Pollen Mixture Research Articles

Coleopteran-specific Cry3Bb Toxin from Transgenic Corn Pollen Does Not Affect the Fitness of a Nontarget Species,Coleomegilla maculataDeGeer (Coleoptera: Coccinellidae)

Coleomegilla maculata DeGeer is a polyphagous predator that is important for suppressing pest populations in corn. We evaluated the impact of Cry3Bb-expressing transgenic corn pollen (event MON863) on C. maculata fitness parameters in the laboratory. C. maculata larvae were fed mixtures of pollen containing 0, 25, 50, 75, or 100% transgenic pollen, aphids, or were not fed; and the duration of each instar and pupal weight were compared among treatments. In a second trial, other C. maculata larvae were reared on one of the pollen mixtures or artificial diet; and the duration of larval and pupal stages, pupal weight, adult mobility, adult survivorship, and female fecundity were compared among treatments. There were no differences in any of the fitness parameters among C. maculata in the treatments fed different mixtures of pollen. Beetles in the pollen mixture treatments had faster larval development times, greater larval survivorship, and greater pupal weight than the beetles fed only aphids or an artificial diet. We conclude that we did not detect any effects on the fitness of C. maculata that ingested pollen from event MON863. However, these results do not necessarily apply to other transgenic crops expressing toxins specific to Coleoptera.

Polinização natural e artificial da cherimóia (Annona cherimola Mill.) no Estado de São Paulo

Para estudar o método de polinização da cherimóia (Annona cherimola Mill.) que produza frutos em maior quantidade e melhor qualidade, instalou-se experimento em Pedra Bela - SP, a 1150 metros de altitude. As plantas eram de pé-franco, de 20 anos de idade. O experimento foi realizado em dois períodos, tendo o primeiro se iniciado em novembro de 1999 e finalizado em junho de 2000; o segundo iniciou-se em novembro de 2000 e encerrado em junho de 2001. A montagem do experimento foi efetuada no delineamento de blocos ao acaso, com 3 tratamentos e 12 repetições. Os tratamentos foram os seguintes: 1) polinização natural; 2) polinização manual cruzada; 3) autopolinização (flores ensacadas). As plantas dos blocos foram polinizadas em diferentes dias. Avaliaram-se o vingamento dos frutos 10 dias após a polinização e a quantidade dos frutos com conformação perfeita ou defeituosa aos 40 dias. Após a colheita, os frutos foram pesados individualmente. Foram também amostrados dois frutos de cada bloco (em 1999) e três (em 2000), para as seguintes determinações: massa das sementes e da polpa, número de sementes por 100 gramas de polpa. A polinização artificial proporcionou maior vingamento de frutos (61,2% e 50,4%) em relação à polinização natural (19,6% e 3,3%), nos anos de 1999 e 2000, respectivamente. Foi mais efetiva quando realizada sob condições de temperatura variando de 17 ºC até 22 ºC e umidade relativa do ar entre 70 e 80 %. Verificou-se também aumento da porcentagem de frutos perfeitos (em 2000), da massa do fruto e do índice de sementes/100 g de polpa dos frutos polinizados artificialmente em relação aos polinizados naturalmente.

Pollen mixture, a recombination alternative in common bean breeding

Recombination can be a limiting factor in recurrent selection programs, especially in species such as common bean where artificial hybridization reduces seed setting. Therefore, the development of new techniques to improve recombination efficiency is important. Experiments were conducted in two sowing periods, July (autumn-winter season) and February (dry growing season), using the ESAL 686 and Small White cultivars as the female parents and the Carioca, Ouro Negro and ESAL 686 or Small White cultivars as male parents, depending on the female parent. Five technicians with different hybridization experience and two hybridization methods (with and without emasculation) were tested. A mixture of pollen from the three male parents was used in the pollinations, which were made in greenhouse. The resulting F 1 seeds were sown in the field and the male parent identified by morphological markers. The greatest pollination success (68%) occurred in the autumnwinter season without emasculation. It was also observed that the technicians differed in efficiency, but no method x technician interaction was detected. The pollen mixture was efficient to improve recombination.

