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.

Confirmation of QTL mapping and marker validation for partial seedling resistance to crown rot in wheat line '2-49'

We have tested the efficacy of putative microsatellite single sequence repeat (SSR) markers, previously identified in a 2-49 (Gluyas Early/Gala) × Janz doubled haploid wheat (Triticum aestivum) population, as being linked to partial seedling resistance to crown rot disease caused by Fusarium pseudograminearum. The quantitative trait loci (QTLs) delineated by these markers have been tested for linkage to resistance in an independent Gluyas Early × Janz doubled haploid population. The presence of a major QTL on chromosome 1DL (QCr.usq-1D1) and a minor QTL on chromosome 2BS (QCr.usq-2B1) was confirmed. However, a putative minor QTL on chromosome 2A was not confirmed. The QTL on 1D was inherited from Gluyas Early, a direct parent of 2-49, whereas the 2B QTL was inherited from Janz. Three other putative QTLs identified in 2-49 × Janz (on 1AL, 4BL, and 7BS) were inherited by 2-49 from Gala and were not able to be confirmed in this study. The screening of SSR markers on a small sample of elite wheat genotypes indicated that not all of the most tightly linked SSR markers flanking the major QTLs on 1D and 1A were polymorphic in all backgrounds, indicating the need for additional flanking markers when backcrossing into some elite pedigrees. Comparison of SSR haplotypes with those of other genotypes exhibiting partial crown rot resistance suggests that additional, novel sources of crown rot resistance are available.

Genotypic variation in seedling root architectural traits and implications for drought adaptation in wheat (Triticum aestivum L.)

Root system characteristics are of fundamental importance to soil exploration and below-ground resource acquisition. Root architectural traits determine the in situ space-filling properties of a root system or root architecture. The growth angle of root axes is a principal component of root system architecture that has been strongly associated with acquisition efficiency in many crop species. The aims of this study were to examine the extent of genotypic variability for the growth angle and number of seminal roots in 27 current Australian and 3 CIMMYT wheat (Triticum aestivum L.) genotypes, and to quantify using fractal analysis the root system architecture of a subset of wheat genotypes contrasting in drought tolerance and seminal root characteristics. The growth angle and number of seminal roots showed significant genotypic variation among the wheat genotypes with values ranging from 36 to 56 (degrees) and 3 to 5 (plant-1), respectively. Cluster analysis of wheat genotypes based on similarity in their seminal root characteristics resulted in four groups. The group composition reflected to some extent the genetic background and environmental adaptation of genotypes. Wheat cultivars grown widely in the Mediterranean environments of southern and western Australia generally had wider growth angle and lower number of seminal axes. In contrast, cultivars with superior performance on deep clay soils in the northern cropping region, such as SeriM82, Baxter, Babax, and Dharwar Dry exhibited a narrower angle of seminal axes. The wheat genotypes also showed significant variation in fractal dimension (D). The D values calculated for the individual segments of each root system suggested that, compared to the standard cultivar Hartog, the drought-tolerant genotypes adapted to the northern region tended to distribute relatively more roots in the soil volume directly underneath the plant. These findings suggest that wheat root system architecture is closely linked to the angle of seminal root axes at the seedling stage. The implications of genotypic variation in the seminal root characteristics and fractal dimension for specific adaptation to drought environment types are discussed with emphasis on the possible exploitation of root architectural traits in breeding for improved wheat cultivars for water-limited environments.

Testing insecticides against Phthorimaea operculella (Zeller) (Lepidoptera: Gelechiidae) using a tomato plant bioassay

