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Abstract Dung beetles are economically important beneficial insects that process dung. To locate this source, they use volatile organic compounds (VOCs). The objectives of the study were to evaluate the attractiveness of ten electrophysiologically-active dung volatiles (phenol, skatole, indole, p-cresol, butanone, butyric acid, eucalyptol, dimethyl sulphide, dimethyl disulphide, and toluene) to dung beetles in the field and to investigate how the composition of volatile blends influences efficacy as lures for use in traps. Six combinations of the compounds were compared with field collected cattle dung bait and a negative control, across three seasons. Both dung and synthetic baits captured all exotic dung beetle species present in the study area. A six-compound mix (M1), comprising major dung volatiles, served as an attractive chemical mixture. The addition of dimethyl sulphide, dimethyl disulphide (M2) and toluene (M4) enhanced attractancy of M1 for dung beetles, while eucalyptol (M3) decreased the attractancy. The degree of attraction by various dung beetle species to synthetic baits varied, but baits proved to be effective, especially for summer trapping. The trap design used in this study presented a convenient and practical way to sample dung beetle and other associated scarabs from open pastures. The attraction of introduced dung beetle species to synthetic baits is documented here for the first time in Australia. In addition, necrophagous Omorgus sp. is reported here for the first time to be attracted to synthetic baits. They showed a significant attraction to the mixture containing dimethyl sulphide and dimethyl disulphide (M2). The current study represents a promising first step towards formulating a synthetic chemical lure for dung beetles, offering a consistent, standardised, and bio-secure trapping method compared to use of naturally occurring dung baits, especially as a multi-species lure.

Ovulation and artificial insemination rates have been observed to decrease in sheep and cows when exposed to dietary phytoestrogens at concentrations greater than 25 mg/kg DM. A grazing trial was undertaken to investigate the effects of coumestrol and other key phytoestrogens on the superovulatory response, embryo numbers and quality in beef cows grazing legume pastures. A 7-week controlled grazing trial was conducted with legume and ryegrass pasture treatments, with cows exposed to legumes at two timed treatments, 4 and 7 weeks. Twenty Angus cows were subjected to a conventional estrus synchronization and superovulation protocol. Embryos were recovered via conventional uterine body flushing 7 days post artificial insemination (AI). Numerous phytoestrogens were identified in both pasture and plasma samples, including coumestrol and formononetin. Concentrations of phytoestrogens in the pasture ranged from 0.001 to 47.5 mg/kg DM and 0 to 2.6 ng/mL in plasma. Approximately 50% of cows produced viable embryos 7 days post AI. A significant interaction between the effect of treatment groups on the embryo stage was observed (p < 0.05). The results suggest that concentrations of >25 mg/kg DM of phytoestrogens less than 20 days preceding AI may negatively affect oocyte developmental competence, reduce progesterone production and thus contribute to early embryonic loss.

Locating sporadically distributed food resources and mate finding are strongly aided by volatile cues for most insects, including dung beetles. However, there is limited information on the olfactory ecology of dung beetles. We conducted a scanning electron microscopy study on the morphology and distribution of the antennal sensilla of three introduced dung beetle species in Australia: Geotrupes spiniger (Coleoptera: Geotrupidae), Bubas bison and Onitis aygulus (Coleoptera: Scarabaeidae). Three main morphological types of antennal sensilla were identified: sensilla trichodea (ST), sensilla basiconica (SB) and sensilla chaetica (SCh). Distinct variations of SB distribution were observed in B. bison and G. spiniger and on different lamellar surfaces in both sexes of all three species. Sexual dimorphism in antennal sensilla distribution or their abundance was not evident. To complement the morphological characterisation of sensilla, electroantennography (EAG) was carried out to construct EAG response profiles of the three species to selected dung volatiles. An initial study revealed that antennae of all species were sensitive to a mix of phenol, skatole, indole, p-cresol, butanone and butyric acid, common components of livestock dung headspace. In addition to these six compounds, dimethyl sulfide, dimethyl disulfide, eucalyptol and toluene were tested for antennal activity. All compounds evoked measurable EAG responses, confirming antennal sensitivity. Geotrupes spiniger exhibited significant responses to all the compounds compared to the control, whereas B. bison and O. aygulus only responded to a subset of compounds. A comparison of relative EAG amplitudes revealed highly significant responses to p-cresol in G. spiniger and to skatole in B. bison. Geotrupes spiniger displayed differential responses to all the compounds. Pooled EAG data suggest highly significant differences in responses among the three species and among compounds. Our findings suggest that a blend of volatiles may offer potential for the trapping of dung beetles, thereby avoiding the use of dung baits that are inconvenient, inconsistent and may pose a threat to farm biosecurity.

Abstract Australia has over 30 Panicum spp. (panic grass) including several non-native species that cause crop and pasture loss and hepatogenous photosensitisation in livestock. It is critical to correctly identify them at the species level to facilitate the development of appropriate management strategies for efficacious control of Panicum grasses in crops, fallows and pastures. Currently, identification of Panicum spp. relies on morphological examination of the reproductive structures, but this approach is only useful for flowering specimens and requires significant taxonomic expertise. To overcome this limitation, we used multi-locus DNA barcoding for the identification of ten selected Panicum spp. found in Australia. With the exception of P. buncei, other native Australian Panicum were genetically separated at the species level and distinguished from non-native species. One nuclear (ITS) and two chloroplast regions (matK and trnL intron-trnF) were identified with varying facility for DNA barcode separation of the Panicum species. Concatenation of sequences from ITS, matK and trnL intron-trnF regions provided clear separation of eight regionally collected species, with a maximum intraspecific distance of 0.22% and minimum interspecific distance of 0.33%. Two of three non-native Panicum species exhibited a smaller genome size compared to native species evaluated, and we speculate that this may be associated with biological advantages impacting invasion of non-native Panicum species in novel locations. We conclude that multi-locus DNA barcoding, in combination with traditional taxonomic identification, provides an accurate and cost-effective adjunctive tool for further distinguishing Panicum spp. at the species level.

