Comparative growth and pathogenicity of geographical isolates of Sclerotinia sclerotiorum on lentil genotypes

Isolates of Sclerotinia sclerotiorum were collected from infected lentil plants from 2 agro-ecological zones of Syria and used to study their comparative growth on culture media and pathogenicity on different lentil genotypes. The growth studies were carried out on Potato Dextrose Agar (PDA) growth media under laboratory conditions. Mycelial radial growth and sclerotial production were the parameters used to compare the isolates. Pathogenicity studies were carried out with selected isolates on 10 lentil genotypes, infected as detached shoots and as whole potted-plants in the plastic house. The isolates showed considerable variation in cultural characteristics through mycelial growth, mycelial pigmentation and sclerotial production in the media plates. There were significant differences in the growth and sclerotial production of most of the isolates, but no apparent correlation between mycelial growth and sclerotial production among the isolates. Genotype by isolate interactions was significant for the isolates tested for pathogenicity. These interactions, however, appeared to be caused by differences in virulence of the isolates and did not suggest the occurrence of distinct pathogenic races of the pathogen isolates.

Host-delivered RNAi: An effective strategy to silence genes in plant parasitic nematodes

Root-knot nematodes (Meloidogyne spp.) are obligate, sedentary endoparasites that infect many plant species causing large economic losses worldwide. Available nematicides are being banned due to their toxicity or ozone-depleting properties and alternative control strategies are urgently required. We have produced transgenic tobacco (Nicotiana tabacum) plants expressing different dsRNA hairpin structures targeting a root-knot nematode (Meloidogyne javanica) putative transcription factor, MjTis11. We provide evidence that MjTis11 was consistently silenced in nematodes feeding on the roots of transgenic plants. The observed silencing was specific for MjTis11, with other sequence-unrelated genes being unaffected in the nematodes. Those transgenic plants able to induce silencing of MjTis11, also showed the presence of small interfering RNAs. Even though down-regulation of MjTis11 did not result in a lethal phenotype, this study demonstrates the feasibility of silencing root-knot nematode genes by expressing dsRNA in the host plant. Host-delivered RNA interference-triggered (HD-RNAi) silencing of parasite genes provides a novel disease resistance strategy with wide biotechnological applications. The potential of HD-RNAi is not restricted to parasitic nematodes but could be adapted to control other plant-feeding pests.

Effect of microwave radiation on seed mortality of rubber vine (Cryptostegia grandiflora R.Br.), parthenium (Parthenium hysterophorous L.) and bellyache bush (Jatropha gossypiifolia L.)

A trial was undertaken to evaluate the effect of microwaves on seed mortality of three weed species. Seeds of rubber vine (Cryptostegia grandiflora R.Br.), parthenium (Parthenium hysterophorous L.) and bellyache bush (Jatropha gossypiifolia L.) were buried at six depths (0, 2.5, 5, 10, 20 and 40 cm) in coarse sand maintained at one of two moisture levels, oven dry or wet (field capacity), and then subjected to one of five microwave radiation durations of (0, 2, 4, 8 and 16 min). Significant interactions between soil moisture level, microwave radiation duration, seed burial depth and species were detected for mortality of seeds of all three species. Maximum seed mortality of rubber vine (88%), parthenium (67%) and bellyache bush (94%) occurred in wet soil irradiated for 16 min. Maximum seed mortality of rubber vine and bellyache bush seeds occurred in seeds buried at 2.5 cm depth whereas that of parthenium occurred in seeds buried at 10 cm depth. Maximum soil temperatures of 114.1 and 87.5°C in dry and wet soil respectively occurred at 2.5 cm depth following 16 min irradiation. Irrespective of the greater soil temperatures recorded in dry soil, irradiating seeds in wet soil generally increased seed mortality 2.9-fold compared with dry soil. Moisture content of wet soil averaged 5.7% compared with 0.1% for dry soil. Results suggest that microwave radiation has the potential to kill seeds located in the soil seed bank. However, many factors, including weed species susceptibility, determine the effectiveness of microwave radiation on buried seeds. Microwave radiation may be an alternative to conventional methods at rapidly depleting soil seed banks in the field, particularly in relatively wet soils that contain long lived weed seeds.

