Siegfried L. Krauss

Accurate gene diversity estimates from amplified fragment length polymorphism (AFLP) markers

Three procedures for the estimation of null allele frequencies and gene diversity from dominant multilocus data were empirically tested in natural populations of the outcrossing angiosperm Persoonia mollis (Proteaceae). The three procedures were the square root transform of the null homozygote frequency, the Lynch & Milligan procedure, and the Bayesian method. Genotypes for each of 116 polymorphic loci generated by amplified fragment length polymorphism (AFLP) were inferred from segregation patterns in progeny arrays. Therefore, for the plus phenotype (band present), heterozygotes were distinguished from homozygotes. In contrast to previous studies, all three procedures produced very similar mean estimates of heterozygosity, which were in turn accurate estimators of the direct value (HO = 0.28). A second population of P. mollis displayed markedly lower levels of heterozygosity (HO = 0.20) but approximately twice as many polymorphic loci (284). These AFLP results show that biases in estimates of average null allele frequency and heterozygosity are largely eliminated in highly polymorphic dominant marker data sets displaying a J‐shaped beta distribution with a high percentage of loci containing more than three null homozygotes and relatively few loci with no null homozygotes. This distribution may be typical of outcrossing angiosperms.

The Central Role of Dispersal in the Maintenance and Persistence of Seagrass Populations

Global seagrass losses parallel significant declines observed in corals and mangroves over the past 50 years. These combined declines have resulted in accelerated global losses to ecosystem services in coastal waters. Seagrass meadows can be extensive (hundreds of square kilometers) and long-lived (thousands of years), with the meadows persisting predominantly through vegetative (clonal) growth. They also invest a large amount of energy in sexual reproduction. In this article, we explore the role that sexual reproduction, pollen, and seed dispersal play in maintaining species distributions, genetic diversity, and connectivity among seagrass populations. We also address the relationship between long-distance dispersal, genetic connectivity, and the maintenance of genetic diversity that may enhance resilience to stresses associated with seagrass loss. Our reevaluation of seagrass dispersal and recruitment has altered our perception of the importance of long-distance dispersal and has revealed extensive dispersal at scales much larger than was previously thought possible.

Anthropogenic disturbance promotes hybridization between Banksia species by altering their biology

Abstract Putative hybrids between Banksia hookeriana and B. prionotes were identified among 12 of 106 populations of B. hookeriana located at or near anthropogenically disturbed sites, mainly roadways, but none in 156 undisturbed populations. Morphometrics and AFLP markers confirmed that a hybrid swarm existed in a selected disturbed habitat, whereas no intermediates were present where the two species co‐occurred in undisturbed vegetation. Individuals of both species in disturbed habitats at 12 sites were more vigorous, with greater size and more flower heads than their counterparts in undisturbed vegetation. These more fecund plants also showed a shift in season and duration of flowering. By promoting earlier flowering of B. hookeriana plants and prolonging flowering of B. prionotes, anthropogenic disturbance broke the phenological barrier between these two species. We conclude that anthropogenic disturbance promotes hybridization through increasing opportunities for gene flow by reducing interpopulation separation, increasing gamete production and, especially, promoting coflowering.

Long‐distance seed dispersal in a metapopulation of Banksia hookeriana inferred from a population allocation analysis of amplified fragment length polymorphism data