Southern Australian farming systems operate predominantly under Mediterranean climatic conditions, which limit the choice of cover crops suitable for enhancement of ground cover and soil moisture retention, erosion control, atmospheric soil nitrogen (N) fixation, and weed suppression between cash crop rotations. Given that the successful establishment of cover crops is climate-driven and also influenced by edaphic factors such as soil pH and salinity, there has been increased interest by southern Australian producers in identifying potential cover crop species well adapted to specific Australian farming systems, which provide vital ecosystem services and sustainable economic benefits through the improvement of soil properties. This review summarises recent findings on cover crop inclusion in diverse farming systems in southern Australia, including continuous and mixed broadacre cropping as well as viticulture and horticulture systems, to identify opportunities and limitations related to their use. Cover crop inclusion in viticulture and pasture systems with lower moisture stress was observed to benefit the subsequent cash crop through enhanced production potential. Long-term, multi-site field experimentation incorporating summer cover crops in winter crop rotations showed that cover crops enhanced ground cover and soil water infiltration in some locations across southern Australia while sometimes increasing winter crop yield, suggesting that soil type and regional climatic conditions greatly influenced the delivery of multiple cover crop benefits. Collectively, these studies have suggested a need for longer-term field evaluations using multiple cover crop species and investigations of termination options under varying environmental and soil conditions to better quantify the legacy effects of cover crops.

Abstract Background Progesterone Receptor Membrane Component 1 (PGRMC1) is expressed in many cancer cells, where it is associated with detrimental patient outcomes. It contains phosphorylated tyrosines which evolutionarily preceded deuterostome gastrulation and tissue differentiation mechanisms. Results We demonstrate that manipulating PGRMC1 phosphorylation status in MIA PaCa-2 (MP) cells imposes broad pleiotropic effects. Relative to parental cells over-expressing hemagglutinin-tagged wild-type (WT) PGRMC1-HA, cells expressing a PGRMC1-HA-S57A/S181A double mutant (DM) exhibited reduced levels of proteins involved in energy metabolism and mitochondrial function, and altered glucose metabolism suggesting modulation of the Warburg effect. This was associated with increased PI3K/AKT activity, altered cell shape, actin cytoskeleton, motility, and mitochondrial properties. An S57A/Y180F/S181A triple mutant (TM) indicated the involvement of Y180 in PI3K/AKT activation. Mutation of Y180F strongly attenuated subcutaneous xenograft tumor growth in NOD-SCID gamma mice. Elsewhere we demonstrate altered metabolism, mutation incidence, and epigenetic status in these cells. Conclusions Altogether, these results indicate that mutational manipulation of PGRMC1 phosphorylation status exerts broad pleiotropic effects relevant to cancer and other cell biology.

Abstract Background Progesterone receptor membrane component 1 (PGRMC1) is often elevated in cancers, and exists in alternative states of phosphorylation. A motif centered on PGRMC1 Y180 was evolutionarily acquired concurrently with the embryological gastrulation organizer that orchestrates vertebrate tissue differentiation. Results Here, we show that mutagenic manipulation of PGRMC1 phosphorylation alters cell metabolism, genomic stability, and CpG methylation. Each of several mutants elicited distinct patterns of genomic CpG methylation. Mutation of S57A/Y180/S181A led to increased net hypermethylation, reminiscent of embryonic stem cells. Pathways enrichment analysis suggested modulation of processes related to animal cell differentiation status and tissue identity, as well as cell cycle control and ATM/ATR DNA damage repair regulation. We detected different genomic mutation rates in culture. Conclusions A companion manuscript shows that these cell states dramatically affect protein abundances, cell and mitochondrial morphology, and glycolytic metabolism. We propose that PGRMC1 phosphorylation status modulates cellular plasticity mechanisms relevant to early embryological tissue differentiation.

The introduction of numerous exotic dung beetles across southern Australia in regions where native dung beetles are not generally efficient in processing livestock dung has resulted in significant reductions in the quantity of such dung on the soil surface in recent years. However, the direct impacts of such ecosystem services on pasture quality and soil nutrient mobility have not yet been investigated in the Riverina region of New South Wales (NSW), an area recognised for prime cattle and sheep production in Australia. Utilising 48 soil columns for lysimetry, we quantified the impact of a common introduced dung beetle (Bubas bison) in this region on water quality after permeation through four different soil types sown to winter annual pastures. Dung beetle treatments included dung plus dung beetles, dung alone and no dung beetles, and no dung and no beetles as a control. Dung beetles and soil type impacted on the performance of improved overseeded annual pastures as measured by biomass accumulation over a four-month growing season. The four soil types, namely, Chromosol, Kandosol, Rudosol, and Vertosol, differed considerably with respect to their water-holding capacity and nutrient profiles, as assessed by initial soil testing and soil leachate evaluation following rainfall plus simulated rainfall events. The concentration of Escherichia coli resulting from cattle dung, cattle dung plus beetles, and the control soils without dung or beetles was assessed in collected leachates over a three-month period. E. coli numbers were significantly increased following B. bison activity, when compared to the dung-only and control treatments. Evaluation of the soil microbiome, by assessing genomic DNA in soils sampled 10 cm below the soil surface where dung beetles remained active following tunnelling, revealed significant differences among soil types with respect to bacterial and fungal communities. Within each soil type, dung beetle activity impacted the fungal community structure, but not the bacterial community. Pasture performance as assessed by biomass accumulation was significantly improved following dung beetle activity in later stages of pasture growth, while E. coli numbers and total coliforms appeared unaffected by beetle presence.