EFFECTS OF FRACTIONS OF TRAFFIC PARTICULATE MATTER ON TH2-CYTOKINES, IgE LEVELS, AND BRONCHIAL HYPERRESPONSIVENESS IN MICE

In recent decades an increased prevalence of allergic conditions has been observed in developed countries. Although lifestyles, exposure to infection, and diet are all likely important factors, many studies have also shown a strong link between industrialization and allergy. The aim of this study was to investigate which extract fractions from traffic particulate matter (TPM, collected in a tunnel in Prague) have the greatest impact on different inflammatory and immunological parameters, such as cytokine production, levels of immunoglobulin E (IgE) antibodies, and bronchial hyperresponsiveness (BHR) in mice, when the extracts are used together with birch pollen for immunization. BP2 mice were immunized with birch pollen and different fractions of TPM (fractions 1-8). They were provoked intranasally with a mixture of pollen and TPM or pollen alone before they were challenged with methacholine. The BHR was evaluated in a whole-body plethysmograph. Th2 cytokines and fibronectin concentrations were measured, and differential cell counts were performed in the bronchioalveolar lavage (BAL) fluid. Sera were collected for determination of antibody titers. The highest titers of IgE and the highest BHR were found in the positive control mice (immunized and provoked with a mixture of pollen and TPM), followed by mice immunized with pollen and fraction 2 (which contains organic acids). Fraction 2 also induced the highest number of eosinophils and increased levels of interleukin 5 (IL-5) in the BAL fluid. The highest levels of IL-5, in BAL fluid and sera, were obtained in mice immunized with fraction 6 (moderately polar compounds), a somewhat surprising result since those mice did not produce any IgE, did not have any eosinophils in their BAL, or showed almost no BHR. Our data demonstrate that fractions 2 (organic acids) and 7 (highly polar compounds) seem to contain potential adjuvants stimulating the IL-5 production, the IgE synthesis, the eosinophil recruitment, and the bronchial hyperreactivity. Further characterization at the molecular level is now necessary to be able to identify the exact nature of those potential adjuvants. This will be of help in the future to improve the quality of the urban air aerosol.

Genotype-environment interactions in pollen competitive ability in an anemophilous tree, Betula pendula Roth.

This study describes genotype-environment interactions in pollen competitive ability expressed as pollen-tube growth rate and seed-siring success in Betula pendula Roth. A factorial crossing design was applied using the same maternal and paternal clones in two different environmental conditions, in a B. pendula seed orchard established in a greenhouse and at an outdoor clone collection. Both single donor and two-donor pollinations were employed. Female inflorescences were collected after a fixed time of germination, pollen-tube lengths were measured for each cross, and paternity of the seeds sired by two-donor pollen mixtures was analyzed using isozyme markers. The pollination site had a significant influence on pollen-tube growth rate and seed-siring success. Significant interactions between pollination site and pollen donor indicated genotype-environment interactions in pollen-tube growth rate and seed-siring success. A highly significant positive correlation between pollen-tube growth rate and seed-siring success was found in the greenhouse but not at the outdoor clone collection. These results suggest that the pollen-tube growth rate can be a predictor of seed-siring success in controlled greenhouse conditions, where differences among maternal plants are mainly of genetic origin, but not in more heterogeneous outdoor conditions. In natural birch stands, environmental maternal effects probably diminish the significance of pollen competition for sexual selection in Betula pendula. At seed orchards, the effects of environmental conditions on pollen competitive ability can have important consequences for the genetic composition of the seed crop.

Effects of birch pollen and traffic particulate matter on Th2 cytokines, immunoglobulin E levels and bronchial hyper-responsiveness in mice.

Health effects due to air pollution arising from motor vehicles are a major public and political concern world-wide. Epidemiological studies have shown that the manifestations of asthma are increased by air pollution in already affected individuals. To investigate the potential role of air-polluted tunnel dust (traffic particulate matter, TPM) or pure carbon core particles in the initiation and persistence of experimental allergic inflammation. BP2 mice were immunized with birch pollen alone (group B) or pollen together with TPM (group A), or with birch pollen and Al(OH)3 (group C), or with birch pollen and carbon core particles (group D). Before methacholine challenge they were challenged intranasally and thereafter bronchial hyper-reactivity (BHR) was evaluated in a whole-body plethysmograph. Levels of Th2 cytokines, fibronectin and lactate dehydrogenase (LDH) were determined, and differential counts were performed in the bronchoalveolar lavage (BAL) fluid. Sera were collected for determination of antibody titres and cytokine levels. Specific IgE titres, BHR, the number of recruited eosinophils and levels of fibronectin and LDH in BAL were increased in mice immunized and challenged with a mixture of birch pollen and TPM. However, mice immunized with birch pollen alone and challenged intranasally with pollen or a mixture of pollen and TPM demonstrated the highest levels of IL-4 and IL-5. This study highlights the importance of the exposure to a combination of particulate matters and pollen allergens, in the induction of allergic disease in the airways, and we have demonstrated that polluted tunnel dust has an effect on both the inflammatory and immunological components of experimental allergy. Immunization and challenge with carbon core particles together with birch pollen increased neither the BHR nor the specific IgE production significantly. Our results therefore strongly suggest that it is most likely to be the organic phase bound to the carbon core of the diesel exhaust particles that might have an important adjuvant effect in the induction of experimental allergy.