A bioassay technique was developed to test the efficacy of insecticides against potato moth (Phthorimaea operculella (Zeller)) on tomatoes. The technique tested efficacy against both larvae in mines and neonate larvae that had not yet penetrated the leaf, and explained the failure of some insecticides to control P. operculella infestations in commercial tomato crops. Neonate larvae placed on leaves of potted plants several days before treatment provided larvae for testing of insecticides against larvae in mines; other neonates were placed on leaves after treatment to test efficacy against larvae yet to penetrate the leaf. The plants were sprayed with the candidate insecticides, held for 5-7 days, and larval mortality assessed. Chlorfenapyr (100, 200 g a.i. ha-1) and abamectin (8.1 g a.i. ha-1) were effective against neonate larvae and larvae in mines. Sulprofos (720 g a.i. ha -1), methomyl (450 g a.i. ha-1) and spinosad (96 g a.i. ha-1) were effective against neonate larvae but not against larvae in mines. Methamidophos (1102 g a.i. ha-1), endosulfan (700 g a.i. ha-1) and Bacillus thuringiensis kurstaki (1000 g ha-1) had some effect against exposed larvae but little against larvae in mines. Thiodicarb (525 g a.i. ha-1), azinphos-ethyl (440 g a.i. ha -1), imidacloprid (59.5 g a.i. ha-1), hexaflumuron (50 g a.i. ha-1), methoxyfenozide (300 g a.i. ha-1) and tebufenozide (200 g a.i. ha-1) were ineffective. A field trial using chlorfenapyr (25, 50, 100, 150 and 200 g a.i. ha-1) and methamidophos (1102 g a.i. ha-1) validated the bioassay technique, with chlorfenapyr effective in reducing the numbers of larvae in mines in leaves.

A detached leaf bioassay to screen Durian cultivars for susceptibility to Phytophthora palmivora

A detached-leaf bioassay was developed and used to screen five durian (Durio zibethinus) cultivars against Phytophthora palmivora isolates from a trunk canker, root and fruit. The fruit isolate was less aggressive than the canker and root isolates. The bioassay using the canker isolate was later used to determine the variation in resistance of D. macarantha and nineteen cultivars of D. zibethinus. The cultivars displayed a range of responses with Parung and Gob being most tolerant, with Gaan Yaow, Chanee and Penang 88 being susceptible. The remaining germplasm fell between Gaan Yaow and Penang 88 in susceptibility. The leaf bioassay was found to be a rapid and reliable method for assessing the susceptibility of durian cultivars.

Lantana (Lantana camara) Seed Bank Dynamics: Seedling Emergence and Seed Survival.

Seed persistence is poorly quantified for invasive plants of subtropical and tropical environments and Lantana camara, one of the world's worst weeds, is no exception. We investigated germination, seedling emergence, and seed survival of two lantana biotypes (Pink and pink-edged red [PER]) in southeastern Queensland, Australia. Controlled experiments were undertaken in 2002 and repeated in 2004, with treatments comprising two differing environmental regimes (irrigated and natural rainfall) and sowing depths (0 and 2 cm). Seed survival and seedling emergence were significantly affected by all factors (time, biotype, environment, sowing depth, and cohort) (P < 0.001). Seed dormancy varied with treatment (environment, sowing depth, biotype, and cohort) (P < 0.001), but declined rapidly after 6 mo. Significant differential responses by the two biotypes to sowing depth and environment were detected for both seed survival and seedling emergence (P < 0.001). Seed mass was consistently lower in the PER biotype at the population level (P < 0.001), but this variation did not adequately explain the differential responses. Moreover, under natural rainfall the magnitude of the biotype effect was unlikely to result in ecologically significant differences. Seed survival after 36 mo under natural rainfall ranged from 6.8 to 21.3%. Best fit regression analysis of the decline in seed survival over time yielded a five-parameter exponential decay model with a lower asymptote approaching −0.38 (% seed survival = [( 55 − (−0.38)) • e (k • t)] + −0.38; R2 = 88.5%; 9 df). Environmental conditions and burial affected the slope parameter or k value significantly (P < 0.01). Seed survival projections from the model were greatest for buried seeds under natural rainfall (11 yr) and least under irrigation (3 yr). Experimental data and model projections suggest that lantana has a persistent seed bank and this should be considered in management programs, particularly those aimed at eradication.

