pollen produced per fig and the quantities of pollen carried by emerging fig wasps suggest that less than 10% of the pollen is transported. Pollinators of F. carica that emerged earlier from figs carried more pollen, and also generated more seeds when introduced into receptive female figs.We show here that all pollinators are not equally valuable and producing more pollinators is not necessarily a good option in terms of Ficus male fitness. Previous results on F. montana figs showed that only around half of the flowers where pollinators lay eggs produced adult offspring. The amount of pollen collected by young female fig wasps may be a major determinant of their reproductive success.     

Effect on in Vitro Pollen Growth of an Isolated Style Glycoprotein Associated with Self-Incompatibility in Nicotiana alata

The effect on in vitro pollen tube growth of an isolated style glycoprotein (S₂-glycoprotein) associated with self-incompatibility in Nicotiana alata was investigated. Tube growth of pollen bearing the S₂-allele was inhibited, but tube growth of pollen bearing other alleles was not affected. Inhibition showed a dose response effect. The percentage of pollen grains that germinated was not significantly affected by the S₂-glycoprotein. Growth of S₂-pollen in the presence of the S₂-glycoprotein resulted in increased binding to the pollen of monoclonal antibody (PCBC3) which has a primary specificity for α-l-arabinofuranosyl residues. Growth of pollen bearing other alleles in the presence of the glycoprotein resulted in no increased binding of the antibody.     

Short-distance pollen dispersal for an outcrossed, wind-pollinated southern beech (Nothofagus nervosa (Phil.) Dim. et Mil.)

The mating system (outcrossing, selfing, and biparental inbreeding) and the extent of pollen flow are two of the most important genetic features that determine the genetic structure of plant populations, and both are crucial for the design of conservation strategies. The objectives here were to estimate mating system parameters and to fit the pollen dispersal kernel for the southern beech, Nothofagus nervosa. We sampled 25 mothers and 372 progeny from two stands in the Tromen Lake region of Argentina. We registered spatial positions of the maternal trees, and genotyped mothers and offspring for five simple sequence repeat markers. We estimated single-locus (t _s?=?0.95) and multilocus (t _m?=?0.99) outcrossing rates and biparental inbreeding (t _m-t _s?=?0.04). The species is strongly outcrossing, but correlated paternity within maternal sibships (r _p?=?0.10) indicates that each maternal parent is sampling a different and restricted array of pollen donors. We used two protocols (twogener and kindist) to fit an exponential power dispersal kernel to the structure of pollen clouds sampled by different mothers. The estimated effective number of pollen donors contributing to a single mother was N _ep?=?9.9. The twogener and kindist analyses yielded slightly different estimates, but both indicated short average distances for pollen dispersal (<35?m), indicating that the dispersal kernel was strongly leptokurtic (???=?0.36). While short-distance pollen dispersal predominates, there remains a nontrivial probability of long-distance dispersal. The results are discussed in the context of ongoing conservation and management programs.     

Extensive pollen flow in a natural fragmented population of Patagonian cypress Austrocedrus chilensis

Habitat fragmentation might significantly affect mating and pollen dispersal patterns in plant populations, contributing to the decline of remnant populations. However, wind-pollinated species are able to disperse pollen at longer distances after opening of the canopy. Our objectives were to characterize the mating system parameters and to estimate the average distance of effective pollen dispersal in the wind-pollinated conifer Austrocedrus chilensis. We sampled 19 “mother trees,” 200 progeny, and 81 additional adults (both male and female), in a fragmented population at the Argentinean Patagonian steppe. We registered the spatial positions of individuals and genotyped all samples with five microsatellite markers. We found a high genetic diversity, a moderated rate of biparental inbreeding (t _m? ??t _s?=?0.105), and a complete absence of correlated paternity (r _p?=??0.015). The effective number of pollen donors contributing to a single mother (N _ep) was 13.9. Applying TWOGENER, we estimated a low but significant differentiation among the inferred pollen pools (Φ_FT?=?0.036, p?=?0.001) and a very large average pollen dispersal distance (d?=?1,032.3 m). The leptokurtic distribution (b?=?0.18) presumes a potential for even larger dispersal distances. The high genetic diversity, the mating patterns, and the extensive pollen dispersal presume that habitat fragmentation did not have a negative impact on pollen movement in this population of A. chilensis. Genetic connectivity among fragmented populations scattered in the Patagonian region is possible, and we stress the need of management policies at the landscape level.     

