The Use of Combining Ability Analysis to Identify Elite Parents for Artemisia annua F1 Hybrid Production

Artemisia annua is an important medicinal crop used for the production of the anti-malarial compound artemisinin. In order to assist in the production of affordable high quality artemisinin we have carried out an A. annua breeding programme aimed at improving artemisinin concentration and biomass. Here we report on a combining ability analysis of a diallel cross to identify robust parental lines for hybrid breeding. The parental lines were selected based on a range of phenotypic traits to encourage heterosis. The general combining ability (GCA) values for the diallel parental lines correlated to the positive alleles of quantitative trait loci (QTL) in the same parents indicating the presence of beneficial alleles that contribute to parental performance. Hybrids generated from crossing specific parental lines with good GCA were identified as having an increase in both artemisinin concentration and biomass when grown either in glasshouse or experimental field trials and compared to controls. This study demonstrates that combining ability as determined by a diallel cross can be used to identify elite parents for the production of improved A. annua hybrids. Furthermore, the selection of material for breeding using this approach was found to be consistent with our QTL-based molecular breeding approach.     

Hydrologic and mechanical control for an invasive wetland plant, <Emphasis Type="Italic">Juncus ingens</Emphasis>, and implications for rehabilitating and managing Murray River floodplain wetlands,Australia

Wetlands are prone to increased invasion by plant species following changes in hydrologic regime, leading to shifts in plant community composition and potentially ecosystem function and health. In this paper, the ecology and potential control of Juncus ingens, a native wetland plant in the Murray-Darling Basin of south-eastern Australia, is investigated. J. ingens has benefited from altered Murray River hydrologic conditions by expanding its range and invading seasonally-flooded grassland and riparian habitats along the Murray River. Here results of complementary glasshouse and field research of seedling and mature J. ingens growth and resilience as influenced by hydrologic regime (moist, saturated, partial inundation and when possible, complete inundation) and mechanical control (i.e., clipping) are presented. A moist hydrologic regime (soil held at field capacity) resulted in the most vigorous seedlings (13.9–73.0% more total biomass), while saturated conditions (flooding maintained level with the soil surface) resulted in the most vigorous mature J. ingens (14.1–98.4% more total biomass). Seedling mortality was greatest under complete and prolonged inundation (60% fatal with remaining 40% showing severe stress), conditions suspected to have limited prior invasion but which currently occur infrequently as a result of reduced flooding magnitude. Clipping was fatal for nearly all seedlings regardless of hydrologic regime but was only fatal for mature plants if coupled with prolonged inundation. Coupling ground-level clipping with strategic flooding may be the most effective means for controlling current populations of J. ingens and limiting further invasion, promoting re-colonisation by displaced species and rehabilitating the health of Barmah Forest and similar Murray River floodplain wetlands.     

Risk Assessment of Transgenic Virus-resistant White Clover: Non-target Plant Community Characterisation and Implications for Field Trial Design

One of the most difficult elements of the ecological risk assessment of transgenic plants is investigation of their potential impact on biodiversity in complex non-target communities. This problem is particularly acute for pasture plants, since many have a track record of invasiveness. In this paper we develop an understanding of some aspects of the ecology of Trifolium repens L. (white clover) in montane vegetation communities in southeastern Australia as part of a larger project investigating potential ecological risks associated with commercial release of newly-developed transgenic virus-resistant T. repens cultivars. We use a combination of floristic surveys and biomass sampling to determine the habitat affinity of T. repens, the structure and composition of associated communities, and the scale at which different abiotic and biotic factors correlate with T. repens abundance. We also compare the abundance of native and exotic species within specific morpho-functional groups and use this to determine the relative significance of T. repens as a community constituent, and to identify native species that would be most at risk by expanding populations of T. repens. We found that T. repens comprises a relatively small component of the total community cover and biomass, but is one of the most abundant herbaceous species in mesic Poa – dominated grasslands and Poa-Eucalyptus woodlands in the study area, and that T. repens abundance is correlated at the within-community scale primarily with soil moisture and fertility. At smaller scales T. repens is limited by the hierarchical dominance of native graminoids and we conclude that competition for inter-tussock space in mesic communities is the most likely mechanism by which expanding populations of T. repens would influence populations of associated native species. These results have significant implications for the manner in which future analysis and risk quantification stages of the risk assessment of virus-resistant T.␣repens and other transgenic pasture plants in complex plant communities are performed.     