Weeds are a major impediment to crop and pasture production and profitability in Australian farming systems, and knowledge of their occurrence is vital in devising weed management solutions. This study examined the relationships between weed and crop species from observations made during surveys of randomly selected fields in grain and/or pasture production. Between 2013 and 2017, 996 fields across New South Wales (NSW) were surveyed to record the occurrence and density of weed species. Annual ryegrass (Lolium rigidum) (69% of fields), wild oats (Avena spp.) (60%), and sowthistle (Sonchus oleraceus) (34%) were the most common species identified, with a total of 65 weed species recorded to be occurring in NSW cropping fields. Densities of occurring weeds were low, at one plant per square metre or less. Multiple species of wild oats were recorded, with their occurrence varying between survey regions; for example, Avena sterilis subsp. ludoviciana was more prevalent in the north and Avena fatua in the south. Differences in weed species occurrence were noted between this survey and those previously conducted in NSW, with marked increases in annual ryegrass and sowthistle, and this is attributed to the change in crop production practices between survey periods. The study of variable distribution patterns of weed species across cropping systems allows for informed decision-making regarding weed management research and development planning.

Legume crops and pastures have a high economic value in Australia. However, legume species commonly used for grazing enterprises have been identified to produce high concentrations of phytoestrogens. These compounds are heterocyclic phenolic, and are similar in structure to the mammalian estrogen, 17β-estradiol. The biological activity of the various phytoestrogen types; isoflavones, lignans and coumestans, are species-specific, although at concentrations of 25 mg/kg of dry matter each of the phytoestrogen types affect reproductive functions in grazing livestock. The impacts upon fertility in grazing livestock such as cattle and sheep, vary greatly over length of exposure time, age and health of animal and the stress stimuli the plant is exposed to. More recently, research into the other effects that phytoestrogens may have upon metabolism, immune capacity and growth and performance of grazing livestock has been conducted. Potential new benefits for using these phytoestrogens, such as daidzein and genistein, have been identified by observing the stimulation of production in lymphocytes and other antibody cells. Numerous isoflavones have also been recognized to promote protein synthesis, increase the lean meat ratio, and increase weight gain in cattle and sheep. In Australia, the high economic value of legumes as pasture crops in sheep and cattle production enterprises requires proactive management strategies to mitigate risk associated with potential loss of fertility associated with inclusion of pasture legumes as forages for grazing livestock.

In this technologically advanced era, with the proliferation of artificial intelligence, many mobile apps are available for plant disease detection, diagnosis, and treatment, each with a variety of features. These apps need to be categorized and reviewed following a proper framework that ensures their quality. This study aims to present an approach to evaluating plant disease detection mobile apps, which includes providing ratings of distinct features of the apps and insights into the exploitation of artificial intelligence used in plant disease detection. The applicability of these apps for pathogen or disease detection, identification, and treatment will be assessed along with significant insights garnered. For this purpose, plant disease detection apps were searched in three prominent app stores (the Google Play store, Apple App store, and Microsoft store) using a set of keywords. A total of 606 apps were found and from them, 17 relevant apps were identified based on inclusion and exclusion criteria. The selected apps were reviewed by three raters using our devised app rating scale. To validate the rater agreements on the ratings, inter-rater reliability is computed alongside their intra-rater reliability, ensuring their rating consistency. Also, the internal consistency of our rating scale was evaluated against all selected apps. User comments from the app stores are collected and analyzed to understand their expectations and views. Following the rating procedure, most apps earned acceptable ratings in software quality characteristics such as aesthetics, usability, and performance but gained poor ratings in AI-based advanced functionality, which is the key aspect of this study. However, most of the apps cannot be used as a complete solution to plant disease detection, diagnosis, and treatment. Only one app, Plantix–your crop doctor, could successfully identify plants from images, detect diseases, maintain a rich plant database, and suggest potential treatments for the disease presented. It also provides a community where plant lovers can communicate with each other to gain additional benefits. In general, all existing apps need to improve functionalities, user experience, and software quality. Therefore, a set of design considerations has been proposed for future app improvements.

Volatile cues can play a significant role in the location and discrimination of food resources by insects. Dung beetles have been reported to discriminate among dung types produced by different species, thereby exhibiting behavioral preferences. However, the role of volatile organic compounds (VOCs) in dung localization and preference remains largely unexplored in dung beetles. Here we performed several studies: firstly, cage olfactometer bioassays were performed to evaluate the behavioral responses of Bubas bison (Coleoptera: Scarabaeidae) to VOCs emanating from fresh horse, sheep, and cattle dung; secondly, concurrent volatilome analysis was performed to characterize volatilomes of these dung types. Bubas bison adults exhibited greater attraction to horse dung and less attraction to cattle dung, and they preferred dung from horses fed a pasture-based diet over dung from those fed lucerne hay. Volatilomes of the corresponding dung samples from each livestock species contained a diverse group of alkanes, alkenes, alkynes, alcohols, aldehydes, ketones, esters, phenols, and sulfurous compounds, but the composition and abundance of annotated VOCs varied with dung type and livestock diet. The volatilome of horse dung was the most chemically diverse. Results from a third study evaluating electroantennogram response and supplementary olfactometry provided strong evidence that indole, butyric acid, butanone, p-cresol, skatole, and phenol, as well as toluene, are involved in the attraction of B. bison to dung, with a mixture of these components significantly more attractive than individual constituents.

