Assisted species migration and hybridization to conserve cold-adapted plants under climate change.

Temperature rise due to climate change is putting many arctic and alpine plants at risk of extinction because their ability to react is outpaced by the speed of climate change. We considered assisted species migration (ASM) and hybridization as methods to conserve cold-adapted species (or the genes thereof) and to minimize the potential perturbation of ecosystems due to climate change. Assisted species migration is the deliberate movement of individuals from their current location to where the species' ecological requirements will be matched under climate projections. Hybridization refers to crossbreeding of closely related species, where for arctic and alpine plants, 1 parent is the threatened cold-adapted and the other its reproductively compatible, warm-adapted sibling. Traditionally, hybridization is viewed as negative and leading to a loss of biodiversity, even though hybridization has increased biodiversity over geological times. Furthermore, the incorporation of warm-adapted genes into a hybrid may be the only means for the persistence of increasingly more maladapted, cold-adapted species. If approached with thorough consideration of fitness-related parameters of the source population and acknowledgement of the important role hybridization has played in shaping current biodiversity, ASM and hybridization could help save partial or whole genomes of key cold-adapted species at risk due to climate change with minimal negative effects on ecosystem functioning.

Migración Asistida e Hibridación de Especies para Conservar Plantas Adaptadas al Frío Ante el Cambio Climático 20--040 Resumen El incremento de la temperatura debido al cambio climático pone en riesgo de extinción a muchas plantas alpinas y árticas debido a que su habilidad para reaccionar está sobrepasada por la velocidad del cambio climático. Consideramos a la migración asistida de especies (MAE) y la hibridación como métodos para conservar las especies adaptadas al frío (los genes de las mismas) y para minimizar la potencial perturbación de los ecosistemas causada por el cambio climático. La migración asistida de especies es el movimiento deliberado de individuos desde su ubicación actual hacia donde se verán igualados los requerimientos ecológicos de la especie bajo las proyecciones de cambio climático. La hibridación se refiere al cruzamiento de especies cercanas, en el caso de las plantas árticas y alpinas un progenitor es la especie amenazada adaptada al frío y el otro su especie hermana adaptada al calor y con compatibilidad reproductiva. Tradicionalmente, la hibridación es vista bajo un concepto negativo que culmina en la pérdida de biodiversidad, a pesar de que la hibridación ha incrementado la biodiversidad a lo largo del tiempo geológico. Más allá de esto, la incorporación de genes adaptados al calor dentro de un híbrido puede ser el único medio para la persistencia de especies adaptadas al frío cada vez más desadaptadas. Si se utilizan bajo un análisis meticuloso de los parámetros relacionados con la aptitud de la población originaria y un reconocimiento del importante papel que la hibridación ha desempeñado en la formación de la biodiversidad actual, la ASM y la hibridación podrían ayudar a rescatar genomas parciales o completos de las especies adaptadas al frío que se encuentran en riesgo debido al cambio climático con unos efectos negativos mínimos sobre el funcionamiento del ecosistema.

Environmental influence on primary sex ratio in a dioecious plant.

The proximity of mates can influence mating opportunities and the quantity and quality of offspring, especially in dioecious plant species. Progeny sex ratios modulated by environmental conditions is one of the most radical ways in which offspring quality may be influenced, yet it has rarely been reported in plants. A mechanism proposed to influence progeny sex ratios in dioecious plants involves competition between female- and male-determining microgametophytes (certation) as a result of variation in pollination intensity. However, the role of selective fertilization in dioecious plants is controversial and has not been demonstrated under field conditions. Here we investigate whether natural variation in the spatial arrangement of females and males influences pollination intensity and progeny sex ratios in the wind-pollinated herb Rumex nivalis. Based on previous experimental manipulation of pollination intensity in this species, we predicted that maternal parents in close proximity to males would produce more strongly female-biased progeny sex ratios. We tested this prediction in six alpine populations in Switzerland by measuring the distance between focal females and neighboring males and assessing pollen loads and seed sex ratios of maternal parents. In four of the six populations, females positioned in close proximity to males captured more pollen and exhibited more female-biased sex ratios. Our results demonstrate that demographic aspects of the maternal mating environment can influence progeny sex ratios. The most probable explanation for biased primary sex ratios in Rumex is selective fertilization resulting from pollen tube competition.

