Environmental influences on akinete germination and development in Nodularia spumigena (Cyanobacteriaceae), isolated from the Gippsland Lakes, Victoria, Australia

This study was carried out to investigate the genesis of N. spumigena blooms by specifically studying the effects of environmental variables (salinity, nitrogen, phosphorus and light) on the germination of N. spumigena akinetes. Optimal conditions for maximum germination and germling growth were determined by exposing akinetes to a range of salinities and nutrient (nitrogen and phosphorus) concentrations under two different irradiances. At pre-determined time periods, treatments were sampled and the percent germination and length of germlings assessed. The results indicated that akinete germination and germling growth were optimal at salinities from 5 to 25 and significantly reduced outside this range. A positive correlation in germination was observed with increasing nutrient (phosphorus and nitrate) concentration. Similarly, germling growth increased with increasing concentrations of both nutrients. Irradiance significantly influenced both germination and growth during salinity experiments, whereas in nutrient addition experiments, irradiance had no effect on germination; however, growth was significantly influenced during phosphorus addition experiments. Consequently, salinity and light appeared to be most critical in the germination process for N. spumigena akinetes, with phosphorus most important for germling growth. The study showed that N. spumigena may be able to germinate under environmental conditions outside its optimal range, but the growth of the germling is significantly reduced, which in turn suggests that its ability to form a bloom outside its optimal environmental conditions would also be greatly reduced.

Germination ecology and seed dispersal of a critically endangered plant: a case study of Pomaderris vacciniifolia (Round-leaf Pomaderris)

Change in ecosystem disturbance regimes from human land-use poses a worldwide problem for management of rare species. Two important types of disturbance influencing the persistence of species in Australian ecosystems are habitat fragmentation and fire. In this study, seed dispersal and the germination ecology of Pomaderris vacciniifolia-a critically endangered, rare endemic Australian shrub-were examined to identify likely influences of fire and fragmentation on the decline of populations. The response of seed germination to simulated effects of wildfire and canopy openings was investigated, as was the unaided dispersal capability of seeds from parent plants. A significant increase in germination rate was observed following 100°C heat treatment to seeds, while smoke and light exposure had little influence. Seed imbibition was strongly influenced by heat treatment. The findings indicate a likely positive post-fire germination response, with implications for recruitment success determined by moisture availability following fire. Unaided seed dispersal was limited, which partly explains the apparent decline of populations. Understanding disturbance requirements for threatened species, and subsequent management of landscapes for disturbance, will aid conservation of rare species throughout the world.

Reduced triacylglycerol mobilization during seed germination and early seedling growth in Arabidopsis containing nutritionally important polyunsaturated fatty acids

There are now several examples of plant species engineered to synthesize and accumulate nutritionally important polyunsaturated fatty acids in their seed triacylglycerols (TAG). The utilization of TAG in germinating seeds of such transgenic plants was unknown. In this study, we examined the TAG utilization efficiency during seed germination in transgenic Arabidopsis seeds containing several examples of these fatty acids. Seed TAG species with native fatty acids had higher utilization rate than the TAG species containing transgenically produced polyunsaturated fatty acids. Conversely, quantification of the fatty acid components remaining in the total TAG after early stages of seed germination revealed that the undigested TAGs tended to contain elevated levels of the engineered polyunsaturated fatty acids (PUFA). LC-MS analysis further revealed asymmetrical mobilization rates for the individual TAG species. TAGs which contained multiple PUFA fatty acids were mobilized slower than the species containing single PUFA. The mobilized engineered fatty acids were used in de novo membrane lipid synthesis during seedling development.

Zostera nigricaulis seed germination experiment

Data from seed germination experiment to determine the impact of temperature, salinity, sediment type and sediment depth on seed germination.

