Allocation to reproduction following experimental defoliation in Platanthera bifolia (Orchidaceae)

The roles of herbivory and pollination success in plant reproduction have frequently been examined, but interactions between these two factors have gained much less attention. In three field experiments, we examined whether artificial defoliation affects allocation to attractiveness to pollinators, pollen production, female reproductive success and subsequent growth in Platanthera bifolia L. (Rich.). We also recorded the effects of inflorescence size on these variables. We studied the effects of defoliation on reproductive success of individual flowers in three sections of inflorescence. Defoliation and inflorescence size did not have any negative effects on the proportion of opened flowers, spur length, nectar production or the weight of pollinia. However, we found that hand-pollination increased relative seed production and defoliation decreased seed set in most cases. Interactions between hand-pollination and defoliation were non-significant indicating that defoliation did not affect female reproductive success indirectly via decreased pollinator attraction. Plants with a large inflorescence produced relatively more seeds than plants with a small inflorescence only after hand-pollination. The negative effect of defoliation on relative capsule production was most clearly seen in the upper sections of the inflorescence. In addition to within season effects of leaf removal, defoliated P. bifolia plants may also have decreased lifetime fitness as a result of lower seed set within a season and because of a lower number of reproductive events due to decreased plant size (leaf area) following defoliation. Our study thus shows that defoliation by herbivores may crucially affect reproductive success of P. bifolia.     

Nutrient versus pollination limitation in Platanthera bifolia and Dactylorhiza incarnata (Orchidaceae)

Availability of resources and pollination services have been demonstrated to have impact on reproductive success in some orchid species, but to our knowledge no studies have examined the effects of nutrient application and pollination limitation in the same experiment. In this study, factors limiting reproductive success were studied with two terrestrial orchid species in Central Finland during 1996 and 1997. In a field experiment using a factorial design, plants of nectar-producing Platanthera bifolia and nectarless Dactylorhiza incarnata were treated with nutrient application and hand-pollination. Inflorescence size was considered as an indicator of attractiveness to pollinators as well as of the general condition of the plant. In 1996, fertilizer treatment increased relative capsule production in P. bifolia plants with small inflorescences, indicating poorer store of resources in the underground corm and dependence on nutrient availability during capsule maturation, which was not found in large plants . Hand-pollination and large inflorescence size in P. bifolia did not affect capsule initiation, but increased the proportion of mature capsules. P. bifolia may thus be regarded as both resource- and pollination-limited within a year. Pollination success was observed to be the only factor limiting reproduction of the nectarless D. incarnata within a year, because hand-pollination increased capsule production, and there were no significant effects of fertilizer treatment or inflorescence size in 1996. Capsule production in 1996 did not affect the probability of P. bifolia flowering in the following year, while high capsule production decreased the subsequent probability of flowering in D. incarnata . Species with different pollination strategies differed in the use of resources. The nectar-producing P. bifolia had a lower fruit/flower ratio than the nectarless D. incarnata , also after hand-pollination.     

Variation and post-pollination changes in floral odours released by Platanthera bifolia (Orchidaceae)

Volatile compounds were collected in the field from individual specimens of un-pollinated and hand-pollinated Platanthera bifolia , and investigated by combined gas chromatography-mass spectrometry (GC-MS). In all, 32 compounds were identified of which five were not previously reported in Platanthera. Pollinated flowers wilted significantly faster than unpollinated flowers. Twentyfour hours after pollination the production of volatile compounds had decreased slightly and there was a small change in their composition. Variation in floral scents within and between populations, as well as effects of changes in floral attractiveness on pollination and reproductive success are discussed.     

Variable selection in Platanthera bifolia (Orchidaceae): phenotypic selection differed between sex functions in a drought year

We estimated selection on three morphological characters in the hermaphroditic, hawkmoth-pollinated orchid Platanthera bifolia and explored selection surfaces through male and female function. The work was carried out in northern Sweden during two flowering seasons (1994 and 1995) in one natural population and one season (1995) in another natural population. Fitness was estimated as number of pollinia removed (male function) and number of fruits produced (female function). We detected directional selection towards larger inflorescence size (flower number) through both sex functions in both populations in 1995. In 1994, with an unusually dry growing season, 78% of the individuals failed to set any fruit, and there was selection for larger inflorescences only through male function. In this year, there was selection towards longer flower spurs, which could be a direct or indirect effect of spurs being shortened by drought. The results demonstrate that selection patterns may vary temporally and spatially, and that the 'male function hypothesis' may be applicable as female function is more resource dependent than male function.     

