Functionally associated molecular genetic marker map construction in perennial ryegrass (Lolium perenne L.)

A molecular marker-based map of perennial ryegrass (Lolium perenne L.) has been constructed through the use of polymorphisms associated with expressed sequence tags (ESTs). A pair-cross between genotypes from a North African ecotype and the cultivar Aurora was used to generate a two-way pseudo-testcross population. A selection of 157 cDNAs assigned to eight different functional categories associated with agronomically important biological processes was used to detect polymorphic EST–RFLP loci in the F1(NA6 × AU6) population. A comprehensive set of EST–SSR markers was developed from the analysis of 14,767 unigenes, with 310 primer pairs showing efficient amplification and detecting 113 polymorphic loci. Two parental genetic maps were produced: the NA6 genetic map contains 88 EST–RFLP and 71 EST–SSR loci with a total map length of 963 cM, while the AU6 genetic map contains 67 EST–RFLP and 58 EST–SSR loci with a total map length of 757 cM. Bridging loci permitted the alignment of homologous chromosomes between the parental maps, and a sub-set of genomic DNA-derived SSRs was used to relate linkage groups to the perennial ryegrass reference map. Regions of segregation distortion were identified, in some instances in common with other perennial ryegrass maps. The EST-derived marker-based map provides the basis for in silico comparative genetic mapping, as well as the evaluation of co-location between QTLs and functionally associated genetic loci.

A simplified PCR test for early detection of dwarf off-types in micropropagated Cavendish bananas (Musa spp. AAA)

The dwarf somaclonal variant is a major problem affecting micropropagation of the banana cultivar Williams (Musa spp. AAA; subgroup Cavendish). This problem arises from genetic changes that occur during the tissue culture process. Early identification of this problem is difficult and propagators must wait until plants are ex vitro in order to visualise the dwarfism phenotype. In this study, we have improved a SCAR-based molecular diagnostic technique, developed by Damasco et al. [Acta Hortic. 461 (1997) 157], for the early identification of dwarf off-types. We have included a positive internal control in a multiplex PCR and adapted the technique for use with small amounts of fresh in vitro leaf material as PCR template. The control product is a 500 bp fragment from 18S rRNA and is amplified in all tissues irrespective of phenotype. The use of small in vitro leaf material removing the need for genomic DNA extraction.

Yield of synthetic backcross-derived lines in rainfed environments of Australia

Wheat is one of the major food crops in the world. It is Australia's largest crop and most important agricultural commodity. In Australia the crop is grown under rainfed conditions with inherently important regional environmental differences; wheat growing areas are characterized by winter dominant rainfall in southern and western Australia and summer rainfall in northern Australia. Maximizing yield potential across these diverse regions is dependent upon managing, either genetically or agronomically, those factors in the environment that limit yield. The potential of synthetic backcross lines (SBLs) to increase yield in the diverse agroecological zones of Australia was investigated. Significant yield advantages were found for many of the SBLs across diverse environments. Depending on the environment, the yield of the SBLs ranged from 8% to 30% higher than the best local check in Australia. Apart from adaptation to semiarid water stressed conditions, some SBLs were also found to be significantly higher yielding under more optimal (irrigated) conditions. The four testing environments were classified into two groups, with the northern and southern environments being in separate groups. An elite group of SBLs was identified that exhibited broad adaptation across all diverse Australian environments included in this study. Other SBLs showed specific adaptation to either northern or southern Australia. This study showed that SBLs are likely to provide breeders with the opportunity to significantly improve wheat yield beyond what was previously possible in a number of diverse production environments.

Designing better on-farm research in Australia using a participatory workshop process

