Stimulating dormancy release and emergence of annual ryegrass (Lolium rigidum) seeds using short-term hydrated storage in darkness

Experiments were performed to determine whether the dormancy release effect of hydrated storage in darkness (dark-stratification) is common amongst annual ryegrass populations and has the potential to occur under field conditions. Dormant seeds from all populations tested (22) became sensitive to light during dark-stratification, enabling them to germinate when subsequently exposed to light. Under controlled temperature (25/15degreesC), light (12-h photoperiod), and hydration (solidified agar-water) conditions, more seeds germinated by 28 days if the first 14 days were in darkness followed by exposure to light for 12 h per day than if they were exposed to light throughout or darkness throughout. Constraint over the conditions imposed during dark-stratification and germination was gradually reduced to investigate whether the dormancy release effect was diminished. Dark-stratification was effective in promoting germination when performed under natural diurnal temperatures, and burial in moist soil provided suitable conditions for dark-stratification to occur. The surface of moist soil, with natural diurnal temperatures and sunlight, was suitable for germination of dark-stratified seeds. Dark-stratification is a quick and effective means to enhance the sensitivity of dormant annual ryegrass seeds to light, enabling the majority of the population to germinate. However, large quantities of light are required to promote germination of dark-stratified seeds, so buried seeds must be moved to the soil surface to allow exposure to adequate light for germination.

Seed dormancy and germination responses of nine Australian fire ephemerals

Fire ephemerals are short-lived plants with seeds that persist in the soil and germinate after a fire or physical soil disturbance. Ex situ germination of many Australian fire ephemerals has previously been difficult. Dormancy was present in most of the nine fire ephemerals examined. Alyogyne hakeifolia (Giord.) Alef. and Alyogyne huegelii (Endl.) Fryxell (Malvaceae) seeds had physical and possibly also physiological dormancy, Actinotus leucocephalus Benth. (Apiaceae) seeds had morphophysiological dormancy, Austrostipa compressa (R.Br.) S.W.L. Jacobs & J. Everett and Austrostipa macalpinei (Reader) S.W.L. Jacobs & J. Everett (Poaceae) seeds were either non-dormant or possessed physiological dormancy, and seeds of all remaining species possessed physiological dormancy. A proportion of the Alyogyne hakeifolia, Alyogyne huegelii, Austrostipa compressa and Austrostipa macalpinei seed populations were non-dormant because some seeds could germinate at the various incubation temperatures without further treatment. At 20 degrees C, artificial methods of inducing germination such as manual or acid scarification were among the optimal treatments for Austrostipa compressa, Austrostipa macalpinei, Alyogyne huegelii, Actinotus leucocephalus and Grevillea scapigera A.S. George (Proteaceae), and gibberellic acid induced maximum germination of Tersonia cyathiflora (Fenzl) J.W. Green (Gyrostemonaceae) seeds. Heat (70 degrees C for 1 h) and smoke water was one of the most effective treatments for germinating Actinotus leucocephalus and Codonocarpus cotinifolius (Desf.) F. Muell. (Gyrostemonaceae) seeds. Germination of Grevillea scapigera, Codonocarpus cotinifolius, Gyrostemon racemiger H. Walter (Gyrostemonaceae) and Tersonia cyathiflora did not exceed 40% and may require other treatments to overcome dormancy. Although the nine fire ephemerals examined require fire to germinate under natural conditions, a range of germination responses and dormancy types was observed.

Dormancy release in Australian fire ephemeral seeds during burial increases germination response to smoke water or heat

Fire ephemerals are short-lived plants that primarily germinate after fire. Fresh and laboratory-stored seeds are difficult to germinate ex situ, even in response to fire-related cues such as heat and smoke. Seeds of eight Australian fire ephemeral species were buried in unburnt and recently burnt sites of natural bushland during autumn. Seeds were exhumed after 6 and 12 months and incubated in water and smoke water, either with or without a heat treatment at 70 degrees C for 1 h. Generally, germination did not increase after 6 months of burial, but after 12 months of burial germination was enhanced in seven of the eight species. Actinotus leucocephalus produced higher germination following 12 months of burial without any further treatment, and smoke water and heat further improved germination. The four Gyrostemonaceae species, Codonocarpus cotinifolius, Gyrostemon racemiger, Gyrostemon ramulosus and Tersonia cyathiflora, only germinated in the presence of smoke water, and their germination was enhanced by burial. Burial improved germination in response to a heat treatment in Grevillea scapigera and Alyogyne huegelii seeds, but did not enhance Alyogyne hakeifolia germination. During concurrent dry laboratory storage of seeds at 15 degrees C, only Actinotus leucocephalus produced increased germination in response to smoke water and heat over time. In summary, soil burial can alter the dormancy status of a number of Australian fire ephemeral seeds, rendering them more responsive to germination cues such as smoke water and heat. The requirement for a period of burial before seeds become responsive to smoke and/or heat would ensure that seeds persist in the soil until a subsequent fire, when there is an increase in nutrients available for growth and reduced competition from other plants.