Microtubule dynamics in compatible and incompatible interactions of soybean hypocotyl cells with Phytophthora sojae

The arrangement of microtubules in soybean (Glycine max) cells was examined during compatible and incompatible interactions of hypocotyls of soybean cv. Harosoy (susceptible) and cv. Haro 1272 (resistant) with race 1 of the soybean-specific pathogen Phytophthora sojae. Both reaction types were similar during the first 3 h after zoospore inoculation in terms of the number of cells penetrated, and depth penetrated into the cortex. By 3 h postinoculation, clear differences had developed between the two interaction types: incompatible interactions were characterized by a hypersensitive response that was confined to single penetrated cells; while compatibly responding cells appeared unchanged. Both types of response were characterized by autofluorescence of cell walls or cytoplasm and, at 6 h after inoculation, complete disorganization of cell cytoplasm. Reorientation and loss of microtubules was seen in the early stages of the incompatible interaction in association with cellular hypersensitivity, but not in compatible responses. In cells adjacent to those that reacted hypersensitively, there was little evidence of change in microtubule orientation. Treatment of hypocotyls with the microtubule depolymerizer oryzalin prior to inoculation did not alter the compatible response, but led to breakdown of the incompatible response. Changes in microtubule orientation and state are thus among the first structural changes that are visible within cells during incompatibility in this system.

Investigation of compounds causing water repellency in the rhizosphere of sandy soils from a wide range of locations.

Although soils are generally considered to wet readily, some are actually water repellent at the surface and in the rhizosphere. This phenomenon occurs at low to moderate moisture contents and has been reported from soils under a range of vegetation types and from many regions around the globe. Water repellency in soils can have serious environmental implications including reduced seed germination and plant growth as well as irrigation efficiency, accelerated soil erosion, and enhanced leaching of agrochemicals through preferential flow. it has been proposed that water repellency is caused by the accumulation of hydrophobic organic compounds released as root exudates, microbial byproducts or from decomposing organic matter, which are deposited on mineral or aggregate surfaces, or are present as interstitial matter, Few studies to date have attempted to isolate and characterize these compounds and their structure is therefore only poorly understood, These studies have generally focussed on only a single soil or a small range of samples, have not included non-repellent soils as a control and have not always been able to demonstrate that the substances isolated are indeed responsible for repellency formation.

This study reports on the first part (extraction procedures) of a research programme addressing these gaps in current knowledge by investigating a wide range of severely repellent and wettable ‘control’ samples from different countries, and by including assessments of extraction efficiency and ability of extracts to cause repellency. Analytical methods include DRIFT (Diffuse Reflectance Infrared Fourier Transform Spectroscopy) of soils and IR (Infrared) analysis of extracts.

Key findings are that (i) soil sample heating after extraction is valuable in assessing the effectiveness of the extraction procedure, (ii) Soxhlet extraction using isopropanol/ ammonia (70/30 v/v) was the most effective method in extracting hydrophobic compounds, while leaving the ability of extracted compounds to induce water repellency virtually unaffected, (iii) wettable control soils also contain hydrophobic substances capable of inducing water repellency, (iv) the amount of organic compounds extracted was poorly related to sample repellency, indicating that compounds responsible for repellency may only represent a small fraction of the extract, (v) differences in extraction efficiency between different samples indicate that the compounds responsible may differ generically and/or in terms of their bonding to minerals, and (vi) the combination of repellency assessments with DRIFT on soils and JR on extracts used with internal standards has considerable potential to allow quantification of CH bearing organic matter in the soil, the efficiency of extraction processes for its removal, and its significance in causing water repellency in soils.

Heterotrimeric G proteins facilitate arabidopsis resistance to necrotrophic pathogens and are involved in jasmonate signaling

Heterotrimeric G proteinshave been previously linked to plant defense; however a role for the Gbg dimer in defense signaling has not been described to date. Using available Arabidopsis (Arabidopsis thaliana) mutants lacking functional Ga or Gb subunits, we show that defense against the necrotrophic pathogens Alternaria brassicicola and Fusarium oxysporum is impaired in Gb-deficient mutants while Ga-deficient mutants show slightly increased resistance compared to wild-type Columbia ecotype plants. In contrast, responses to virulent (DC3000) and avirulent (JL1065) strains of Pseudomonas syringae appear to be independent of heterotrimeric G proteins. The induction of a number of defense-related genes in Gb-deficient mutants were severely reduced in response to A. brassicicola infection. In addition, Gb-deficient mutants exhibit decreased sensitivity to a number of methyl jasmonate-induced responses such as induction of the plant defensin gene PDF1.2, inhibition of root elongation, seed germination, and growth of plants in sublethal concentrations of methyl jasmonate. In all cases, the behavior of the Ga-deficient mutants is coherent with the classic heterotrimeric mechanism of action, indicating that jasmonic acid signaling is influenced by the Gbg functional subunit but not by Ga. We hypothesize that Gbg acts as a direct or indirect enhancer of the jasmonate signaling pathway in plants.

