A marker-assisted backcross approach for developing submergence-tolerant rice cultivars

Submergence stress regularly affects 15 million hectares or more of rainfed lowland rice areas in South and Southeast Asia. A major QTL on chromosome 9, Sub1, has provided the opportunity to apply marker assisted backcrossing (MAB) to develop submergence tolerant versions of rice cultivars that are widely grown in the region. In the present study, molecular markers that were tightly linked with Sub1, flanking Sub1, and unlinked to Sub1 were used to apply foreground, recombinant, and background selection, respectively, in backcrosses between a submergence-tolerant donor and the widely grown recurrent parent Swarna. By the BC2F2 generation a submergence tolerant plant was identified that possessed Swarna type simple sequence repeat (SSR) alleles on all fragments analyzed except the tip segment of rice chromosome 9 that possessed the Sub1 locus. A BC3F2 double recombinant plant was identified that was homozygous for all Swarna type alleles except for an approximately 2.3-3.4 Mb region surrounding the Sub1 locus. The results showed that the mega variety Swarna could be efficiently converted to a submergence tolerant variety in three backcross generations, involving a time of two to three years. Polymorphic markers for foreground and recombinant selection were identified for four other mega varieties to develop a wider range of submergence tolerant varieties to meet the needs of farmers in the flood-prone regions. This approach demonstrates the effective use of marker assisted selection for a major QTL in a molecular breeding program.

Buoyancy and germination of pond apple (Annona glabra L.) propagules in fresh and salt water

Stephen Setter, Melissa Setter, Michael Graham and Joe Vitelli recently published their paper 'Buoyancy and germination of pond apple (Annona glabra L.) propagules in fresh and salt water' in Proceedings of the 16th Australian Weeds Conference. Stephen also presented this paper at the conference. Pond apple is an aggressive woody weed which has invaded many wetlands, drainage lines and riparian systems across the Wet Tropics bioregion of Far North Queensland. Most fruit and seed produced by pond apple during the summer wet season fall directly into creeks, river banks, flood plains and swamps from where they are dispersed. They reported that pond apple seeds can float for up to 12 months in either fresh or salt water, with approximately 38% of these seeds germinating in a soil medium once removed from the experimental water tanks at South Johnstone. Their study suggested that the removal of reproductive trees from areas adjacent to creeks and rivers will have an immediate impact on potential spread of pond apple by limiting seed input into flowing water bodies.

Amenity grasses for salt-affected parks in coastal Australia

In February 2004, Redland Shire Council with help from a Horticulture Australia research project was able to establish a stable grass cover of seashore paspalum (Paspalum vaginatum) on a Birkdale park where the soil had previously proved too salty to grow anything else. Following on from their success with this small 0.2 ha demonstration area, Redland Shire has since invested hundreds of thousands of dollars in successfully turfing other similarly “impossible” park areas with seashore paspalum. Urban salinity can arise for different reasons in different places. In inland areas such as southern NSW and the WA wheatbelt, the usual cause is rising groundwater bringing salt to the surface. In coastal sites, salt spray or periodic tidal inundation can result in problems. In Redland Shire’s case, the issue was compacted marine sediments (mainly mud) dug up and dumped to create foreshore parkland in the course of artificial canal developments. At Birkdale, this had created a site that was both strongly acid and too salty for most plants. Bare saline scalds were interspersed by areas of unthrifty grass. Finding a salt tolerant grass is no “silver bullet” or easy solution to salinity problems. Rather, it buys time to implement sustainable long-term establishment and maintenance practices, which are even more critical than with conventional turfgrasses. These practices include annual slicing or coring in conjunction with gypsum/dolomite amendment and light topdressing with sandy loam soil (to about 1 cm depth), adequate maintenance fertiliser, weed control measures, regular leaching irrigation was applied to flush salts below the root zone, and irrigation scheduling to maximise infiltration and minimise run off. Three other halophytic turfgrass species were also identified, each of them adapted to different environments, management regimes and uses. These have been shortlisted for larger-scale plantings in future work.

Recovery of fish and crustacean communities during remediation of tidal wetlands affected by leachate from acid sulfate soils in north-eastern Australia.