Impact of control strategies on bellyache bush (Jatropha gossypiifolia L.) mortality, seedling recruitment, population dynamics, pasture yield and cost analysis

Bellyache bush (Jatropha gossypiifolia L.) is an invasive weed that has the potential to greatly reduce biodiversity and pasture productivity in northern Australia’s rangelands. This paper reports an approach to develop best practice options for controlling medium to dense infestations of bellyache bush using combinations of control methods. The efficacy of five single treatments including foliar spraying, slashing, stick raking, burning and do nothing (control) were compared against 15 combinations of these treatments over 4 successive years. Treatments were evaluated using several attributes, including plant mortality, changes in population demographics, seedling recruitment, pasture yield and cost of treatment. Foliar spraying once each year for 4 years proved the most cost-effective control strategy, with no bellyache bush plants recorded at the end of the study. Single applications of slashing, stick raking and to a lesser extent burning, when followed up with foliar spraying also led to significantly reduced densities of bellyache bush and changed the population from a growing one to a declining one. Total experimental cost estimates over 4 successive years for treatments where burning, stick raking, foliar spraying, and slashing were followed with foliar spraying were AU$408, AU$584, AU$802 and AU$789 ha–1, respectively. Maximum pasture yield of 5.4 t ha–1 occurred with repeated foliar spraying. This study recommends that treatment combinations using either foliar spraying alone or as a follow up with slashing, stick raking or burning are best practice options following consideration of the level of control, changes in pasture yield and cost effectiveness.

Comparison of bioassay responses to the potential fungal biopesticide Metarhizium anisopliae in Rhipicephalus(Boophilus) microplus and Lucilia cuprina.

Quantal response bioassays were conducted with cattle ticks and sheep blowflies with three different isolates of Metarhizium anisopliae and different methods of inoculation. Ticks were either topically dosed with 2 mu l or immersed in the conidial preparations. Blowflies were either topically dosed with 2 mu l of the conidial preparation or fed on conidia mixed with sugar. Probit analyses were carried out on the mortality data to compare the virulence of these isolates to ticks and blowflies and look for indications of different virulence mechanisms employed by M. anisopliae isolates when invading these hosts. One isolate (ARIM16) showed high virulence to both hosts killing 95% of ticks after 2 days and 88 (+/- 2)% of blowflies after 4 days. Strikingly different mortality patterns indicated that virulence is dependent on different mechanisms in ticks and blowflies. The pattern of mortality seen with ticks suggested that the number of conidia adhering per unit area of the cuticle was more important for rapid tick death than the total number of conidia contacting the entire tick surface. Blowflies fed conidia mixed with food died rapidly after an initial lag phase regardless of dose.

Identification of intersectional Corymbia hybrids based on seedling morphology improves with parental divergence

Differences in morphology have provided a basis for detecting natural interspecific hybridisation in forest trees for decades but have come to prominence again more recently as a means for directly measuring gene flow from planted forests. Here we examined the utility of seedling morphology for hybrid discrimination in three hybrid groups relevant to the monitoring of gene flow from plantings of Corymbia (L.D. Pryor & L.A.S. Johnson ex Brooker) taxa in subtropical Australia. Thirty leaf and stem characters were assessed on 907 8-month old seedlings from four parental and six hybrid taxa grown in a common garden. Outbred F1 hybrids between spotted gums (Corymbia citriodora subspecies variegata, C. citriodora subspecies citriodora and Corymbia henryi) tended to more closely resemble their maternal Corymbia torelliana parent and the most discriminating characters were the ratio of blade length to maximum perpendicular width, the presence or absence of a lignotuber, and specific leaf weight. Assignment of individuals into genealogical classes based on a multivariate model limited to a set of the more discriminating and independent characters was highest in the hybrid group, where parental taxa were genetically most divergent. Overall power to resolve among outbred F1 hybrids from both parental taxa was low to moderate, but this may not be a limitation to its likely major application of identifying hybrids in seedlots from native spotted gum stands. Advanced generation hybrids (outbred F2 and outbred backcrosses) were more difficult to resolve reliably due to the higher variances of hybrid taxa and the tendency of backcrosses to resemble their recurrent parents. Visual assessments of seedling morphology may provide a filter allowing screening of the large numbers needed to monitor gene flow, but will need to be combined with other hybrid detection methods to ensure hybrids are detected.