Heterotrimeric G protein γ subunits provide functional selectivity in Gβγ dimer signaling in arabidopsis

The Arabidopsis thaliana heterotrimeric G protein complex is encoded by single canonical Galpha and Gbeta subunit genes and two Ggamma subunit genes (AGG1 and AGG2), raising the possibility that the two potential G protein complexes mediate different cellular processes. Mutants with reduced expression of one or both Ggamma genes revealed specialized roles for each Ggamma subunit. AGG1-deficient mutants, but not AGG2-deficient mutants, showed impaired resistance against necrotrophic pathogens, reduced induction of the plant defensin gene PDF1.2, and decreased sensitivity to methyl jasmonate. By contrast, both AGG1- and AGG2-deficient mutants were hypersensitive to auxin-mediated induction of lateral roots, suggesting that Gbetagamma1 and Gbetagamma2 synergistically inhibit auxin-dependent lateral root initiation. However, the involvement of each Ggamma subunit in this root response differs, with Gbetagamma1 acting within the central cylinder, attenuating acropetally transported auxin signaling, while Gbetagamma2 affects the action of basipetal auxin and graviresponsiveness within the epidermis and/or cortex. This selectivity also operates in the hypocotyl. Selectivity in Gbetagamma signaling was also found in other known AGB1-mediated pathways. agg1 mutants were hypersensitive to glucose and the osmotic agent mannitol during seed germination, while agg2 mutants were only affected by glucose. We show that both Ggamma subunits form functional Gbetagamma dimers and that each provides functional selectivity to the plant heterotrimeric G proteins, revealing a mechanism underlying the complexity of G protein-mediated signaling in plants.

Ecology of Pittosporum undulatum Vent. (Pittosporaceae) an environmental weed in south east Australia

Pittosporum undulatum Vent. (Sweet Pittosporum) is a densely foliaged tall shrub or small tree, native to the wet forests of south east Australia, This species now functions as a serious environmental weed in a range of habitats in Australia and on other continents and islands throughout the temperate, sub-tropical and tropical zones. This study investigated some of the ecological causes and consequences of P. undulatum invasion across a range of habitat types in south east Australia. Key aspects of P. undulatum biology and ecology investigated in the current study include; patterns of morphological variation across the range of habitats occupied (as a measure of the species’ plasticity), dispersal ecology and seed germinability, population structure and spatial pattern, community relationships and the ecological impacts of invasion. Phenotypic plasticity is considerable in P. undulatum. No clear patterns of geographic variation emerged from a study of leaf morphological attributes across the current range of this species on mainland south east Australia. The pattern of morphological variation is particularly complex in Victoria, where the invasion of this species is most advanced. The species’ adaptability to a range of environments and environmental conditions will likely promote further range expansion. The abundant winter fruit crop produced by functionally female P. undulatum plants attracts a suite of generalist opportunistic frugivores, which feed on P. undulatum fruits and seeds at various stages of fruit dehiscence, thereby enhancing dispersal opportunities for this species. P. undulatum seed collected from natural and invasive populations, at two stages of fruit maturity and from the scats and pellets of dispersal agents, displayed high germinability. European Blackbirds and Pied Currawongs are implicated as the main avian dispersal agents of P undulatum in south east Australia. The broader ecological implications of developing relationships between invasive fleshy-fruited bird-dispersed plant species and adaptive frugivores are likely to be considerable. The distribution of P. undulatutn seedlings was significantly negatively correlated with adult conspecifics and significantly positively correlated with trees and shrubs of other genera. This pattern reflects the importance of both firugivorous dispersal agents and the species’ germination and establishment requirements, in shaping the contagious distribution pattern typical of this species. These analyses suggest that recruitment opportunities for conspecific seedlings are limited beneath the canopy of adult conspecifics. Densities of P. undulatum were on average, 2.7 times higher in invaded populations, compared to the natural populations sampled. A male-bias was evident in all populations and no relationships between reproductive activity and the density of seedlings and juveniles were evident. Invading populations of P. undulatum impose substantial changes on ecosystem-level properties and functions. Mean species richness and cover-abundance declined notably once P. undulatum cover-abundance exceeded 20% at the invaded sites and 60% at the natural sites sampled. The natural communities sampled displayed comparatively greater resilience to the competitive effects of P. undulatum, but community attributes were affected at high densities and cover-abundance of this species. The cover-abundance of herbs and grasses declined most substantially with increasing P. undulatum at invaded sites, whereas, at the natural sites sampled, the species’ structural analogues appeared to be most affected by increasing P. undulatum cover-abundance. This study has demonstrated that the ecological consequences of P. undulatum population expansion are substantial and contribute to changes in the composition and successional trajectory of affected communities. These processes ultimately lead to the loss and simplification of biodiversity values and the homogenisation of affected habitats. P. undulatum has the potential to emerge as one of south east Australia's most serious environmental weed species.