Aberrant Classopollis pollen reveals evidence for unreduced (2n) pollen in the conifer family Cheirolepidiaceae during the Triassic–Jurassic transition

Polyploidy (or whole-genome doubling) is a key mechanism for plant speciation leading to new evolutionary lineages. Several lines of evidence show that most species among flowering plants had polyploidy ancestry, but it is virtually unknown for conifers. Here, we study variability in pollen tetrad morphology and the size of the conifer pollen type Classopollis extracted from sediments of the Triassic–Jurassic transition, 200 Ma. Classopollis producing Cheirolepidiaceae were one of the most dominant and diverse groups of conifers during the Mesozoic. We show that aberrant pollen Classopollis tetrads, triads and dyads, and the large variation in pollen size indicates the presence of unreduced (2n) pollen, which is one of the main mechanisms in modern polyploid formation. Polyploid speciation may explain the high variability of growth forms and adaptation of these conifers to different environments and their resistance to extreme growth conditions. We suggest that polyploidy may have also reduced the extinction risk of these conifers during the End-Triassic biotic crisis.     

The weak effects of climatic change on Plantago pollen concentration: 17 years of monitoring in Northwestern Spain

Plantago L. species are very common in nitrified areas such as roadsides and their pollen is a major cause of pollinosis in temperate regions. In this study, we sampled airborne pollen grains in the city of León (NW, Spain) from January 1995 to December 2011, by using a Burkard® 7-day-recording trap. The percentage of Plantago pollen compared to the total pollen count ranged from 11 % (1997) to 3 % (2006) in the period under study. Peak pollen concentrations were recorded in May and June. Our 17-year analysis failed to disclose significant changes in the seasonal trend of plantain pollen concentration. In addition, there were no important changes in the start dates of pollen release and the meteorological parameters analyzed did not show significant variations in their usual trends. We analyzed the influence of several meteorological parameters on Plantago pollen concentration to explain the differences in pollen concentration trends during the study. Our results show that temperature, sun hours, evaporation, and relative humidity are the meteorological parameters best correlated to the behavior of Plantago pollen grains. In general, the years with low pollen concentrations correspond to the years with less precipitation or higher temperatures. We calculated the approximate Plantago flowering dates using the cumulative sum of daily maximum temperatures and compared them with the real bloom dates. The differences obtained were 4 days in 2009, 3 days in 2010, and 1 day in 2011 considering the complete period of pollination.     

Effects of Previous Pollination and Stylar Ethylene on Pollen Tube Growth in Petunia hybrida Styles

The effect of ethylene on the growth rate of pollen tubes in styles of Petunia hybrida was examined. Apart from its strong inhibition of pollination-induced ethylene synthesis, aminoethoxyvinylglycine, placed on the stigma, did not impede tube growth. The inhibitors of the action of ethylene, silver thiosulfate and 2,5-norbornadiene, were similarly ineffective. Application of the ethylene precursor, 1-amino-cyclopropane-1-carboxylic acid, onto the stigma at different intervals prior to pollination evoked synthesis of ethylene, but was without effect on tube growth. However, prepollination (by 24 hours) with Nicotiana tabacum pollen, significantly enhanced tube growth of Petunia pollen. This enhancement was not counteracted by the pretreatment of stigmas with aminoethoxy-vinylglycine. It is concluded that the ethylene associated with pollination is without effect on pollen tube growth in the style, but that other pollination-induced factors may lead to an acceleration of growth.     

Coflowering invasive plants and a congener have neutral effects on fitness components of a rare endemic plant

