Dynamics of island populations of Cypripedium calceolus in the Biebrza river valley (north-east Poland)

Three Cypripedium calceolus populations were observed in the Biebrza National Park (north-east Poland) for 11 years (1989–99). The populations differed in the number of genets and ramets and in their dynamics. Differences were in the proportion of flowering ramets, number of flowers, fruiting and recruitment. Fruiting and recruitment are low in populations of Cypripedium calceolus . Fruiting is probably pollinators limited, and recruitment is limited by environmental conditions. Changes in the size of populations of this species are caused by the following processes: changes in the number of genets (their appearance and death), changes in the size of clones (growth of individuals and their disintegration, death of individual ramets), dormancy of genets, and animal pressure. © 2002 The Linnean Society of London, Botanical Journal of the Linnean Society , 2002, 139 , 67–77.     

Cost of reproduction in the pink lady's slipper orchid (Cypripedium acaule, Orchidaceae): an eleven-year experimental study of three populations

An 11-yr experimental study of the cost of reproduction in three wild populations of the perennial orchid Cypripedium acaule contrasted experimental plants that were repeatedly hand-pollinated and often made fruits with control plants that were not hand-pollinated and only rarely made fruits. Repeated flowering without subsequent fruit production resulted in no detectable reduction in either plant size or probability of flowering in subsequent years. A cost of fruit production was evident in experimental plants in all three populations in terms of a reduced probability of flowering and smaller leaf area in subsequent years, but was not evident in terms of mortality rate. Experimental effects of fruit production reached maximum values at 3-7 yr, depending on the population. The probability of remaining dormant below ground in a given year was strongly dependent on plant size in the previous year. Furthermore, the length of the dormancy period (one to several years) was a significant and inverse function of plant size just prior to dormancy. Sample sizes and the consequent ability to detect experimental effects declined over time as more plants died or stopped flowering. Four to seven years appears to be an optimal duration for studies of the cost of reproduction in perennial herbs similar to this species. Studies lasting less than 4 yr may be too brief to reveal experimental effects, whereas those lasting more than 7 yr may fail to reveal new insights.     

Factors affecting germinable seed yield in Cypripedium calceolus var. pubescens (Willd.) Correll and Epipactis helleborine (L.) Crantz (Orchidaceae)

One of the challenges facing orchid conservationists is to get a better understanding of seed parent fecundity. This can be especially laborious for orchids whose seed is difficult to germinate. Mature seeds ofCypripedium calceolusvar.pubescens(Willd.) Correll (Orchidaceae) have been reported to require 8 weeks pre-chilling at 4°C and a further month at 20°C to induce germination, but seeds prematurely harvested at 7–8 weeks post-pollination germinate asymbiotically without chilling in less than 1 month. Mature seeds ofEpipactis helleborine(L.) Crantz (Orchidaceae) have also presented a challenge but the identification of plants whose seeds are quick and easy to germinate has provided us with a tool to study patterns of production of this seed class. Preliminary field experiments suggest that timing of pollination in these two taxa is critical for the development of seed. When seeds from capsules ofC. calceolusvar.pubescensharvested 7 weeks after pollination were examined, the proportion of fully mature embryos increased with increasing anthesis-pollination interval over the first 10 days. Paired flowers from five genets xenogamously hand-pollinated at 2-day intervals from the second day after anthesis until the flowers wilted were significantly less likely to produce fruits as the flowers aged but the flower age at which this occurred varied between genets. The germinability of seeds from different pollen and seed parent combinations was tested on eight different media formulations and over a range of post-anthesis and post-pollination intervals. Different aged seeds germinated variably, the post-anthesis interval being the most reliable predictor of germination outcomes. Intercrossing of plants ofC. calceolusvar.pubescenssuggests that seed production and germinability are maternally mediated traits. WithE. helleborine, germinable seed can be produced both by self- and by cross-pollination of selected plants but we have observed an influence of flower position in the inflorescence on self-compatibility and on the production of the easy-to-germinate seed class. The timing of pollination in these two unrelated orchids has a considerable influence upon seed production and seed quality in plants of established fecundity. These findings suggest that flower age at pollination must be considered when interpreting similar studies in the Orchidaceae.     

