SSR analysis of cultivated groundnut (Arachis hypogaea L.) germplasm resistant to rust and late leaf spot diseases

Cultivated groundnut (Arachis hypogaea L.) is an agronomically and economically important oilseed crop grown extensively throughout the semi-arid tropics of Asia, Africa and Latin America. Rust (Puccinia arachidis) and late leaf spot (LLS, Phaseoisariopsis personata) are among the major diseases causing significant yield loss in groundnut. The development of varieties with high levels of resistance has been constrained by adaptation of disease isolates to resistance sources and incomplete resistance in resistant sources. Despite the wide range of morphological diversity observed in the cultivated groundnut gene pool, molecular marker analyses have thus far been unable to detect a parallel level of genetic diversity. However, the recent development of simple sequence repeat (SSR) markers presents new opportunities for molecular diversity analysis of cultivate groundnut. The current study was conducted to identify diverse disease resistant germplasm for the development of mapping populations and for their introduction into breeding programs. Twenty-three SSRs were screened across 22 groundnut genotypes with differing levels of resistance to rust and LLS. Overall, 135 alleles across 23 loci were observed in the 22 genotypes screened. Twelve of the 23 SSRs (52%) showed a high level of polymorphism, with PIC values ≥0.5. This is the first report detecting such high levels of genetic polymorphism in cultivated groundnut. Multi-dimensional scaling and cluster analyses revealed three well-separated groups of genotypes. Locus by locus AMOVA and Kruskal-Wallis one-way ANOVA identified candidate SSR loci that may be valuable for mapping rust and LLS resistance. The molecular diversity analysis presented here provides valuable information for groundnut breeders designing strategies for incorporating and pyramiding rust and late leaf spot resistances and for molecular biologists wishing to create recombinant inbred line populations to map these traits.

Laboratory information management software for genotyping workflows: Applications in high throughput crop genotyping

Background: With the advances in DNA sequencer-based technologies, it has become possible to automate several steps of the genotyping process leading to increased throughput. To efficiently handle the large amounts of genotypic data generated and help with quality control, there is a strong need for a software system that can help with the tracking of samples and capture and management of data at different steps of the process. Such systems, while serving to manage the workflow precisely, also encourage good laboratory practice by standardizing protocols, recording and annotating data from every step of the workflow Results: A laboratory information management system (LIMS) has been designed and implemented at the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT) that meets the requirements of a moderately high throughput molecular genotyping facility. The application is designed as modules and is simple to learn and use. The application leads the user through each step of the process from starting an experiment to the storing of output data from the genotype detection step with auto-binning of alleles; thus ensuring that every DNA sample is handled in an identical manner and all the necessary data are captured. The application keeps track of DNA samples and generated data. Data entry into the system is through the use of forms for file uploads. The LIMS provides functions to trace back to the electrophoresis gel files or sample source for any genotypic data and for repeating experiments. The LIMS is being presently used for the capture of high throughput SSR (simple-sequence repeat) genotyping data from the legume (chickpea, groundnut and pigeonpea) and cereal (sorghum and millets) crops of importance in the semi-arid tropics. Conclusions: A laboratory information management system is available that has been found useful in the management of microsatellite genotype data in a moderately high throughput genotyping laboratory. The application with source code is freely available for academic users and can be downloaded from http://www.icrisat.org/bt-software-d-lims.htm

DNA markers linked to the R(2) rust resistance gene in sunflower (Helianthus annuus L.) facilitate anticipatory breeding for this disease variant.

Pre-emptive breeding for host disease resistance is an effective strategy for combating and managing devastating incursions of plant pathogens. Comprehensive, long-term studies have revealed that virulence to the R (2) sunflower (Helianthus annuus L.) rust resistance gene in the line MC29 does not exist in the Australian rust (Puccinia helianthi) population. We report in this study the identification of molecular markers linked to this gene. The three simple sequence repeat (SSR) markers ORS795, ORS882, and ORS938 were linked in coupling to the gene, while the SSR marker ORS333 was linked in repulsion. Reliable selection for homozygous-resistant individuals was efficient when the three markers, ORS795, ORS882, and ORS333, were used in combination. Phenotyping for this resistance gene is not possible in Australia without introducing a quarantinable race of the pathogen. Therefore, the availability of reliable and heritable DNA-based markers will enable the efficient deployment of this gene, permitting a more effective strategy for generating sustainable commercial cultivars containing this rust resistance gene.