Production of medicinal tubers of Rehmannia glutinosa is severely hindered by replanting issues. However, a mechanistic understanding of the plant-soil factors associated with replant problems is currently limited. Thus, we aimed to identify the R. glutinosa root exudates, evaluate their potential phytotoxicity and profile the interactions between the plant and its associated rhizobiome. Stereomicroscopy and liquid chromatography coupled to a quadrupole/time of flight mass spectrometer were used to monitor and identify secreted metabolites, respectively. Seedling bioassays were used to evaluate the phytotoxicity of R. glutinosa root exudates. Two complimentary experiments were performed to investigate allelochemical fate in rhizosphere soil and profile the associated microbiota. Root specific microbes were further isolated from R. glutinosa rhizosphere. Impacts of isolated strains were evaluated by co-cultivation on plate and on seedlings in tissue culture, with a focus on their pathogenicity. Interactions between key R. glutinosa root exudates and isolated rhizobiomes were investigated to understand the potential for plant-soil feedbacks. Quantification and phytotoxic analysis of metabolites released from R. glutinosa indicated catalpol was the most abundant and bioactive metabolite in root exudates. Subsequent microbial profiling in soil containing accumulated and ecologically significant levels of catalpol identified several taxa (e.g., Agromyces, Lysobacter, Pseudomonas, Fusarium) that were specifically shifted. Isolation of R. glutinosa rhizobiomes obtained several root specific strains. A significant antagonistic effect between strain Rh7 (Pseudomonas aeruginosa) and two pathogenic strains Rf1 (Fusarium oxysporum) and Rf2 (Fusarium solani) was observed. Notably, the growth of strain Rh7 and catalpol concentration showed a hormesis-like effect. Field investigation further indicated catalpol was increasingly accumulated in the rhizosphere of replanted R. glutinosa, suggesting that interactions of biocontrol agents and pathogens are likely regulated by the presence of bioactive root exudates and in turn impact the rhizo-ecological process. In summary, this research successfully monitored the release of R. glutinosa root exudates, identified several abundant bioactive R. glutinosa secreted metabolites, profiled associated root specific microbes, and investigated the plant-soil feedbacks potentially regulated by catalpol and associated rhizobiomes. Our findings provide new perspectives toward an enhanced understanding R. glutinosa replant problems.

The use of competitive crops in successional rotations has been shown to reduce the growth and establishment of annual weeds by the depletion of the weed seedbank in broadacre cropping systems. However, the impact of specific crop rotational sequences contributing to weed seedbank density has not been quantified in the Riverina region of southern Australia. Trials were established in two locations in 2014–2018 to quantify the impact of selected annual rotations featuring grain, pulse, and pasture crops on weed infestation and seedbank dynamics with a focus on winter and summer annual weeds. The weed seedbank dynamics were evaluated by a twice-annual soil sampling regime (at planting and harvest), followed by soil sample screening for weed propagule germination and seedling establishment in a subsequent controlled-environment screening performed from 2014–2020. The weed seedling density decreased in the cereal rotations in years experiencing average to above-average rainfall, as crops established a dense canopy leading to reduced weed establishment and fecundity. Several rotational treatments were effective in suppressing the weed propagule numbers over time, including those using dual-purpose cereals only or a canola break-crop along with the cereals. Rotational selection can be an important and cost-effective tool in integrated weed management systems when applied over multiple growing seasons.

Weed competitiveness in wheat (Triticum aestivum L.) has previously been shown to be positively associated with shoot biomass. This study evaluated the impact of increased early shoot vigour on the weed competitiveness of Australian wheats. Breeding lines generated for early shoot vigour were top-crossed with two commercial wheat cultivars (Yitpi and Wyalkatchem) and the resulting high vigour lines (HV lines) were assessed for early growth and weed pressure in the field. These lines were directly compared with their parental lines, other commercial cultivars, and the tall heritage cultivar, Federation. Moreover, rye (Secale cereale L.) or triticale (× Triticosecale) was included in each trial as a positive control for vigour. The association between shoot growth and vigour and weed suppression was evaluated over 3 years in the cereal belt of south-eastern Australia during contrasting seasons. The HV lines consistently displayed greater leaf area, ground cover, and canopy light interception in both dry and wet seasons and suppressed weed growth significantly in contrast to commercial cultivars. Light interception at the first tiller stage, and ground cover at the end of tillering were identified as the most important variables for predicting weed suppression. This study demonstrated the enhancement of competitiveness in commercial wheat through the selection for early vigour, and identified traits that best predicted weed suppression.

The potential of multi-purpose barley (Hordeum vulgare L.) cultivars to suppress weeds while maintaining optimal yield and grain quality has been reported but not recently evaluated in replicated field trials performed under southern Australian field conditions. Therefore, to investigate this potential, aboveground competitive traits were assessed in nine genetically diverse commercial barley cultivars in 2015, 2016 and 2017, in two locations in the Riverina region of NSW in replicated field trials performed in the absence of pre-emergent herbicide treatment. Crop and weed establishment, early vigour, leaf area index, photosynthetically active radiation (PAR) and biomass were assessed at various crop growth stages, including early growth, vegetative, flowering, grain fill and harvest. Cultivar differences in crop and weed biomass accumulation at ~50, 100 and 150 days after planting were noted at both locations. Early barley biomass accumulation was inversely related to weed biomass in both locations and most years, suggesting strong (over 90%) potential for heritable competitive barley interference against weeds. The current study also observed a positive relationship between PAR light interception and crop biomass in all three years at both locations, suggesting that PAR light interception contributed positively to crop biomass accumulation by directly increasing photosynthesis (50–70%) and growth or indirectly influencing weed biomass accumulation (10–15%) and weed interference (50–75). Partial least squares modelling was performed with 2015 and 2016 datasets to assess the interactions between crop developmental traits and weed suppression. Cultivars exhibiting enhanced early vigour and PAR light interception were generally more weed suppressive under optimal higher soil moisture conditions. Our results indicate that the choice of barley cultivar has a significant impact on weed establishment, fecundity and seedbank dynamics. The use of competitive barley genotypes is, thus, a cost-effective strategy to reduce weed seedbank numbers over time and may reduce potential herbicide use.

