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Recruitment of CRABS CLAW to promote nectary development within the eudicot clade.

Nectaries are secretory organs that are widely present in flowering plants that function to attract floral pollinators. Owing to diversity in nectary positions and structures, they are thought to have originated multiple times during angiosperm evolution, with their potential contribution to the diversification of flowering plants and pollinating animals being considerable. We investigated the genetic basis of diverse nectary forms in eudicot angiosperm species using CRABS CLAW (CRC), a gene required for nectaries in Arabidopsis. CRC expression is conserved in morphologically different nectaries from several core eudicot species and is required for nectary development in both rosids and asterids, two major phylogenetic lineages of eudicots. However, in a basal eudicot species, no evidence of CRC expression in nectaries was found. Considering the phylogenetic distribution of nectary positions and CRC expression analyses in eudicots, we propose that diverse nectaries in core eudicots share conserved CRC gene regulation, and that derived nectary positions in eudicots have altered regulation of CRC. As the ancestral function of CRC lies in the regulation of carpel development, it may have been co-opted as a regulator of nectary development within the eudicots, concomitant with the association of nectaries with reproductive organs in derived lineages.

Arabidopsis↗

Expression of the cell cycle in sperm of Arabidopsis: implications for understanding patterns of gametogenesis and fertilization in plants and other eukaryotes.

The relationship between developmental events and the cell cycle was examined in sperm of Arabidopsis thaliana. Sperm of Arabidopsis rapidly enter the S (synthesis) phase of the cell cycle after inception from mitosis of the generative cell. Sperm in pollen grains within anthers continue to synthesize DNA, and at the time of pollination, contain approximately 1.5C DNA. Following pollination, sperm continue through the S phase of the cell cycle during pollen tube growth. By the time pollen tubes reach the ovary, sperm nuclei contain approximately 1.75C DNA. Just prior to double fertilization, sperm nuclei within embryo sacs contain the 2C quantity of DNA. These data indicate that molecular programs associated with the G1-S transition and the S phase of the cell cycle are expressed in sperm cells of developing pollen grains and pollen tubes in Arabidopsis. This pattern of prefertilization S phase activity in the sperm of a flowering plant stands in marked contrast to all other non-plant eukaryotes (from ciliates to yeast to sea urchins to mammals) where sperm remain in G1 during development, prior to the initiation of gametic fusion. In addition, when patterns of cell cycle activity in sperm of Arabidopsis and other flowering plants are compared, developmental analysis reveals that heterochronic alterations (changes in the relative timing of ontogenetic events) in cell cycle activity are a central cause of the diversification of patterns of gametogenesis in higher plants. Finally, comparative analysis of the patterns of cell cycle activity in Arabidopsis and other angiosperms may be used to predict which flowering plants will be amenable to development of successful in vitro fertilization techniques.

Arabidopsis↗

Pollen-stigma interactions in Brassica oleracea. I. Ultrastructure and physiology of the stigmatic papillar cells.

The osmotic potential (psi pi) of the stigmatic papillar cells of Brassica oleracea is -14.8 bars. In laboratory conditions each cell transpires water at rates within the range from 3 X 10(-5) to 5 X 10(-5) mm3h-1. A small increase in transpiration rate is detected following cross-(compatible) but not self-(incompatible)pollination. No significant changes in psi pi occur following pollinations of either compatibility. Electron microscopy reveals an active papillar cytoplasm apparently secreting proteins into the cell wall via small vesicles. The cuticle is discontinuous and freeze-fracture techniques indicate that channels transverse the cell wall, suggesting a possible pathway for the movement of protein molecules of high molecular weight from the cytoplasm to the stigma surface. Analysis of electron-microscopic autoradiographs of mature, self-incompatible papillae following pulse-chase experiments with L-[3H]leucine and treatment with cycloheximide shows that protein molecules secreted into the cell wall may return to the cytoplasm at a later stage. The significance of these results is discussed in terms of current models of the pollen-stigma interaction in Brassica.

