Abstract: We conducted a comparative analysis to determine life history and ecological correlates of pollen limitation among 224 species of animal-pollinated flowering plants. To test predictions concerning the occurrence of pollen limitation, species were classified categorically for six life-history traits (self-compatible vs. self-incompatible, autogamous vs. non-autogamous, specialized vs. unspecialized floral morphology, nectariferous vs. nectarless, monocarpic vs. polycarpic, herbaceous vs. woody) and two ecological conditions (presence in open vs. forested habitats and temperate vs. tropical biomes). Pollen limitation of species in contrasting categories was compared using non-parametric tests (TIPs analysis) and phylogenetically-independent contrasts (PICs). The results of TIPs and PICs analyses were generally congruent, although fewer significant contrasts were evident with PICs, probably because of low statistical power. Overall the results suggest that the influence of phylogenetic history on the intensity of pollen limitation was not particularly strong. Nonetheless, significant variation in the degree of pollen limitation was demonstrated among seven angiosperm families suggesting some phylogenetic component to the phenomenon. With both TIPs and PICs, pollen limitation was less intense in self-compatible and autogamous species. TIPs analysis demonstrated that herbaceous, nectariferous, and temperate species were less likely to be pollen-limited, but using PICs this could only be corroborated for those that were self-incompatible. None of the traits were singularly unambiguous predictors of pollen limitation, possibly reflecting the stochastic nature of pollinator service.

Abstract: Pollen ranges from 25% to 61% protein content Most pollen proteins are likely to be enzymes that function during pollen tube growth and subsequent fertilization, but the vast range of protein quantity may not reflect only pollen–pistil interactions Because numerous vertebrate and invertebrate floral visitors consume pollen for protein, protein content may influence floral host choice Additionally, many floral visitors pollinate their host plants If protein content influences pollinator visitation, then pollinators are hypothesized to select for increased protein content of host plants We analyzed or gleaned from the literature crude pollen protein concentrations of 377 plant species from 93 plant families Using this database, we compared pollen protein concentration with (1) pollination mode, (2) pollen collection by bees, and (3) distance from stigma to ovule, after accounting for phylogeny through paired phylogenetic comparisons and a nested ANOVA including taxonomic rank We found that pollen pr

Abstract: The present article reviews the various definitions and terminology of pollen viability and longevity as well as the various tests of its assessment. We compare the advantages and the disadvantages of each method and suggest some practical implications as revealed by the extensive data. We recognize eight main hypotheses concerning the ecology and the evolution of pollen longevity and critically evaluated them according to the literature. The hypotheses are grouped as follows: (1) Desiccation risk-carbohydrate content; (2) Pollen packaging; (3) Pollen competitive ability; (4) Pollinator activity-stigma receptivity duration; (5) Self-pollination chance; (6) Pollen exposure schedule; (7) Pollen travel distance, and (8) Pollen removal chance.

Abstract: Very long chain lipids contribute to the hydrophobic cuticle on the surface of all land plants and are an essential component of the extracellular pollen coat in the Brassicaceae. Mutations in Arabidopsis CER genes eliminate very long chain lipids from the cuticle surface and, in some cases, from the pollen coat. In Arabidopsis, the loss of pollen coat lipids can disrupt interactions with the stigma, inhibiting pollen hydration and causing sterility. We have positionally cloned CER6 and demonstrate that a wild-type copy complements the cer6-2 defect. In addition, we have identified a fertile, intragenic suppressor, cer6-2R, that partially restores pollen coat lipids but does not rescue the stem wax defect, suggesting an intriguing difference in the requirements for CER6 activity on stems and the pollen coat. Importantly, analysis of this suppressor demonstrates that low amounts of very long chain lipids are sufficient for pollen hydration and germination. The predicted CER6 amino acid sequence resembles that of fatty acid–condensing enzymes, consistent with its role in the production of epicuticular and pollen coat lipids >28 carbons long. DNA sequence analysis revealed the nature of the cer6-1, cer6-2, and cer6-2R mutations, and segregation analysis showed that CER6 is identical to CUT1, a cDNA previously mapped to a different chromosome arm. Instead, we have determined that a new gene, CER60, with a high degree of nucleotide and amino acid similarity to CER6, resides at the original CUT1 locus.

