Analysing the effects of distance, taxon and biomass on vertebrate detections using bulk-collected carrion fly iDNA.

Invertebrate-derived DNA (iDNA) metabarcoding from carrion flies is a powerful, non-invasive tool that has value for assessing vertebrate diversity. However, unknowns exist around the factors that influence vertebrate detections, such as spatial limits to iDNA signals or if detections are influenced by taxonomic class or estimated biomass of the vertebrates of interest. Using a bulk-collection method, we captured flies from within a zoo and along transects extending 4 km away from this location. From 920 flies, we detected 28 vertebrate species. Of the 28 detected species, we identified 9 species kept at the zoo, 8 mammals and 1 bird, but no reptiles. iDNA detections were highly geographically localized, and only a few zoo animals were detected outside the zoo setting. However, due to the low number of detections in our dataset, we found no influence of the taxonomic group or the estimated biomass of animals on their detectability. Our data suggest that iDNA detections from bulk-collected carrion flies, at least in urban settings in Australia, are predominantly determined by geographic proximity to the sampling location. This study presents an important step in understanding how iDNA techniques can be used in biodiversity monitoring.

Growers' perceptions and attitudes towards fungicide resistance extension services.

Agricultural extension services have been successful in promoting knowledge sharing and innovation in agriculture. However, the adoption of new agricultural practices, particularly in integrated pest management, has been slow. Using a case study with a co-designed survey instrument, this research aimed to understand how growers in the Southwest Western Australian Grainbelt access information and resources to manage fungicide resistance. We found that the growers rely on a combination of paid agronomists, government or research institutes, local grower groups, and field days for fungicide resistance information. Growers seek information from trusted experts who can simplify complex research, value easy-to-understand communication and prefer resources tailored to their local context. Additionally, growers valued information regarding new fungicide developments and having access to rapid fungicide resistance diagnostic services. These findings highlight the importance of providing growers with effective agricultural extension services to manage the risk of fungicide resistance.

Spatial Dependency in Stubble-Borne Pyrenophora teres f. teres and Influence of Sample Support Size on DNA Concentration and Fungicide Resistance Frequency.

Fungicide resistance in foliar fungal pathogens is an increasing challenge to crop production. Yield impacts due to loss of fungicide efficacy may be reduced through effective surveillance and appropriate management intervention. For stubble-borne pathogens, off-season crop residues may be used to monitor fungicide resistance to inform pre-planting decisions; however, appropriate sampling strategies and support sizes for crop residues have not previously been considered. Here, we used Pyrenophora teres f. teres (Ptt) with resistance to demethylase inhibitor fungicides as a model system to assess spatial dependency and to compare the effects of different sampling strategies and support sizes on pathogen density (Ptt DNA concentration) and the frequency of fungicide resistance mutation. The results showed that sampling strategies (hand-picked versus raked) did not affect estimates of pathogen density or fungicide resistance frequency; however, sample variances were lower from raked samples. The effects of differing sample support size, as the size of the collection area (1.2, 8.6, or 60 m2), on fungicide resistance frequency were not evident (P > 0.05). However, measures of pathogen density increased with area size (P < 0.05); the 60 m2 area yielded the highest Ptt DNA concentration and produced the lowest number of pathogen-absent samples. Sample variances for pathogen density and fungicide resistance frequency were generally homogeneous between area sizes. The pattern of pathogen density was spatially independent; however, spatial dependency was identified for fungicide resistance frequency, with a range of 110 m, in one of the two fields surveyed. Collectively, the results inform designs for monitoring of fungicide resistance in stubble-borne pathogens.

The pursuit of genetic gain in agricultural crops through the application of machine-learning to genomic prediction.

Current practice in agriculture applies genomic prediction to assist crop breeding in the analysis of genetic marker data. Genomic selection methods typically use linear mixed models, but using machine-learning may provide further potential for improved selection accuracy, or may provide additional information. Here we describe SelectML, an automated pipeline for testing and comparing the performance of a range of linear mixed model and machine-learning-based genomic selection methods. We demonstrate the use of SelectML on an in silico-generated marker dataset which simulated a randomly-sampled (mixed) and an unevenly-sampled (unbalanced) population, comparing the relative performance of various methods included in SelectML on the two datasets. Although machine-learning based methods performed similarly overall to linear mixed models, they performed worse on the mixed dataset and marginally better on the unbalanced dataset, being more affected than linear mixed models by the imposed sampling bias. SelectML can assist in the training, comparison, and selection of genomic selection models, and is available from https://github.com/darcyabjones/selectml.

Spatiotemporal patterns of wheat response to Pyrenophora tritici-repentis in asymptomatic regions revealed by transcriptomic and X-ray fluorescence microscopy analyses.