Growth and species interactions of Eucalyptus pellita in a mixed and monoculture plantation in the humid tropics of north Queensland

This study investigated whether mixed-species designs can increase the growth of a tropical eucalypt when compared to monocultures. Monocultures of Eucalyptus pellita (E) and Acacia peregrina (A) and mixtures in various proportions (75E:25A, 50E:50A, 25E:75A) were planted in a replacement series design on the Atherton Tablelands of north Queensland, Australia. High mortality in the establishment phase due to repeated damage by tropical cyclones altered the trial design. Effects of experimental designs on tree growth were estimated using a linear mixed-effects model with restricted maximum likelihood analysis (REML). Volume growth of individual eucalypt trees were positively affected by the presence of acacia trees at age 5 years and this effect generally increased with time up to age 10 years. However, the stand volume and basal area increased with increasing proportions of E. pellita, due to its larger individual tree size. Conventional analysis did not offer convincing support for mixed-species designs. Preliminary individual-based modelling using a modified Hegyi competition index offered a solution and an equation that indicates acacias have positive ecological interactions (facilitation or competitive reduction) and definitely do not cause competition like a eucalypt. These results suggest that significantly increased in growth rates could be achieved with mixed-species designs. This statistical methodology could enable a better understanding of species interactions in similarly altered experiments, or undesigned mixed-species plantations.

Selecting for increased barley grain size

In this study, 120–144 commercial varieties and breeding lines were assessed for grain size attributes including plump grain (>2.8 mm) and retention (>2.5 mm+>2.8 mm). Grain samples were produced from replicated trials at 25 sites across four years. Climatic conditions varied between years as well as between sites. Several of the trial sites were irrigated while the remaining were produced under dryland conditions. A number of the dryland sites suffered severe drought stress. The grain size data was analysed for genetic (G), environmental (E) and genotype by environment (G×E) interactions. All analyses included maturity as a covariate. The genetic effect on grain size was greater than environmental or maturity effects despite some sites suffering terminal moisture stress. The model was used to calculate heritability values for each site used in the study. These values ranged from 89 to 98% for plump grain and 88 to 96% for retention. The results demonstrated that removing the sources of non-heritable variation, such as maturity and field effects, can improve genetic estimates of the retention and plump grain fractions. By partitioning all variance components, and thereby having more robust estimates of genetic differences, plant breeders can have greater confidence in selecting barley genotypes which maintain large, stable grain size across a range of environments.

Short-term responses of leaf growth rate to water deficit scale up to whole-plant and crop levels: An integrated modelling approach in maize.

Physiological and genetic studies of leaf growth often focus on short-term responses, leaving a gap to whole-plant models that predict biomass accumulation, transpiration and yield at crop scale. To bridge this gap, we developed a model that combines an existing model of leaf 6 expansion in response to short-term environmental variations with a model coordinating the development of all leaves of a plant. The latter was based on: (1) rates of leaf initiation, appearance and end of elongation measured in field experiments; and (2) the hypothesis of an independence of the growth between leaves. The resulting whole-plant leaf model was integrated into the generic crop model APSIM which provided dynamic feedback of environmental conditions to the leaf model and allowed simulation of crop growth at canopy level. The model was tested in 12 field situations with contrasting temperature, evaporative demand and soil water status. In observed and simulated data, high evaporative demand reduced leaf area at the whole-plant level, and short water deficits affected only leaves developing during the stress, either visible or still hidden in the whorl. The model adequately simulated whole-plant profiles of leaf area with a single set of parameters that applied to the same hybrid in all experiments. It was also suitable to predict biomass accumulation and yield of a similar hybrid grown in different conditions. This model extends to field conditions existing knowledge of the environmental controls of leaf elongation, and can be used to simulate how their genetic controls flow through to yield.