Use of phosphite as a control for Phytophthora cinnamomi in southeastern Victorian vegetation communities

One of the major aims of the research presented in this thesis was to assist managers of native vegetation communities in southeastern Australia in understanding the dynamics of P. cinnamomi with an important ecological species, Xanthorrhoea australis. It trialed the use of phosphite in large-scale field applications to establish the usefulness of this management option for the first time on Victorian flora. This thesis describes the process of disease development within mature X. Australia plants. For the first time it was shown that within X. australis plants, secondary disease symptoms are related to the percentage of stem that has been infested by the disease. It was evident that after initial invasion the pathogen moves via root xylem and throughout the plant within vascular to the stem, especially within the desmium. The research shows that the pathogen could not be isolated consistently even though it was considered to be responsible for disease symptoms. Trials of a control fungicide (Foli-R-fos 200) shows that protection occurs in many susceptible plants when 2 and 6g a.i./L phosphite is applied. Phytotoxicity occurred in native plants at Anglesea and within controlled environment trials when using ≥ 6g a.i./L. It will be shown that 2g a.i./L phosphite controls disease in sprayed plots within heathlands at Anglesea and a recently burnt coastal woodland community at Wilson’s Promontory. The proportion of healthy X. australis plants treated with phosphite was significantly higher than the proportion in control plots without phosphite. The research shows that phosphite was recovered from leaves of three species treated with Foli-R-fos 200 in the field. For the first time it has been shown that seed germination was reduced in two species when high concentrations of phosphite were applied. The first documentation of the effect that phosphite has on soil properties showed that nitrogen and oxidised organic carbon were the only parameters to alter significantly. This thesis provides answers to some important questions, answers that can now be used by managers in formulating better policies and actions at an operational level. There has been a dire need in Victoria to address many issues regarding P. cinnamomi and this thesis provides relevant and informative approaches to disease control, and a better understanding of the disease progress.

Influence of stocking rate and mixed grazing of Angora goats and Merino sheep on animal and pasture production in southern Australia : 1 : botanical composition, sward characteristics and availability of components of annual temperate pastures

The effects of animal species (AS; Angora goats, Merino sheep or goats and sheep mixed grazed together at ratio 1:1) and stocking rate (SR; 7.5, 10 and 12.5 animals/ha) on the availability, botanical composition and sward characteristics of annual temperate pastures under continuous grazing were determined in a replicated experiment from 1981 to 1984. AS and SR had significant effects on pasture availability and composition and many AS SR interactions were detected. The pastures grazed by sheep had significantly reduced content and proportion of subterranean clover and more undesirable grasses compared with those grazed by goats. There were no differences in dry matter availabilities between goat- and sheep-grazed pastures at 7.5/ha, but at 10 and 12.5/ha goat pastures had significantly increased availabilities of green grass, dead and green clover and less weeds compared with sheep pastures. There was a significant AS SR interaction for the density of seedlings in May following pasture germination. Between July and January, the height of pastures was greater under goats than sheep but from January to March pasture height declined more on goat-grazed than on sheep-grazed pastures. There was an AS SR interaction for incidence of bare ground. Increasing the SR increased bare ground in pastures grazed by sheep but no change occurred on pastures grazed by goats. Changes in pasture characteristics due to increased SR were minimised on pastures grazed by goats but the grazing of sheep caused larger and faster changes and the pastures were damaged at the highest SR. Goats did not always select the same herbage material as sheep, changed their selection between seasons and were not less selective than sheep. Angora goats were flexible grazers and continually adapted their grazing behaviour to changing herbage conditions. Goat grazing led to an increase in subterranean clover, an accumulation of dead herbage at the base of the sward, reduced bare ground, taller pastures in spring and a more stable botanical composition. Mixed-grazed pasture characteristics were altered with SR. With careful management Angora goats on sheep farms may be used to manipulate pasture composition, to speed up establishment of subterranean clover, to decrease soil erosion and to reduce weed invasion.