Chromosome complements in the spermatogenesis of two penaeid prawns, Penaeus merguiensis and P. esculentus.

Although the fisheries for, and mariculture of, penaeid prawns are of major commercial importance, there has been relatively little research undertaken on the chromosome number, structure and composition in the Penaeidae. One reason for this is due to the relatively small size and large number of chromosomes, which makes production of histological material difficult. In this paper, we report a simple and effective technique for determining chromosome complements during spermatogenesis in two species of penaeid prawns, Penaeus merguiensis and P. esculentus in Australia. The first estimates of the number of chromosomes in these species are given.

Effect of nitrogen on the skin colour and other quality attributes of ripe ‘Kensington Pride’ mango (Mangifera indica L.) fruit

Near-ripe ‘Kensington Pride’ mango (Mangifera indica L.) fruit with green skin colour generally return lower wholesale and retail prices. Pre-harvest management, especially nitrogen (N) nutrition, appears to be a major causal factor. To obtain an understanding of the extent of the problem in the Burdekin district (dry tropics; the major production area in Australia), green mature ‘Kensington Pride’ mango fruit were harvested from ten orchards and ripened at 20 ± 0.5 O C. Of these orchards, 70% produced fruit with more than 25% of the skin surface area green when ripe. The following year, the effect of N application on skin colour and other quality attributes was investigated on three orchards, one with a high green (HG) skin problem and two with a low green (LG) skin problem. N was applied at pre-flowering and at panicle emergence at the rate of 0,75,150,300 g per tree (soil applied) or 50 g per tree as foliar N for the HG orchard, and 0,150,300,450 g per tree (soil applied) or 50 g per tree (foliar) for the LG orchards. In all orchards the proportion of green colour on the ripe fruit was significantly (P<0.05) higher with soil applications of 150 g N or more per tree. Foliar sprays resulted in a higher proportion of green colour than the highest soil treatment in the HG orchard, but not in the LG orchards. Anthracnose disease severity was significantly (P<0.05) higher with 300 g of N per tree or foliar treatment in the HG orchard, compared with no additional N. Thus, N can reduce mango fruit quality by increasing green colour and anthracnose disease in ripe fruit.

Egg parasitoids of the diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae), from south-east Queensland

Field surveys of egg parasitoids of the diamondback moth, Plutella xylostella, were conducted at Redlands and Gatton, south-east Queensland. Eggs of P. xylostella were present all year round in both localities, and parasitized eggs were consistently found between late spring and early winter. Percent parasitism in the range 30–75% was recorded on many occasions, although rates less than 10% were more common. The major parasitoids included Trichogrammatoidea bactrae Nagaraja and Trichogramma pretiosum Riley. Laboratory evaluation showed that the T. pretiosum from Gatton has a high capacity to parasitize P. xylostella eggs under suitable conditions. This study represents the first record of egg parasitoids of P. xylostella from Australia.

A comparison of the performance and biological efficiency of new knapsack sprayers and a controlled droplet application (CDA) sprayer for the control of the diamondback moth, Plutella xylostella L. (Lepidoptera: Plutellidae)

Field trials and laboratory bioassays were undertaken to compare the performance and efficacy (mortality of diamondback moth larvae) of insecticides applied to cabbages with three high volume hydraulic knapsack sprayers (NS-16, PB-20 and Selecta 12V) and a controlled droplet application (CDA) sprayer. In field experiments, the high volume knapsack sprayers (application rate 500-600 L ha-') provided better spray coverage on the upper and lower surfaces of inner leaves, the upper surfaces of middle and outer leaves, and greater biological efficacy than the CDA sprayer (application rate 20~40 L ha-'). The PB-20 provided better spray coverage on the upper surface of middle leaves and both Surfaces of outer leaves when compared with the Selecta I2V. However, its biological efficacy in the field was not significantly different from that of the other high volume sprayers. Increasing the application rate from 20 to 40 L ha - ' for the CDA sprayer significantly increased droplet density but had no impact on test insect mortality. Laboratory evaluations of biological efficacy yielded higher estimates than field evaluations and there was no significant difference between the performance of the PB-20 and the CDA sprayer. Significant positive relationships were detected between insect mortality and droplet density deposited for both the PB-20 and the CDA sprayers