Re-establishment of the lady's slipper orchid (Cypripedium calceolus L.) in Britain

The Sainsbury Orchid Conservation Project (SOCP), based in the Micropropagation Unit of the Royal Botanic Gardens, Kew, has worked for many years with English Nature, and the Species Recovery Programme in particular, in raising plants of the lady's slipper orchid for re-establishment. This is a collaborative venture with the involvement of a large number of organizations and individuals at national and local level.Cypripedium calceolusis one of Britain's rarest plants. Thought at one time to be extinct, its decline is due to overcollection by botanists for herbarium specimens and by gardeners and it is found now on a single, fragile, natural site. It has been wardened since 1970s because of concerns about long-term safety and is protected under Schedule 8 of the Wildlife and Countryside Act 1981. Natural pollination has not been observed and hand pollination is carried out to ensure seed set. Some seed is scattered on site resulting in a few seedlings and the remaining capsules are sent to the Sainsbury Orchid Conservation Project. In addition, some plants of native origin are held in cultivation and molecular techniques are being carried out at Kew to ascertain how these are related. The best pollination strategy to increase genetic diversity is co-ordinated by English Nature and SOCP, as is the optimum collection time post-pollination to achieve maximum germination. Seedlings are propagated in the laboratory using immature green capsules sown on a range of nutrient media. No symbiotic fungus has yet been found for this species. Germination occurs after 6 weeks in the dark and the plants chilled when roots are well developed and shoots beginning to form. Plants potted up at Kew and near the native site have produced leaves and buds and extensive planting trials have now commenced. The main objective is to increase the number of localities whereCypripediumoccurs through re-establishment. As its decline is due to overcollection rather than habitat loss, many old sites have changed very little. Observation of continental populations is important in assessing suitability of potential sites as the extant clone may not be in optimum conditions. An initial planting trial on the wild site in winter 1989/90 resulted in a 75% survival rate of seedlings planted out. However, establishing seedlings is a long process and careful monitoring is required.     

Genetic diversity, compatibility patterns and seed quality in isolated populations of Cypripedium calceolus (Orchidaceae)

Cypripedium calceolus has suffered an alarming decline, and today mainly occurs in small and isolated populations. In Denmark there are only two populations, close to each other and situated far from other European stands. One population is stagnant or in slow decline, whereas the other is in rapid increase. We examined the levels of genetic diversity and compatibility and seed quality following experimental crosses. No genetic variation could be detected in plastid and nuclear markers within or between the two populations—in contrast to results previously reported from other European populations of C. calceolus. This may indicate a founder effect in both populations, but it could also be the outcome of prolonged inbreeding or reflect a genetic bottleneck after the populations were established. According to fruit dimensions and frequency of fully developed seeds there was full self-compatibility in the stagnant population, and partial late-acting self-incompatibility in the proliferating population. In combination with previous reports from other countries, this suggests that several self-incompatibility systems may occur in C. calceolus. Seeds from the older and stagnant population performed more poorly in germination tests in vitro than seeds from the thriving population. The difference needs not be genetically based, but could be due to environmental differences during seed maturation, producing different seed quality or dormancy characteristics. However, low level of genetic diversity within the populations may affect their ability to adapt and the possibility of inbreeding depression should be investigated.     

Allozyme differentiation and genetic structure of the Lady's slipper (Cypripedium calceolus) island populations in north-east Poland

The population genetic structure in the rare and endangered perennial plant, Cypripedium calceolus , from north-east Poland was investigated. A total of 11 loci were analysed, of which five were polymorphic for the species and in every population. The 11 loci resolved gave a total of 19 alleles with the mean number of 1.73 alleles per locus. The proportion of polymorphic loci (P) was the same for the species and in every population, 45.5%. The average observed and expected heterozygosities for the total Lady's slipper data set were 0.156 and 0.184, respectively. The average expected heterozygosity for the populations under study ranged from 0.176 to 0.188 and did not differ significantly among populations. Genetic diversity within C. calceolus populations from the Biebrza Valley is relatively high compared with rare taxa and taxa with similar life histories. The genetic diversity among populations of C. calceolus studied ( F _ST = 0.016) is much smaller than the genetic diversity within populations. In the species studied, there was no significant correlation between genetic and geographical distances ( r = −0.73, Mantel test). This may indicate the relatively recent origin of the populations studied, or a high level of gene flow among populations. © 2002 The Linnean Society of London, Botanical Journal of the Linnean Society , 2002, 138 , 433–440.     