Abstract: The literature is reviewed and new evidence presented that pollen produces odors, which serve multiple functions in pollination and defense. Pollen odor, which originates from pollenkitt, comprises volatiles that belong to the same chemical classes found in flower scents, that are in species-specific mixtures, and that contrast with odors of other floral parts. Pollen can also take up volatiles from surrounding floral odors, but this adsorption is selective and varies among species. Pollen odors are more pronounced in insect- than bird- or wind-pollinated plants, suggesting that volatile emission evolved in part under selection to attract pollinators. Pollen-feeding insects can perceive pollen odor and use it to discriminate between different pollen types and host plants. Pollen odor influences bee foraging, including the location of pollen sources, discrimination of flowers with different amounts of pollen, and host-plant recognition by pollen-specialist species. In the few wind-pollinated plants studied, odors of male flowers or pollen are comparatively high in a-methyl alcohols and ketones; these volatiles may serve in pollen defense, with some known to repel insects. Pollen odor often includes chemicals with documented defense activity, which is probably aimed mainly at nonpollinator pollen-feeding insects and pathogens; an involvement in pollen allelopathy is also possible. Pollen volatiles comprise chemically diverse compounds that may play multiple roles, and their emission in pollen odor undoubtedly evolved under the principle, and often conflicting, selective pressures to both protect the male gametophyte and increase its dispersal by animals.

Abstract: In central New South Wales, Australia, flowers of Acacia brachybotrya and Eremophila glabra plants growing in linear vegetation remnants received less pollen than conspecifics in nearby reserves. Pollen supplementation increased fruit production by both species, indicating pollen limitation of fruit set. Together these observations explain why fruit production by these species was depressed in linear-strip populations relative to nearby reserves. This study confirms that habitat fragmentation can lead to decline in pollination and subsequent fruit set in wild plant populations. Disrupted pollination interactions of the kind documented in this study may offer a substantial challenge to the conservation of biodiversity in fragmented landscapes.

Abstract: We tested pollen viability of eight species using four vital dyes, a new peroxidase test together with three other established methods (MTT, Baker’s and X-Gal), to determine their potential to differentiate fresh pollen from pollen heated for 2 h and 24 h at 80°C (killed pollen) and compared the results with in vitro germination. We found that two of three dyes previously employed to determine viability also stained killed pollen, while the new peroxidase test and MTT did not. We suggest that the latter two are the best methods to test pollen viability, since they do not normally stain either killed or aborted pollen.

Abstract: Bees require pollen for their reproduction and pollen comprises the basic larval food for bees. Most bees acquire pollen passively during flower visitation, but female bees may also collect pollen actively with the aid of various structural and behavioral adaptations. Most bees have evolved adaptations to concentrate pollen into discrete loads and transport pollen back to their nests. The various structural and behavioral adaptations of female bees for acquiring and transporting pollen are the basis of this review.

Abstract: Many flowering plants avoid inbreeding through a genetic mechanism termed self-incompatibility. An extremely polymorphic S-locus controls the gametophytic self-incompatibility system that causes pollen rejection (that is, active arrest of pollen tube growth inside the style) when an S-allele carried by haploid pollen matches one of the S-alleles present in the diploid style. The only known product of the S-locus is an S-RNase expressed in the mature style. The pollen component to this cell-cell recognition system is unknown and current models propose that it either acts as a gatekeeper allowing only its cognate S-RNase to enter the pollen tube, or as an inhibitor of non-cognate S-RNases. In the latter case, all S-RNases are presumed to enter pollen tubes; thus, the two models make diametrically opposed predictions concerning the entry of S-RNases into compatible pollen. Here we use immunocytochemical labelling of pollen tubes growing in styles to show accumulation of an S-RNase in the cytoplasm of all pollen-tube haplotypes, thus providing experimental support for the inhibitor model.