Heterologous signals allow efficient targeting of a nuclear-encoded fusion protein to plastids and endoplasmic reticulum in diverse plant species

Approximately 30% of plant nuclear genes appear to encode proteins targeted to the plastids or endoplasmic reticulum (ER). The signals that direct proteins into these compartments are diverse in sequence, but, on the basis of a limited number of tests in heterologous systems, they appear to be functionally conserved across species. To further test the generality of this conclusion, we tested the ability of two plastid transit peptides and an ER signal peptide to target green fluorescent protein (GFP) in 12 crops, including three monocots (barley, sugarcane, wheat) and nine dicots (Arabidopsis, broccoli, cabbage, carrot, cauliflower, lettuce, radish, tobacco, turnip). In all species, transient assays following microprojectile bombardment or vacuum infiltration using Agrobacterium showed that the plastid transit peptides from tomato DCL (defective chloroplast and leaves) and tobacco RbcS [ribulose bisphosphate carboxylase (Rubisco) small subunit] genes were effective in targeting GFP to the leaf plastids. GFP engineered as a fusion to the N-terminal ER signal peptide from Arabidopsis basic chitinase and a C-terminal HDEL signal for protein retention in the ER was accumulated in the ER of all species. The results in tobacco were confirmed in stably transformed cells. These signal sequences should be useful to direct proteins to the plastid stroma or ER lumen in diverse plant species of biotechnological interest for the accumulation of particular recombinant proteins or for the modification of particular metabolic streams.

Simulating crop-parasitic weed interactions using APSIM: Model evaluation and application

The parasitic weed Orobanche crenata inflicts major damage on faba bean, lentil, pea and other crops in Mediterranean environments. The development of methods to control O. crenata is to a large extent hampered by the complexity of host-parasite systems. Using a model of host-parasite interactions can help to explain and understand this intricacy. This paper reports on the evaluation and application of a model simulating host-parasite competition as affected by environment and management that was implemented in the framework of the Agricultural Production Systems Simulator (APSIM). Model-predicted faba bean and O. crenata growth and development were evaluated against independent data. The APSIM-Fababean and -Parasite modules displayed a good capability to reproduce effects of pedoclimatic conditions, faba bean sowing date and O. crenata infestation on host-parasite competition. The r(2) values throughout exceeded 0.84 (RMSD: 5.36 days) for phenological, 0.85 (RMSD: 223.00 g m(-2)) for host growth and 0.78 (RMSD: 99.82 g m(-2)) for parasite growth parameters. Inaccuracies of simulated faba bean root growth that caused some bias of predicted parasite number and host yield loss may be dealt with by more flexibly simulating vertical root distribution. The model was applied in simulation experiments to determine optimum sowing windows for infected and non-infected faba bean in Mediterranean environments. Simulation results proved realistic and testified to the capability of APSIM to contribute to the development of tactical approaches in parasitic weed control.

Development of key soil health indicators for the Australian banana industry

To improve the sustainability and environmental accountability of the banana industry there is a need to develop a set of soil health indicators that integrate physical, chemical and biological soil properties. These indicators would allow banana growers, extension and research workers to improve soil health management practices. To determine changes in soil properties due to the cultivation of bananas, a paired site survey was conducted comparing soil properties under conventional banana systems to less intensively managed vegetation systems, such as pastures and forest. Measurements were made on physical, chemical and biological soil properties at seven locations in tropical and sub-tropical banana producing areas. Soil nematode community composition was used as a bioindicator of the biological properties of the soil. Soils under conventional banana production tended to have a greater soil bulk density, with less soil organic carbon (C) (both total C and labile C), greater exchangeable cations, higher extractable P, greater numbers of plant-parasitic nematodes and less nematode diversity, relative to less intensively managed plant systems. The organic banana production systems at two locations had greater labile C, relative to conventional banana systems, but there was no significant change in nematode community composition. There were significant interactions between physical, chemical and nematode community measurements in the soil, particularly with soil C measurements, confirming the need for a holistic set of indicators to aid soil management. There was no single indicator of soil health for the Australian banana industry, but a set of soil health indicators, which would allow the measurement of soil improvements should include: bulk density, soil C, pH, EC, total N, extractable P, ECEC and soil nematode community structure.