Introgression in black poplar (Populus nigra L. ssp. nigra) and its transmission

: Introgression was studied in the offspring of P. nigra and P. × canadensis female trees using 3 enzymatic patterns (6-PGD, LAP and GPI). Our investigations were aimed at the offspring from open pollination and controlled pollination by the pollen mixture from 3 clones of P. nigra and 3 clones of P. × canadensis. The rate of introgression in <br /> P. nigra offspring from open pollination was 0.67%. Introgression in 3 offsprings of P. nigra from controlled pollination was 7.14%. In P. × canadensis the heterozygous: homozygous phenotype ratio in 2 offsprings from open pollination was 1:1; it documents prevailing fertilization by P. nigra pollen. After fertilization by the pollen mixture this segregation ratio was 1:1 (6-PGD; GPI) or 3:1 (LAP). The offspring of P. × canadensis maternal components are assumed to have a major share in introgression.

Pollen competition and seed-siring success in Picea abies.

The aim of the present work was to study pollen-tube competition in Picea abies. Controlled crossings were performed with pollen mixtures including pairs of pollen lots with fast and slowly elongating pollen-tubes. Paternity analysis using isozyme markers was performed on the progenies in order to study whether the in vitro pollen-germination vigour corresponds to the proportion of seeds sired by the pollen donor. Paternal success was found to be unequal, 15 out of 23 crossings producing progeny that differed significantly from the hypothetical ratio of 1:1. The paternal contribution in the majority of the crossings was as expected: the pollen parent with more-vigorous in vitro germination sired more seeds than the less-vigorous pollen. In the case of two pollen mixtures, however, the seed-siring success summed over the maternal trees was the opposite to the expected value. Despite these aberrations, the results support the hypothesis that pollen-tube competition is one of the factors contributing to male fitness in P. abies.However, when all the other factors affecting pollination and seed set under natural conditions are taken into account, it is clear that the seed-siring success of a particular paternal genotype cannot be predicted reliably by measuring only the in vitro pollen vigour.

Pollen competition between cultivated and wild rice species (Oryza sativaandO. rufipogon)

Summary Post‐pollination competition is reported here in cultivated rice (Oryza sativa) and a perennial wild rice (O. rufipogon) to investigate the occurrence of crop‐to‐wild gene flow. Wild and cultivated rice (variety Minghui‐63) were grown in a common garden in Hunan province, China, and crop‐specific genetic markers were used to detect hybridization following hand‐pollinations. Using 11 sequential pollination treatments, the effects of the relative timing of pollination on the success of foreign pollen was investigated. Foreign pollen from the crop resulted in lower pollen germination, fewer pollen tubes per style, and a significant reduction of seed set, demonstrating a disadvantage of foreign pollen even in the absence of pollen competition. When 1 : 1 pollen mixtures were applied, only 2% of the resulting seeds were hybrids, revealing a much stronger disadvantage of foreign pollen when competing with conspecific pollen. Testing the effects of the relative timing of pollination on the success of foreign pollen suggested that conspecific pollen is often more successful than foreign pollen. Nonetheless, hybridization is possible following the deposition of pollen mixtures, especially when foreign pollen arrives earlier than conspecific pollen. Pollen competition between wild and cultivated rice could slow the rate of crop‐to‐wild gene flow, but even if pollen competition was ubiquitous it would not prevent gene flow from the crop.