Sheep rumen digestion and transmission of weedy Malva parviflora seeds

The effect of sheep digestion and mastication on Malva parviflora L. seed transmission, viability and germination was investigated. Mature M. parviflora seeds were subjected to 2 seed treatments: 'scarified', where the hard seed coat was manually cut to allow inhibition, and 'unscarified', where the hard seed coat was not cut. Seeds were placed directly into the rumen of fistulated sheep and removed at 0, 12, 24, 36 and 48 h of rumen digestion. After 12 h of in sacco exposure to digestion in the rumen, the germination of seeds that were initially scarified dropped from 99.2 to 1.4% and longer exposure periods produced no germinable seeds. In contrast, seeds that were unscarified when placed in the rumen produced over 92% germination regardless of in sacco digestion time, although manual scarification after retrieval was essential to elicit germination. In a second experiment, unscarified seeds (29000) were fed in a single meal to fistulated sheep and feces were collected at regular intervals between 6 and 120 h after feeding. Fecal subsamples were taken to determine number of seeds excreted, seed germination on agar and seed germination from feces. Major seed excretion in the feces commenced after 12 h and continued until 144 h, with peaks between 36 and 72 h after consumption. Although mastication and gut passage killed the majority of unscarified seeds, about 20% were recovered intact and over 90% of these recovered seeds were viable and could, thus, potentially form an extensive seed bank. A few excreted seeds (1%) were able to germinate directly from feces, which increased to a maximum of 10% after subsequent dry summer storage (3 months). Through information gained in this study, there is a potential to utilise livestock in an integrated weed management program for the control of M. parviflora, provided additional measures of weed control are in place such as holding periods (> 7 days) for movement of livestock from weed infested areas.

Seed dormancy mechanisms in warm season grass species

Available evidence suggests that there are at least two locations for dormancy mechanisms in primary dormant seeds: mechanisms based within the embryo covering structures, and mechanisms based within the embryo. Mechanisms within the covering structures may involve mechanical, permeability and chemical barriers to germination. Mechanisms within the embryo may involve the expression of certain genes, levels of certain plant growth regulators, the activity of important respiratory pathways or the mobilisation and utilisation of food reserves. In addition, some embryos may be too immature to germinate immediately and must undergo a further growth phase before germination is possible. An individual species could have one or several of these various dormancy mechanisms and these mechanisms need to be understood when selecting treatments to overcome dormancy. The way in which certain dormancy breaking agents are thought to work is discussed and practical applications of such agents in field situations are explained.

Towards the clonal propagation of coconut

Past studies from our laboratory have shown that whole immature, or mature sliced, zygotic embryos are a very good starting explant for coconut somatic embryogenesis. The highest rate of somatic embryogenesis was obtained when certain polyamines were added into the culture medium as well as activated charcoal (AC) to absorb unwanted phenolics. These past studies also showed that the development and maturation of the somatic embryos produced could be improved by the addition of abscisic acid (ABA), alone or with one of several osmotically active agents, into the culture medium. In the present study this well characterised somatic embryogenic system for zygotic tissues is being modified and applied to somatic tissues. This recent approach should be a better method for the rapid production of clonal, true-to-type coconut palms. The present research approach is focused on young leaf section explants which have been found to be very responsive to callus production. Young leaf sections produced optimum callus when cultured on media containing 2,4-D (150 μM) and the amount produced could be increased by soaking the sections in sterile water (15 to 60 minutes) or ascorbic acid (15 to 30 minutes) prior to culturing. Further improvement in callus production, as well as a reduction in the time taken for callogenesis was obtained when casein hydrolysate and/or certain polyamines were added to the callus induction medium. The development of the somatic embryos was improved by using ABA and polyethylene glycol (PEG) in the maturation medium. Despite these initial successes in improving coconut somatic embryogenesis, further studies are now being considered to shorten the time to achieve somatic embryogenesis, to better germinate somatic embryos and to improve the rate of somatic seedling conversion into plantlets.