Effect of Lighting Conditions on the Reproductive Success of Cypripedium calceolus L. (Orchidaceae,Liliopsida)

Biology Bulletin - Abstract—The influence of lighting conditions on the reproductive characteristics of Cypripedium calceolus L. was studied on the territory of the Komi Republic, where the...     

Pollination of Cypripedium plectrochilum (Orchidaceae) by Lasioglossum spp. (Halictidae): the roles of generalist attractants versus restrictive floral architecture

The pollination of Cypripedium plectrochilum Franch. was studied in the Huanglong Nature Reserve, Sichuan, China. Although large bees (Bombus, Apis), small bees (Ceratina, Lasioglossum), ants (Formica sp.), true flies (Diptera) and a butterfly were all found to visit the flowers, only small bees, including three Lasioglossum spp. (L. viridiclaucum, L. sichuanense and L. sp.; Halictidae) and one Ceratina sp., carried the flower's pollen and contacted the receptive stigma. Measurements of floral architecture showed that interior floral dimensions best fit the exterior dimensions of Lasioglossum spp., leading to the consistent deposition and stigmatic reception of dorsally-placed, pollen smears. The floral fragrance was dominated by one ketone, 3-methyl-Decen-2-one. The conversion rate of flowers into capsules in open (insect) pollinated flowers at the site was more than 38%. We conclude that, while pigmentation patterns and floral fragrance attracted a wide variety of insect foragers, canalization of interior floral dimensions ultimately determined the spectrum of potential pollinators in this generalist, food-mimic flower. A review of the literature showed that the specialised mode of pollination-by-deceit in C. plectrochilum, limiting pollinators to a narrow and closely related guild of 'dupes' is typical for other members of this genus.     

Dynamics of Typha latifolia L. populations in treatment wetlands in Estonia

The aim of this paper is to evaluate and compare broadleaved cattail (Typha latifolia L.) biomass production and the nitrogen (N) and phosphorus (P) content in phytomass in three treatment wetland systems and to propose suitable areas for treatment wetlands in Estonia for raw material production. The biomass samples (roots/rhizomes, shoots with leaves and spadixes) and litter were collected from 1 m × 1 m plots—15 plots in the Tänassilma semi-natural wetland, 15 plots in the Põltsamaa free water surface (FWS) constructed wetland (CW), and 10 plots in the Häädemeeste FWS CW. The average aboveground biomass of T. latifolia varied from 0.37 to 1.76 kg DW m^?2 in autumn and from 0.33 to 1.38 kg DW m^?2 in winter. The greatest average nitrogen (22,950 mg N kg^?1) concentration was found in spadixes in 2002, and the phosphorus (6500 mg P kg^?1) concentration was measured in roots–rhizomes in 2003. Average standing stock of nitrogen and phosphorus was higher in aboveground than belowground phytomass. In FWS CWs with high hydraulic and nutrient loadings, however, the harvesting of aboveground biomass is not an effective means for the removal of nutrients. Cattail biomass is a valuable insulation material, whereas the fibre from spadixes mixed with clay gives elasticity to clay plasters. According to our estimates, about 5412 km² could be used for Typha cultivation in Estonia.     

银杏传粉生物学研究进展

银杏(Ginkgo biloba L.)是最古老裸子植物之一,其传粉受精过程表现出许多特殊的原始性状和特征,在种子植物的系统演化上具有重要研究价值;同时,银杏种实经济价值高,但栽培上由于授粉受精不良而导致落花落果现象严重,因此了解银杏传粉生物学特性对于其种实的优质高产具有实际意义.本文从银杏雌雄株开花物候学、雌雄花形态结构特征、花粉和胚珠发育进程、花粉和胚珠生物学特性、传粉机制及花粉在胚珠内萌发生长进程等方面,对银杏传粉生物学相关研究进行综述,为银杏的系统演化、授粉受精和种实发育研究提供参考.     