Widening the genetic basis of virus resistance in tomato

New and old accessions of Lycopersicon chilense (LA 1932, LA 1938 and LA 1963), L. peruvianum (PI-143679 and PI-126944), and L. hirsutum (UPV-16910) recently reported as resistant to TSWV, TYLCV and PepMV, viral agents responsible for the most severe economic losses in tomato crop in Spain, were selected as male parents for crosses with L. esculentum in order to widen the genetic basis of virus resistance in tomato. After the analysis of the nature and severity of the barriers found in each case, crossability barriers between wild and cultivated species were circumvented by using several techniques. Crosses with pollen mixture (1:1, wild:cultivated) were successful in obtaining interspecific hybrids with the two L. chilense accessions (LA 1932 and LA 1963) that exhibited more relaxed post-cigotic barriers. Embryo rescue was also successful in crosses with these accessions and also in crosses with L. peruvianum accessions in which embryos beyond the globular stage were found (PI-143679). However, a combination of two or more strategies was necessary in crosses with those accessions exhibiting more severe crossability barriers. Mixture pollen crosses (10:1, wild:cultivated), combined with embryo rescue, allowed for the recovery of hybrids with LA 1938. Stigma and pistil complementation with H 3BO 3 and GA 3, followed by immature seed culture was found to be an effective method for the production of novel interspecific hybrids with PI-126944, the accession with the highest level of resistance to TSWV and TYLCV. By using the best combination of techniques we also backcrossed these interspecific hybrids to tomato to generate the BC1 progeny. The new genes introgressed will be useful to obtain new tomato varieties resistant to TSWV, TYLCV and PepMV.

Trade-offs between male and female reproduction associated with allozyme variation in phosphoglucoisomerase in an annual plant (Clarkia unguiculata: Onagraceae).

The genotype of an individual for allozymes such as phosphoglucoisomerase (Pgi) is often not neutral with regard to fitness. Studies of several taxa have found consistent fitness differences among Pgi genotypes expressing different allozymes. We conducted a greenhouse experiment with Clarkia unguiculata to determine whether allelic variation at the Pgi-C1 locus may affect components of male and female function. We found significant differences in siring success between pollen donors homozygous for different Pgi alleles. When a mixture of pollen was applied to stigmas under conditions of gametophytic competition (more pollen deposited on stigmas than there are ovules available to fertilize), donors homozygous for the C allele of Pgi sired more seeds per fruit than B-allele donors. Differences between genotypes with respect to female fertility per fruit contrasted with the male advantage associated with the C allele. Recipients homozygous for the C allele produced fruits with more aborted seeds and fewer viable seeds than recipients homozygous for the B allele. These results suggest that allelic variation at a single locus may have opposing effects on male and female reproductive success in C. unguiculata, and that trade-offs between the two types of reproductive success could contribute to the maintenance of variation at the Pgi-C1 locus.

Effects of mycorrhizal infection and soil phosphorus availability on in vitro and in vivo pollen performance in Lycopersicon esculentum (Solanaceae)

The effects of mycorrhizal infection and soil P availability on in vitro and in vivo pollen performance were studied in two cultivars of tomato (Lycopersicon esculentum). In the first study, plants were grown in a greenhouse under three treatment combinations: nonmycorrhizal, low P (NMPO); nonmycorrhizal, high P (NMP3); and mycorrhizal, low P (MPO). Mycorrhizal infection and high soil P conditions significantly increased in vitro pollen tube growth rates but not percentage of germination. In addition, pollen from NMP3 and MPO plants sired significantly more seeds than pollen from NMPO plants in pollen mixture studies. In the second study, plants were grown initially in a greenhouse under two treatment combinations: NMPO and MPO. After all plants began to flower, they were placed in experimental arrays in the field. Under open pollination, pollen from MPO plants sired significantly more seeds than pollen from NMPO plants. This result was primarily attributed to increased flower production (and thus pollen production) in MPO plants. Thus, mycorrhizal infection and high soil P conditions can increase pollen quality (in vitro and in vivo pollen performance) as well as pollen quantity, thereby enhancing fitness through the male function. Anthocyanin production (used to determine paternity) also affected pollen performance.

Competition for pollination: effects of pollen of an invasive plant on seed set of a native congener.