Improved germination of the Australian natives: Hibbertia commutata, Hibbertia amplexicaulis (Dilleniaceae), Chameascilla corymbosa (Liliaceae) and Leucopogon nutans (Epacridaceae)

Hibbertia commutata (Steudel), H. amplexicaulis (Steudel), Chameascilla corymbosa [(R.Br.) F.Muell. Ex Benth.] and Leucopogon nutans (E. Pritzel) are four Australian species that are difficult to germinate during mine-site rehabilitation. Laboratory germination trails were conducted to identify dormancy mechanisms and to improve germination response. Treatments applied to all species included scarification and scarification followed by soaking seeds in smoke water (1, 5 or 10%) or gibberellic-acid solution (50, 200 or 1000 muM). Additional treatments with kinetin solution (50, 200 or 1000 muM) and smoke water (50 or 100%) were applied to scarified or unscarified seeds of C. corymbosa. Thermal-shock treatment was applied to L. nutans fruit, some of which were subsequently scarified and subjected to both smoke water (10%) and gibberellic-acid solution (1000 muM). Significant germination increases were obtained by using dormancy-breaking treatments on H. commutata ( from 12.8 to 76.0%), H. amplexicaulis (from 6.8 to 55.1%) and C. corymbosa (from 48.5 to 86.4%). Scarification alone increased germination of both Hibbertia species, suggesting that these species display a physical seed coat-imposed dormancy mechanism. Germination of H. amplexicaulis was further increased by the application of gibberellic-acid solution, indicating a possible embryo-imposed dormancy mechanism. Scarification followed by the application of smoke water produced the highest germination response for C. corymbosa seeds. Scarification alone did not significantly increase germination, inferring the existence of a smoke-responsive embryo dormancy mechanism. Seeds of L. nutans, although viable, failed to germinate and are thought to display complex seed coat- and embryo-imposed dormancy mechanisms.

Climatic regulation of seed dormancy and emergence of diverse Malva parviflora populations from a Mediterranean-type environment

Malva parviflora L. (Malvaceae) is rapidly becoming a serious weed of Australian farming systems. An understanding of the variability of its seed behaviour is required to enable the development of integrated weed management strategies. Mature M. parviflora seeds were collected from four diverse locations in the Mediterranean-type climatic agricultural region of Western Australia. All of the seeds exhibited physical dormancy at collection; manual scarification or a period of fluctuating summer temperatures (50/20 degrees C or natural) were required to release dormancy. When scarified and germinated soon (1 month) after collection, the majority of seeds were able to germinate over a wide range of temperatures (5-37 degrees C) and had no light requirement. Germination was slower for seeds stored for 2 months than seeds stored for 2 years, suggesting the presence of shallow physiological dormancy. Seed populations from regions with similar annual rainfall exhibited similar dormancy release patterns; seeds from areas of low rainfall (337-344mm) were more responsive to fluctuating temperatures, releasing physical dormancy earlier than those from areas of high rainfall (436-444mm). After 36 months, maximum seedling emergence from soil in the field was 60%, with buried seeds producing 13-34% greater emergence than seeds on the surface. Scanning electron microscopy of the seed coat revealed structural differences in the chalazal region of permeable and impermeable seeds, suggesting the importance of this region in physical dormancy breakdown of M. parviflora seeds. The influence of rainfall during plant growth in determining dormancy release, and hence, germination and emergence timing, must be considered when developing management strategies for M. parviflora.

The effects of temperature and salinity on Acacia harpophylla (brigalow) (Mimosaceae) germination

Acacia harpophylla F. Muell. (brigalow) used to naturally occur over a range of about 50 000 km(2) in Queensland and New South Wales, Australia. Large scale clearing for agriculture has reduced the area to less than 20 000 km(2) and it is estimated that 20-25% of vertebrate fauna living in brigalow communities will become locally extinct as a result of the current clearing induced loss of habitat. Some coal mining companies in central Queensland have become interested in providing habitat for the endangered bridle nail-tailed wallaby that lives in brigalow vegetation. However, there is little known about establishment techniques for brigalow on mine sites and other disturbed ground; an understanding of brigalow biology and ecology is required to assist in the conservation of this threatened vegetation community and for re-creation of bridled nail-tail wallaby habitat in the post mining landscape. Brigalow is an unusual species of Acacia because it is not hard-seeded and germinates readily without the need to break seed-coat imposed dormancy. Germination trials were undertaken to test the ability of brigalow seed to germinate with a range of temperatures and salinity levels similar to those experienced in coal mine spoil. Optimum germination was found to occur at temperatures from 15 to 38 degrees C and no germination was recorded at 45 degrees C. Brigalow was very tolerant of high salt levels and germinated at percentages greater than 50% up to the highest salinity tested, 30 dS/m. Germination of greater than 90% occurred up to an electrical conductivity of 20 dS/m. The results indicate brigalow seed can be sown in summer when rains are most likely to occur, however, shading of the seed with extra soil or mulch may ensure the ground surface does not become too hot for germination. Because of its ability to germinate at high salinity levels, brigalow may be suitable for use in saline mine wastes which are common on sites to be rehabilitated after mining.