Abstract: We present the first evidence that transgenic Bacillus thuringiensis (Bt) corn pollen naturally deposited on Asclepias syriaca; common milkweed, in a corn field causes significant mortality of Danaus plexippus L (Lepidoptera: Danaidae) larvae Larvae feeding for 48 h on A syriaca plants naturally dusted with pollen from Bt corn plants suffered significantly higher rates of mortality at 48 h (20±3%) compared to larvae feeding on leaves with no pollen (3±3%), or feeding on leaves with non-Bt pollen (0%) Mortality at 120 h of D plexippus larvae exposed to 135 pollen grains/cm2 of transgenic pollen for 48 h ranged from 37 to 70% We found no sub-lethal effects on D plexippus adults reared from larvae that survived a 48-h exposure to three concentrations of Bt pollen Based on our quantification of the wind dispersal of this pollen beyond the edges of agricultural fields, we predict that the effects of transgenic pollen on D plexippus may be observed at least 10 m from transgenic field borders However, the highest larval mortality will likely occur on A syriaca plants in corn fields or within 3 m of the edge of a transgenic corn field We conclude that the ecological effects of transgenic insecticidal crops need to be evaluated more fully before they are planted over extensive areas

Abstract: Although environmental factors such as precipitation and temperature are recognized as influencing pollen production, the impact of rising atmospheric carbon dioxide concentration ([CO2]) on the potential growth and pollen production of hay-fever-inducing plants is unknown. Here we present measurements of growth and pollen production of common ragweed (Ambrosia artemisiifolia L.) from pre-industrial [CO2] (280 mol mol–1) to current concentrations (370 mol mol–1) to a projected 21st century concentration (600 mol mol–1). We found that exposure to current and elevated [CO2] increased ragweed pollen production by 131 and 320%, respectively, compared to plants grown at pre-industrial [CO2]. The observed stimulations of pollen production from the pre-industrial [CO2] were due to an increase in the number (at 370 mol mol–1) and number and size (at 600 mol mol–1) of floral spikes. Overall, floral weight as a percentage of total plant weight decreased (from 21% to 13%), while investment in pollen increased (from 3.6 to 6%) between 280 and 600 mol mol–1 CO2. Our results suggest that the continuing increase in atmospheric [CO2] could directly influence public health by stimulating the growth and pollen production of allergy-inducing species such as ragweed.

Abstract: Flowering plants possess specialized extracellular matrices in the female organs of the flower that support pollen tube growth and sperm cell transfer along the transmitting tract of the gynoecium. Transport of the pollen tube cell and the sperm cells involves a cell adhesion and migration event in species such as lily that possess a transmitting tract epidermis in the stigma, style, and ovary. A bioassay for adhesion was used to isolate from the lily stigma/stylar exudate the components that are responsible for in vivo pollen tube adhesion. At least two stylar components are necessary for adhesion: a large molecule and a small (9 kD) protein. In combination, the two molecules induced adhesion of pollen tubes to an artificial stylar matrix in vitro. The 9-kD protein was purified, and its corresponding cDNA was cloned. This molecule shares some similarity with plant lipid transfer proteins. Immunolocalization data support its role in facilitating adhesion of pollen tubes to the stylar transmitting tract epidermis.