Competition for pollination can be an important factor in plant reproduction, but little attention has been given to the effect of the growing number of invasive plant species on pollination of native species. As a first step in understanding this threat, we used hand pollination to investigate the effects of pollen from an invasive species (Lythrum salicaria) on seed set in a sympatric and co-flowering native congener (L. alatum). Dispersal of fluorescent dyes in the field confirms that pollinators (bumble bees and honey bees) transfer pollen between species. To determine the potential effect of such interspecific pollen transfer on seed set of the native, we pollinated 773 flowers on 20 plants with one of three treatments: legitimate conspecific pollen, a mixture of conspecific and foreign pollen, and foreign pollen. The mixed-pollen treatment resulted in 28.8% lower seed set relative to conspecific pollination. Foreign crosses resulted in extremely low seed set. Observations of pollen germination indicate that events at the stigmatic surface contribute to the reduction in seed set for mixed pollination. Our results indicate that the impacts of invasive species may extend beyond vegetative competition to include competition for pollination.

Terrestrial organic matter in surface sediments of the Baltic Sea, Northwest Europe, as determined by CuO oxidation

We studied the distribution and composition of terrestrial organic matter in sediments of the Baltic Sea (Northwest Europe). To this end, surface sediments from all basins of the Baltic Sea were analyzed for their lignin oxidation product yields and compositions after CuO oxidation. Lignin oxidation product yields depend on the concentration of organic carbon and range from 0.4 to 10.2 mg g −1 total organic carbon (TOC). On the basis of an average of 13 mg g −1 TOC in two river sediments, we estimate that the upper limit of terrestrial organic matter in Baltic Sea sediments is 30% of TOC. The contribution of terrestrial organic matter differed between the individual basins, depending on the distance from runoff discharge areas and on the area occupied by each submarine catchment. Lignin composition showed a relative decrease of angiosperm tissue from the Southwest to the Northeast, reflecting the shift from temperate to boreal vegetation type. The Gotland and the Bornholm Seas, which have no significant river input, were characterized by high relative contributions of nonwoody, strongly altered material. The source may either be a mixture of pollen and peat being eroded from geologically older strata at the seafloor or laterally advected material from the other basins. However, the pronounced compositional differences between the basins indicated that interbasin transport of terrestrial organic matter is less important than direct river input, although river signals can only be traced at a few places in the Baltic Sea.

TRADE-OFFS BETWEEN MALE AND FEMALE REPRODUCTION ASSOCIATED WITH ALLOZYME VARIATION IN PHOSPHOGLUCOISOMERASE IN AN ANNUAL PLANT (CLARKIA UNGUICULATA: ONAGRACEAE)

The genotype of an individual for allozymes such as phosphoglucoisomerase (Pgi) is often not neutral with regard to fitness. Studies of several taxa have found consistent fitness differences among Pgi genotypes expressing different allozymes. We conducted a greenhouse experiment with Clarkia unguiculata to determine whether allelic variation at the Pgi-C1 locus may affect components of male and female function. We found significant differences in siring success between pollen donors homozygous for different Pgi alleles. When a mixture of pollen was applied to stigmas under conditions of gametophytic competition (more pollen deposited on stigmas than there are ovules available to fertilize), donors homozygous for the C allele of Pgi sired more seeds per fruit than B-allele donors. Differences between genotypes with respect to female fertility per fruit contrasted with the male advantage associated with the C allele. Recipients homozygous for the C allele produced fruits with more aborted seeds and fewer viable seeds than recipients homozygous for the B allele. These results suggest that allelic variation at a single locus may have opposing effects on male and female reproductive success in C. unguiculata, and that trade-offs between the two types of reproductive success could contribute to the maintenance of variation at the Pgi-C1 locus.Corresponding Editor: O. Savolainen

Effect of pollen load size and source (self, outcross) on seed and fruit production in highbush blueberry cv. ‘Bluecrop’ (VACCINIUM CORYMBOSUM; Ericaceae)