There is currently a poor understanding of the nature and extent of long‐distance seed dispersal, largely due to the inherent difficulty of detection. New statistical approaches and molecular markers offer the potential to accurately address this issue. A log‐likelihood population allocation test (aflpop) was applied to a plant metapopulation to characterize interpopulation seed dispersal. Banksia hookeriana is a fire‐killed shrub, restricted to sandy dune crests in fire‐prone shrublands of the Eneabba sandplain, southwest Australia. Population genetic variation was assessed for 221 individuals sampled from 21 adjacent dune‐crest populations of B. hookeriana using amplified fragment length polymorphism. Genetic diversity was high, with 175 of 183 (96%) amplified fragment length polymorphism markers polymorphic. Of the total genetic diversity, 8% was partitioned among populations by amova and FST. There was no relationship between genetic diversity within populations and population demographic parameters such as population size and sample size. A population allocation test on these data unambiguously assigned 177 of 221 (80.1%) individuals to a single population. Of these, 171 (77.4% of total) were assigned to the population from which they were sampled and 6 (2.7% of total) were assigned to a known population other than the one from which they were sampled. A further 9 (4.1% of total) were assigned to outside the sampled metapopulation area, and 35 individuals (15.8%) could not be assigned unambiguously to any particular population. These results suggest that both the extent [15 of 221 (6.8%) individuals originating from a population other than the one in which they occur] and distance (1.6 to > 2.5 km), of seed dispersal between dune‐crest populations is greater than expected from previous studies. The extent of long‐distance interpopulation seed dispersal observed provides a basis for explaining the survival of populations of the fire‐killed B. hookeriana in a landscape experiencing frequent fire, where local extinctions and recolonizations may be a regular occurrence.

Conservation genetics of the rare and endangered Leucopogon obtectus (Ericaceae)

Leucopogon obtectus Benth. is a declared rare species found in the kwongan vegetation in Western Australia. Plants on a mineral sand mine and the rehabilitation area are subject to disturbance. Genetic diversity was examined within and among all known populations using random amplified polymorphic DNA (RAPD) and amplified fragment length polymorphisms (AFLPs) for conservation. Both molecular markers revealed a high percentage (> 89%) of polymorphic markers and a high mean genetic distance among individuals (D = 0.3). Analysis of molecular variance showed that 86.7% (RAPD) and 89.7% (AFLP) of variability was partitioned among individuals within populations. Exact tests showed no significant population differentiation. The analyses indicated that L. obtectus exhibits high levels of genetic diversity despite small population sizes. The high levels of variability among individuals and the lack of clear population differentiation suggest that this species comprises a single, genetically diverse group. Conservation and management of L. obtectus should concentrate on maintaining the high levels of genetic variability through mixing genotypes and promoting outcrossing.

The movement ecology of seagrasses

A movement ecology framework is applied to enhance our understanding of the causes, mechanisms and consequences of movement in seagrasses: marine, clonal, flowering plants. Four life-history stages of seagrasses can move: pollen, sexual propagules, vegetative fragments and the spread of individuals through clonal growth. Movement occurs on the water surface, in the water column, on or in the sediment, via animal vectors and through spreading clones. A capacity for long-distance dispersal and demographic connectivity over multiple timeframes is the novel feature of the movement ecology of seagrasses with significant evolutionary and ecological consequences. The space–time movement footprint of different life-history stages varies. For example, the distance moved by reproductive propagules and vegetative expansion via clonal growth is similar, but the timescales range exponentially, from hours to months or centuries to millennia, respectively. Consequently, environmental factors and key traits that interact to influence movement also operate on vastly different spatial and temporal scales. Six key future research areas have been identified.

Genetic fidelity and viability of Anigozanthos viridis following tissue culture, cold storage and cryopreservation

Abstract The effects of long-term storage conditions on the viability and genetic fidelity of plant somatic tissues are poorly known. In this study, the effects of three storage methods (tissue culture, cold storage and cryostorage) on genetic fidelity and shoot apex viability were evaluated for Anigozanthos viridis subspp. terraspectans (Haemodoraceae), a threatened plant from south west Australia. Genetic fidelity was assessed following 12 months of storage using the PCR-based multi-locus DNA fingerprinting technique Amplified Fragment Length Polymorphism (AFLP). Shoot apex viability was evaluated at 0, 3, 6 and 12 months for cryogenically stored material. The AFLP technique generated a total of 95 fragments for three primer pairs, and no differences were detected across treatments. Post-cryostorage viability was high (mean=85%) and not significantly different across storage times. These results show that genetic fidelity and shoot apex viability (for cryopreserved material) were maintained following tissue culture, cold storage and cryostorage of A. viridis subspp. terraspectans for up to 12 months.