Autoradiography↗

In vitro propagation by asymbiotic seed germination and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity studies of tissue culture raised plants of three medicinally important species of dendrobium.

A simple and efficient plant propagation system has been developed by asymbiotic germination of seeds in three medicinally important Dendrobium species, namely, Dendrobium tosaense, Dendrobium moniliforme, and Dendrobium linawianum. Plants obtained from natural habitats were grown in the greenhouse. The flowers were hand pollinated. Seeds of the capsules derived after 12 weeks of hand-pollination germinated asymbiotically (50-74%) on half strength Murashige and Skoog's (MS) basal medium with 3% sucrose and solidified with 0.9% Difco agar. Active growth in the germinated seedlings was achieved by re-culturing on full strength MS basal medium supplemented with 8% banana homogenate, 8% potato homogenate, 8% coconut water, 1.5% sucrose and 0.9% Difco agar. Healthy plantlets, transferred to plastic trays containing moss or moss and tree fern, successfully acclimatized (84-100%) in the greenhouse. A marked varied response was observed in the free radical scavenging activity of methanolic extracts of in vitro propagated plants, on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical using a UV spectrophotometer assay. Methanolic extracts were prepared by dissolving the powdered plant material, obtained from six months old in vitro propagated plants, each about 5 g, in boiling methanol. The percentage of scavenging effect of D. tosaense extract was 95.9% at 0.4 mg/ml concentration, whereas D. monoliforme, and D. linawianum extracts scavenged 83.4% and 92.3%, respectively, at a concentration of 0.4 mg/ml. All the extracts scavenged DPPH radical significantly in a concentration dependent manner.

Biphenyl Compounds↗

Allozyme diversity and population structure of Japanese and Korean populations of wild radish, Raphanus sativus var. hortensis f. raphanistroides (Brassicaceae).

Raphanus satiuus var. hortensis f. raphanistroides (wild radish: Brassicaceae) is an insect-pollinated wild plant that grows mainly on beaches in East Asia. Starch gel electrophoresis was used to investigate the allozyme diversity and genetic structure of 25 Japanese and 9 Korean populations of this plant. Although the Korean populations were small, isolated, and patchily distributed, they maintained a high level of genetic diversity; the average percentage of polymorphic loci was 63.1%, the mean number of alleles per locus was 2.27, and the average heterozygosity was 0.278. The corresponding estimates for these parameters in the Japanese populations were 53.3%, 2.26, and 0.278. These estimates are considerably higher than those from species with similar life history and ecological characteristics, but they are lower than those from R. raphanistrum, the wild radish that grows in Europe and the U.S.A. The combination of an insect-pollinated, outcrossing breeding system, large population sizes, gene flow from cultivated radish population, and a propensity for high fecundity may explain the high level of genetic diversity within wild populations.

Alleles↗

Alteration of the self-incompatibility phenotype in Brassica by transformation of the antisense SLG gene.

Self-incompatible (SI) Brassica rapa (syn. B. campestris) was transformed with an antisense SLG gene by using SLG8 cDNA isolated from the B. campestris S8 homozygote. Two transformed lines were obtained and analyzed. Northern blot and Western blot analyses revealed that endogenous SLG and SRK were greatly reduced of the transcriptional and translational levels in the transformant. Pollination experiments confirmed that their SI phenotype had broken down. In addition, the progeny with the antisense SLG gene, resulting from self- or cross-pollination of the transgenic plant, also showed the self-compatible phenotype. The breakdown of SI in the tranformants was due to the change in property of the stigma and not of the pollen. These results provide strong evidence that SLG and/or SRK is implicated in the pollen-stigma recognition of SI and that they act only as stigmatic factors.

Brassica↗

Conservation planning for ecosystem services.