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

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

Spatial genetic structure and clonal diversity of island populations of lady's slipper (Cypripedium calceolus) from the Biebrza National Park (northeast Poland)

Three populations of the rare and endangered plant species Cypripedium calceolus were included in a study of genetic diversity and spatial genetic structure in the Biebrza National Park, northeast Poland. Analysis of 11 allozyme loci indicate that the populations of this species contained high genetic variability (P = 45.5%, A= 1.73). On the other hand, the genetic differentiation (FST = 0.014, P < 0.05) among C. calceolus populations was very low when compared to other species with similar life history characteristics. The observed high rate of gene flow (Nm = 18) may suggest that the populations studied derived from each other in the recent past. Five polymorphic allozyme markers identified 109 multilocus genotypes in three populations and the majority of them (67%) were population-specific. One of the populations studied, characterized by particularly extensive vegetative reproduction, showed the lowest clonal diversity (G/N = 0.15) and heterozygosity (HO = 0.111) values and the highest FIS(0.380), when compared to other two populations (G/N = 0.26-0.27, HO= 0.166-178, FIS = 0.024-0.055). This may indicate that clonal reproduction has an important influence on the genetic structure of C. calceolus populations. The longevity of genets, the out-crossing breeding system and the presence of recruitment from seeds are factors maintaining genetic diversity in C. calceolus.     

Fruit-set of unpollinated ovaries of Pisum sativum L.

The influence of removing the apical shoot and different leaves above and below the flower on the fruit-set of unpollinated pea ovaries (Pisum sativum L. cv. Alaska) has been studied. Unpollinated ovaries were induced to set and develop either by topping or by removing certain developing leaves of the shoot. Topping had a maximum effect when carried out before or on the day of anthesis, and up to four consecutive ovaries were induced to set in the same plant. The inhibition of fruit-set was due to the developing leaves and not to the apex. The third leaf above the first flower, which had a simultaneous development to the ovary, had the stronger inhibitory effect on parthenocarpic fruit-set. The application of different plant-growth regulators (indoleacetic acid, naphthylacetic acid, 2,4-dichlorophenoxyacetic acid, gibberellic acid, benzyladenine and abscisic acid) did not mimic the negative effect of the shoot.Abbreviations CCC (2-chloroethyl)trimethylammonium chloride - MH maleic hydrazide - IAA indole-3-acetic acid - NAA 1-naphthaleneacetic acid - 2,4-D 2,4-dichlorophenoxyacetic acid - GA₃ gibberellic acid - 6-BAP benzyladenine - ABA abscisic acid     

Pollination of the Lady's slipper orchid (Cypripedium calceolus) in Scandinavia – taxonomic and conservational aspects

The Lady's slipper orchid Cypripedium calceolus L. is considered one of the most beautiful orchids of Europe. Consequently, the species has suffered from over-collecting and is now critically endangered in many countries. Although pollination success is suspected to influence the long-term survival of Cypripedium calceolus , relatively little is known about the identity of its pollinators in mainland Sweden – a region that comprises the largest European populations. In order to identify which species pollinate eight representative populations in mainland Sweden, we observed and sampled visitors to flowers using a standardized protocol. Specimens were identified and any pollen smear found on their body was examined for the presence of Cypripedium pollen. Nine species were recognized as effective pollen vectors ( Andrena cineraria, A. carantonica, A. haemorrohoa, A. helvola, A. nigroaenea, A. praecox, Colletes cunicularius, Lasioglossum fratellum and L. fulvicorne ), four of them for the first time in Scandinavia. This is the first time that a species of Colletes is reported to carry pollen of Cypripedium in this region. All but one specimens were females. Our results suggest a taxonomically heterogeneous pollinator fauna for Cypripedium calceolus and are discussed in light of the management of this species.     