In the 1970s, acid sulfate soils (ASS) distributed within about 720 ha of predominantly mangrove and salt pan wetlands at East Trinity in north Queensland were developed after the area was isolated from tidal flooding by a surrounding seawall and the installation of tidal gates on major drainage creeks. Following drainage and oxidation of these estuarine acidic sediments, resultant acid leachate caused considerable, ongoing environmental problems including regular fish kills. A rehabilitation program covering much of these former tidal wetlands commenced in 2000 using a lime-assisted tidal exchange management regime. Changes in the established populations of estuarine fish and crustaceans were monitored in the two creeks (Firewood and Hills Creeks) where tidal flows were reinstated. In Firewood Creek between 2001 and 2005, there was a progressive increase in fish species richness, diversity and abundance. The penaeid prawn Fenneropenaeus merguiensis was a major component of the cast net catches in the lower sections of both Firewood and Hills Creeks but its relative abundance decreased upstream of the tidal gates on the seawall. Well established stocks of predominantly juvenile, male Scylla serrata resident upstream of the tidal gates indicated suitable habitats with acceptable water and sediment quality and adequate availability of food. The regular fish kills that occurred prior to the management regime abated and, overall, the implementation of the rehabilitation program is yielding positive benefits for the local fisheries.

Polychaete-assisted sand filters

The objective of this study was to investigate the productivity and functionality of sand filters stocked with marine worms for wastewater treatment at mariculture facilities. Medium bedding sand which is commonly available in coastal sedimentary deposits and nereidid polychaetes (Perinereis nuntia and P. helleri) from Moreton Bay in southeast Queensland were combined and studied in down-flow sand filtration beds. This combination appears to provide a new option for brackish wastewater treatment whereby the activities of the worms help to prevent sand filters from blocking with organic debris and their biomass offers a valuable by-product. Phytoplankton-rich pond waters percolating through sand-worm beds were reliably treated in several useful ways: suspended solids and chlorophyll a levels were consistently reduced by >50% by the process, and nutrients were converted into bio-available dissolved forms. Dissolved oxygen, redox and pH levels were also lowered significantly by the process. Water treatment rates of approx 1500 L m-2 d-1 were routinely achieved. P. nuntia appeared more suitable than P. helleri for stocking directly into sand filtration beds as nectochaetes, but generally exhibited slower growth. Survival and growth were influenced by stocking density. Sand-filter beds stocked with juvenile worms and fed only with eutrophic pond water demonstrated polychaete production capacities in the order of 300-400 g m-2 (eg. P. helleri: 328 g m-2 in 16 weeks). These results show how nereidid polychaetes can be reliably produced within simple, low-maintenance sand filters, and provide data necessary for the functional integration of this novel wastewater treatment system into contemporary seafood farming systems.

Polychaete-assisted sand filters – prawn farm wastewater remediation trial

Medium bedding sand which is commonly available in coastal sedimentary deposits, and a marine polychaete-worm species from Moreton Bay recently classified as Perinereis helleri (Nereididae), were deployed in a simple low-maintenance sand filter design that potentially has application at large scale. Previous work had shown that this physical and biological combination can provide a new option for saline wastewater treatment, since the worms help to prevent sand filter blocking with organic debris and offer a profitable by-product. To test the application of this new concept in a commercial environment, six 1.84 m2 Polychaete-assisted sand filters were experimentally tested for their ability to treat wastewater from a semi-intensive prawn culture pond. Polychaetes produced exclusively on the waste nutrients that collected in these gravity-driven sand filters