Sexing pollen reveals female bias in a dioecious plant.

* Information on angiosperm sex ratios has largely been restricted to surveys of flowering individuals. These often deviate from equality, with male bias more commonly reported. Female-biased sex ratios are concentrated in a few taxa and have been linked to the possession of heteromorphic sex chromosomes and bias introduced during the gametophytic stage of the life cycle. It has been proposed that differences in gamete quantity and quality could give rise to female bias, although there is no direct evidence with which to evaluate this possibility. * Here, we use flow cytometry to investigate microgametophytic 'sex ratios' in a flowering plant. We demonstrate that differences in DNA content between the sexes in Rumex nivalis, a species with heteromorphic sex chromosomes, make it possible to distinguish female- vs male-determining pollen nuclei. * We found a small but significant female bias in microgametophytes produced by males (mean 0.515) with significant variation among family means (range 0.463-0.586), and 18 of 22 families averaging > 0.50. * The observed female bias at the gametophytic stage of the life cycle is consistent with the direction of bias previously reported for seeds and vegetative and reproductive plants in wild populations of R. nivalis, but is insufficient to fully explain the degree of bias.

Pollination intensity influences sex ratios in dioecious Rumex nivalis, a wind-pollinated plant.

Determining the mechanisms governing sex-ratio variation in dioecious organisms represents a central problem in evolutionary biology. It has been proposed that in plants with sex chromosomes competition between pollen tubes of female- versus male-determining microgametophytes (certation) causes female-biased primary sex ratios. Experimental support for this hypothesis is limited and recent workers have cast doubt on whether pollen-tube competition can modify sex ratios in dioecious plants. Here we investigate the influence of variation in pollination intensity on sex ratios in Rumex nivalis, a wind-pollinated alpine herb with strongly female-biased sex ratios. In a garden experiment, we experimentally manipulated pollination intensity using three concentric rings of female recipient plants at different distances from a central group of male pollen donors. This design enabled us to test the hypothesis that increasing pollen load size, by intensifying gametophyte competition, promotes female-biased sex ratios in R. nivalis. We detected a significant decline in pollen load at successive distance classes with concomitant reductions in seed set. Sex ratios of progeny were always female biased, but plants at the closest distance to male donors exhibited significantly greater female bias than more distant plants. The amount of female bias was positively correlated with the seed set of inflorescences. Hand pollination of stigmas resulted in approximately 100-fold higher stigmatic pollen loads than wind-pollinated stigmas and produced exceptionally female-biased progenies (female frequency = 0.96). Our results are the first to demonstrate a functional relation between stigmatic pollen capture, seed set, and sex ratio and suggest that certation can contribute towards female-biased sex ratios in dioecious plants.

Spatial ecology of mating success in a sexually polymorphic plant.

The spatial context of reproduction is of crucial importance to plants because of their sessile habit. Since pollen and seed dispersal is often restricted, mating success is likely to depend on the quantity and quality of mates in local neighbourhoods. Here we use neighbourhood models to investigate the spatial ecology of pollination and mating in Narcissus assoanus, a sexually polymorphic plant with two mating morphs that differ in style length. By mapping individuals in eight populations from southwestern France, we investigated the influence of the density and morph identity of plants at different spatial scales on variation in female fertility. By using inferences on the expected patterns of pollen transfer based on floral morphology, we were able to predict the quantitative relations between local morph ratios and variation in fertility. Our analyses revealed differences in the spatial clustering of morphs and in their response to plant density and morph identity within local neighbourhoods. Mating success in N. assoanus was characterized by both density- and frequency-dependent processes, a condition that may be a general feature of the spatial ecology of plant mating.

Mechanisms governing sex-ratio variation in dioecious Rumex nivalis.