Abstract Background Primary photosensitisation (PS) subsequent to ingestion of the pasture legume Biserrula pelecinus L. (biserrula) has recently been confirmed in grazing livestock. Given the potential utility of this pasture species in challenging climates, a grazing trial was undertaken to examine if both varieties ‘Casbah’ and ‘Mauro’ were able to cause photosensitisation in livestock, and if this could be mitigated by grazing in winter, or in combination with other common pasture species. Results A controlled grazing trial was undertaken in winter in Australia with plots containing a dominant pasture of Biserrula pelecinus L. cv. ‘Casbah’ or ‘Mauro’, or mixed biserrula/perennial ryegrass populations. A photosensitisation grading system was established. 167 prime meat ewe lambs were introduced to the plots and monitored twice daily. Mild clinical signs were observed at 72 h on pasture. All animals were removed from biserrula dominant stands at this point. Four animals grazing ‘Casbah’ dominant pasture rapidly proceeded to severe photosensitisation in the following 12 h. Animals remaining on mixed biserrula/ryegrass stands did not exhibit severe PS but showed an 89% incidence of mild to moderate photosensitisation over the following 14 days. Animals on mixed lucerne showed significantly lower PS score than animals grazing biserrula varieties of any composition. The trial was halted at 14 days as only plots with low biserrula proportion still contained unaffected animals. Necropsy revealed severe multifocal erythematous ulcerations and alopecia of the ear pinnae, severe bilateral periorbital and conjunctival oedema and variably severe subcutaneous facial oedema. No evidence of hepatopathy was present. A diagnosis of acute unseasonal primary photosensitisation caused by biserrula ingestion with no other underlying pathology was confirmed. Conclusions We report an unseasonal outbreak of acute photosensitisation in sheep grazing Biserrula pelecinus L cvs.’Casbah’ and ‘Mauro’ with exceedingly high morbidity. A grading system is also proposed as a tool for objective and consistent clinical appraisal of future PS outbreaks. This finding expands our definition of seasonal and temporal risk periods for biserrula photosensitisation, and is the first to identify that both commercial cultivars of biserrula can cause primary photosensitisation in sheep.

Weeds are an increasingly significant issue inhibiting agricultural production worldwide. Forage conservation could form part of an integrated weed management program if ensiling killed weed seeds. In Experiment 1, seeds of five grass (Hordeum spp., Bromus diandrus, Bromus hordeaceum, Lolium rigidum and Vulpia spp.) and two broad-leaved temperate weed species (Echium spp. and Raphanus raphanistrum), that were either untreated, ensiled in pasture (Trifolium subterranean/Lolium rigidum mixture) forage for a minimum of three months, underwent 48 h in sacco digestion in steers or ensiled prior to digestion were tested for germinability and viability. In Experiment 2, seeds of eight tropical weed species (Cenchrus ciliaris, Rumex spp., Bidens pilosa, Sorghum halepense, Urochloa panicaoides, Paspalum dilatatum, Brachiara eruciformis and Choris truncata) were ensiled in Sorghum bicolor forage. In Experiment 3, L. rigidum and R. raphanistrum seeds were ensiled in either Medicago sativa forage wilted to 336.9, 506.5 or 610.7 g/kg dry matter; or in chaff to which water or water plus acid was added at rates to achieve 350, 450 or 550 g/kg dry matter content with lactic plus acetic acid added in the ratio of 3:2 at 80, 45 or 10 g/kg DM, respectively. In Experiment 4, L. rigidum and R. raphanistrum seeds were ensiled in cotton wool to which water or water plus acid was added at the same rates as in Experiment 3. Germinability of all seeds following ensiling was substantially reduced or nil. The extent of the reduction varied with species and experiment. In sacco digestion reduced germinability in Experiment 1, but to a lesser extent than ensiling; while ensiling plus digestion reduced germination rates to 0%.

Phytoestrogens are plant-produced bioactive secondary metabolites known to play an integral role in plant defense that frequently accumulate in times of stress and/or microbial infection. Phytoestrogens typically belong to two distinct chemical classes; flavonoids (isoflavones) and non-flavonoids (lignans and coumestans). Upon consumption by livestock, high concentrations of phytoestrogens can cause long-term disruption in reproduction due to structural similarities with mammalian estrogens and their tendency to bind estrogen receptors. Wide variation in phytoestrogen concentration has been reported in pasture legumes and corresponding silage or hay. Lucerne is a common perennial pasture legume in temperate climates, but information on phytoestrogen production or accumulation in grazing livestock is currently limited. Therefore, metabolic profiling using UHPLC-MS-QToF was performed to identify and quantitate key phytoestrogens in both fresh and dried lucerne fodder from replicated field or controlled glasshouse environments. Phytoestrogens were also profiled in the blood plasma of Angus cattle grazing field-grown lucerne. Results revealed that phytoestrogens varied quantitatively and qualitatively among selected lucerne cultivars grown under glasshouse conditions. Fresh lucerne samples contained higher concentrations of coumestans and other phytoestrogenic isoflavones than did dried samples for all cultivars profiled, with several exceeding desirable threshold levels for grazing cattle. Coumestans and isoflavones profiled in plasma of Angus heifers grazing lucerne increased significantly over a 21-day sampling period following experimental initiation. Currently, threshold concentrations for phytoestrogens in plasma are unreported. However, total phytoestrogen concentration exceeded 300 mg·kg−1 in fresh and 180 mg·kg−1 in dried samples of selected cultivars, suggesting that certain genotypes may upregulate phytoestrogen production, while others may prove suitable sources of fodder for grazing livestock.