Despite increasing attention to the human dimension of conservation projects, a rigorous, systematic methodology for planning for ecosystem services has not been developed. This is in part because flows of ecosystem services remain poorly characterized at local-to-regional scales, and their protection has not generally been made a priority. We used a spatially explicit conservation planning framework to explore the trade-offs and opportunities for aligning conservation goals for biodiversity with six ecosystem services (carbon storage, flood control, forage production, outdoor recreation, crop pollination, and water provision) in the Central Coast ecoregion of California, United States. We found weak positive and some weak negative associations between the priority areas for biodiversity conservation and the flows of the six ecosystem services across the ecoregion. Excluding the two agriculture-focused services-crop pollination and forage production-eliminates all negative correlations. We compared the degree to which four contrasting conservation network designs protect biodiversity and the flow of the six services. We found that biodiversity conservation protects substantial collateral flows of services. Targeting ecosystem services directly can meet the multiple ecosystem services and biodiversity goals more efficiently but cannot substitute for targeted biodiversity protection (biodiversity losses of 44% relative to targeting biodiversity alone). Strategically targeting only biodiversity plus the four positively associated services offers much promise (relative biodiversity losses of 7%). Here we present an initial analytical framework for integrating biodiversity and ecosystem services in conservation planning and illustrate its application. We found that although there are important potential trade-offs between conservation for biodiversity and for ecosystem services, a systematic planning framework offers scope for identifying valuable synergies.

Biodiversity↗

Molecular identification of Hymenopteran insects collected by using Malaise traps from Hazarganji Chiltan National Park Quetta, Pakistan.

The order Hymenoptera holds great significance for humans, particularly in tropical and subtropical regions, due to its role as a pollinator of wild and cultivated flowering plants, parasites of destructive insects and honey producers. Despite this importance, limited attention has been given to the genetic diversity and molecular identification of Hymenopteran insects in most protected areas. This study provides insights into the first DNA barcode of Hymenopteran insects collected from Hazarganji Chiltan National Park (HCNP) and contributes to the global reference library of DNA barcodes. A total of 784 insect specimens were collected using Malaise traps, out of which 538 (68.62%) specimens were morphologically identified as Hymenopteran insects. The highest abundance of species of Hymenoptera (133/538, 24.72%) was observed during August and least in November (16/538, 2.97%). Genomic DNA extraction was performed individually from 90/538 (16.73%) morphologically identified specimens using the standard phenol-chloroform method, which were subjected separately to the PCR for their molecular confirmation via the amplification of cytochrome c oxidase subunit 1 (cox1) gene. The BLAST analyses of obtained sequences showed 91.64% to 100% identities with related sequences and clustered phylogenetically with their corresponding sequences that were reported from Australia, Bulgaria, Canada, Finland, Germany, India, Israel, and Pakistan. Additionally, total of 13 barcode index numbers (BINs) were assigned by Barcode of Life Data Systems (BOLD), out of which 12 were un-unique and one was unique (BOLD: AEU1239) which was assigned for Anthidium punctatum. This indicates the potential geographical variation of Hymenopteran population in HCNP. Further comprehensive studies are needed to molecularly confirm the existing insect species in HCNP and evaluate their impacts on the environment, both as beneficial (for example, pollination, honey producers and natural enemies) and detrimental (for example, venomous stings, crop damage, and pathogens transmission).

Humans↗

Production of interspecific hybrid plants in Primula.

The methods of production of inter-specific hybrids in Primula are categorized into four steps: (1) emasculation, (2) pollination, (3) rescue culture of immature embryo, and (4) confiration of hybridity and ploidy level of the regenerated plants. Although most of the Primula pecies have a heteromorphic self-incompatibility system, an emasculation step is usually needed to avoid self-pollination since self-incompatibility is not always complete. At the resue culture step, addition of plant hormones (e.g., auxin, cytokinin, and gibberellin) to the culture medium is proved to be effective. The hybridity of the plants is efficiently confirmed at seedling stage by DNA analysis in addition to the comparison of morphological characters. The analysis of relative DNA contents by flow cytometry is easy and rapid technique to confirm hybridity and to estimate ploidy level and genomic combination.