Pathogen attacks elicit dynamic and widespread molecular responses in plants. While our understanding of plant responses has advanced considerably, little is known of the molecular responses in the asymptomatic 'green' regions adjoining lesions. Here, we explore gene expression data and high-resolution elemental imaging to report the spatiotemporal changes in the asymptomatic green region of susceptible and moderately resistant wheat cultivars infected with a necrotrophic fungal pathogen, Pyrenophora tritici-repentis. We show, with improved spatiotemporal resolution, that calcium oscillations are modified in the susceptible cultivar, resulting in 'frozen' host defence signals at the mature disease stage, and silencing of the host's recognition and defence mechanisms that would otherwise protect it from further attacks. In contrast, calcium accumulation and a heightened defence response were observed in the moderately resistant cultivar in the later stage of disease development. Furthermore, in the susceptible interaction, the asymptomatic green region was unable to recover after disease disruption. Our targeted sampling technique also enabled detection of eight previously predicted proteinaceous effectors in addition to the known ToxA effector. Collectively, our results highlight the benefits of spatially resolved molecular analysis and nutrient mapping to provide high-resolution spatiotemporal snapshots of host-pathogen interactions, paving the way for disentangling complex disease interactions in plants.

Optimized Sample Processing Pipeline for PCR-Based Fungicide Resistance Quantification of Stubble-Borne Fungal Pathogens.

Globally, yield losses associated with failed crop protection due to fungicide-resistant pathogens present an increasing problem. For stubble-borne pathogens, assessment of crop residues during the off-season could provide early fungicide resistance quantification for informed management decisions to mitigate yield losses. However, stubble assessment is hampered by assay inhibitors that are derived from decaying organic matter. To overcome assay inhibition from weathered stubble samples, we used a systems approach to quantify the frequency of resistance to demethylase inhibitor fungicides of the barley pathogen Pyrenophora teres f. teres. The system canvassed (i) 10 ball-milling conditions; (ii) four DNA extraction methodologies; and (iii) three column purification techniques for the provision of sufficient yield, quality, and purity of fungal DNA for a PCR-based fungicide resistance assay. Results show that DNA quantity and purity differed within each of the above three categories, with the optimized pipeline being (i) ball-milling samples in a 50-ml stainless steel canister for 5 min using a 20-mm ball at 30 revolutions s-1; (ii) a modified Brandfass method (extracted 64% more DNA than other methods assessed); and (iii) use of silica resin columns for the highest DNA concentration with optimal DNA purity. The chip-digital PCR assay, which quantified fungicide resistance from field samples, was unaffected by the DNA extraction method or purification technique, provided that thresholds of template quantity and purity were satisfied. In summary, this study has developed molecular pipeline options for pathogen fungicide resistance quantification from cereal stubbles, which can guide management for improved crop protection outcomes.

DNA metabarcoding identifies urban foraging patterns of oligolectic and polylectic cavity-nesting bees.

Urbanisation modifies natural landscapes resulting in built-up space that is covered by buildings or hard surfaces and managed green spaces that often substitute native plant species with exotics. Some native bee species have been able to adapt to urban environments, foraging and reproducing in these highly modified areas. However, little is known on how the foraging ecology of native bees is affected by urbanised environments, and whether impacts vary among species with different degrees of specialisation for pollen collection. Here, we aim to investigate the responses of native bee foraging behaviour to urbanisation, using DNA metabarcoding to identify the resources within nesting tubes. We targeted oligolectic (specialist) and polylectic (generalist) cavity-nesting bee species in residential gardens and remnant bushland habitats. We were able to identify 40 families, 50 genera, and 23 species of plants, including exotic species, from the contents of nesting tubes. Oligolectic bee species had higher diversity of plant pollen in their nesting tubes in residential gardens compared to bushland habitats, along with significantly different forage composition between the two habitats. This result implies a greater degree of forage flexibility for oligolectic bee species than previously thought. In contrast, the diversity and composition of plant forage in polylectic bee nesting tubes did not vary between the two habitat types. Our results suggest a complex response of cavity-nesting bees to urbanisation and support the need for additional research to understand how the shifts in foraging resources impact overall bee health.

Impact of different processing techniques on the key volatile profile, sensory, and consumer acceptance of black truffle (Tuber melanosporum Vittadini).