Pheromone of the banana-spotting bug, Amblypelta lutescens lutescens Distant (Heteroptera: Coreidae): Identification, synthesis, and field bioassay

The banana-spotting bug, Amblypelta lutescens lutescens Distant (Heteroptera: Coreidae), is one of the principal pests of tree fruits and nuts across northern and eastern Australia. Apart from visual damage assessment, there are currently no reliable methods for monitoring bug activity to aid management decisions. An attractant pheromone for this species that could be used as a trap lure could potentially fill this void. Earlier, two male-specific compounds were identified in airborne extracts from A. lutescens lutescens, (E,E)-α-farnesene and (R,E)-nerolidol; an unknown compound with a molecular weight 220 was also detected. We now report the identification of this hitherto unknown compound as (R,E,E)-α-farnesene-10,11-oxide. Synthesis of this epoxide was conducted using a regioselective asymmetric dihydroxylation of a sulfolene. A blend mimicking the natural proportions of (E,E)-α-farnesene, (R,E)-nerolidol, and (R,E,E)-α-farnesene-10,11- oxide attracted male and female A. lutescens lutescens as well as nymphs in the field, verifying that the aggregation pheromone comprises or is contained within this group of compounds. Copyright © 2012 Ashot Khrimian et al.

Markers for seedling and adult plant crown rot resistance in four partially resistant bread wheat sources

Key message: QTLidentified for seedling and adult plant crown rot resistance in four partially resistant hexaploid wheat sources. PCR-based markers identified for use in marker-assisted selection. Abstract: Crown rot, caused by Fusarium pseudograminearum, is an important disease of wheat in many wheat-growing regions globally. Complete resistance to infection by F. pseudograminearum has not been observed in a wheat host, but germplasm with partial resistance to this pathogen has been identified. The partially resistant wheat hexaploid germplasm sources 2-49, Sunco, IRN497 and CPI133817 were investigated in both seedling and adult plant field trials to identify markers associated with the resistance which could be used in marker-assisted selection programs. Thirteen different quantitative trait loci (QTL) conditioning crown rot resistance were identified in the four different sources. Some QTL were only observed in seedling trials whereas others appeared to be adult plant specific. For example while the QTL on chromosomes 1AS, 1BS, and 4BS contributed by 2-49 and on 2BS contributed by Sunco were detected in both seedling and field trials, the QTL on 1DL present in 2-49 and the QTL on 3BL in IRN497 were only detected in seedling trials. Genetic correlations between field trials of the same population were strong, as were correlations between seedling trials of the same population. Low to moderate correlations were observed between seedling and field trials. Flanking markers, most of which are less than 10 cM apart, have now been identified for each of the regions associated with crown rot resistance.

Complete genome sequence and integrated protein localization and interaction map for alfalfa dwarf virus, which combines properties of both cytoplasmic and nuclear plant rhabdoviruses

Summary We have determined the full-length 14,491-nucleotide genome sequence of a new plant rhabdovirus, alfalfa dwarf virus (ADV). Seven open reading frames (ORFs) were identified in the antigenomic orientation of the negative-sense, single-stranded viral RNA, in the order 3′-N-P-P3-M-G-P6-L-5′. The ORFs are separated by conserved intergenic regions and the genome coding region is flanked by complementary 3′ leader and 5′ trailer sequences. Phylogenetic analysis of the nucleoprotein amino acid sequence indicated that this alfalfa-infecting rhabdovirus is related to viruses in the genus Cytorhabdovirus. When transiently expressed as GFP fusions in Nicotiana benthamiana leaves, most ADV proteins accumulated in the cell periphery, but unexpectedly P protein was localized exclusively in the nucleus. ADV P protein was shown to have a homotypic, and heterotypic nuclear interactions with N, P3 and M proteins by bimolecular fluorescence complementation. ADV appears unique in that it combines properties of both cytoplasmic and nuclear plant rhabdoviruses.