A blowfly strike vaccine requires an understanding of host-pathogen interactions

The phase-out of Mulesing by 2010 means the Australian wool industry requires immediate and viable alternatives for the control and prevention of blowfly strike, an economically important parasitic disease of sheep. In this review we have analysed previous research aimed toward the development of a vaccine against blowfly strike and the reasons why the approaches taken were unsuccessful at the time. Close scrutiny has provided new insight into this host-parasite interaction and identified new opportunities for the development of a vaccine. Here we propose that addressing immunosuppression together with the induction of cellular immunity is likely to result in an anti-blowfly strike vaccine, as opposed to the use of "standard" approaches aimed at inducing humoral immunity.

Gene interactions constrain the course of evolution of phosphine resistance in the lesser grain borer, Rhyzopertha dominica

Phosphine, a widely used fumigant for the protection of stored grain from insect pests, kills organisms indirectly by inducing oxidative stress. High levels of heritable resistance to phosphine in the insect pest of stored grain, Rhyzopertha dominica have been detected in Asia, Australia and South America. In order to understand the evolution of phosphine resistance and to isolate the responsible genes, we have undertaken genetic linkage analysis of fully sensitive (QRD14), moderately resistant (QRD369) and highly resistant (QRD569) strains of R. dominica collected in Australia. We previously determined that two loci, rph1 and rph2, confer high-level resistance on strain QRD569, which was collected in 1997. We have now confirmed that rph1 is responsible for the moderate resistance of strain QRD369, which was collected in 1990, and is shared with a highly resistant strain from the same geographical region, QRD569. In contrast, rph2 by itself confers only very weak resistance, either as a heterozygote or as a homozygote and was not discovered in the field until weak resistance (probably due to rph1) had become ubiquitous. Thus, high-level resistance against phosphine has evolved via stepwise acquisition of resistance alleles, first at rph1 and thereafter at rph2. The semi-dominance of rph2 together with the synergistic interaction between rph1 and rph2 would have led to rapid selection for homozygosity. A lack of visible fitness cost associated with alleles at either locus suggests that the resistance phenotype will persist in the field.

Development of a small-plant bioassay to assess banana grown from tissue culture for consistent infection by Fusarium oxysporum f. sp. cubense

Two reliable small-plant bioassays were developed using tissue-cultured banana, resulting in consistent symptom expression and infection by Fusarium oxysporum f. sp. cubense (Foc). One bioassay was based on providing a constant watertable within a closed pot and the second used free-draining pots. Culture medium for spore generation influenced infectivity of Foc. Inoculation of potted banana by drenching potting mix with a conidial suspension, consisting mostly of microconidia, few macroconidia and no chlamydospores, generated from one-quarter-strength potato dextrose agar + streptomycin sulfate, resulted in inconsistent infection. When a conidial suspension that consisted of all three spore types, microconidia, macroconidia and chlamydospores, prepared from spores generated on carnation leaf agar was used, all plants became infected, indicating that the spore type present in conidial suspensions may contribute to inconsistency of infection. Inconsistency of infection was not due to loss of virulence of the pathogen in culture. Millet grain precolonised by Foc as a source of inoculum resulted in consistent infection between replicate plants. Sorghum was not a suitable grain for preparation of inoculum as it was observed to discolour roots and has the potential to stunt root growth, possibly due to the release of phytotoxins. For the modified closed-pot system, a pasteurised potting mix consisting of equal parts of bedding sand, perlite and vermiculite plus 1 g/L Triabon slow release fertiliser was suitable for plant growth and promoted capillary movement of water through the potting mix profile. A suitable potting mix for the free-draining pot system was also developed.