were assessed for their production levels and nutritional contents. Water parameters studied included temperature, salinity, pH, dissolved oxygen (DO), oxidation/ reduction potential (redox), suspended solids, chlorophyll a, biological oxygen demand (BOD), and common forms of nitrogen and phosphorus. Pond water which had percolated through the sand bed had significantly lower pH, DO and redox levels compared with inflow water. Suspended solids and chlorophyll a levels were consistently more than halved by the process. Reductions in BOD appeared dependant on regular subsurface flows. Only marginal reductions in total nitrogen and phosphorus were documented, but their forms were altered in a potentially useful way: dissolved forms (ammonia and orthophosphate) were generated by the process, and this remineralisation also seemed to be accentuated by intermittent flow patterns. Flow rates of approximately 1,500 L m-2 d-1 were achieved suggesting that a 1 ha polychaete bed of this nature could similarly treat the discharge from a 10 ha semi-intensive prawn farm. Sixteen weeks after stocking sand beds with one-month-old P. helleri, over 3.6 kg of polychaete biomass (wet weight) was recovered from the trial. Production on a sand bed area basis was 328 g m-2. Similar (P>0.05) overall biomass production was found for the two stocking densities tested (2000 and 6000 m-2; n = 3), but survival was lower and more worms were graded as small (<0.6 g) when produced at the higher density (28.2 ± 1.5 % and approx. 88 %, respectively) compared with the lower density (46.8 ± 4.4 % and approx. 76 %, respectively). When considered on a weight for weight basis, about half of the worm biomass produced was generally suitable for use as bait. The nutritional contents of the worms harvested were analysed for different stocking densities and graded sizes. These factors did not significantly affect their percentages of dry matter (DM) (18.23 ± 0.57 %), ash (19.77 ± 0.80 % of DM) or gross energy 19.39 ± 0.29 MJ kg-1 DM) (n = 12). Although stocking density did not affect the worms’ nitrogen and phosphorus contents, small worms had a higher mean proportion of nitrogen and phosphorus (10.57 ± 0.17 % and 0.70 ± 0.01 % of DM, respectively) than large worms (9.99 ± 0.12 % and 0.65 ± 0.01 % of DM, respectively) (n = 6). More lipid was present in large worms grown at the medium density (11.20 ± 0.19 %) compared with the high density (9.50 ± 0.31 %) and less was generally found in small worms (7.1-7.6 % of DM). Mean cholesterol and total phospholipid levels were 5.24 ± 0.15 mg g-1 and 13.66 ± 2.15 mg g-1 DM, respectively (n = 12). Of the specific phospholipids tested, phosphatidyl-serine or sphingomyelin were below detection limits (<0.05 mg g-1), whilst mean levels of phosphatidyl-ethanolamine, phosphatidyl-inositol, phosphatidyl-choline and lysophosphatidyl-choline were 6.89 ± 1.09, 0.89 ± 0.26, 4.04 ± 1.17 and 1.84 ± 0.37 mg g-1, respectively (n = 12). Culture density generally had a more pronounced effect on phospholipid contents than did size of worms. By contrast, worm size had a more pronounced effect on total fatty acid contents, with large worms containing significantly higher (P<0.001) levels on a DM basis (46.88 ± 2.46 mg g-1) than smaller worms (27.76 ± 1.28 mg g-1). A very broad range of fatty acids were detected with palmitic acid being the most heavily represented class (up to 14.23 ± 0.49 mg g-1 DM or 27.28 ± 0.22 % of total fatty acids). Other heavily represented classes included stearic acid (7.4-8.8 %), vaccenic acid (6.8-7.8 %), arachidonic acid (3.5-4.4 %), eicosapentaenoic acid (9.9-13.8 %) and docosenoic acid (5.7-7.0 %). Stocking density did not affect (P>0.05) the levels of amino acids present in polychaete DM, but there was generally less of each amino acid tested on a weight per weight basis in large worms than in small worms. This difference was significant (P<0.05) for the most heavily represented classes being glutamic acid (73-77 mg g-1), aspartic acid (50-54 mg g-1), and glycine (46-53 mg g-1). These results demonstrate how this polychaete species can be planted and sorted at harvest according to various strategies aimed at providing biomass with specific physical and nutritional qualities for different uses.