Markers for seedling and adult plant crown rot resistance in four partially resistant bread wheat sources

QTL identified for seedling and adult plant crown rot resistance in four partially resistant hexaploid wheat sources. PCR-based markers identified for use in marker-assisted selection. Crown rot, caused by Fusarium pseudograminearum, is an important disease of wheat in many wheat-growing regions globally. Complete resistance to infection by F. pseudograminearum has not been observed in a wheat host, but germplasm with partial resistance to this pathogen has been identified. The partially resistant wheat hexaploid germplasm sources 2-49, Sunco, IRN497 and CPI133817 were investigated in both seedling and adult plant field trials to identify markers associated with the resistance which could be used in marker-assisted selection programs. Thirteen different quantitative trait loci (QTL) conditioning crown rot resistance were identified in the four different sources. Some QTL were only observed in seedling trials whereas others appeared to be adult plant specific. For example while the QTL on chromosomes 1AS, 1BS, and 4BS contributed by 2-49 and on 2BS contributed by Sunco were detected in both seedling and field trials, the QTL on 1DL present in 2-49 and the QTL on 3BL in IRN497 were only detected in seedling trials. Genetic correlations between field trials of the same population were strong, as were correlations between seedling trials of the same population. Low to moderate correlations were observed between seedling and field trials. Flanking markers, most of which are less than 10 cM apart, have now been identified for each of the regions associated with crown rot resistance.

Dynamics of plant populations in Heteropogon contortus(black speargrass) pastures on a granite landscape in southern Queensland. 2. Seed production and soil seed banks of H. contortus

Seed production and soil seed hanks of H. contortus were studied in a subset of treatments within an extensive grazing study conducted in H. contortus pasture in southern Queensland between 1990 and 1996. Seed production of H. contortus in autumn ranged from 260 to 1800 seeds/m2 with much of this variation due to differences in rainfall between years. Seed production was generally higher in the silver-leaved ironbark than in the narrow-leaved ironbark land class and was also influenced by a consistent stocking rate x pasture type interaction. Inflorescence density was the main factor contributing to the variable seed production and was related to the rainfall received during February. The number of seeds per inflorescence was unaffected by seasonal rainfall, landscape position, stocking rate or legume oversowing. Seed viability was related to the rainfall received during March. Soil seed banks in spring varied from 130 to 520 seeds/m2 between 1990 and 1995 with generally more seed present in the silver-leaved ironbark than in the narrow-leaved ironbark land class. There were poor relationships between viable seed production and the size of the soil seed bank, and between the size of the soil seed bank and seedling recruitment. This study indicates that H. contortus has the potential to produce relatively large amounts of seed and showed that the seasonal pattern of rainfall plays a major role in achieving this potential

Dynamics of plant populations in Heteropogon contortus(black speargrass) pastures on a granite landscape in southern Queensland. 1. Dynamics of H. contortus populations

The dynamics of Heteropogon contortus (black speargrass) populations were measured in a subset of treatments contained within an extensive grazing study conducted between 1990 and 1996 in H. contortus pasture in southern Queensland. This subset included 2 landscape positions and 3 stocking rates in both native pasture and legume-oversown native pasture. Severe drought conditions throughout much of the study necessitated ongoing adjustments to the original stocking rates and, as a result, drought was the major influence on the dynamics of H. contortus populations. Plant density and basal area in the silver-leaved ironbark landscape were consistently higher than those in the narrow-leaved ironbark landscape. There was limited evidence of any impact by either light or moderate stocking rate but there was evidence of an impact at the heaviest stocking rate. There was minimal impact of legume oversowing. Relatively large fluctuations in plant density occurred during this study resulting from the death of existing plants, due mainly to drought, and seedling recruitment. Similarly, there were relatively large fluctuations in basal area caused mainly by changes in plant size. Rates for turnover of plant numbers were relatively high whereas plant turnover rates of basal areas were relatively low. Regular seedling recruitment appeared necessary to ensure the persistence of this species. Despite the high turnover, populations were maintained at reasonable levels indicating the overall resilience of H. contortus.