Barley grass (Hordeum spp.) is a short-lived annual weed which competes with preferred crop and pasture species and frequently contaminates wool and carcasses, and irritates the ears, eyes and noses of sheep. Barley grass, annual ryegrass (Lolium rigidum L.) and wild radish (Raphanus raphanistrum L.) are annual winter crop weeds which reduce grain yield through competition. In three consecutive years (2015 to 2017) cereal, legume and cereal/legume forage crops were grown and harvested in early October, late October or early November consistent with an early silage harvest (ES), late silage harvest (LS) or hay cut (H). A spring wheat cultivar was sown over each site in the following year (2016 to 2018). Weed density was recorded during the forage crop and wheat phases. Forage crop weed populations varied between years. Late paddock preparation for sowing in 2015 effectively eradicated barley grass from all forage crops; however, the competitiveness of legume and cereal/legume crops against annual ryegrass was reduced. In contrast, legume and cereal/legume mixtures tended to have higher barley grass densities than cereal crops in 2016 and 2017, when paddock preparation was earlier. Cutting in October 2015 reduced annual ryegrass and wild radish populations in 2016 wheat by 92.0% and 86.7%, respectively. In 2017 and 2018, regrowth and subsequent seed set following cutting appeared to negate cutting time effects for all crop and crop/legume combinations. Late paddock preparation, an early October forage harvest and effective regrowth control provided the best opportunity for barley grass, annual ryegrass and wild radish control in a single year.

Herbicide resistance in weeds restricts control options, thereby escalating economic loss and threatening agricultural sustainability in cereal production. Field evaluation of the crop performance, competitive traits, and consequent weed suppressive potential of 13 commercial winter wheat (Triticum aestivum L.) cultivars was performed in central NSW Australia with a focus on the evaluation and modelling of above-ground interactions. In 2015 and 2016, replicated field trials were established with genetically diverse commercial wheat genotypes under moderate to low rainfall conditions in Wagga Wagga (572 mm) and Condobolin (437 mm) New South Wales, respectively. The heritage cultivar Federation and a commercial cultivar of winter cereal rye (Secale cereale L.) were included as known weed-suppressive controls. Crop and weed growth, as well as early vigour, leaf area index, and photosynthetically active radiation, were monitored at various crop phenological stages including early growth, vegetative, flowering, grain fill, and harvest. Significant differences between wheat cultivar and location were observed for crop biomass, early vigour, leaf area index, weed number, weed biomass, canopy architecture, and yield in both 2015 and 2016. Differences in weed establishment were largely impacted first by rainfall and season and secondly by crop architecture (i.e., height, size, canopy) and phenology (i.e., growth stages). Early vigour and early canopy closure were instrumental in suppressing weed establishment and growth. Cultivar performance and competition with weeds were also clearly influenced by both environmental factors and genotype, as evidenced by differences in early cultivar performance, yield, and weed suppression by season and location. Specifically, Federation, Condo, and Janz wheat cultivars were superior performers in terms of weed suppression in both locations and years; however, Federation produced up to 55% lower yield than recently introduced cultivars. Partial least squares (PLS) regression was performed to develop a predictive linear model for weed competition in commercial wheat cultivars based on weed dry biomass as the response variable and selected aboveground crop canopy traits as predictors. In 2015, the model differed in accordance with crop growth stage, but the impact of predictors on weed biomass at both locations was not significant. In 2016, under local above average rainfall conditions, the model showed a significant negative correlation (p < 0.001) of most predictors on weed biomass (r2 = 0.51 at Condobolin, r2 = 0.62 at Wagga Wagga), suggesting the most influential factors in reducing weed numbers and establishment as crop vigour, biomass, and height. Our results indicate the establishment of competitive wheat cultivars in the absence of post-emergent herbicides resulted in a two to five-fold increased weed suppression over less suppressive genotypes, without significant yield penalties. Therefore, cultivar choice constitutes a cost-effective and sustainable weed management tool, particularly when weed pressure is significant.

Parthenium weed (Parthenium hysterophorus L.) is an invasive plant species in around 50 countries and a ‘Weed of National Significance’ in Australia. This study investigated the relative toxicity of the leaf, shoot and root extracts of two geographically separate and morphologically distinct biotypes of parthenium weed in Queensland, Australia. Parthenium weed exhibited higher phytotoxic, cytotoxic and photocytotoxic activity in leaf tissue extracts in contrast to shoot and root. The germination and seedling growth of a dicot species (garden cress) were inhibited more than those of a monocot species (annual ryegrass) using a phytotoxicity bioassay. The cytotoxicity of leaf extracts was assessed in a mouse fibroblast cell suspension assay and increased under high ultraviolet A(UV-A) radiation. A major secondary metabolite, parthenin, was found in abundance in leaf extracts and was positively correlated with cytotoxicity but not with photocytotoxicity or phytotoxicity. Ambrosin and chlorogenic acid were also detected and were positively correlated with germination inhibition and the inhibition of radicle elongation, respectively. In addition, other currently unidentified compounds in the leaf extracts were positively correlated with phytotoxicity, cytotoxicity and photocytotoxicity with two to three molecules strongly correlated in each case. Both parthenium weed biotypes investigated did not differ with respect to their relative toxicity, despite their reported differences in invasive potential in the field. This suggests that secondary chemistry plays a limited role in their invasion success.