Sex ratios of flowering individuals in dioecious plant populations are often close to unity, or are male biased owing to gender-specific differences in flowering or mortality. Female-biased sex ratios, although infrequent, are often reported in species with heteromorphic sex chromosomes. Two main hypotheses have been proposed to account for female bias: (1) selective fertilization resulting from differential pollen-tube growth of female- versus male-determining microgametophytes (certation); (2) differences in the performance and viability of the sexes after parental investment. Here we investigate these hypotheses in Rumex nivalis (Polygonaceae), a European alpine herb with female-biased sex ratios in which females possess XX, and males XY1Y2, sex chromosomes. Using field surveys and a glasshouse experiment we investigated the relation between sex ratios and life-history stage in 18 populations from contrasting elevations and snowbed microsites and used a male-specific SCAR-marker to determine the sex of nonflowering individuals. Female bias among flowering individuals was one of the highest reported for populations of a dioecious species (mean female frequency = 0.87), but males increased in frequency at higher elevations and in the center of snowbeds. Female bias was also evident in nonflowering individuals (mean 0.78) and in seeds from open-pollinated flowers (mean 0.59). The female bias in seeds was weakly associated with the frequency of male flowering individuals in populations in the direction predicted when certation occurs. Under glasshouse conditions, females outperformed males at several life-history stages, although male seeds were heavier than female seeds. Poor performance of Y1Y2 gametophytes and male sporophytes in R. nivalis may be a consequence of the accumulation of deleterious mutations on Y-sex chromosomes.

Implications of ITS sequences and RAPD markers for the taxonomy and biogeography of the Oxytropis campestris and O. arctica (Fabaceae) complexes in Alaska.

Taxonomic consensus is lacking on the Oxytropis arctica and O. campestris species complexes, two polyploid complexes found in the interior and arctic areas of Alaska. One classification has emphasized flower size, whereas flower color is considered a key diagnostic character in another classification. Our analyses of internal transcribed spacer (ITS) sequences and random amplified polymorphic DNA (RAPD) markers provided no support for either classification system. The trees generated from ITS sequences and the phenogram derived from RAPD markers suggest that most recognized taxa in the two complexes are probably polyphyletic, including O. arctica var. barnebyana, which is listed as threatened in Alaska. The only consistent pattern detected by both types of molecular markers was a geographic split dividing the northeastern arctic populations from most other populations (48.60-55.03% in AMOVA analyses). This genetic subdivision probably reflects a Pleistocene barrier formed by the northern coastal ice shield. Our molecular data, in conjunction with the previously reported variation of ploidy levels in these groups, suggest a scenario of recent and multiple origins of polyploidy. It is possible that most Alaskan populations of these two complexes are best referred to as a single taxonomic species despite morphological differentiation within the complexes.

Glacial history of the alpine herb Rumex Nivalis (Polygonaceae): a comparison of common phylogeographic methods with nested clade analysis.

The glacial history of the alpine herb Rumex nivalis was investigated using amplified fragment length polymorphisms (AFLPs) and restriction fragment length polymorphisms with polymerase chain reaction (PCR-RFLPs) of cpDNA. Both traditional statistical methods widely applied in phylogeographic research and nested clade analysis were used. The AFLPs indicated little geographic structure probably due to the wind-pollinated reproductive system of the dioecious R. nivalis. Because cpDNA haplotypes exhibited distinct distributional patterns, correlation between AFLPs and PCR-RFLPs was low. The results of common statistical methods and of nested clade analysis were largely congruent. Both supported in situ survival of one group of common haplotypes in the Central Alps. For another group of common haplotypes, classical phylogeographic analyses gave strong evidence for survival in peripheral refugia at the northern alpine border, whereas this conclusion was not as clearly supported in the nested clade analysis. Nested clade analysis provided several detailed insights on past and ongoing populational demographic processes. Thus, it is a valuable tool in the phylogeographical analysis of haplotype data, but it should preferably be combined with other statistical analyses. In situations with low genetic variation in cpDNA, classical phylogeographic analytical tools on nuclear DNA will still be the methods of choice.

Estimation of the relative importance of sexual and vegetative reproduction in the clonal woodland herb Anemone nemorosa.

Anemone nemorosa is a perennial rhizomatous plant of European woodlands. The "probability of clonal identity" method estimated the relative proportion of sexual to vegetative reproduction in this species to be 4.4% from allozyme genotype distributions. This result is congruent with investigations on the germination, short-term demography, population genetics, and breeding system of this species, and supports the hypothesis that even low levels of seedling recruitment can maintain considerable intrapopulational genetic diversity.