Cell Culture Techniques↗

Exploitation of Erythrina dominguezii Hassl. (Fabaceae) nectar by perching birds in a dry forest in western Brazil.

Among the vertebrate pollinated plants, the genus Erythrina includes tree species in which birds are the pollen vectors. Two groups in this genus may be distinguished: a) the hummingbird, and b) the perching bird pollinated species. Erythrina dominguezii is included in the second group and occurs in deciduous/semi-deciduous forests in the southwestern neotropics. I studied the exploitation of Erythrina dominguezii nectar by perching birds in a dry forest in western Brazil. Six perching bird species from two distinct groups (Psittacidae: Brotogeris chiriri, Nandayus nenday, Aratinga acuticaudata; Icterinae: Psarocolius decumanus, Icterus cayanensis, I. icterus) consumed its nectar. The two most important consumers were Brotogeris chiriri (51.5% of the flowers visited by birds) and Psarocolius decumanus (20%). While B. chiriri was a flower predator, P. decumanus removed the nectar without damaging the flowers which it opened by inserting its large bill between the standard and the keel. Nandayus nenday, Aratinga acuticaudata, and I. icterus exploited the nectar like P. decumanus, and presumably also contributed to pollen transfer. As the flowering in E. dominguezii was intense and synchronous during the dryest period of the year, and its nectar was highly consumed by birds, the present data suggest that the nectar of this species may be important as an alternative resource to frugivorous/omnivorous birds when other resources are scarce.

Animals↗

Effect of local mate competition on fig wasp sex ratios.

In fig wasps, mating takes place among the offspring of one or a few foundress mothers inside the fig from which mated females disperse to found new broods. Under these conditions, related males will compete with each other for mating and several studies have shown female bias in brood sex ratios as a response to Local Mate Competition (LMC). Studying Pegoscapus tonduzi which pollinates Ficus citrifolia in Brazil, we analysed the effect of LMC (number of foundresses) on the sex ratio of the offspring of pollinating wasps. The relationship between the foundress number and brood sex ratio qualitatively followed the theory, however the empirical sex ratio was more female biased than expected from theoretical values. The model for an optimal sex ratio considers that each foundress wasp contributes the same number of eggs to be bred and that the violation of this assumption may explain the lack of adjustment in relation to the theory.

Animals↗

Hazards of insecticides to the bumble bees Bombus impatiens (Hymenoptera: Apidae) foraging on flowering white clover in turf.

Insecticides used on turf are sometimes applied to areas with flowering weeds that attract honey bees and native pollinators. We tested residual effects of such treatments on colony vitality and behavior of the bumble bees Bombus impatiens Cresson foraging on turf containingwhite clover, Trifolium repens L. Imidacloprid, a syst emic chloronicotinyl used for preventive control of root-feeding grubs, was applied as granules, followed by irrigation, or sprayed as a wettable powder, with or without irrigation. Hives were confined on the plots in large field cages after residues had dried and colony vitality (i.e., numbers of brood, workers, and honey pots, and weights of queens, workers, and whole colonies with hives) was evaluated after 28-30 d. Workers' foraging activity and defensive response to an aggressive stimulus also were evaluated. In another test, weedy turf was sprayed with chlorpyrifos, carbaryl, or cyfluthrin at labeled rates for surface-feeding pests. Bee colonies were confined on the plots after residues had dried, with effects on colony vitality evaluated after 14 d. Finally, foraging activity of wild bumble bees was monitored on open plots to determine if insecticide-treated areas were avoided. Imidacloprid granules, and imidacloprid sprays applied with posttreatment irrigation, had no effect on colony vitality or workers' behavior, suggesting that such treatments pose little systemic or residual hazard to bumble bees. In contrast, exposure to dry nonirrigated residues of all of the aforementioned insecticides had severe impact on colony vitality. Foraging workers did not avoid insecticide-treated areas. Means by which turf managers can reduce hazards of insecticide applications to pollinators are discussed.