There are renewed calls for end-user participation and the integration of local knowledge in agricultural research. In Australia, the response has included an increased emphasis on participatory on-farm research with farmers and commercial agronomists that tests accepted principals to answer practical local farming questions. However, this pursuit of greater relevance has often led to compromises in research designs, unclear results and frustration amongst farmers, commercial agronomists and Research Development and Extension (RDE) agency researchers. This paper reports on a series of pre-season planning workshops from `Doing successful on-farm research', a workshop-based initiative that provides guidelines and a series of interactive activities to plan better participatory on-farm research. The workshop approach helps people design on-farm research that is appropriate to their own needs and local conditions. It assists them to clearly identify their issues, develop specific research questions and decide the best approach to answer those questions with the appropriate rigour for their own situations. These `Doing successful on-farm research' workshops address four potential deficiencies in on-farm research and farming systems RDE more generally in Australia: (1) variable participation of scientists and farmers in on-farm research; (2) the lack of clear guidelines for effective participatory practice and on-farm research; (3) limited support for on-farm research beyond the intensive investigations conducted by RDE agencies and (4) limited support for industry and farmers to contextualise information and research outcomes for specific individual circumstances and faster adaptation of technology. This may be a valuable contribution to balancing the demands for both relevance and rigour in on-farm research in Australia. In "Ground–breaking Stuff’- Proceedings of the 13th Australian Society of Agronomy Conference, 10-14 September 2006, Perth, Western Australia.

An assessment of the genetic relationship between sweet and grain sorghums, within Sorghum bicolor ssp. bicolor (L.) Moench, using AFLP markers

Compared to grain sorghums, sweet sorghums typically have lower grain yield and thick, tall stalks which accumulate high levels of sugar (sucrose, fructose and glucose). Unlike commercial grain sorghum (S. bicolor ssp. bicolor) cultivars, which are usually F1 hybrids, commercial sweet sorghums were selected as wild accessions or have undergone limited plant breeding. Although all sweet sorghums are classified within S. bicolor ssp. bicolor, their genetic relationship with grain sorghums is yet to be investigated. Ninety-five genotypes, including 31 sweet sorghums and 64 grain sorghums, representing all five races within the subspecies bicolor, were screened with 277 polymorphic amplified fragment length polymorphism (AFLP) markers. Cluster analysis separated older sweet sorghum accessions (collected in mid 1800s) from those developed and released during the early to mid 1900s. These groups were emphasised in a principle component analysis of the results such that sweet sorghum lines were largely distinguished from the others, particularly by a group of markers located on sorghum chromosomes SBI-08 and SBI-10. Other studies have shown that QTL and ESTs for sugar-related traits, as well as for height and anthesis, map to SBI-10. Although the clusters obtained did not group clearly on the basis of racial classification, the sweet sorghum lines often cluster with grain sorghums of similar racial origin thus suggesting that sweet sorghum is of polyphyletic origin within S. bicolor ssp. bicolor.

A digital imaging technique to quantify the incidence of internal browning in potato (Solanum tuberosum) tubers

Internal browning disorders, including brown fleck (BF), in potato (Solanum tuberosum) tubers greatly reduce tuber quality, but the causes are not well understood. This is due, in part, to the highly variable data provided by visual value-based rating systems. A digital imaging technique was developed to quantify accurately the incidence of internal browning in potato tubers. Images of tuber sections were scanned using a flatbed scanner and digitally enhanced to highlight tuber BF lesions, and the area of affected tissue calculated using pixel quantification software. Digital imaging allowed for the determination of previously unused indices of the incidence and severity of internal browning in potato tubers. Statistical analysis of the comparison between digitally derived and visual-rating BF data from a glasshouse experiment showed that digital data greatly improved the delineation of treatment effects. The F-test probability was further improved through square root or logarithmic data transformations of the digital data, but not of the visual-rating data. Data from a field experiment showed that the area of tuber affected by BF and the number of small BF lesions increased with time and with increase in tuber size. The results from this study indicate that digital imaging of internal browning disorders of potato tubers holds much promise in determining their causes that heretofore have proved elusive.

Modelling environmental effects on phenology and canopy development of diverse sorghum genotypes