Network analyses rarely include fitness components, such as germination, to tie invasive plants to population‐level effects on the natives. We address this limitation in a previously studied network of flower visitors around a suite of native and invasive plants that includes an endemic plant at Badlands National Park, South Dakota, USA. Eriogonum visheri coflowers with two abundant invasive plants, Salsola tragus and Melilotus officinalis, as well as a common congener, E. pauciflorum. Network analyses had suggested strong linkages between E. visheri and S. tragus and E. pauciflorum, with a weaker link to M. officinalis. We measured visitation, pollen deposited on stigmas, achene weight and germination over three field seasons (two for germination) in four populations (two in the final season) of E. visheri and applied in situ pollen treatments to E. visheri, adding pollen from other flowers on the same plant; flowers on other E. visheri plants; S. tragus, M. officinalis, or E. pauciflorum; open pollination; or excluding pollinators. Insect visitation to E. visheri was not affected by floral abundance of any of the focal species. Most visitors were halictid bees; one of these (Lasioglossum packeri) was the only identified species to visit E. visheri all three years. Ninety‐seven percent of pollen on collected E. visheri stigmas was conspecific, but 22% of flowers had >1 grain of E. pauciflorum pollen on stigmas and 7% had >1 grain of S. tragus pollen; <1% of flowers had M. officinalis pollen on stigmas. None of the pollen treatments produced significant differences in weight or germination of E. visheri achenes. We conclude that, in contrast to the results of the network analysis, neither of the invasive species poses a threat, via heterospecific pollen deposition, to pollination of the endemic E. visheri, and that its congener provides alternative pollen resources to its pollinators.     

Shared pollinators and pollen transfer dynamics in two hybridizing species, Rhinanthus minor and R. angustifolius

Gene flow between hybridizing plant species depends strongly on pollinator behaviour, which affects pollen transfer among floral types and reproductive isolation. We examined bumblebee behaviour and pollen transfer between two hybridizing Rhinanthus species that are very similar in ecology and floral traits. The two species, Rhinanthus minor and R. angustifolius, shared similar pollinator guilds and assemblages, but pollinator recruitment and flower visitation rates were higher in R. angustifolius sites, probably because of its higher reward levels and better visibility. When presented with Rhinanthus flowers, bumblebees that previously foraged on R. angustifolius were less prone to visit R. minor inflorescences, while R. minor foragers accepted both species in similar proportions. Although Rhinanthus has been cited as a case of mechanical isolation resulting from interactions between bee behaviour and differences in stigma and anther placement, we found no support for efficient mechanical reproductive isolation. Bumblebees that foraged on R. minor flowers carried more pollen, but pollen placement on their bodies was similar to that of bees that visited R. angustifolius, and cross-specific stigmatic pollen deposition was similar in both directions. However, the asymmetry in pollinator handling time between the two species, due to dissimilar pollen rewards, may have lowered relative heterospecific pollen receipt on R. angustifolius, suggesting that net gene flow resulting from pollen transfer dynamics is more likely towards R. minor, although this effect remains weak and will be most likely counterbalanced by context-based labile pollinator preference.     

Annexin5 Plays a Vital Role in Arabidopsis Pollen Development via Ca2+-Dependent Membrane Trafficking

The regulation of pollen development and pollen tube growth is a complicated biological process that is crucial for sexual reproduction in flowering plants. Annexins are widely distributed from protists to higher eukaryotes and play multiple roles in numerous cellular events by acting as a putative “linker” between Ca²⁺ signaling, the actin cytoskeleton and the membrane, which are required for pollen development and pollen tube growth. Our recent report suggested that downregulation of the function of Arabidopsis annexin 5 (Ann5) in transgenic Ann5-RNAi lines caused severely sterile pollen grains. However, little is known about the underlying mechanisms of the function of Ann5 in pollen. This study demonstrated that Ann5 associates with phospholipid membrane and this association is stimulated by Ca²⁺ in vitro. Brefeldin A (BFA) interferes with endomembrane trafficking and inhibits pollen germination and pollen tube growth. Both pollen germination and pollen tube growth of Ann5-overexpressing plants showed increased resistance to BFA treatment, and this effect was regulated by calcium. Overexpression of Ann5 promoted Ca²⁺-dependent cytoplasmic streaming in pollen tubes in vivo in response to BFA. Lactrunculin (LatB) significantly prohibited pollen germination and tube growth by binding with high affinity to monomeric actin and preferentially targeting dynamic actin filament arrays and preventing actin polymerization. Overexpression of Ann5 did not affect pollen germination or pollen tube growth in response to LatB compared with wild-type, although Ann5 interacts with actin filaments in a manner similar to some animal annexins. In addition, the sterile pollen phenotype could be only partially rescued by Ann5 mutants at Ca²⁺-binding sites when compared to the complete recovery by wild-type Ann5. These data demonstrated that Ann5 is involved in pollen development, germination and pollen tube growth through the promotion of endomembrane trafficking modulated by calcium. Our results provide reliable molecular mechanisms that underlie the function of Ann5 in pollen.