Jennifer K. Rowntree

CONSERVATION IN VITRO OF THREATENED PLANTS – PROGRESS IN THE PAST DECADE

SummaryIn vitro techniques have found increasing use in the conservation of threatened plants in recent years and this trend is likely to continue as more species face risk of extinction. The Micropropagation Unit at Royal Botanic Gardens, Kew, UK (RBG Kew) has an extensive collection of in vitro plants including many threatened species from throughout the world. The long history of the unit and the range of plants cultured have enabled considerable expertise to be amassed in identifying the problems and developing experimental strategies for propagation and conservation of threatened plants. While a large body of knowledge is available on the in vitro culture of plants, there are limited publications relating to threatened plant conservation. This review highlights the progress in in vitro culture and conservation of threatened plants in the past decade (1995–2005) and suggests future research directions. Works on non-threatened plants are also included wherever methods have applications in rare plant conservation. Recalcitrant plant materials collected from the wild or ex situ collections are difficult to grow in culture. Different methods of sterilization and other treatments to establish clean material for culture initiation are reviewed. Application of different culture methods for multiplication, and use of unconventional materials for rooting and transplantation are reviewed. As the available plant material for culture initiation is scarce and in many cases associated with inherent problems such as low viability and endogenous contamination, reliable protocols on multiplication, rooting, and storage methods are very important. In this context, photoautotrophic micropropagation has the potential for development as a routine method for the in vitro conservation of endangered plants. Long-term storage of material in culture is challenging and the potential applications of cryopreservation are significant in this area. Future conservation biotechnology research and its applications must be aimed at conserving highly threatened, mainly endemic, plants from conservation hotspots.

Forward from the crossroads of ecology and evolution

Community genetics is a synthesis of community ecology and evolutionary biology. It examines how genetic variation within a species affects interactions among species to change ecological community structure and diversity. The use of community genetics approaches has greatly expanded in recent years and the evidence for ecological effects of genetic diversity is growing. The goal of current community genetics research is to determine the circumstances in which, and the mechanisms by which community genetic effects occur and is the focus of the papers in this special issue. We bring a new group of researchers into the community genetics fold. Using a mixture of empirical research, literature reviews and theoretical development, we introduce novel concepts and methods that we hope will enable us to develop community genetics into the future.

Climate Change and Eco-Evolutionary Dynamics in Food Webs

Abstract The analysis of the causes of selection is in essence a problem in ecology Wade and Kalisz 1990 Past evolution determines the genetically determined available phenotypes in populations which affect ecological dynamics in communities, shaping in turn the selective pressures that further model phenotypes. Because an increase in temperature increases metabolic rates and encounter rates, climate change may have profound eco-evolutionary effects, possibly affecting the future persistence and functioning of food webs. We introduce a semi-spatially explicit individual-based model (IBM) framework to study functional eco-evolutionary dynamics in food webs. Each species embedded in the web includes 13 genetically determined and multidimensionally variable traits (the G matrix), 4 of which are flexible physiological and behavioural (personality) traits that respond to temperature. An increase in temperature and stronger correlation among traits leads to stronger trophic cascades but higher stochasticity, with higher probability of extinction for some trophic levels. A combination of the abiotic (temperature) and biotic (predators’ presence/absence) matrix of selective agents (the O matrix) generates differential selection for activation energies for metabolic rates and several instances of correlational selection (selection in one trait changes with the levels of another), suggesting how global warming might favour certain trait combinations. Our results and the future prospects of this IBM approach open new avenues for climate change research.

Genetic variation changes the interactions between the parasitic plant-ecosystem engineer Rhinanthus and its hosts

Within-species genetic variation is a potent factor influencing between-species interactions and community-level structure. Species of the hemi-parasitic plant genus Rhinanthus act as ecosystem engineers, significantly altering above- and below-ground community structure in grasslands. Here, we show the importance of genotypic variation within a single host species (barley—Hordeum vulgare), and population-level variation among two species of parasite (Rhinanthus minor and Rhinanthus angustifolius) on the outcome of parasite infection for both partners. We measured host fitness (number of seeds) and calculated parasite virulence as the difference in seed set between infected and uninfected hosts (the inverse of host tolerance). Virulence was determined by genetic variation within the host species and among the parasite species, but R. angustifolius was consistently more virulent than R. minor. The most tolerant host had the lowest inherent fitness and did not gain a fitness advantage over other infected hosts. We measured parasite size as a proxy for transmission ability (ability to infect further hosts) and host resistance. Parasite size depended on the specific combination of host genotype, parasite species and parasite population, and no species was consistently larger. We demonstrate that the outcome of infection by Rhinanthus depends not only on the host species, but also on the underlying genetics of both host and parasite. Thus, genetic variations within host and parasite are probably essential components of the ecosystem-altering effects of Rhinanthus.

Pleiotropic effects of environment‐specific adaptation in Arabidopsis thaliana

Local adaptation may be important for the preservation of genetic diversity and the promotion of speciation. However, local adaptation may also constrain establishment in different environments. The consequences of local adaptation depend strongly on the pleiotropic effects of the genes involved in adaptation. Here, we investigated the pleiotropic effects of the genetic response to selection in outbred lines of Arabidopsis artificially selected to flower earlier under both winter- and spring-annual simulated conditions. The consequences of adaptation were evaluated by reciprocally transplanting selected and control lines between the two conditions. Selected lines always flower earlier than their controls, independent of growing conditions. However, selected lines, growing in the same condition in which they were selected, flower earlier than plants selected in the alternative environment. Plants selected to flower earlier in spring produce more fruits than controls when growing in the spring, and less fruits when growing in the winter; indicating that local adaptation has negative pleiotropic effects in another environment. Our results indicate that local adaptation can arise even when selection targets the same trait in the same direction. Furthermore, it suggests that adaptation under the two different environments can generate fitness trade-offs that can maintain genetic variation for flowering time.