Reproductive fitness of a plant is ultimately determined by both number and quality of seed offspring. This is determined by sexual selection of pollen microspores and ovules during pollination and fertilization. These processes may include pollen competition and seed abortion, which reduce the number of microspores and ovules available for final seed production. Thus, even an excess of pollen microspores to ovules does not result in fertile seeds equal to ovule number. We investigated pollen requirements of highbush blueberry (Vaccinium corymbosum cultivar 'Bluecrop') for maximal seed production and how fertile seed number translates into fruit quality, since fruit quality would ultimately determine the dispersal of its offspring. We demonstrate that individual blueberry flowers with a mean of 106 ovules reach their maximum fruit set and mass and minimum time to ripen when 125 outcross pollen tetrads pollinate a flower, compared to 10 or 25. Three hundred tetrads resulted in the increase of fertile seeds, but did not result in a further increase of fruit mass or fruit set, or decrease in time to ripen. We also examined the effect of pure and mixed loads of self and outcross pollen (25 and 125 tetrads), and found no differences in fertile seed number, fruit mass, or percentage fruit set when pollen loads were either 25 self or outcross pollen tetrads, although number of days to ripen was significantly shorter by 8 d with 25 outcross tetrads. When the pollen load of 125 tetrads consisted of self or a 50:50 mixture of self and outcross pollen, fruit mass, days to ripen, and percentage fruit set were not different from loads of 125 outcross pollen. In addition, a pollen load of 25 outcross tetrads resulted in fertile seed number and fruit quality in between that of 25 self, and 125 self, 125 mixed, or 125 outcross tetrads. Large, small, and flat seed types were identified, and only large seeds (length = 1.7 mm) were fertile. These results improve our understanding of pollen load size and source requirements of a crop plant and the limits to pollen transfer when translated to fruit growth.

COMPONENTS OF REPRODUCTIVE SUCCESS IN TWO DIOECIOUS FIG SPECIES,FICUS EXASPERATAANDFICUS HISPIDA

We studied components of reproductive success in two dioecious fig species, Ficus exasperata and F. hispida, at a deciduous and an evergreen site in south India, over a two-year period. Unlike monoecious figs, which produce a mixture of wasps, seeds, and pollen in every fruit, dioecious fig species have hermaphrodite (functionally male) trees, which produce wasps and pollen only, and female trees, which produce seeds only. Di- oecious fig species are free of the seed-wasp trade-off involved within each monoecious fig fruit; wasp and seed production in dioecious species reflect selective pressures acting on each sex to maximize offspring production. We studied specific predictions of the reproductive behavior of dioecious fig species, predictions that were made based on studies of monoecious figs. Pollinator (and associated parasitic wasp) arrivals at trees bearing receptive inflores- cences were closely linked to the seasonal variation in receptive fig numbers. More wasps arrived at female trees of F. exasperata than at trees, apparently because of syn- chronized wasp release by male trees during the time of synchronous female tree receptivity. Wasp numbers arriving at male and female trees of F. hispida did not differ significantly, apparently because inflorescence production on male and female trees was more evenly spread over time. Female inflorescences had more pollinators than male inflorescences in F. hispida; female and male inflorescences had approximately equal numbers of pollinators in F. exasperata. Hand-pollination experiments introducing one, three, or eight pollinators into figs of F. hispida showed that foundress number was a significant predictor of resultant wasp or seed number. In both fig species, female inflorescences initiated more flowers than male inflorescences, and produced more seeds than male inflorescences produced wasps. Female fruits were not larger than male fruits at maturity in either fig species. Only in F. exasperata were male fruits significantly smaller in the dry than in the wet site. Both species had a relatively high average level of pollination (>70%). The mean percentage of fruit matured was 56% in F. hispida, and a surprisingly high 78% in F. exasperata. Both fig species exhibited strong sex differences in foundress numbers, numbers of flowers initiated, and numbers of wasps or seeds matured, suggesting strong selection on components of reproductive success within each sex after dioecy evolved. Our quantitative analyses elucidate how some of the many factors in plant-pollinator relationships affect plant reproductive success in the fig-wasp mutualism.

Interspecific Hybridization in Hazelnut (Corylus)