Annual legumes from the Mediterranean region are receiving attention in Australia as alternatives to traditional pasture species. The current study employed novel metabolic profiling approaches to quantify key secondary metabolites including phytoestrogens to better understand their biosynthetic regulation in a range of field-grown annual pasture legumes. In addition, total polyphenol and proanthocyanidins were quantified using Folin–Ciocalteu and vanillin assays, respectively. Metabolic profiling coupled with biochemical assay results demonstrated marked differences in the abundance of coumestans, flavonoids, polyphenols, and proanthocyanidins in annual pasture legume species. Genetically related pasture legumes segregated similarly from a chemotaxonomic perspective. A strong and positive association was observed between the concentration of phytoestrogens and upregulation of the flavonoid biosynthetic pathway in annual pasture legumes. Our findings suggest that evolutionary differences in metabolic dynamics and biosynthetic regulation of secondary metabolites have logically occurred over time in various species of annual pasture legumes resulting in enhanced plant defense.

Barley grass (Hordeum murinum subsp. glaucum.) is an annual weed associated with grain revenue loss and sheep carcass damage in southern Australia. Increasing herbicide resistance led to a recent investigation into effective integrated weed management strategies for barley grass in southern Australia. Field studies in Wagga Wagga, New South Wales (NSW) during 2016 and 2017 examined the effect of post-emergent herbicide applications and strategic defoliation by mowing on barley grass survival and seed production in a mixed legume pasture. Statistically significant differences between herbicide-only treatments in both years showed propaquizafop to be more than 98% effective in reducing barley grass survival and seed production. Paraquat was not effective in controlling barley grass (58% efficacy), but led to a 36% and 63.5% decrease in clover and other weed biomass, respectively, after 12 months and increased lucerne biomass by over three-fold after 24 months. A single repeated mowing treatment resulted in a 46% decline in barley grass seedling emergence after 12 months and, when integrated with herbicide applications, reduced other weed biomass after 24 months by 95%. Resistance to acetyl-CoA carboxylase (ACCase)-inhibiting herbicides observed in local barley grass populations led to additional and more focused investigation comparing the efficacy of other pre- and post-emergent herbicides for barley grass management in legume pastures. Haloxyfop-R + simazine or paraquat, applied at early tillering stage, were most efficacious in reducing barley grass survival and fecundity. Impact of defoliation timing and frequency on barley grass seedlings was also evaluated at various population densities, highlighting the efficacy of repeated post-inflorescence defoliations in reducing plant survival and seed production. Results highlight the importance of optimal environmental conditions and application timing in achieving efficacious control of barley grass and improving pasture growth and biomass accumulation.

Metabolic profiling allows for simultaneous and rapid annotation of biochemically similar organismal metabolites. An effective platform for profiling of toxic pyrrolizidine alkaloids (PAs) and their N-oxides (PANOs) was developed using ultra high pressure liquid chromatography quadrupole time-of-flight (UHPLC-QTOF) mass spectrometry. Field-collected populations of invasive Australian weeds, Echium plantagineum and E. vulgare were raised under controlled glasshouse conditions and surveyed for the presence of related PAs and PANOs in leaf tissues at various growth stages. Echium plantagineum possessed numerous related and abundant PANOs (>17) by seven days following seed germination, and these were also observed in rosette and flowering growth stages. In contrast, the less invasive E. vulgare accumulated significantly lower levels of most PANOs under identical glasshouse conditions. Several previously unreported PAs were also found at trace levels. Field-grown populations of both species were also evaluated for PA production and highly toxic echimidine N-oxide was amongst the most abundant PANOs in foliage of both species. PAs in field and glasshouse plants were more abundant in the more widely invasive species, E. plantagineum, and may provide competitive advantage by increasing the plant’s capacity to deter natural enemies in its invaded range through production of novel weapons.

Photosensitivity in animals is defined as a severe dermatitis that results from a heightened reactivity of skin cells and associated dermal tissues upon their exposure to sunlight, following ingestion or contact with UV reactive secondary plant products. Photosensitivity occurs in animal cells as a reaction that is mediated by a light absorbing molecule, specifically in this case a plant-produced metabolite that is heterocyclic or polyphenolic. In sensitive animals, this reaction is most severe in non-pigmented skin which has the least protection from UV or visible light exposure. Photosensitization in a biological system such as the epidermis is an oxidative or other chemical change in a molecule in response to light-induced excitation of endogenous or exogenously-delivered molecules within the tissue. Photo-oxidation can also occur in the plant itself, resulting in the generation of reactive oxygen species, free radical damage and eventual DNA degradation. Similar cellular changes occur in affected herbivores and are associated with an accumulation of photodynamic molecules in the affected dermal tissues or circulatory system of the herbivore. Recent advances in our ability to identify and detect secondary products at trace levels in the plant and surrounding environment, or in organisms that ingest plants, have provided additional evidence for the role of secondary metabolites in photosensitization of grazing herbivores. This review outlines the role of unique secondary products produced by higher plants in the animal photosensitization process, describes their chemistry and localization in the plant as well as impacts of the environment upon their production, discusses their direct and indirect effects on associated animal systems and presents several examples of well-characterized plant photosensitization in animal systems.

Sweet potato (Ipomoea batatas (L.) Lam.) is a logical candidate crop to suppress invasive plants, but additional information is needed to support its potential application as a suppressive ground cover. The current study utilized a de Wit replacement series incorporating five ratios of sweet potato grown in the field in combination with one of three invasive plants (Ageratum conyzoides L., Bidens pilosa L., and Galinsoga parviflora Cav.) in replicated 9 m2 plots. Stem length, total biomass, and leaf area were higher for monoculture-grown sweet potato than these parameters for any of the invasive plants grown in monoculture. In mixed culture, the plant height, branch, leaf, inflorescence, seed, and biomass of all invasive plants were suppressed by sweet potato. The relative yield parameter indicated that intraspecific competition was greater than interspecific competition for sweet potato, while the reverse was true for invasive species. The net photosynthetic rate was higher for sweet potato than for B. pilosa and G. parviflora but not A. conyzoides. Superoxide dismutase and peroxidase activities of each of the three invasive plants were reduced in mixture with sweet potato. Our results demonstrated that these three invasive plants were significantly suppressed by sweet potato competition due to the rapid growth and phenotypic plasticity of sweet potato.