Wetland plant growth under contrasting water regimes associated with river regulation and drought: implications for environmental water management

An important characteristic of many wetland plants in semi-arid regions is their capacity to withstand fluctuations between extended dry phases and floods. However, anthropogenic river regulation can reduce natural flow variability in riverine wetlands, causing a decline in the frequency and duration of deep flooding as well as extended droughts, and an increase in shallow flooding and soil saturation. Our aim in this paper was to use an experimental approach to examine whether reductions in flooding and drought disadvantage species adapted to both these extremes, and favours those with water requirements that match the new regime of frequent low-level flooding. We compared the growth characteristics and biomass allocation of three native Australian aquatic macrophytes (Pseudoraphis spinescens, Juncus ingens and Typha domingensis), which co-occur at Barmah Forest, south-eastern Australia, under three water treatments: drought, soil saturation and deep flooding. The responses of species to the treatments largely reflected changes in their relative abundance at Barmah Forest since river regulation. Typha domingensis, which has remained uncommon, performed relatively poorly in all treatments, while J. ingens, which has increased its range, exhibited more vigorous growth under soil saturation. Pseudoraphis spinescens, which was once widespread but has declined markedly in its distribution, grew strongly under all water treatments. These findings suggest that a return to more natural, variable river flow regimes can potentially be an important conservation and restoration strategy in ecosystems characterised by species that have adaptations to extreme hydrological growing conditions.     

Ecological risk assessment of transgenic pasture plants: a community gradient modelling approach

The extension of transgene technology to pasture species presents new challenges for ecological risk assessment because, unlike most crops, pasture plants are well adapted for persistence in mixed plant communities and are an important source of invasive species worldwide. Despite this, the impact of transgenic pasture plants on native ecosystems remains relatively unstudied. Here we use a community gradient modelling approach to investigate the performance of subterranean clover containing a nutrition‐enhancing transgene over a range of ecologically important grassland communities at a study site in south‐eastern Australia. Our data, which were collected over a full annual growing season, show that the transgenic clover line has different seedling survival and seed dormancy breakdown rates than the commercial non‐transgenic line, which suggests that transgenic populations will decline faster than non‐transgenic ones in perennial native grassland communities but could, under optimal climatic conditions, have potentially higher growth rates in grazed annual grasslands. These results demonstrate that changes to ecologically important demographic parameters can occur in plants containing transgenes that are not typically considered fitness enhancing, and that genotype by environment interactions and management regimes that impact on these parameters may influence the invasiveness of transgenic plants. Our study also demonstrates the utility of incorporating plant community gradient modelling into ecological risk assessment protocols designed for transgenic pasture plants.     

Polyploidy affects the seed,dormancy and seedling characteristics of a perennial grass,conferring an advantage in stressful climates

• Polyploidy (the state of having more than two genome copies) is widely distributed in flowering plants and can vary within species, with polyploid races often associated with broad ecological tolerances. Polyploidy may influence within‐species variation in seed development, germination and establishment. We hypothesized that interactions between polyploidy and the seed developmental environment would affect subsequent dormancy, germination and early growth traits, particularly in stressful environments.
• Using seeds developed in a common garden under ambient and warmed conditions, we conducted germination trials under drought and temperature stress, and monitored the subsequent growth of seedlings. The study species, Themeda triandra, is a widespread, keystone, Australian native grass and a known polyploid complex.
• Tetraploid plants produced heavier, more viable seeds than diploids. Tetraploids were significantly more dormant than diploids, regardless of seed developmental environment. Non‐dormant tetraploids were more sensitive to germination stress compared to non‐dormant diploids. Finally, tetraploid seedlings were larger and grew faster than diploids, usually when maternal plants were exposed to developmental temperatures atypical to the source environment.
• Seed and seedling traits suggest tetraploids are generally better adapted to stressful environments than diploids. Because tetraploid seeds of T. triandra are more dormant they are less likely to germinate under stress, and when they do germinate, seedling growth is rapid and independent of seed developmental environment. These novel results demonstrate that polyploidy, sometimes in interaction with developmental environment and possibly also asexuality, can have within‐species variation in seed and seedling traits that increase fitness in stressful environments.

     

Self-compatibility and plant invasiveness: Comparing species in native and invasive ranges

A longstanding hypothesis (“Baker's rule”) is that plant invasiveness is facilitated by floral self compatibility rather than self incompatibility. Extending this idea, invasive species whose individuals vary in degree of self compatibility within the native range might be self compatible in invading or weedy populations, due to natural selection on the mating system. We compared mating system between native and invasive ranges for two major world invasives, one annual (Echium plantagineum) and one perennial (Solanum elaeagnifolium). For an additional annual species (Centaurea solstitialis) we compared non-weedy and weedy populations in the native range. No species was strongly spontaneously self pollinating, but the degree of self compatibility after hand pollination varied dramatically. Both annuals were self incompatible in native or non-weedy populations but self compatible in invasive or weedy ones; the reverse was true for the perennial. Individuals within populations of all three species also varied in their degree of self compatibility, suggesting a basis for natural selection, and populations of the same species sharing a status (native/non-weedy, invading/weedy) varied in average self compatibility. These results support the hypothesis that differential selection of progeny during invasion can result in self-compatible populations derived from ancestrally self-incompatible ones, but that this process may be less important in perennial species, which experience multiple opportunities for sexual reproduction. Overall, however, mating system may not operate alone and its contributions to invasiveness may be conditional on other attributes of a species including physiology, morphology, and life history.