Abstract: My objective is the examination of selective forces that affect pollen number. Rela- tionships among other floral traits of animal- pollinated plants, including pollen size, stigma area and depth, and the pollen-bearing area of the pollinator may affect pollen number and also provide a model to examine how change in one trait may elicit change in other traits. The model provides a conceptual framework for appreciating intra- and inter-specific differences in these traits. An equivalent model is presented for wind-polli- nated plants. For these plants the distance between putative mates may be the most important factor affecting pollen number. I briefly consider how many pollen grains must reach a stigma to assure fruit set. I use pollen-ovule ratios (P/Os) to examine how breeding system, sexual system, pollen vector, and dispersal unit influence pollen grain number. I also compare the P/Os of plants with primary and secondary pollen presentation and those that pro- vide only pollen as a reward with those that provide nectar as part or all of the reward. There is a substantial decrease in P/O from xenogamy to facultative xenogamy to autogamy. Relative to homoecious species the P/Os of species with most other sexual systems are higher. This suggests that there is a cost associated with changes in sexual system. The P/Os of wind-pollinated plants are substantially higher than those of animal-polli- nated plants, and the available data suggest there is little difference in the pollination efficiency of the various animal vectors. The P/Os of plants whose pollen is dispersed in tetrads, polyads, or pollinia are substantially lower than those of species whose pollen is dispersed as monads. There was no difference in the P/Os of plants with primary and secondary pollen presentation. The P/Os of plants that provide only pollen as a reward were higher than those that provide nectar as a reward. All of these conclusions merit additional testing as they are based on samples that are relatively small and/ or systematically biased.

Abstract: Newly-emerged honey bees were placed in cages and fed sucrose syrup and one of the fol- lowing single-pollen diets: Malus domesticaBorkh., Brassica campestrisL., Phacelia tanacetifolia L., Melilotus officinalis(L.) Pall., Helianthus annuus L., Pinus banksiana(Lamb.), artificial supplement (Bee-Pro ® ) or nothing. Hypopharyngeal gland protein was determined at intervals of 0, 3, 8 and 14 days and ovary development was visually scored on day 14. The development of hypopharyngeal glands and ovaries varied with diet and, collectively, proved to be sensitive measures of protein uti- lization and pollen quality. For workers fed 1-year-old Phacelia pollen, protein was utilized in a differential fashion, promoting the development of ovaries over that of hypopharyngeal glands. Development of glands and ovaries was strongly correlated with the amount of protein workers con- sumed from pollen diets, and to a lesser extent, the crude protein content of diets. Storing pollen for 1 year by freezing did not affect gland or ovary development. Apis mellifera / hypopharyngeal gland / ovary / nutrition / protein / pollen

Abstract: The transport and capture of pollen in ∼ 20% of all angiosperm families occurs in air and water. In other words, pollination is abiotic and occurs via the fluid media, not an animal vector. Whereas some early concepts considered abiotic pollination to be largely a stochastic phenomenon, there is sufficient evidence to indicate that wind pollination (i.e. anemophily) and water pollination (i.e. hydrophily) have deterministic features and are sophisticated fluid dynamic solutions to the problem of pollen release, dispersal, and capture. An abiotic pollination syndrome is defined in which there is spatial or temporal separation of carpellate and staminate flowers, which are drab, a reduction in perianth parts, stigmas and anthers are exposed to the fluid, and typically unclumped pollen may be produced in large amounts. Separate pollination syndromes are defined for anemophilous (i.e. wind-pollinated), ephydrophilous (i.e. surface-pollinated), and hydrophilous (i.e. submarine-pollinated) plants. Distinctions are based on habitat and physical conditions for pollination, pollen size, shape, and ultrastructure, morphology and ultrastructure of stigmas, and outcrossing rates. For example, anemophilous pollen are spherical and small, ephydrophilous pollen are spherical or reniform and large, while hydrophilous pollen are filiform (i.e. filamentous) or functionally filiform. The pollination mechanisms and mechanics associated with these syndromes reveals a strong evolutionary relationship between plant morphology and fluid dynamics.