Establishment and management of salt-tolerant amenity grasses to reduce urban salinity effect

This project built upon the successful outcomes of a previous project (TU02005) by adding to the database of salt tolerance among warm season turfgrass cultivars, through further hydroponic screening trials. Hydroponic screening trials focussed on new cultivars or cultivars that were not possible to cover in the time available under TU02005, including: 11 new cultivars of Paspalum vaginatum; 13 cultivars of Cynodon dactylon; six cultivars of Stenotaphrum secundatum; one accession of Cynodon transvaalensis; 12 Cynodon dactylon x transvaalensis hybrids; two cultivars of Sporobolus virginicus; five cultivars of Zoysia japonica; one cultivar of Z. macrantha, one common form of Z. tenuifolia and one Z. japonica x tenuifolia hybrid. The relative salinity tolerance of different turfgrasses is quantified in terms of their growth response to increasing levels of salinity, often defined by the salt level that equates to a 50% reduction in shoot yield, or alternatively the threshold salinity. The most salt tolerant species in these trials were Sporobolus virginicus and Paspalum vaginatum, consistent with the findings from TU02005 (Loch, Poulter et al. 2006). Cynodon dactylon showed the largest range in threshold values with some cultivars highly sensitive to salt, while others were tolerant to levels approaching that of the more halophytic grasses. Coupled with the observational and anecdotal evidence of high drought tolerance, this species and other intermediately tolerant species provide options for site specific situations in which soil salinity is coupled with additional challenges such as shade and high traffic conditions. By recognising the fact that a salt tolerant grass is not the complete solution to salinity problems, this project has been able to further investigate sustainable long-term establishment and management practices that maximise the ability of the selected grass to survive and grow under a particular set of salinity and usage parameters. Salt-tolerant turf grasses with potential for special use situations were trialled under field conditions at three sites within the Gold Coast City Council, while three sites, established under TU02005 within the Redland City Council boundaries were monitored for continued grass survival. Several randomised block experiments within Gold Coast City were established to compare the health and longevity of seashore paspalum (Paspalum vaginatum), Manila grass (Zoysia matrella), as well as the more tolerant cultivars of other species like buffalo grass (Stenotaphrum secundatum) and green couch (Cynodon dactylon). Whilst scientific results were difficult to achieve in the field situation, where conditions cannot be controlled, these trials provided valuable observational evidence of the likely survival of these species. Alternatives to laying full sod such as sprigging were investigated, and were found to be more appropriate for areas of low traffic as the establishment time is greater. Trials under controlled and protected conditions successfully achieved a full cover of Paspalum vaginatum from sprigs in a 10 week time frame. Salt affected sites are often associated with poor soil structure. Part of the research investigated techniques for the alleviation of soil compaction frequently found on saline sites. Various methods of soil de-compaction were investigated on highly compacted heavy clay soil in Redlands City. It was found that the heavy duplex soil of marine clay sediments required the most aggressive of treatments in order to achieve limited short-term effects. Interestingly, a well constructed sports field showed a far greater and longer term response to de-compaction operations, highlighting the importance of appropriate construction in the successful establishment and management of turfgrasses on salt affected sites. Fertiliser trials in this project determined plant demand for nitrogen (N) to species level. This work produced data that can be used as a guide when fertilising, in order to produce optimal growth and quality in the major turf grass species used in public parkland. An experiment commenced during TU02005 and monitored further in this project, investigated six representative warm-season turfgrasses to determine the optimum maintenance requirements for fertiliser N in south-east Queensland. In doing so, we recognised that optimum level is also related to use and intensity of use, with high profile well-used parks requiring higher maintenance N than low profile parks where maintaining botanical composition at a lower level of turf quality might be acceptable. Kikuyu (Pennisetum clandestinum) seemed to require the greatest N input (300-400 kg N/ha/year), followed by the green couch (Cynodon dactylon) cultivars ‘Wintergreen’ and ‘FLoraTeX’ requiring approximately 300 kg N/ha/year for optimal condition and growth. ‘Sir Walter’ (Stenotaphrum secundatum) and ‘Sea Isle 1’ (Paspalum vaginatum) had a moderate requirement of approximately 200 kg/ha/year. ‘Aussiblue’ (Digitaria didactyla)maintained optimal growth and quality at 100-200 kg N/ha/year. A set of guidelines has been prepared to provide various options from the construction and establishment of new grounds, through to the remediation of existing parklands by supporting the growth of endemic grasses. They describe a best management process through which salt affected sites should be assessed, remediated and managed. These guidelines, or Best Management Practices, will be readily available to councils. Previously, some high salinity sites have been turfed several times over a number of years (and Council budgets) for a 100% failure record. By eliminating this budgetary waste through targeted workable solutions, local authorities will be more amenable to investing appropriate amounts into these areas. In some cases, this will lead to cost savings as well as resulting in better quality turf. In all cases, however, improved turf quality will be of benefit to ratepayers, directly through increased local use of open space in parks and sportsfields and indirectly by attracting tourists and other visitors to the region bringing associated economic benefits. At the same time, environmental degradation and erosion of soil in bare areas will be greatly reduced.

Salt-tolerant amenity grasses to reduce salt effect

Assess establishment and management of salt tolerant turf grasses on salt affected parklands.

Marker Assisted Breeding Support for Tropical Horticulture and Forestry in far north Queensland

Molecular marker development for tropical fruit and forestry species.

Comparison of compost and a sandy loam as turf underlay materials on salt-affected parkland

Turfgrasses range from extremely salt sensitive to highly salt tolerant. However, the selection of a salt tolerant turf is not a 'silver bullet' solution to successful turf growth on salt-affected parklands. Interactions between factors such as cultivar, construction practices, establishment, and maintenance can be complex and should not be considered in isolation of one another. Taking this holistic approach, a study investigating cultivar evaluation for salt-affected sites also included a comparison of topsoil materials as turf underlay, as well as pre-treatment of the sod. The turf species and cultivars used in the study were: Cynodon dactylon, cultivar 'Oz Tuff (I) '; Paspalum vaginatum, cultivars 'Sea Isle 1 (I) ' and 'Velvetene (I) '; Zoysia matrella cultivar 'A-1 (I) '; and Zoysia japonica, cultivar 'Empire (I) '. The two underlay materials were compost (100%) or a sandy clay topsoil each applied above a coastal sand profile to a depth of 10 cm. Rooting depth or root dry weight did not significantly differ among turf cultivars. Compost profile treatment had significantly greater root mass than the topsoil among all turf cultivars. This higher root production was reflected by improved quality of all turf at the final evaluation. Turfgrass grown on compost had a higher normalised difference vegetation index (NDVI), regardless of whether full sod or bare-rooted turfgrass was used. The use of a quality underlay was paramount to the successful growth of the turf cultivars investigated. While each cultivar had superior performance in sub-optimal conditions, the key to success was the selection of the right species and cultivar for each situation combined with proper establishment and maintenance of each turf grass.