Small molecule discovery has benefitted from the development of technologies that have aided in the culture and identification of soil microorganisms and the subsequent analysis of their respective metabolomes. We report herein on the use of both culture dependent and independent approaches for evaluation of soil microbial diversity in the rhizosphere of canola, a crop known to support a diverse microbiome, including plant growth promoting rhizobacteria. Initial screening of rhizosphere soils showed that microbial diversity, particularly bacterial, was greatest at crop maturity; therefore organismal recovery was attempted with soil collected at canola harvest. Two standard media (Mueller Hinton and gellan gum) were evaluated following inoculation with soil aqueous suspensions and compared with a novel “rhizochip” prototype buried in a living canola crop rhizosphere for microbial culture in situ. Following successful recovery and identification of 375 rhizosphere microbiota of interest from all culture methods, isolates were identified by Sanger sequencing and/or characterization using morphological and biochemical traits. Three bacterial isolates of interest were randomly selected as case studies for intensive metabolic profiling. After successful culture in liquid media and solvent extraction, individual extracts were subjected to evaluation by UHPLC-DAD-QToF-MS, resulting in the rapid characterization of metabolites of interest from cultures of two isolates. After evaluation of key molecular features, unique or unusual bacterial metabolites were annotated and are reported herein.

In contrast to plant-animal interactions, the conceptual framework regarding the impact of secondary metabolites in mediating plant-plant interference is currently less well defined. Here, we address hypotheses about the role of chemically-mediated plant-plant interference (i.e. allelopathy) as a driver of Mediterranean forest dynamics. Growth and defense abilities of a pioneer (Pinus halepensis) and a late-successional (Quercus pubescens) Mediterranean forest species were evaluated under three different plant interference conditions: i) allelopathy simulated by application of aqueous needle extracts of Pinus, ii) resource competition created by the physical presence of a neighboring species (Pinus or Quercus), and iii) a combination of both allelopathy and competition. After 24 months of experimentation in simulated field conditions, Quercus was more affected by plant interference treatments than was Pinus, and a hierarchical response to biotic interference (allelopathy < competition < allelopathy + competition) was observed in terms of relative impact on growth and plant defense. Both species modulated their respective metabolic profiles according to plant interference treatment and thus their inherent chemical defense status, resulting in a physiological trade-off between plant growth and production of defense metabolites. For Quercus, an increase in secondary metabolite production and a decrease in plant growth were observed in all treatments. In contrast, this trade-off in Pinus was only observed in competition and allelopathy + competition treatments. Although Pinus and Quercus expressed differential responses when subjected to a single interference condition, either allelopathy or competition, species responses were similar or positively correlated when strong interference conditions (allelopathy + competition) were imposed.

Metabolic profiling can be successfully implemented to analyse a living system’s response to environmental conditions by providing critical information on an organism’s physiological state at a particular point in time and allowing for both quantitative and qualitative assessment of a specific subset(s) of key metabolites. Shikonins are highly reactive chemicals that affect various cell signalling pathways and possess antifungal, antibacterial and allelopathic activity. Based on previous bioassay results, bioactive shikonins, are likely to play important roles in the regulation of rhizosphere interactions with neighbouring plants, microbes and herbivores. An effective platform allowing for rapid identification and accurate profiling of numerous structurally similar, difficult-to-separate bioactive isohexenylnaphthazarins (shikonins) was developed using UHPLC Q-TOF MS. Root periderm tissues of the invasive Australian weeds Echium plantagineum and its congener E. vulgare were extracted overnight in ethanol for shikonin profiling. Shikonin production was evaluated at seedling, rosette and flowering stages. Five populations of each species were compared for qualitative and quantitative differences in shikonin formation. Each species showed little populational variation in qualitative shikonin production; however, shikonin was considerably low in one population of E. plantagineum from Western New South Wales. Seedlings of all populations produced the bioactive metabolite acetylshikonin and production was upregulated over time. Mature plants of both species produced significantly higher total levels of shikonins and isovalerylshikonin > dimethylacrylshikonin > shikonin > acetylshikonin in mature E. plantagineum. Although qualitative metabolic profiles in both Echium spp. were nearly identical, shikonin abundance in mature plant periderm was approximately 2.5 times higher in perennial E. vulgare extracts in comparison to those of the annual E. plantagineum. These findings contribute to our understanding of the biosynthesis of shikonins in roots of two related invasive plants and their expression in relation to plant phenological stage.

Post-rice irrigated soils offer several potential advantages for the growth of subsequent crops, but Australian producers have often been reluctant to grow grain legumes immediately following a rice crop due to physico-chemical constraints. A field experiment was thus conducted to explore the potential for producing grain legumes following rice in comparison to those following a fallow during 2012 and 2013. Two grain legumes, field pea and faba bean, were sown 5, 7 and 12 weeks after rice harvest in 2013 at Yanco, NSW, and plant growth indicators and grain yield were compared. Early sowing of field pea following rice gave the best outcome, with plants flowering three weeks earlier and yielding 1330 kg·ha−1 more grain than after fallow. In contrast, faba bean yield was 35 kg·ha−1 less after rice than after fallow across the three sowing dates. Higher pea yield was consistent with the early emergence of seedlings, higher light interception and overall greater plant growth following rice. Post-rice crops also had 10-fold less weed infestation than crops in a similarly-established fallow treatment and, thus, required far less weed management. Legume crops sown at the later seeding date had significantly reduced (~50%–60%) yields compared to those of the first two sowings; this is most likely a reflection of reduced temperatures and day lengths experienced during vegetative and reproductive growth phases.