Abstract: A single laboratory study on monarch butterflies has prompted widespread concern that corn pollen, engineered to express Bacillus thuringiensis (Bt) endotoxin, might travel beyond corn fields and cause mortality in nontarget lepidopterans. Among the lepidopterans at high potential risk from this technology is the black swallowtail butterfly, Papilio polyxenes, whose host plants in the midwestern U. S. are located principally in narrow strips between roads and crop fields. A field study was performed to assess whether mortality of early instar black swallowtails was associated either with proximity to a field of Bt corn or by levels of Bt pollen deposition on host plants. Potted host plants were infested with first instar black swallowtails and placed at intervals from the edge of a field of Bt corn (Pioneer 34R07 containing Monsanto event 810) at the beginning of anthesis. We confirmed by ELISA that pollen from these plants contained Cry1Ab endotoxin (2.125 +/- 0.289 ng/g). Although many of the larvae died during the 7 days that the experiments were run, there was no relationship between mortality and proximity to the field or pollen deposition on host plants. Moreover, pollen from these same plants failed to cause mortality in the laboratory at the highest pollen dose tested (10,000 grains/cm(2)), a level that far exceeded the highest pollen density observed in the field (200 grains/cm(2)). We conclude that Bt pollen of the variety tested is unlikely to affect wild populations of black swallowtails. Thus, our results suggest that at least some potential nontarget effects of the use of transgenic plants may be manageable.

Abstract: Background: Type I allergic symptoms in the oropharyngeal mucosa upon contact with plant-derived food in patients with pollen allergies have been termed oral allergy syndrome (OAS). IgE cross-reactivity between pollen and food allergens represents the molecular basis for this phenomenon. The sensitizing allergen source (pollen or plant food) in OAS is a controversial issue. Objective: We sought to determine the primary sensitizing molecules in patients with OAS. Methods: We used recombinant birch pollen (rBet v 1 and rBet v 2) and plant food allergens (apple, rMal d 1; celery, rApi g 1; and carrot, rDau c 1), as well as natural pollen (birch and timothy grass) and plant food (apple, peach, kiwi, hazelnut, celery, and carrot) allergens, to identify cross-reactive allergens by using qualitative immunoblot inhibitions. In addition, we determined the percentage of plant food–specific IgE that can be preadsorbed with recombinant and natural pollen allergens by quantitative RAST inhibitions by using sera from 71 patients with OAS. Results: Preincubation of sera with recombinant and natural pollen allergens led to an almost complete inhibition of IgE binding to plant food allergens in Western blots, as well as in RAST inhibition experiments. In contrast, recombinant plant food allergens poorly inhibited IgE binding to Bet v 1. Conclusion: Most IgE epitopes in plant food recognized by patients with OAS are resembled by pollen allergens. Thus pollen allergens may be responsible for the elicitation and maintenance of OAS. (J Allergy Clin Immunol 2000;105:116-25.)

Abstract: By offering pollen and/or nectar as a food resource, angiosperms exploit flower visitors for pollen transport. Pollen thus acts not only as a means for transportation of male gametes, but also as a food reward for potential pollinators. Many findings provide compelling evidence that pollen acts, in addition, as a visual signal. The present contribution reviews several strategies that angiosperms have evolved to attract potential pollinators to the site of reward. We here consider evolutionary, ecological, sensory-physiological, and behavioural aspects of flower-pollinator interactions that are correlated with visual signals provided by pollen and pollen-producing organs, or imitations thereof.

Abstract: The development of the inflorescence, microspores and anthesis were well synchronized among individuals or in the panicles of barley under controlled environmental conditions. To study the effects of high-temperature stress on the development of pollen mother cells (PMCs) and microspores, the plants were subjected to high temperature treatment at different stages of reproductive growth. When plants were exposed to high temperature for five days at the early differentiation stage of the panicle, pollen grains had apparently normal exine but no or little cytoplasm. At the pre-meiotic stage of PMCs, high temperature caused subsequent development of short anthers possessing no pollen grains. When plants were exposed to high temperature during meiosis of PMCs, all pollen grains possessed exine and were swollen but showed little starch accumulation. In these plants treated at high temperature, the panicles at the heading stage had a normal appearance, but their seeds were virtually sterile. These results indicated that there are at least three stages of reproductive growth hypersensitive to high temperature, which resulted in abnormal terminal phenotypes different from one another.