Agriculture↗

Effect of temperature on Osmia lignaria (Hymenoptera: Megachilidae) prepupa-adult development, survival, and emergence.

The solitary bee Osmia lignaria Say continues to be developed as an orchard pollinator in the United States and southern Canada. Female bees are active during the early spring and construct nests consisting of a linear series of unlined cells delimited by mud partitions. Cells are provisioned with a pollen/nectar mass on which an egg is deposited, and nests are sealed with a mud plug. In 1997, we initiated two experiments on the development, mortality, and emergence of O. lignaria at selected laboratory temperature regimes and outdoors. In the first experiment (published previously), we compared temperature treatments for their adequacy in maintaining healthy O. lignaria populations. In a second experiment (reported here), we determined the relationship between rearing temperatures and prepupa-adult development rates as well as emergence time and longevity after wintering and incubation the following spring. We observed important differences in O. lignaria prepupa versus pupa responses to selected temperature treatments. The relationship between temperature and days to pupa was U-shaped, with additional time to transit the prepupa-pupa interval at temperatures above and below 26 degrees C. The negative relationship between temperature and the length of the pupa to adult interval contrasts with the U-shaped thermal response observed for prepupae. Thus, with each increase in thermal heat units over the range of temperature treatments tested, we observed an additional reduction in the pupa-adult interval. Individuals reared at constant 18 degrees C required 2.4 times as many days to transit the pupa-adult interval compared with those at constant 32 degrees C. Our results suggest that there is a need for the development of regionally adapted, latitude-specific breeding populations of this outstanding orchard crop pollinator.

Animals↗

Orchid bees don't need orchids: evidence from the naturalization of an orchid bee in Florida.

Almost 200 species of orchid bees are the exclusive pollinators of nearly 700 specialized orchids in the neotropics. This well-known mutualism involves orchids, called perfume orchids, which produce species-specific blends of floral fragrances, and male orchid bees, which collect and use these fragrance compounds during their courtship. We report here the naturalization of an orchid bee, Euglossa viridissima, in southern Florida, USA, where perfume orchids are absent. Chemical analysis of the contents of the fragrance storage organs in the hind tibias of 59 male bees collected in Florida identified 55 fragrance compounds, including 27 known from the perfumes of nine species of E. viridissima's orchid mutualists in Mesoamerica. Aromatic leaves, such as basil, were found to be important surrogate sources of needed fragrance compounds in Florida. The bee's ability to live and become abundant in the absence of its orchid mutualists suggests that the orchid bee-perfume orchid mutualism may be facultative for the bees, even though it is obligatory for the orchids. This invasive bee visits and potentially pollinates the flowers of many plants in Florida, behavior that could promote the abundance of selected exotic and native species.

Animal Communication↗

Consequences of vegetative herbivory for maintenance of intermediate outcrossing in an annual plant.

Given the occurrence of mixed mating systems among plants, a general mechanism explaining the evolution and maintenance of this condition is needed. Although numerous theoretical models predict mixed mating to be evolutionarily stable, conditions favoring intermediate selfing are often stringent and have limited applicability. Here we investigated the role of vegetative herbivory, a ubiquitous biotic factor limiting plant reproduction, in the mating system expression of Impatiens capensis (Balsaminaceae), a species with an obligate mixed-mating system (individuals produce both selfing, cleistogamous, and facultatively outcrossing, chasmogamous flowers). Herbivory reduced proportional chasmogamous reproduction partially, but not entirely, through a reduction in plant size and the strength of this effect varied among replicates. Herbivory decreased geitonogamous selfing in chasmogamous flowers via several mechanisms including reduced chasmogamous flower display size and pollinator visitation rate and altered pollinator composition. Overall, herbivory caused a decrease in whole-plant outcrossing, indicating that the effects of herbivory on proportional chasmogamous reproduction, which favor selfing, outweigh the effects on chasmogamous outcrossing rate, which favor outcrossing. Not only do our findings unravel the mechanisms underlying herbivore-mediated changes in the mating system, but they also point to the role of natural enemies in contributing to the maintenance of a mixed mating system.