Characteristics and distribution of intermediates between Platanthera bifolia and P. chlorantha (Orchidaceae) in the Nordic countries

In areas of sympatry Plantanthera bifolia (L.) L. C. Rich. and P. chlorantha (Cust.) Rchb. occasionally produce hybrids. These plants exhibit intermediate floral morphology. The present study presents the distribution, floral morphology, and general characteristics of these intermediates in the Nordic area. The most useful and reliable diagnostic characters for hybrid plants were (1) the distance between viscidia (normally about 1.4 mm), (2) the length of the caudicle of pollinia (about 0.7 mm), and (3) the presence of a distinct pedicel at the connection between caudicle and viscidium. Comparative chemical analyses of the floral fragrances are reported. Material in the principal Nordic herbaria indicated a generally low production of hybrids (0.46%) and that the main area of occurrence is SE Sweden and Alandia, Finland. Intermediates seem more frequent in provinces where P. bifolia is relatively long-spurred. Very few were recorded from Denmark and Norway (hybrids in the latter country here reported for the first time). Aggregations of intermediates displaying patterns of introgressive hybridization were found only on Öland and Gotland. Occurrence of hybrids outside the Nordic area is also discussed and remarks are made on the morphologically similar P. metabifolia Maek. in E Asia. All known Nordic intermediates are listed.     

Volatiles from flowers of Platanthera bifolia (Orchidaceae) attractive to the silver Y moth, Autographa gamma (Lepidoptera: Noctuidae)

We examined the attractiveness of a natural headspace sample of Platanthera bifolia blossoms, synthetic blends and single compounds to the silver Y moth, Autographa gamma, in a flight tunnel. The synthetic blend consisted of previously identified electrophysiologically active compounds from P. bifolia : benzyl benzoate, benzyl salicylate, cinnamyl alcohol, lilac aldehydes, methyl benzoate and methyl salicylate. This blend had a similar attractivity as the natural headspace sample. Subtraction of lilac aldehydes significantly decreased attractiveness of the synthetic blend. When a mixture of lilac aldehydes was tested alone, it showed attractiveness similar to that of the synthetic blend. One or a mixture of lilac aldehydes accounts for the attraction of moths to P. bifolia . All other compounds elicited significantly lower responses. Results are discussed in relation to the pollination biology of P. bifolia .     

Processes of isolation and introgressive interplay between Platanthera bifolia (L.) Rich and P. chlorantha (Custer) Reichb. (Orchidaceae)

NILSSON, L. A., 1983. Processes of isolation and introgressive interplay between Platanthera bifolia L. Rich, and P. chlorantha (Custer; Reichb. (Orchidaceae). Platanthera bifolia and P. chlorantha are highly specialized for pollination by crepuscular and nocturnal Lepidoptera. The plants are largely sympatric, have overlapping flowering-times, and are completely interfertile. Principal differences exist in floral fragrance and column morphology. Isolation is effected by a set of integrated morphological, ethological, and seasonal mechanisms. Odour-based constancy by moths, differences in column structure, attachment of pollinaria, spurlength, nectar availability, and flowering-peaks reduce interspecific pollen transfer. The plants form a functional species pair in relation to their pollination environment and locally share the same main pollinator species.
Hybrids, which infrequently occur in areas of sympatry, emit a blend of the species-specific fragrance compounds and have intermediate column structure. They are frequently visited by-moths but suffer a serious reduction in seed production due to imperfect interaction vis-à-vis the pollen vectors. Hybrid viscidia, in an intermediate position on the column, are touched by moths' scaly and hairy palps and get insufficient attachment, evidently causing hybrids to act solely as female recipient. First backcross individuals largely join the functional range of either species which promotes rapid introgression. The two gene-pools interplay, although always at a low level. Patterns of introgressive hybridization were exemplified.     