Abstract: The pollen competition hypothesis predicts that when the number of pollen grains deposited onto stigmas exceeds the number of ovules, selection can operate in the time frame between deposition and fertilization. Moreover, because of the overlap in gene expression between the two phases of the life cycle, selection on microgametophytes may alter the resulting sporophytic generation. The extent to which pollen competition occurs in nature has been unclear, because tests of the predictions of the pollen competition hypothesis have used cultivars and/or artificial growth conditions and hand-pollination techniques. In this study we used a wild species, Cucurbita foetidissima, in its natural habitat (southern New Mexico) to determine the amount and timing of the arrival of pollen onto stigmas, the relationship between pollen deposition and seed number, and the effects of the intensity of pollen competition on progeny vigor. We found that ;900 pollen grains are necessary for full seed set and that a single visit by a pollinator results in the deposition of 653.0 6 101.8 pollen grains. About 29% of the flowers receiving a single pollinator visit had 900 or more pollen grains on its stigma. Moreover, within 2 h of anthesis, .4000 pollen grains were deposited onto a typical stigma, indicating that multiple pollinator visits must have occurred. Fruits produced by multiple visits had greater seed numbers (206 vs. 147) than fruits produced by a single visit. Finally, the progeny produced by multiple pollinator visits were more vigorous than those produced by single visits with respect to five measures of vegetative growth (MANCOVA, Wilks’ lambda 5 0.96, F6,370 5 2.54, P , 0.02. These data demonstrate that conditions for pollen competition exist in nature and support the prediction that pollen competition enhances offspring vigor.

Abstract: Background: The allergic reaction is characterized by an inflammatory response, which is correlated to the allergen exposure, and is detectable in mite allergic patients, even when symptoms are absent. Objective: The study was aimed at assessing the presence of inflammation in patients with pollen allergy during a long observation period. Methods: Six patients, sensitized only to Betula alba, were enrolled. Evaluated parameters were (1) nasal symptoms, (2) inflammatory markers (ie, neutrophil and eosinophil number and intercellular adhesion molecule-1 expression on nasal epithelial cells), and (3) pollen count. Patients were examined during the pollen season every 4 days for 40 days and were observed 3 times after the pollen season. Results: A significant inflammatory reaction was evident throughout the pollen season, even during the days with a low pollen count and low or absent symptoms. Conclusions: The results of this study indicate that the global therapeutic strategy for allergic rhinitis should be revised and targeted to inflammatory phenomena rather than to symptoms alone. (J Allergy Clin Immunol 2000;105:54-7.)

Abstract: Background: We assessed the efficacy of preseasonal local allergoid immunotherapy in a group of children with asthma and/or rhinitis and/or rhinoconjunctivitis due to grass pollen. Methods: We randomly assigned 24 children allergic to grass pollen to receivelocal allergoid immunotherapy for 3 months before the pollen season and 24 such patients to receive identically appearing placebo. The immunotherapy consisted of tablets of monomeric allergoid grass pollen allergens held in the mouth until they dissolved and then swallowed. The study was double-blind. Symptoms and medications were scored on diary cards during the pollen season. Nasal eosinophil cationic protein levels were measured by the monoclonal antibodies EG1 and EG2 outside the pollen season and at low and at high pollen concentration during the pollen season. Results: The active-treatment group had a statistically significant reduction of total symptoms (P<0.05), especially bronchial symptoms (P<0.05), in comparison with the placebo group. Immunotherapy was well tolerated and compliance was good. Nasal levels of EG2 and EG1 increased significantly during the pollen season, but there was no difference between groups. EG2/EG1 increased significantly only in the placebo group during natural allergen exposure (P<0.01). Conclusions: Our results suggest that this immunotherapy is effective for the treatment of asthma due to grass pollen in children.