Next generation restoration genetics: Applications and opportunities

Restoration ecology is a young scientific discipline underpinning improvements in the rapid global expansion of ecological restoration. The application of molecular tools over the past 20 years has made an important contribution to understanding genetic factors influencing ecological restoration success. Here we illustrate how recent advances in next generation sequencing (NGS) methods are revolutionising the practical contribution of genetics to restoration. Novel applications include a dramatically enhanced capacity to measure adaptive variation for optimal seed sourcing, high-throughput assessment and monitoring of natural and restored biological communities aboveground and belowground, and gene expression analysis as a measure of genetic resilience of restored populations. Challenges remain in data generation, handling and analysis, and how best to apply NGS for practical outcomes in restoration.

Contrasting impacts of pollen and seed dispersal on spatial genetic structure in the bird-pollinated Banksia hookeriana

In plants, pollen- and seed-dispersal distributions are characteristically leptokurtic, with significant consequences for spatial genetic structure and nearest-neighbour mating. However, most studies to date have been on wind- or insect-pollinated species. Here, we assigned paternity to quantify effective pollen dispersal over 9 years of mating, contrasted this to seed dispersal and examined their effects on fine-scale spatial genetic structure, within the bird-pollinated shrub Banksia hookeriana (Proteaceae). We used 163 polymorphic amplified fragment length polymorphism markers to assess genetic structure and pollen dispersal in a spatially discrete population of 112 plants covering 0.56 ha. Spatial autocorrelation analysis detected spatial genetic structure in the smallest distance class of 0–5 m (r=0.025), with no significant structure beyond 8 m. Experimentally quantified seed-dispersal distances for 337 seedlings showed a leptokurtic distribution around a median of 5 m, reaching a distance of 36 m. In marked contrast, patterns of pollen dispersal for 274 seeds departed strikingly from typical near-neighbour pollination, with a distribution largely corresponding to the spatial distribution of plants. We found very high multiple paternity, very low correlated paternity and an equal probability of siring for the 50 closest potential mates. Extensive pollen carryover was demonstrated by multiple siring in 83 of 86 (96.5%) two-seeded fruits. Highly mobile nectar-feeding birds facilitate this promiscuity through observed movements that were effectively random. As the incidence of bird-pollination is markedly greater in the Southwest Australian Floristic Region than elsewhere, our results have broad and novel significance for the evolution and conservation for many species in Gondwanan lineages.

Low Rate of Between-Population Seed Dispersal Restricts Genetic Connectivity and Metapopulation Dynamics in a Clonal Shrub

Clonal species normally have low seed production, low recruitment rates and long lifespans, and it is expected that the rates of long-distance dispersal (LDD) of seeds will be low as well. Banksia candolleana is a clonal shrub in Mediterranean-type, fire-prone sclerophyll shrublands of southwestern Australia, whose reproductive biology and population dynamics contrast with those of co-occurring nonclonal congeneric species, all of which are restricted to a mosaic of sand dunes set within a matrix of inhospitable swales. Using microsatellite markers, we genotyped 499 plants in all 15 populations of B. candolleana within a 12-km2 area, assessed population genetic differentiation, and quantified the effective rate of interpopulation seed dispersal through genetic assignment of individuals to populations. We measured life history, reproductive and demographic attributes, and compared these with two co-occurring Banksia species, a non-clonal resprouter and a nonsprouter. B. candolleana has much higher levels of population genetic differentiation, and one-third the rate of interpopulation seed migration, as the other two species (2.2% vs 5.5−6.8% of genotyped plants inferred to be immigrants), though distances reached by LDD are comparable (0.3−2.3 km). The low rate of interpopulation dispersal was supported by an analysis of the age structure of three populations that suggests a mean interdune migration rate of <800 m in 200 years, and 60% of suitable dunes remain uninhabited. Thus, B. candolleana has poor properties for promoting long-distance dispersal. It is unclear if these are idiosyncratic to this species or whether such properties are to be expected of clonal species in general where LDD is less critical for species survival.