Predicting mortality of phosphine-resistant adults of Sitophilus oryzae (L) (Coleoptera: Curculionidae) in relation to changing phosphine concentration

Adults of a phosphine-resistant strain of Sitophilus oryzae (L) were exposed to constant phosphine concentrations of 0.0035-0.9 mg litre(-1) for periods of between 20 and 168 h at 25 °C, and the effects of time and concentration on mortality were quantified. Adults were also exposed to a series of treatments lasting 48, 72 or 168 h at 25 °C, during which the concentration of phosphine was varied. The aim of this study was to determine whether equations from experiments using constant concentrations could be used to predict the efficacy of changing phosphine concentrations against adults of S oryzae. A probit plane without interaction, in which the logarithms of time (t) and concentration (C) were variables, described the effects of concentration and time on mortality in experiments with constant concentrations. A derived equation of the form C^nt = k gave excellent predictions of toxicity when applied to data from changing concentration experiments. The results suggest that for resistant S oryzae adults there is nothing inherently different between constant and changing concentration regimes, and that data collected from fixed concentrations can be used to develop equations for predicting mortality in fumigations in which phosphine concentration changes. This approach could simplify the prediction of efficacy of typical fumigations in which concentrations tend to rise and then fall over a period of days.

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.

Simulation of the supply capability of sheep on the Mitchell grass downs of north west Queensland

Sheep in western Queensland have been predominantly reared for wool. When wool prices became depressed interest in the sheep meat industry, increased. For north west Queensland producers, opportunities may exist to participate in live sheep and meat export to Asia. A simulation model was developed to determine whether this sheep producing area has the capability to provide sufficient numbers of sheep under variable climatic conditions while sustaining the land resources. Maximum capacity for sustainability of resources (as described by stock numbers) was derived from an in-depth study of the agricultural and pastoral potential of Queensland. Decades of sheep production and climatic data spanning differing seasonal conditions were collated for analysis. A ruminant biology model adapted from Grazplan was used to simulate pregnancy rate. Empirical equations predict mortalities, marking rates, and weight characteristics of sheep of various ages from simple climatic measures, stocking rate and reproductive status. The initial age structure of flocks was determined by running the model for several years with historical climatic conditions. Drought management strategies such as selling a proportion of wethers progressively down to two-tooth and oldest ewes were incorporated. Management decisions such as time of joining, age at which ewes were cast-for-age, wether turn-off age and turning-off rate of lambs vary with geographical area and can be specified at run time. The model is run for sequences of climatic conditions generated stochastically from distributions based on historical climatic data correlated in some instances. The model highlights the difficulties of sustaining a consistent supply of sheep under variable climatic conditions.

Peanut stripe potyvirus resistance in peanut (Arachis hypogaea L.) plants carrying viral coat protein gene sequences

Peanut (Arachis hypogaea L.) lines exhibiting high levels of resistance to peanut stripe virus (PStV) were obtained following microprojectile bombardment of embryogenic callus derived from mature seeds. Fertile plants of the commercial cultivars Gajah and NC7 were regenerated following co-bombardmentwith the hygromycin resistance gene and one of two forms of the PStV coat protein (CP) gene, an untranslatable, full length sequence (CP2) or a translatable gene encoding a CP with an N-terminal truncation (CP4). High level resistance to PStV was observed for both transgenes when plants were challenged with the homologous virus isolate. The mechanism of resistance appears to be RNA-mediated, since plants carrying either the untranslatable CP2 or CP4 had no detectable protein expression, but were resistant or immune (no virus replication). Furthermore, highly resistant, but not susceptible CP2 T0 plants contained transgene-specific small RNAs. These plants now provide important germplasm for peanut breeding, particularly in countries where PStV is endemic and poses a major constraint to peanut production.