Eight Corylus L. (hazelnut) species were intercrossed in all possible combinations to reveal genetic relationships. Pollinations were made on either individually bagged branches or trees covered entirely with polyethylene using mixtures of pollen of five genotypes to minimize low cluster set due to single incompatible combinations. Percent cluster set, seed germination, and hybrid seedling survival were determined. Hybridity of seedlings was verified by inspection of morphological traits. Based on percent cluster set, seed germination, and hybrid seedling survival along with observed morphological similarities, Corylus species were placed in three groups: 1) the tree hazels C. colurna L. (turkish tree hazel) and C. chinensis Franchet (chinese tree hazel), 2) the bristle-husked shrub species C. cornuta Marshall (beaked hazel), C. californica (A.DC.) Rose (california hazel), and C. sieboldiana Blume (manchurian hazel), and 3) the leafy-husked shrub species C. avellana L. (european hazel), C. americana Marshall (american hazel), C. heterophylla Fischer (siberian hazel), and C. heterophylla Fischer var. sutchuensis Franchet (sichuan hazel). The two tree hazel species crossed with each other readily, as did the three bristle-husked shrub species. The frequency of blanks was low (<20%) for crosses of the tree hazels, and <50% for interspecific crosses within the group of bristle-husked species. The leafy-husked shrub species could be crossed with each other in all directions, although cluster set on C. heterophylla was low. For crosses of species belonging to different groups, set was generally low and the frequency of blanks high. Nevertheless, a few hybrid seedlings were obtained from several combinations. When used as the female parent, C. californica set nuts when crossed with all other species, indicating possible value as a bridge species. Crosses involving C. avellana were more successful when it was the pollen parent. In crosses with C. avellana pollen, cluster set on C. chinensis was better than on C. colurna and the frequency of blanks was much lower, indicating that it might be easier to transfer nonsuckering growth habit from C. chinensis than from C. colurna. Reciprocal differences in the success of crosses was observed. The following crosses were successful C. californica × C. avellana, C. chinensis × C. avellana, C americana × C. heterophylla, C. cornuta × C heterophylla, C. californica × C. colurna, and C. americana × C. sieboldiana, but the reciprocals were not.

Absence of conspecific pollen advantage in the dynamics of anIpomopsis(Polemoniaceae) hybrid zone

The frequency of hybrid formation in angiosperms depends on how often heterospecific pollen is transferred to the stigma and on the success of that heterospecific pollen at fertilizing ovules. Even if heterospecific pollen is capable of effecting fertilization it may perform poorly when conspecific pollen is also available on the stigma. We applied pollen mixtures to stigmas to determine how pollen interactions affect siring success and the frequency of hybrid formation between two species of Ipomopsis (Polemoniaceae) in Colorado. Plants of both parental species and natural hybrids were pollinated with I. aggregata and I. tenuituba pollen in ratios of 100:0, 80:20, 50:50, 20:80, and 0:100 by mass. Plants were homozygous for different alleles at an isozyme marker, allowing us to distinguish the type of pollen parent for 2166 viable seeds from 273 fruits. In contrast to studies of many other hybridizing taxa, there was no evidence of an advantage to conspecific pollen, nor did composition of the stigmatic pollen load affect seed set. Instead, the frequency of seeds sired by a given species was proportional to its representation in the pollen load. In this hybrid zone, both the frequency of first-generation hybrid formation and the relative male fitness of the two parental species should be predictable from the rates of pollen transfer to stigmas.

Floral biology and breeding system of pohutukawa (Metrosideros excelsa, Myrtaceae)

The floral biology and breeding system of pohutukawa (Metrosideros excelsa, Myrtaceae), a mass‐flowering tree of northern New Zealand coastlines, were examined. Trees flower over a peak period of 2 weeks, and compound inflorescences contain an average of 14.3 showy, hermaphrodite, red brush flowers that remain open for 7 days. A brief female flower stage (mean duration 1.3 d) is followed by the main hermaphrodite phase that lasts for 4 days. Neither dichogamy nor herkogamy is important in preventing pollen and stigma interference. Pollen is highly viable (93.6%), and stigma receptivity extends for at least 9 days, as indicated by peroxidase activity, pollen germination, pollen tube length 24 h after pollination, and seed set. Stigmatic exudate production appears to increase up to 5 days post‐anthesis. On average, flowers produce 46 p1 nectar per day, containing 18% (w/v) sucrose. Floral design and display of pohutukawa are consistent with high levels of autogamous and geitonogamous self‐pollination. Controlled pollination experiments were used to assess the effect of self‐ and cross‐pollen and a pollen mixture from five unrelated parents on capsule and seed production, and on pollen tube growth in seven trees. Three trees in the experiment were self‐incompatible, as quantified by the index of self‐incompatibility (ISI), indicating that natural populations may consist of a mosaic of self‐compatible and incompatible individuals. Self‐in‐compatibility is late‐acting as pollen tubes from selfs and crosses reached the ovary simultaneously at 10–15 d after pollination. In common with other Myrtaceae, the seed/ovule ratio in pohutukawa is low and this is likely to be genetically determined rather than limited by stigmatic pollen load. Germination of fertile seeds from all pollination treatments was equally high (98.4%), indicating that no inbreeding depression is acting at this stage of the life cycle. The pollen/ovule ratio of 462.5 (s.e. ±43.3) places the breeding system of pohutukawa between facultative selfing and facultative outcrossing.