Influence of stocking rate and mixed grazing of Angora goats and Merino sheep on animal and pasture production in southern Australia : 2 : liveweight, body condition score, carcass yield and mortality

The effects of animal species (AS; Angora goats, Merino sheep, mixed-grazed goats and sheep at the ratio of 1:1) and stocking rate (SR; 7.5, 10 and 12.5 animals/ha) on the liveweight, body condition score, carcass yield and mortality of goats and sheep were determined in a replicated experiment on improved annual temperate pastures in southern Australia from 1981 to 1984. The pattern of liveweight change was similar for both species with growth from pasture germination in autumn until maturation in late spring followed by weight loss. In winter, sheep grew faster than goats (65 versus 10 g/day, P < 0.05). In mixed-grazed treatments between November and December goats either grew when sheep were losing weight or goats lost less weight than sheep (P < 0.01). Both AS (P < 0.001) and SR (P < 0.001) affected liveweight of sheep and an AS SR interaction (P <  0.05) affected liveweight of goats. Mixed-grazed sheep were heavier than separately grazed sheep at all SR with a mean difference at 10 and 12.5/ha of 4.6 kg. Mixed-grazed goats at 10/ha were heavier than separately grazed goats from the end of the second year of the experiment, but at 12.5/ha, separately grazed goats maintained an advantage over mixed-grazed goats, with a 9.4-kg mean difference in December (P < 0.05). Body condition scores of goats and sheep declined with increasing SR; they were highest in late spring and were highly correlated with liveweight (r² > 0.8). Both AS and SR affected (P < 0.001) carcass weight and GR tissue depth as a direct result of differences in liveweight. Adjusting for differences in carcass weight negated AS effects on GR tissue depth. The carcass weights of sheep and goats increased by similar amounts for each 1-kg increase in liveweight. Mortality of sheep (3.1% p.a.) was unaffected by AS or SR. An AS SR interaction indicated mortality of separately grazed goats at 12.5/ha and mixed-grazed goats at 10 and 12.5/ha were higher (P < 0.05) than all other goat (29 versus 9%) and sheep treatments, primarily because of increased susceptibility to cold stress. Disease prevalence differed between sheep and goats. Mixed grazing of Merino sheep and Angora goats produced complementary and competitive effects depending upon the SR. Goats used summer pasture better but winter pasture less well for liveweight gain than sheep. Angora goats should not be grazed alone or mixed grazed with sheep on annual temperate pastures at SR greater than that recommended for Merino sheep and the evidence indicates a lower SR will reduce risks associated with mortality.

Small seed size increases the potential for dispersal of wetland plants by ducks

1. Long-distance dispersal (LDD) is important in plants of dynamic and ephemeral habitats. For plants of dynamic wetland habitats, waterfowl are generally considered to be important LDD vectors. However, in comparison to the internal (endozoochorous) dispersal of terrestrial plants by birds, endozoochorous dispersal of wetland plants by waterfowl has received little attention. We quantified the capacity for endozoochorous dispersal of wetland plants by waterfowl and identified the mechanisms underlying successful dispersal, by comparing the dispersal capacities of a large number of wetland plant species.

2. We selected 23 common plant species from dynamic wetland habitats and measured their seed characteristics. We fed seeds of all species to mallards (Anas platyrhynchos), a common and highly omnivorous duck species, and quantified seed gut survival, gut passage speed and subsequent germination. We then used a simple model to calculate seed dispersal distances.

3. In total 21 of the 23 species can be dispersed by mallards, with intact seed retrieval and subsequent successful germination of up to 32% of the ingested seeds. The species that pass fastest through the digestive tract of the mallards are retrieved in the greatest numbers (up to 54%) and germinate best (up to 87%). These are the species with the smallest seeds. Seed coat thickness plays only a minor role in determining intact passage through the mallard gut, but determines if ingestion enhances or reduces germination in comparison to control seeds.