Abstract: Pollen studies make important contributions nature, into three main themes: pollen strucs to our knowledge in many interdisciplinary ture and constituents, pollen evolutionary arenas. Pollen identification is widely used in ecology and the pollen-pollinator interface. reconstruction of, e.g., vegetation, the climate Several papers overlap somewhat or are of the past, and plant biodiversity. Studies perhaps even somewhat contradictory and concerning pollen structure, size and form are reflect the author's own ideas and experience. key issues in basic sciences, as, e.g., plant Some could be understood more deeply by taxonomy and evolution, but are also of consulting other closely related articles. The importance in applied fields as, e.g., plant reader is strongly referred to the respective breeding. In pollination studies pollen is literature list of each article. generally used specifically to identify food ofanther ripening and pollen The last steps development (Pacini) and the mature pollen sources of visitors and to reconstruct their foraging routes. Fewer have been devoted to wall structure (Hesse) are key factors to pollen collection mechanisms and to the strucs understand pollen dispersal mechanisms in ture and content of pollen in relation to its biotic pollination (Stroo) as well as abiotic pollination (Ackerman). Pollen size, shape, function.

Abstract: In self-incompatible plants, interference by self pollen or genetically related pollen can potentially exacerbate pollen limitation, although this has rarely been demonstrated. We examined the breeding system, pollen limitation, and pollen interference using self- and cross- pollinations and pollen supplementations in Burchardia umbellata, an insect-pollinated lilioid monocot. Ovule fertilization and seed set were less following selfing than crossing (22 vs. 78% and 4 vs. 73%, respectively), indicating partial self-incompatibility. Flowers were partially protandrous, and flowers opened concurrently on plants potentially allowing self pollen interference. Natural seed set was pollen limited and varied within and among years, probably due to variation in flowering plant density. Interference by self or genetically related pollen caused pollen limitation as evidenced by increased seed set of bagged cross-pollinated plants compared to unbagged pollen-supplemented plants in two years. In 1996, both fertilization and seed set increased in response to cross-pollination, indicating that interference occurred in the style and ovary. In 1997, only seed set increased after cross-pollination indicating that interference occurred in the ovary. Inappropriate pollen deposition may contribute to pollen limitation more often than previously recognized and should select for floral traits that decrease deposition of self or related pollen. interference. Many flowering plants rely on pollinators to deposit sufficient compatible pollen onto stigmas for ovule fertilization and seed production. Pollen limitation occurs when seed production is less than would be achieved if overall quantity or quality of pollen deposited onto stigmas were increased. Pollen quantity may be limiting if

Abstract: Adhesion of pollen grains to the stigmatic surface is a critical step during sexual reproduction in plants. In Brassica, S locus-related glycoprotein 1 (SLR1), a stigma-specific protein belonging to the S gene family of proteins, has been shown to be involved in this step. However, the identity of the interacting counterpart in pollen and the molecular mechanism of this interaction have not been determined. Using an optical biosensor immobilized with S gene family proteins, we detected strong SLR1-binding activity in pollen coat extracts of Brassica campestris. Two SLR1-binding proteins, named SLR1-BP1 and SLR1-BP2, were identified and purified by the combination of SLR1 affinity column chromatography and reverse-phase HPLC. Sequence analyses revealed that these two proteins (i) differ only in that a proline residue near the N terminus is hydroxylated in SLR1-BP1 but not in SLR1-BP2, and (ii) are members of the class A pollen coat protein (PCP) family, which includes PCP-A1, an SLG (S locus glycoprotein)-binding protein isolated from Brassica oleracea. Kinetic analysis showed that SLR1-BP1 and SLR1-BP2 specifically bound SLR1 with high affinity (K(d) = 5.6 and 4.4 nM, respectively). The SLR1-BP gene was specifically expressed in pollen at late stages of development, and its sequence is highly conserved in Brassica species with the A genome.