The cost of inbreeding in Platanthera leucophaea (Orchidaceae)

Fragmentation and isolation are expected to have a considerable impact on viability and recruitment in populations of rare species. Platanthera leucophaea (Orchidaceae), a rare orchid, currently exists in a fragmented landscape of its natural habitat. Floral morphology suggests this species is predominantly outcrossing, but surveys of allozyme diversity suggest high, variable levels of inbreeding in populations (F(IS) = -0.078 to 1.0). This study examines the potential cost of inbreeding and the extent to which inbreeding depression can vary temporally and in populations of different size and genetic structure. Flowers were pollinated by hand in one large population and one small population over three seasons. Seed set, seed mass, and seed viability were compared among self-, outcross-, and open-pollinated fruits. Seed set was greater than 50% in both populations for all years of study. High levels of inbreeding depression were detected in seed viability but not in seed mass in both populations. However, the magnitude of inbreeding depression differed over years and between populations, a pattern that reflects differing environmental conditions and variable evolutionary and demographic histories. Consequently, conservation of this species will be most successful if outcrossing is promoted in populations by maximizing population size and genetic variability.     

Floral longevity and nectar secretion of Platanthera chlorantha (Custer) Rchb. (Orchidaceae)

Flowering and nectar secretion were studied in Platanthera chlorantha in two years. Nectar was secreted and accumulated in this orchid's spur, originating from part of the labellum. The nectary spur was, on average, 32 mm long. It produced 6.86 micro l nectar in 1999 and 7.84 micro l in 2000. The number of flowers per inflorescence and the volume of nectar secreted per flower were not correlated. Nectar secretion and flower longevity differed depending on pollination and flower position in the inflorescence. Among pairs of pollinated and unpollinated flowers there was no difference in the volume of nectar produced; however, the life span of pollinated flowers was shorter than that of unpollinated ones. Within an inflorescence, the lowest-positioned flowers had the largest nectar production and the longest life compared with flowers positioned higher up.     

黄花杓兰云南中甸居群遗传结构及克隆多样性的分析

采用AFLP分子标记方法对云南中甸的6个黄花杓兰( Cypripedium flavum )居群进行了遗传多样性水平、遗传结构及克隆多样性研究,其中两个居群用样方取样法分析其克隆空间结构.POPGENE软件分析结果表明:两组引物共产生104个位点,其中86个为多态位点,多态位点百分率为82.69%.黄花杓兰具有丰富的遗传变异(物种水平上, He=0.2884, Ho=0.4312;居群水平上,P=64.59%, He=0.2449, Ho=0.3606),黄花杓兰居群的遗传分化不大( Gst=0.154),居群间基因交流较为频繁( Nm=2.7460).居群的克隆多样性水平高( D=0.9638-0.9960,G/N=0.83-0.96),同一克隆的分株相邻,克隆生长延伸范围狭窄.黄花杓兰居群之所以保持较高水平的遗传多样性,可能与其既能通过实生幼苗增加基因的杂合度,又能通过无性分株把杂合体固定下来有关.     

Genetic Diversity Patterns and Functional Traits of Bradyrhizobium Strains Associated with Pterocarpus officinalis Jacq. in Caribbean Islands and Amazonian Forest (French Guiana)

Pterocarpus officinalis Jacq. is a legume tree native to the Caribbean islands and South America growing as a dominant species in swamp forests. To analyze (i) the genetic diversity and (ii) the symbiotic properties of its associated nitrogen-fixing soil bacteria, root nodules were collected from P. officinalis distributed in 16 forest sites of the Caribbean islands and French Guiana. The sequencing of the 16S-23S ribosomal RNA intergenic spacer region (ITS) showed that all bacteria belonged to the Bradyrhizobium genus. Bacteria isolated from insular zones showed very close sequence homologies with Bradyrhizobium genospecies V belonging to the Bradyrhizobium japonicum super-clade. By contrast, bacteria isolated from continental region displayed a larger genetic diversity and belonged to B. elkanii super-clade. Two strains from Puerto Rico and one from French Guiana were not related to any known sequence and could be defined as a new genospecies. Inoculation experiments did not show any host specificity of the Bradyrhizobium strains tested in terms of infectivity. However, homologous Bradyrhizobium sp. strain-P. officinalis provenance associations were more efficient in terms of nodule production, N acquisition, and growth than heterologous ones. The dominant status of P. officinalis in the islands may explain the lower bacterial diversity compared to that found in the continent where P. officinalis is associated with other leguminous tree species. The specificity in efficiency found between Bradyrhizobium strains and host tree provenances could be due to a coevolution process between both partners and needs to be taken in consideration in the framework of rehabilitation plantation programs.     