The ability to predict phenology and canopy development is critical in crop models used for simulating likely consequences of alternative crop management and cultivar choice strategies. Here we quantify and contrast the temperature and photoperiod responses for phenology and canopy development of a diverse range of elite Indian and Australian sorghum genotypes (hybrid and landrace). Detailed field experiments were undertaken in Australia and India using a range of genotypes, sowing dates, and photoperiod extension treatments. Measurements of timing of developmental stages and leaf appearance were taken. The generality of photo-thermal approaches to modelling phenological and canopy development was tested. Environmental and genotypic effects on rate of progression from emergence to floral initiation (E-FI) were explained well using a multiplicative model, which combined the intrinsic development rate (Ropt), with responses to temperature and photoperiod. Differences in Ropt and extent of the photoperiod response explained most genotypic effects. Average leaf initiation rate (LIR), leaf appearance rate and duration of the phase from anthesis to physiological maturity differed among genotypes. The association of total leaf number (TLN) with photoperiod found for all genotypes could not be fully explained by effects on development and LIRs. While a putative effect of photoperiod on LIR would explain the observations, other possible confounding factors, such as air-soil temperature differential and the nature of model structure were considered and discussed. This study found a generally robust predictive capacity of photo-thermal development models across diverse ranges of both genotypes and environments. Hence, they remain the most appropriate models for simulation analysis of genotype-by-management scenarios in environments varying broadly in temperature and photoperiod.

Diel variation in mangrove fish abundances and trophic guilds of northeastern Australian estuaries with a proposed trophodynamic model

Diel activity patterns of tropical fish assemblages in turbid, mangrove-dominated estuaries remain largely undocumented, leading to uncertainty about ecological processes in these systems. To capture active fishes by day and night, gill nets were set perpendicular to mangrove shorelines, in six northeastern Australian estuaries during 13 bimonthly trips. Fish were sampled with eight large mesh (102-151 mm) nets, set for 6 hrs (1500-2100), and checked hourly (1146 day, 635 dusk, 872 night checks). Four smaller mesh (19-51 mm) nets were also set for 1 hr before and after sunset (77 day, 78 night checks). Of 157 total species, 22 were netted exclusively before sunset and 47 exclusively after sunset. All of the top 26 species were present both day and night, but of these, 46% were primarily nocturnal (diel index > 0.65). An average of 77.2 fish hr−1 were netted by day vs 171.4 by night. Within the 400 km coastal region, assemblages differed between two northern wave-dominated (WD) estuaries and four southern tide-dominated ('I'D) estuaries. In all six estuaries Lates calcarifer (Bloch, 1790) dominated night assemblages. In 'I'D estuaries, night assemblages were also dominated by Thryssa hamiltoni Gray, 1835 and Eleutheronema tetradactylum (Shaw, 1804); while in WD estuaries Herklotsichthys castelnaui (Ogilby, 1897), Leiognathus equulus (Forsskål, 1775), and Megalops cyprinoids (Broussonet, 1782) were dominant at night. Nocturnal species included planktivores and carnivores, while daytime assemblages were dominated by detritivores (Mugillidae). Higher night catch rates are attributed to increased activity by mobile fishes moving from mangrove to adjacent habitats to forage, especially immediately post-sunset. Although day-night diets and forage resources have yet to be compared in mangrove systems, previously unrecognized trophic relationships involving variation in diel activity among important fishery species (Centropomidae, polynemidae, Carangidae) and their prey may be key ecological processes in these tropical mangrove estuaries. A proposed hypothesis explaining diel variation in mangrove fish assemblages of tropical estuaries is presented through a conceptual model.

An identification tool for the Australian weedy Sporobolus species based on random amplified polymorphic DNA (RAPD) profiles

Based on morphological features alone, there is considerable difficulty in identifying the 5 most economically damaging weed species of Sporobolus [viz. S. pyramidalis P. Beauv., S. natalensis (Steud.) Dur and Schinz, S. fertilis (Steud.) Clayton, S. africanus (Poir.) Robyns and Tourney, and S. jacquemontii Kunth.] found in Australia. A polymerase chain reaction (PCR)-based random amplified polymorphic DNA (RAPD) technique was used to create a series of genetic markers that could positively identify the 5 major weeds from the other less damaging weedy and native Sporobolus species. In the initial RAPD profiling experiment, using arbitrarily selected primers and involving 12 species of Sporobolus, 12 genetic markers were found that, when used in combination, could consistently identify the 5 weedy species from all others. Of these 12 markers, the most diagnostic were UBC51490 for S. pyramidalis and S. natalensis; UBC43310.2000.2100 for S. fertilis and S. africanus; and ORA20850 and UBC43470 for S. jacquemontii. Species-specific markers could be found only for S. jacquemontii. In an effort to understand why there was difficulty in obtaining species-specific markers for some of the weedy species, a RAPD data matrix was created using 40 RAPD products. These 40 products amplified by 6 random primers from 45 individuals belonging to 12 species, were then subjected to numerical taxonomy and multivariate system (NTSYS pc version 1.70) analysis. The RAPD similarity matrix generated from the analysis indicated that S. pyramidalis was genetically more similar to S. natalensis than to other species of the 'S. indicus complex'. Similarly, S. jacquemontii was more similar to S. pyramidalis, and S. fertilis was more similar to S. africanus than to other species of the complex. Sporobolus pyramidalis, S. jacquemontii, S. africanus, and S. creber exhibited a low within-species genetic diversity, whereas high genetic diversity was observed within S. natalensis, S. fertilis, S. sessilis, S. elongates, and S. laxus. Cluster analysis placed all of the introduced species (major and minor weedy species) into one major cluster, with S. pyramidalis and S. natalensis in one distinct subcluster and S. fertilis and S. africanus in another. The native species formed separate clusters in the phenograms. The close genetic similarity of S. pyramidalis to S. natalensis, and S. fertilis to S. africanus may explain the difficulty in obtaining RAPD species-specific markers. The importance of these results will be within the Australian dairy and beef industries and will aid in the development of integrated management strategy for these weeds.