Animals↗

Temporal dynamics of antagonism and mutualism in a geographically variable plant-insect interaction.

Variation among sites and years in the local ecological outcome of interspecific interactions can generate a geographic mosaic of coevolution, as indicated by recent mathematical models. We evaluated whether local temporal dynamics of ecological outcome in the interaction between the moth Greya politella (Prodoxidae) and its host plant Lithophragma parviflorum (Saxifragaceae) are likely to mitigate or magnify geographic differences in ecological outcome found in earlier studies. The moths are highly host-specific pollinating floral parasites, and the mutualism can be swamped in some populations by the presence of effective co-pollinators. Hence, differing community contexts can shift the outcome of the interaction from mutualism to commensalism or antagonism. During each of four years, we evaluated the effect of Greya oviposition on seed development through a paired design that controlled for plant genotype and microenvironment. At Turnbull National Wildlife Refuge in Washington State, the interaction was significantly mutualistic in all four years. Mutualism in this population was indicated by a higher probability of development of capsules visited by ovipositing Greya than capsules not visited by Greya on the same plant. At Rapid River, Idaho, the interaction was commensalistic in three years and antagonistic in one year. Antagonism in this population was indicated by selective withering of capsules containing Greya eggs. Overall, the results suggest stable geographic differences in the range of ecological outcomes in this plant-insect interaction under different community contexts.

Animals↗

Complex responses within a desert bee guild (Hymenoptera: Apiformes) to urban habitat fragmentation.

Urbanization within the Tucson Basin of Arizona during the past 50+ years has fragmented the original desert scrub into patches of different sizes and ages. These remnant patches and the surrounding desert are dominated by Larrea tridentata (creosote bush), a long-lived shrub whose flowers are visited by > 120 native bee species across its range. Twenty-one of these bee species restrict their pollen foraging to L. tridentata. To evaluate the response of this bee fauna to fragmentation, we compared species incidence and abundance patterns for the bee guild visiting L. tridentata at 59 habitat fragments of known size (0.002-5 ha) and age (up to 70 years), and in adjacent desert. The 62 bee species caught during this study responded to fragmentation heterogeneously and not in direct relation to their abundance or incidence in undisturbed desert. Few species found outside the city were entirely absent from urban fragments. Species of ground-nesting L. tridentata specialists were underrepresented in smaller fragments and less abundant in the smaller and older fragments. In contrast, cavity-nesting bees (including one L. tridentata specialist) were overrepresented in the habitat fragments, probably due to enhanced nesting opportunities available in the urban matrix. Small-bodied bee species were no more likely than larger bodied species to be absent from the smaller fragments. The introduced European honey bee, Apis mellifera, was a minor faunal element at > 90% of the fragments and exerted little if any influence on the response of native bee species to fragmentation. Overall, bee response to urban habitat fragmentation was best predicted by ecological traits associated with nesting and dietary breadth. Had species been treated as individual units in the analyses, or pooled together into one analysis, these response patterns may not have been apparent. Pollination interactions with this floral host are probably not adversely affected in this system because of its longevity and ability to attract diverse pollinators but will demand careful further study to understand.

Animals↗

[Natural history of flowers and gravity].

Many flowers have coevolved with their pollinator animals. Gravity has been one of selection pressure for the evolution of flowers. Gravity rules morphology and other features of flowers in many aspects. Pair matching between the flower and its specific pollinator is one of factors that determine the fitness of both sides. Evolution of flower morphology and its molecular basis are reviewed briefly. Anemophilous flowers are also under the influence of gravity. Shape and other features of entomophilous flowers have been highly diversed. Gravitropic response and its mechanism are summarized. Recent findings on gravitropism and phototropism of pistils and stamens are presented in this article.

Biological Evolution↗