Molecular phylogenetics and morphological reappraisal of the Platanthera clade (Orchidaceae: Orchidinae) prompts expansion of the generic limits of Galearis and Platanthera

Background and Aims

The Platanthera clade dominates the North American orchid flora and is well represented in eastern Asia. It has also generated some classic studies of speciation in Platanthera sections Platanthera and Limnorchis. However, it has proved rich in taxonomic controversy and near-monotypic genera. The clade is reviewed via a new molecular phylogenetic analysis and those results are combined with brief reconsideration of morphology in the group, aiming to rationalize the species into a smaller number of larger monophyletic genera and sections.

Methods

Nuclear ribosomal internal transcribed spacer (ITS) sequences were obtained from 86 accessions of 35 named taxa, supplemented from GenBank with five accessions encompassing a further two named taxa.

Key Results

Using Pseudorchis as outgroup, and scoring indels, the data matrix generated 30 most-parsimonious trees that differed in the placement of two major groups plus two closely related species. Several other internal nodes also attracted only indifferent statistical support. Nonetheless, by combining implicit assessment of morphological divergence with explicit assessment of molecular divergence (when available), nine former genera can be rationalized into four revised genera by sinking the monotypic Amerorchis, together with Aceratorchis and Chondradenia (neither yet sequenced), into Galearis, and by amalgamating Piperia, Diphylax and the monotypic Tsaiorchis into the former Platanthera section Platanthera. After further species sampling, this section will require sub-division into at least three sections. The present nomenclatural adjustments prompt five new combinations.

Conclusions

Resolution of major groups should facilitate future species-level research on the Platanthera clade. Recent evidence suggests that ITS sequence divergence characterizes most species other than the P. bifolia group. The floral differences that distinguished Piperia, Diphylax and Tsaiorchis from Platanthera, and Aceratorchis and Chondradenia from Galearis, reflect various forms of heterochrony (notably paedomorphosis); this affected both the perianth and the gynostemium, and may have proved adaptive in montane habitats. Floral reduction was combined with lateral expansion of the root tubers in Piperia and Diphylax (including Tsaiorchis), whereas root tubers were minimized in the putative (but currently poorly supported) Neolindleya–Galearis clade. Allopolyploidy and/or autogamy strongly influenced speciation in Platanthera section Limnorchis and perhaps also Neolindleya. Reproductive biology remains an important driver of evolution in the clade, though plant–pollinator specificity and distinctness of the species boundaries have often been exaggerated.Key words: Aceratorchis, Amerorchis, Chondradenia, Diphylax, Galearis, generic delimitation, internal transcribed spacer, Neolindleya, orchid, Piperia, phylogeny, Platanthera, Pseudorchis, speciation, Tsaiorchis     

Pollution Breaks Down the Genetic Architecture of Life History Traits in Caenorhabditis elegans

When pollution occurs in an environment, populations present suffer numerous negative and immediate effects on their life history traits. Their evolutionary potential to live in a highly stressful environment will depend on the selection pressure strengths and on the genetic structure, the trait heritability, and the genetic correlations between them. If expression of this structure changes in a stressful environment, it becomes necessary to quantify these changes to estimate the evolutionary potential of the population in this new environment. We studied the genetic structure for survival, fecundity, and early and late growth in isogenic lines of a Caenorhabditis elegans population subject to three different environments: a control environment, an environment polluted with uranium, and a high salt concentration environment. We found a heritability decrease in the polluted environments for fecundity and early growth, two traits that were the most heritable in the control environment. The genetic structure of the traits was particularly affected in the uranium polluted environment, probably due to generally low heritability in this environment. This could prevent selection from acting on traits despite the strong selection pressures exerted on them. Moreover, phenotypic traits were more strongly affected in the salt than in the uranium environment and the heritabilities were also lower in the latter environment. Consequently the decrease in heritability was not proportional to the population fitness reduction in the polluted environments. Our results suggest that pollution can alter the genetic structure of a C. elegans population, and thus modify its evolutionary potential.     

Life History Traits of Two Chaetonotids (Gastrotricha) Under Different Experimental Conditions

Summary

A marine and a freshwater species of Chaetonotida (Gastrotricha) were reared under laboratory conditions. Their life tables and principal demographic parameters were determined at 2 different temperatures (20°and 25°C). At 25°C the data relative to the marine species were collected from 5 cohorts kept at 5 different salinity levels (15, 25, 35, 45, 55°/oo).