Nectary structure and nectar secretion in Maxillaria coccinea (Jacq.) L.O. Williams ex Hodge (Orchidaceae)

BACKGROUND AND AIMS: It had previously been assumed that Maxillaria spp. produce no nectar. However, nectar has recently been observed in Maxillaria coccinea (Jacq.) L.O. Williams ex Hodge amongst other species. Furthermore, it is speculated that M. coccinea may be pollinated by hummingbirds. The aim of this paper is to investigate these claims further. METHODS: Light microscopy, histochemistry, scanning and transmission electron microscopy. KEY RESULTS: This is the first detailed account of nectar secretion in Maxillaria Ruiz & Pav. A 'faucet and sink' arrangement occurs in M. coccinea. Here, the nectary is represented by a small protuberance upon the ventral surface of the column and nectar collects in a semi-saccate reservoir formed by the fusion of the labellum and the base of the column-foot. The nectary comprises a single-layered epidermis and three or four layers of small subepidermal cells. Beneath these occur several layers of larger parenchyma cells. Epidermal cells lack ectodesmata and have a thin, permeable, reticulate cuticle with associated swellings that coincide with the middle lamella between adjoining epidermal cells. Nectar is thought to pass both along the apoplast and symplast and eventually through the stretched and distended cuticle. The secretory cells are collenchymatous, nucleated and have numerous pits with plasmodesmata, mitochondria, rough ER and plastids with many plastoglobuli but few lamellae. Subsecretory cells have fewer plastids than secretory cells. Nectary cells also contain large intravacuolar protein bodies. The floral morphology of M. coccinea is considered in relation to ornithophily and its nectary compared with a similar protuberance found in the entomophilous species M. parviflora (Poepp. & Endl.) Garay. CONCLUSIONS: Flowers of M. coccinea produce copious amounts of nectar and, despite the absence of field data, their morphology and the exact configuration of their parts argue strongly in favour of ornithophily.     

Floret specialization, seed production and gender in Artemisia vulgaris L. (Asteraceae, Anthemideae)

Flowering and fruiting behaviour of female and hermaphrodite florets is described and assessed in samples from three populations from Denmark, England and Sweden. Between 25 and 50% of the florets in capitula are female, and flowering gender varies little among plants in each population. Fruiting gender of individuals, G (femaleness), varies from 0 to 0.85, because of variation in fruit set and fruit abortion. Variation in fruiting gender was correlated with plant size parameters in two populations, but not in the third. The data suggest that post-anthesis regulation of maternal investment may be operating. Florets of A. vulgaris are either totally specialized for pollen receipt (female florets) or largely specialized for pollen donation (hermaphrodite florets), and show adaptations for avoiding interference with each other in these functions. Movement of capitula from a pendent position at flowering to an erect position at fruiting optimizes positions for dissemination of pollen and of seeds respectively.     

Critical stages in the recruitment process of Rhamnus alaternus L

BACKGROUND AND AIMS: Rhamnus alaternus is a Mediterranean shrub commonly used in reforestation programs. Although several aspects of its reproductive biology have been studied, little is known about the importance of the different recruitment stages in the overall regeneration process of this species, which limits its proper use in Mediterranean forests and shrubland management. The aim of the present work was to quantify the importance of the different recruitment stages in the regeneration process of R. alaternus. METHODS: Two populations of Rhamnus alaternus on the island of Mallorca that differ in climatic conditions, type of habitat and sex ratio were studied. The importance of seed production, seed dispersal and predation, seedling emergence and seedling survival for the regeneration of this species were quantified. KEY RESULTS: In both populations, fruit set and fruit removal by animals were not critical stages, since almost half of the flowers became mature fruits and 90% of those were dispersed. Most seeds were deposited under female conspecifics (86 and 47%, at Lloret and Esporles, respectively), and very few were found in open inter-spaces (1 and 5%). Post-dispersal seed predation (mostly by ants and rodents) was very high in both populations. Seedling emergence took place during autumn and early winter and it ranged from 31 to 68% depending upon year and microhabitat. The majority of emerged seedlings died during the first year, mainly due to desiccation; such mortality was influenced by rainfall and differed among microhabitats (varying from 67 to 100%). The general spatial distribution of seed rain was concordant with the seedling emergence and survival pattern in both populations. CONCLUSIONS: The recruitment of Rhamnus alaternus appeared to be mainly limited by seed and seedling survival, regardless of the type of habitat in which the species is found.