Rickettsia-like-organisms and phytoplasmas associated with diseases in Australian strawberries

Strawberry lethal yellows (SLY) disease in Australia is associated with the phytoplasmas Candidatus Phytoplasma australiense and tomato big bud, and a rickettsia-like-organism (RLO). Ca. P. australiense is also associated with strawberry green petal (SGP) disease. This study investigated the strength of the association of the different agents with SLY disease. We also documented the location of SLY or SGP plants, and measured whether they were RLO or phytoplasma positive. Symptomatic strawberry plants collected from south-east Queensland (Australia) between January 2000 and October 2002 were screened by PCR for both phytoplasmas and the RLO. Two previously unreported disease symptoms termed severe fruit distortion (SFD) and strawberry leaves from fruit (SLF) were observed during this study but there was no clear association between these symptoms and phytoplasmas or the RLO. Only two SGP diseased plants were observed and collected, compared with 363 plants with SLY disease symptoms. Of the 363 SLY samples, 117 tested positive for the RLO, 67 tested positive for Ca. P. australiense AGY strain and 11 plants tested positive for Ca. P. australiense PYL variant strain. On runner production farms at Stanthorpe, Queensland the RLO was detected in SLY diseased plants more frequently than for the phytoplasmas. On fruit production farms on the Sunshine Coast, Queensland, Ca. P. australiense was detected in SLY disease plants more frequently than the RLO.

Recovery of intravenously infused chromium EDTA and lithium sulphate in the urine of cattle and their use as markers to measure urine volume

A series of metabolism experiments investigated the recovery of continuous-, intravenously infused chromium complexed with ethylenediamine tetra-acetic acid (CrEDTA) and lithium sulphate in the urine of cattle with a view to using the markers to estimate urine and metabolite output in grazing cattle. The recovery of Cr in urine from these infusions was similar (90%) in metabolism trials when cattle consumed three very contrasting diets: high-grain formulated pellet, lucerne hay (Medicago sativa) or low-quality native grass hay (predominantly Heteropogon contortus). By contrast, Li recovery in urine averaged 46.3 +/- 0.40% and 72.6 +/- 0.43% for native pasture and lucerne hays, respectively, but was not constant across days. There was negligible transfer of Cr from CrEDTA in blood serum to the rumen or faeces, whereas appreciable quantities of infused Li were found in both. The ratio of urine volume estimated by spot samples and marker dilution of Cr, to urine volume measured gravimetrically, was 1.05. In grazing studies using rumen-fistulated (RF) steers grazing seven different tropical and temperate grass and legume pastures, the ratio of concentrations of purine derivatives (PD) to Cr in spot samples of urine was shown to vary diurnally in the range of 49% to 157% of the average 24 h value. This finding indicated the need for regular sampling of urine to achieve an accurate average value for the PD: Cr ratio in urine for use in estimating urinary PD excretion and hence microbial protein production in the rumen. It was concluded that continuous, intravenous infusion of CrEDTA resulted in a constant recovery of Cr in the urine of cattle across diets and, provided an intensive sampling regime was followed to account for diurnal variation, it would be suitable as a marker to estimate urine volume and urinary output of PD in grazing cattle.