A higher temperature increases reproductive activity while shortening its duration in both species, whereas the length of the lifespan remains unaffected. Extreme salinity values (15 and 55°/oo) reduce the maximum longevity of the marine species and have opposite effects on reproductive activity, which is higher at low salinity and becomes lower at high salinity. The postparthenogenetic phase is remarkably long relative to the life cycle: this was observed in all experimental conditions and may be related to the existence of a second reproductive phase, which is hermaphroditic and follows the parthenogenetic one, as recently postulated from morphological data.     

Maternal History of Oceania from Complete mtDNA Genomes: Contrasting Ancient Diversity with Recent Homogenization Due to the Austronesian Expansion

Archaeology, linguistics, and existing genetic studies indicate that Oceania was settled by two major waves of migration. The first migration took place approximately 40 thousand years ago and these migrants, Papuans, colonized much of Near Oceania. Approximately 3.5 thousand years ago, a second expansion of Austronesian-speakers arrived in Near Oceania and the descendants of these people spread to the far corners of the Pacific, colonizing Remote Oceania. To assess the female contribution of these two human expansions to modern populations and to investigate the potential impact of other migrations, we obtained 1,331 whole mitochondrial genome sequences from 34 populations spanning both Near and Remote Oceania. Our results quantify the magnitude of the Austronesian expansion and demonstrate the homogenizing effect of this expansion on almost all studied populations. With regards to Papuan influence, autochthonous haplogroups support the hypothesis of a long history in Near Oceania, with some lineages suggesting a time depth of 60 thousand years, and offer insight into historical interpopulation dynamics. Santa Cruz, a population located in Remote Oceania, is an anomaly with extreme frequencies of autochthonous haplogroups of Near Oceanian origin; simulations to investigate whether this might reflect a pre-Austronesian versus Austronesian settlement of the island failed to provide unequivocal support for either scenario.     

Increased Extinction Potential of Insular Fish Populations with Reduced Life History Variation and Low Genetic Diversity

Theoretical work has shown that reduced phenotypic heterogeneity leads to population instability and can increase extinction potential, yet few examples exist of natural populations that illustrate how varying levels expressed diversity may influence population persistence, particularly during periods of stochastic environmental fluctuation. In this study, we assess levels of expressed variation and genetic diversity among demographically independent populations of tidewater goby (Eucyclogobius newberryi), show that reductions in both factors typically coincide, and describe how low levels of diversity contribute to the extinction risk of these isolated populations. We illustrate that, for this annual species, continuous reproduction is a safeguard against reproductive failure by any one population segment, as natural, stochastically driven salinity increases frequently result in high mortality among juvenile individuals. Several study populations deviated from the natural pattern of year-round reproduction typical for the species, rendering those with severely truncated reproductive periods vulnerable to extinction in the event of environmental fluctuation. In contrast, demographically diverse populations are more likely to persist through such periods through the continuous presence of adults with broader physiological tolerance to abrupt salinity changes. Notably, we found a significant correlation between genetic diversity and demographic variation in the study populations, which could be the result of population stressors that restrict both of these diversity measures simultaneously, or suggestive of a causative relationship between these population characteristics. These findings demonstrate the importance of biocomplexity at the population level, and assert that the maintenance of diversity contributes to population resilience and conservation of this endangered species.     

Spatial genetic structure and frequency of interspecific hybridization in Platanthera aquilonis and P. dilatata (Orchidaceae) occurring in sympatry

Knowledge of genetic structure at different scales is necessary for evaluating the importance of interactions between the genome and environment and for inferring underlying processes that bring about evolutionary diversification. Here, genetic and morphological variation was assessed for 154 individuals of Platanthera aquilonis and P. dilatata in Maine, using RAPD and PCR-RFLP markers and measurements of five morphological traits to determine the prevalence of interspecific hybrids and underlying spatial genetic structure of the population. Both species co-mingled in the population, but clumping was observed. Platanthera aquilonis was more abundant, but genetic variation was lower (polymorphic loci [40%], expected heterozygosity [0.137]) than that of P. dilatata (P(p) = 72%; H(e) = 0.245). Interspecific hybrids were rare (eight individuals), and morphology was not consistently reliable for determining hybrid status even though the species differed significantly in several traits. Spatial autocorrelation analyses showed significant genetic structure at small distances for both species, most likely due to restricted seed dispersal. Platanthera aquilonis did not exhibit a pattern of spatial genetic structure consistent with other selfing species. This suggests that the species is facultative autogamous, which allows for limited production of hybrid individuals and reduces the strength of spatial genetic structure relative to P. dilatata.