Exploring profit - Sustainability trade-offs in cropping systems using evolutionary algorithms

Models that implement the bio-physical components of agro-ecosystems are ideally suited for exploring sustainability issues in cropping systems. Sustainability may be represented as a number of objectives to be maximised or minimised. However, the full decision space of these objectives is usually very large and simplifications are necessary to safeguard computational feasibility. Different optimisation approaches have been proposed in the literature, usually based on mathematical programming techniques. Here, we present a search approach based on a multiobjective evaluation technique within an evolutionary algorithm (EA), linked to the APSIM cropping systems model. A simple case study addressing crop choice and sowing rules in North-East Australian cropping systems is used to illustrate the methodology. Sustainability of these systems is evaluated in terms of economic performance and resource use. Due to the limited size of this sample problem, the quality of the EA optimisation can be assessed by comparison to the full problem domain. Results demonstrate that the EA procedure, parameterised with generic parameters from the literature, converges to a useable solution set within a reasonable amount of time. Frontier ‘‘peels’’ or Pareto-optimal solutions as described by the multiobjective evaluation procedure provide useful information for discussion on trade-offs between conflicting objectives.

Climate information needs of Gascoyne-Murchison pastoralists: a representative study of the Western Australian grazing industry

The Gascoyne-Murchison region of Western Australia experiences an arid to semi-arid climate with a highly variable temporal and spatial rainfall distribution. The region has around 39.2 million hectares available for pastoral lease and supports predominantly catle and sheep grazing leases. In recent years a number of climate forecasting systems have been available offering rainfall probabilities with different lead times and a forecast period; however, the extent to which these systems are capable of fulfilling the requirements of the local pastoralists is still ambiguous. Issues can range from ensuring forecasts are issued with sufficient lead time to enable key planning or decisions to be revoked or altered, to ensuring forecast language is simple and clear, to negate possible misunderstandings in interpretation. A climate research project sought to provide an objective method to determine which available forecasting systems had the greatest forecasting skill at times of the year relevant to local property management. To aid this climate research project, the study reported here was undertaken with an overall objective of exploring local pastoralists' climate information needs. We also explored how well they understand common climate forecast terms such as 'mean', median' and 'probability', and how they interpret and apply forecast information to decisions. A stratified, proportional random sampling was used for the purpose of deriving the representative sample based on rainfall-enterprise combinations. In order to provide more time for decision-making than existing operational forecasts that are issued with zero lead time, pastoralists requested that forecasts be issued for May-July and January-March with lead times counting down from 4 to 0 months. We found forecasts of between 20 and 50 mm break-of-season or follow-up rainfall were likely to influence decisions. Eighty percent of pastoralists demonstrated in a test question that they had a poor technical understanding of how to interpret the standard wording of a probabilistic median rainfall forecast. this is worthy of further research to investigate whether inappropriate management decisions are being made because the forecasts are being misunderstood. We found more than half the respondents regularly access and use weather and climate forecasts or outlook information from a range of sources and almost three-quarters considered climate information or tools useful, with preferred methods for accessing this information by email, faxback service, internet and the Department of Agriculture Western Australia's Pastoral Memo. Despite differences in enterprise types and rainfall seasonality across the region we found seasonal climate forecasting needs were relatively consistent. It became clear that providing basic training and working with pastoralists to help them understand regional climatic drivers, climate terminology and jargon, and the best ways to apply the forecasts to enhance decision-making are important to improve their use of information. Consideration could also be given to engaging a range of producers to write the climate forecasts themselves in the language they use and understand, in consultation with the scientists who prepare the forecasts.

Near infrared reflectance as a rapid and inexpensive surrogate measure for fatty acid composition and oil content in peanuts (Arachis hypogaea L.)

The fatty acid composition of ground nuts (Arachis hypogaea L.) commonly known as peanuts, is an important consideration when a new variety is being released. The composition impacts on nutrition and, importantly, self-life of peanut products. To select for suitable breeding material, it was necessary to develop a rapid, non-derstructive and cost-efficient method. Near infrared spectroscopy was chosen as that methodology. Calibrations were developed for two major fatty-acid components, oleic and linoleic acids and two minor components, palmitic and stearic acids, as well as total oil content. Partial least squares models indicated a high level of precision with a squared multiple correlation coefficient of greater than 0.90 for each constitutent. Standard errors for prediction for oleic, linoleic, palmitic, stearic acids and total oil content were 6.4%, 4.5%, 0.8%, 0.9% and 1.3% respectively. The results demonstrated that reasonable calibrations could be developed to predict oil composition and content of peanuts for a breeding programme.