The intrinsic plasticity of farm businesses and their resilience to change. An Australian example.

This paper examines the idea that plasticity in farm management introduces resilience to change and allows farm businesses to perform when operating in highly variable environments. We also argue for the need to develop and apply more integrative assessments of farm performance that combine the use of modelling tools with deliberative processes involving farmers and researchers in a co-learning process, to more effectively identify and implement more productive and resilient farm businesses. In a plastic farming system, farm management is highly contingent on environmental conditions. In plastic farming systems farm managers constantly vary crops and inputs based on the availability of limited and variable resources (e.g. land, water, finances, labour, machinery, etc.), and signals from its operating environment (e.g. climate, markets), with the objective of maximising a number of, often competing, objectives (e.g. maximise profits, minimise risks, etc.). In contrast in more rigid farming systems farm management is more calendar driven and relatively fixed sequences of crops are regularly followed over time and across the farm. Here we describe the application of a whole farm simulation model to (i) compare, in silico, the sensitivity of two farming systems designs of contrasting levels of plasticity, operating in two contrasting environments, when exposed to a stressor in the form of climate change scenarios;(ii) investigate the presence of interactions and feedbacks at the field and farm levels capable of modifying the intensity and direction of the responses to climate signals; and (iii) discuss the need for the development and application of more integrative assessments in the analysis of impacts and adaptation options to climate change. In both environments, the more plastic farm management strategy had higher median profits and was less risky for the baseline and less intensive climate change scenarios (2030). However, for the more severe climate change scenarios (2070), the benefit of plastic strategies tended to disappear. These results suggest that, to a point, farming systems having higher levels of plasticity would enable farmers to more effectively respond to climate shifts, thus ensuring the economic viability of the farm business. Though, as the intensity of the stress increases (e.g. 2070 climate change scenario) more significant changes in the farming system might be required to adapt. We also found that in the case studies analysed here, most of the impacts from the climate change scenarios on farm profit and economic risk originated from important reductions in cropping intensity and changes in crop mix rather than from changes in the yields of individual crops. Changes in cropping intensity and crop mix were explained by the combination of reductions in the number of sowing opportunities around critical times in the cropping calendar, and to operational constraints at the whole farm level i.e. limited work capacity in an environment having fewer and more concentrated sowing opportunities. This indicates that indirect impacts from shifts in climate on farm operations can be more important than direct impacts from climate on the yield of individual crops. The results suggest that due to the complexity of farm businesses, impact assessments and opportunities for adaptation to climate change might also need to be pursued at higher integration levels than the crop or the field. We conclude that plasticity can be a desirable characteristic in farming systems operating in highly variable environments, and that integrated whole farm systems analyses of impacts and adaptation to climate change are required to identify important interactions between farm management decision rules, availability of resources, and farmer's preference.

The conservation status of the world’s reptiles

Effective and targeted conservation action requires detailed information about species, their distribution, systematics and ecology as well as the distribution of threat processes which affect them. Knowledge of reptilian diversity remains surprisingly disparate, and innovative means of gaining rapid insight into the status of reptiles are needed in order to highlight urgent conservation cases and inform environmental policy with appropriate biodiversity information in a timely manner. We present the first ever global analysis of extinction risk in reptiles, based on a random representative sample of 1500 species (16% of all currently known species). To our knowledge, our results provide the first analysis of the global conservation status and distribution patterns of reptiles and the threats affecting them, highlighting conservation priorities and knowledge gaps which need to be addressed urgently to ensure the continued survival of the world’s reptiles. Nearly one in five reptilian species are threatened with extinction, with another one in five species classed as Data Deficient. The proportion of threatened reptile species is highest in freshwater environments, tropical regions and on oceanic islands, while data deficiency was highest in tropical areas, such as Central Africa and Southeast Asia, and among fossorial reptiles. Our results emphasise the need for research attention to be focussed on tropical areas which are experiencing the most dramatic rates of habitat loss, on fossorial reptiles for which there is a chronic lack of data, and on certain taxa such as snakes for which extinction risk may currently be underestimated due to lack of population information. Conservation actions specifically need to mitigate the effects of human-induced habitat loss and harvesting, which are the predominant threats to reptiles.