Plant genotype mediates the effects of nutrients on aphids

Soil nutrients, and factors which influence their concentrations and bioavailability, form a basic component of bottom–up control of ecosystem processes, including plant–herbivore interactions. Increased nutrient levels are linked, through plant defence theory, with increased levels of herbivore susceptibility. The focal point of many ecological experiments examining this link is at the species level, where the response of single species is the average of many different genotypes. Here, we focus on the genetic basis of indirect ecological interactions. We investigated the effects of nutrient concentration on the population growth of an aphid herbivore across multiple genotypes of barley in relation to plant growth rate. In general, both aphid population size and plant growth rate increased with nutrient concentration. However, they were both dependent on the interaction between nutrient concentration and barley genotype. Our data raise the testable possibility of differential defense responses between genotypes of barley, for example divergent, fixed and inducible defences against aphids. We provide evidence that the indirect effects of soil nutrients on aphid population size are mediated by the genetics of the host plant.

How bryophytes came out of the cold: successful cryopreservation of threatened species.

The use of ex situ techniques for the conservation of threatened plants has been increasing over the past decades. Cryopreservation is often used for the long-term storage of plant germplasm where conventional methods (i.e. seedbanking) are inappropriate. Simple encapsulation–dehydration protocols were developed for the cryopreservation of bryophytes at The Royal Botanic Gardens, Kew, as part of an ex situ project for the conservation of UK threatened species. The applicability of these methods was tested on 22 species with a broad range of ecological requirements and found to be highly successful across the board. Regeneration rates from frozen material were >68% for all species tested and half had regeneration rates of 100%. The high regeneration rate and broad applicability of the protocols across a range of species was attributed to a combination of the inherent totipotency of bryophytes and the in-built recovery periods in the pre-treatment protocol. In conclusion, bryophytes are well suited to cryopreservation and such techniques would be applicable for the long-term storage of similar conservation collections across the globe.

Correlated response in plasticity to selection for early flowering in Arabidopsis thaliana

Phenotypic plasticity is an important strategy for coping with changing environments. However, environmental change usually results in strong directional selection, and little is known empirically about how this affects plasticity. If genes affecting a trait value also affect its plasticity, selection on the trait should influence plasticity. Synthetic outbred populations of Arabidopsis thaliana were selected for earlier flowering under simulated spring‐ and winter‐annual conditions to investigate the correlated response of flowering time plasticity and its effect on family‐by‐environment variance (Vg×e) within each selected line. We found that selection affected plasticity in an environmentally dependent manner: under simulated spring‐annual conditions, selection increased the magnitude of plastic response but decreased Vg×e; selection under simulated winter‐annual conditions reduced the magnitude of plastic response but did not alter Vg×e significantly. As selection may constrain future response to environmental change, the environment for crop breeding and ex situ conservation programmes should be carefully chosen. Models of species persistence under environmental change should also consider the interaction between selection and plasticity.

The effect of multiple host species on a keystone parasitic plant and its aphid herbivores

1. The exploitation of shared resources by diverse organisms underpins the structure of ecological communities. Hemiparasitic plants and the insect herbivores feeding on them both rely, directly and indirectly, on the resources supplied by the parasites host plant. Therefore, the identity and number of host plant species providing these resources is likely to be critical for parasite and herbivore performance. 2. We tested the effect of single and multiple host species on the biomass of the generalist parasitic plant Rhinanthus minor and the abundance of its aphid (Aphis gossypii) herbivores. 3. Parasite biomass was proportional to the number of haustorial connections to host roots and was determined by host species identity rather than host functional group. Host species identity was also an important influence on aphid population size, and parasites attached to Lotus corniculatus experienced a considerable reduction in aphid herbivory. 4. The effects on the parasite attaching to multiple hosts depended on the combination of species present. However, host mixtures generally benefitted aphids by diluting the negative effects of particular host species. 5. Our findings suggest that the specificity of host attachment alters the impact of this keystone parasitic plant on its own herbivores and, potentially, on the wider plant and herbivore community.

Which moss is which? Identification of the threatened moss Orthodontium gracile using molecular and morphological techniques

Taxonomic misidentification has potentially serious consequences for the management of threatened species. Closely related moss species are often difficult to distinguish from each other using morphological characteristics. Here we compared the use of molecular (DNA barcoding of the trnL-F intron, AFLPs) and morphological techniques to demonstrate that ex situ cultures, held for re-introduction trails, of the UK critically endangered moss Orthodontium gracile were contaminated with the potentially invasive species O. lineare. Barcoding techniques and AFLPs were both successful in determining Orthodontium species identity. There was some discrepancy between determinations from molecular and morphological techniques and some individuals were misidentified using morphological characteristics alone. When species identity is critical, for example prior to re-establishment or re-introduction programmes, we recommend that identity of mosses and other bryophytes be established by molecular techniques, in particular barcoding of the trnL-F intron.