4. Model calculations estimate that whereas the largest seeds can hardly be dispersed by mallards, most seeds can be dispersed up to 780 km, and the smallest seeds up to 3000 km, by mallards during migration.

5. Synthesis. This study demonstrates the mechanism underlying successful endozoochorous dispersal of wetland plant seeds by mallards: small seed size promotes rapid, and hence intact and viable, passage through the mallard gut. Mallards can disperse wetland plant seeds of all but the largest-seeded species successfully in relatively large numbers (up to 32% of ingested seeds) over long distances (up to thousands of kilometres) and are therefore important dispersal vectors.

The effect of seed morphology on the potential dispersal of aquatic macrophytes by the common carp (Cyprinus carpio)

1. The potential for seed dispersal by fish (ichthyochory) will vary among aquatic plants because of differences in seed size and morphology.

2. To examine how seed morphology influences the probability of dispersal by the common carp (Cyprinus carpio), we studied seed ingestion, retention time and subsequent egestion and germination of seeds of Sparganium emersum and Sagittaria sagittifolia, two aquatic plant species with similar sized but morphologically different seeds.

3. We compared dispersal probabilities between the two plant species, in which the probability of dispersal is assumed to be a function of the probabilities of seed ingestion, egestion and germination, and the dispersal distance is assumed to be a function of seed egestion rate over time.

4. We found that, although the soft seeds of S. sagittifolia had an approximately 1.5 times higher probability of being ingested by the carp than the hard seeds of S. emersum (83.15% ± 1.8% versus 56.16% ± 2.7%, respectively), the latter had an almost twofold higher probability of surviving the passage through the digestive tract (38.58% ± 2.7% versus 20.97% ± 1.5%, respectively). Patterns of seed egestion over time did not differ between the two plant species, despite the difference in seed morphology. Gut passage had a different effect on seed germination between plant species. Compared with non-ingested controls, seeds of S. emersum showed a 12.6% increase in germination and a 2.1 day acceleration in germination rate, whereas seeds of S. sagittifolia displayed a 47.3% decrease and 5.1 day delay, respectively.

5. Our results suggest that seed morphology affects the dispersal probability and postdispersal establishment, but not the dispersal distance, of aquatic plants that are dispersed by fish.

Genetic control of male fertility in Arabidopsis thaliana : structural analysis of postmeiotic developmental mutants

Seven new male-sterile mutants (ms7–ms13) of Arabidopsis thaliana (L.) Heynh. (ecotype columbia) are described that show a postmeiotic defect of microspore development. In ms9 mutants, microspores recently released from the tetrad appear irregular in shape and are often without exines. The earliest evidence of abnormality in ms12 mutants is degeneration of microspores that lack normal exine sculpturing, suggesting that the MS12 product is important in the formation of pollen exine. Teratomes (abnormally enlarged microsporocytes) are also occasionally present and each has a poorly developed exine. In ms7 mutant plants, the tapetal cytoplasm disintegrates at the late vacuolate microspore stage, apparently causing the degeneration of microspores and pollen grains. With ms8 mutants, the exine of the microspores appears similar to that of the wild type. However, intine development appears impaired and pollen grains rupture prior to maturity. In ms11 mutants, the first detectable abnormality appears at the mid to late vacuolate stage. The absence of fluorescence in the microspores and tapetal cells after staining with 4′,6-diamidino-2-phenylindole (DAPI) and the occasional presence of teratomes indicate degradation of DNA. Viable pollen from ms10 mutant plants is dehisced from anthers but appears to have surface abnormalities affecting interaction with the stigma. Pollen only germinates in high-humidity conditions or during in-vitro germination experiments. Mutant plants also have bright-green stems, suggesting that ms10 belongs to the eceriferum (cer) class of mutants. However, ms10 and cer6 are non-allelic. The ms13 mutant has a similar phenotype to ms10, suggesting is also an eceriferum mutation. Each of these seven mutants had a greater number of flowers than congenic male-fertile plants. The non-allelic nature of these mutants and their different developmental end-points indicate that seven different genes important for the later stages of pollen development have been identified.