Melaleuca densispicata in Currawinya National Park, South-West Queensland: Ecology and preliminary implications for management of a rare plant species

Melaleuca densispicata Byrnes is an uncommon species with a limited distribution, comprising disjunct populations in inland southern Queensland and northern New South Wales, Australia. It is a dense, woody shrub, 2–4 m in height, which exhibits a marked 'clumping' growth habit. It has thick, papery bark and displays many white flowers during spring or early summer. Although it has long been known to exist, M. densispicata was only formally described in 1984, and very little is currently known about its ecology or specific management requirements. There are only seven known subpopulations of the species across its range. A major population at the western limit of its distribution occurs on Currawinya National Park (28°52'S, 144°30'E). Here, it is locally abundant and listed as a noteworthy plant species under the Management Plan (Queensland Parks & Wildlife Service 2001). This study aimed to identify patterns in the distribution of M. densispicata in Currawinya National Park, describe its ecological niche and role, and provide management recommendations for the species within the study area. Recent anecdotal observations of recruitment failure in south-western Queensland (Peter McRae, QPWS, October 2004, pers. comm.; Dick O'Connell, local grazier, July 2005 pers. comm.) caused additional emphasis to be placed on the examination of recruitment and recruitment factors.

Beyond chemical dependency for managing plant-parasitic nematodes: Examples from the banana, pineapple and vegetable industries of tropical and subtropical Australia

Plant-parasitic nematodes are important pests of horticultural crops grown in tropical and subtropical regions of Australia. Burrowing nematode (Radopholus similis) is a major impediment to banana production and root-knot nematodes (predominantly Meloidogyne javanica and M. incognita) cause problems on pineapple and a range of annual vegetables, including tomato, capsicum, zucchini, watermelon, rockmelon, potato and sweet potato. In the early 1990s, nematode control in these industries was largely achieved with chemicals, with methyl bromide widely used on some subtropical vegetable crops, ethylene dibromide applied routinely to pineapples and non-volatile nematicides such as fenamiphos applied up to four times a year in banana plantations. This paper discusses the research and extension work done over the last 15 years to introduce an integrated pest management approach to nematode control in tropical and subtropical horticulture. It then discusses various components of current integrated pest management programs, including crop rotation, nematode monitoring, clean planting material, organic amendments, farming systems to enhance biological suppression of nematodes and judicious use of nematicides. Finally, options for improving current management practices are considered.

The environmental fate of diuron under a conventional production regime in a sugarcane farm during the plant cane phase

BACKGROUND: Field studies of diuron and its metabolites 3-(3,4-dichlorophenyl)-1-methylurea (DCPMU), 3,4-dichlorophenylurea (DCPU) and 3,4-dichloroaniline (DCA) were conducted in a farm soil and in stream sediments in coastal Queensland, Australia. RESULTS: During a 38 week period after a 1.6 kg ha^-1 diuron application, 70-100% of detected compounds were within 0-15 cm of the farm soil, and 3-10% reached the 30-45 cm depth. First-order t1/2 degradation averaged 49 ± 0.9 days for the 0-15, 0-30 and 0-45 cm soil depths. Farm runoff was collected in the first 13-50 min of episodes lasting 55-90 min. Average concentrations of diuron, DCPU and DCPMU in runoff were 93, 30 and 83-825 µg L^-1 respectively. Their total loading in all runoff was >0.6% of applied diuron. Diuron and DCPMU concentrations in stream sediments were between 3-22 and 4-31 µg kg^-1 soil respectively. The DCPMU/diuron sediment ratio was >1. CONCLUSION: Retention of diuron and its metabolites in farm topsoil indicated their negligible potential for groundwater contamination. Minimal amounts of diuron and DCMPU escaped in farm runoff. This may entail a significant loading into the wider environment at annual amounts of application. The concentrations and ratio of diuron and DCPMU in stream sediments indicated that they had prolonged residence times and potential for accumulation in sediments. The higher ecotoxicity of DCPMU compared with diuron and the combined presence of both compounds in stream sediments suggest that together they would have a greater impact on sensitive aquatic species than as currently apportioned by assessments that are based upon diuron alone.