Fresh truffles which include black truffle (Tuber melanosporum Vittadini) deteriorate and lose aroma rapidly after harvest; therefore, postharvest processing via freeze-drying or encapsulation is an option to preserve truffle aroma for extended supply. However, the aroma profile that directly affects the truffle quality and consumer acceptance is influenced by processing and producers require processing options that balance processing feasibility with retention of a suitable aroma profile. This study aimed to determine the impact of freeze-drying and encapsulation on the profile of key volatiles, consumer discrimination, and overall sensory impression (aroma intensity, liking, and acceptability) of processed truffle products compared to the starting material (positive control). The study combined experimental-scale processing with GC-MS analysis and consumer sensory evaluation to compare and optimize postharvest processing options. Based on the results, some volatile changes were detected in the processed truffle products compared to the positive control which were aligned with the consumer discrimination (triangle test) and the aroma intensity score (consumer sensory test). Despite some chemical and sensory differences detected, the consumer panel did not have any preference for processed truffle products compared to the positive control. The overall finding indicates the potential value of processing truffles into a natural flavoring ingredient for food application via freeze-drying or encapsulation, which should be of great interest for the truffle and food industry. According to the correlation analysis, the consumer acceptance of a truffle product may be increased by retaining 1-octen-3-ol and methional, while reducing the amount of p-cresol in the product. PRACTICAL APPLICATION: The postharvest process of turning truffles into a food flavoring ingredient may cause undesirable volatile changes that would directly impact the aroma quality and consumer acceptance of the processed truffle products. Hence, the impacts of freeze-drying and encapsulation on the chemical and sensory profile of truffles were evaluated in this study. Overall, the results of the concurrent instrument and sensory analysis demonstrated that both freeze-drying and encapsulation are potential options for processing.

Comparative evaluation of encapsulation using ß-cyclodextrin versus freeze-drying for better retention and stabilizing of black Périgord truffle (Tuber melanosporum) aroma.

This study aimed to develop a novel technique to retain and stabilize compounds contributing to truffle aroma by encapsulation using ß-cyclodextrin. Two experiments were conducted. In the first experiment, the key volatile profile and microbial population of products resulting from three different encapsulation methods, namely direct mixing method (M1), direct mixing followed by ethanol addition method (M2), and paste method (M3), were compared with untreated truffles (positive control) over a 90-day period. The M2-derived product was the least optimal for retaining key volatile compounds despite showing the lowest microbial population. There was no significant difference in the volatile profile of products derived from M1 and M3 on day 0. However, it was observed that the M3-derived product could retain its volatile profile better than the M1-derived product by day 90. M3 was compared with freeze-drying in the second experiment. Freeze-dried truffles showed an overall higher relative percentage of volatiles than the M3-derived product on day 0. However, by day 90, some volatile changes occurred in the freeze-dried truffles but not in the M3-derived product. The findings indicate that while freeze-drying could adequately conserve truffle volatiles, the encapsulation of volatile compounds in ß-cyclodextrin could improve the volatile stability of truffle products and allow for longer storage times. Microbes were found in all encapsulated truffle products and freeze-dried truffles on days 0 and 90, suggesting the need to explore the possibility of incorporating a decontamination step in the process prior to either encapsulation or freeze-drying. PRACTICAL APPLICATION: A technique to capture and stabilize compounds responsible for truffle aroma by encapsulation using ß-cyclodextrin was developed and compared with freeze-drying in this study. The overall finding suggests that while freeze-drying of truffle could sufficiently preserve volatiles, encapsulating truffle volatiles with ß-cyclodextrin may improve its stability, extending its shelf life, which can be applied in the development of a natural truffle ingredient that can be applied in food product development.

Applications of environmental DNA (eDNA) in agricultural systems: Current uses, limitations and future prospects.

Global food production, food supply chains and food security are increasingly stressed by human population growth and loss of arable land, becoming more vulnerable to anthropogenic and environmental perturbations. Numerous mutualistic and antagonistic species are interconnected with the cultivation of crops and livestock and these can be challenging to identify on the large scales of food production systems. Accurate identifications to capture this diversity and rapid scalable monitoring are necessary to identify emerging threats (i.e. pests and pathogens), inform on ecosystem health (i.e. soil and pollinator diversity), and provide evidence for new management practices (i.e. fertiliser and pesticide applications). Increasingly, environmental DNA (eDNA) is providing rapid and accurate classifications for specific organisms and entire species assemblages in substrates ranging from soil to air. Here, we aim to discuss how eDNA is being used for monitoring of agricultural ecosystems, what current limitations exist, and how these could be managed to expand applications into the future. In a systematic review we identify that eDNA-based monitoring in food production systems accounts for only 4 % of all eDNA studies. We found that the majority of these eDNA studies target soil and plant substrates (60 %), predominantly to identify microbes and insects (60 %) and are biased towards Europe (42 %). While eDNA-based monitoring studies are uncommon in many of the world's food production systems, the trend is most pronounced in emerging economies often where food security is most at risk. We suggest that the biggest limitations to eDNA for agriculture are false negatives resulting from DNA degradation and assay biases, as well as incomplete databases and the interpretation of abundance data. These require in silico, in vitro, and in vivo approaches to carefully design, test and apply eDNA monitoring for reliable and accurate taxonomic identifications. We explore future opportunities for eDNA research which could further develop this useful tool for food production system monitoring in both emerging and developed economies, hopefully improving monitoring, and ultimately food security.

Methods used for extraction of plant volatiles have potential to preserve truffle aroma: A review.

Truffles are considered one of the world's most highly prized foods mainly due to their desirable organoleptic properties and rarity. However, truffles are seasonal (harvested mostly in winter from June to August in the Southern Hemisphere and from December to February in the Northern Hemisphere) and extremely perishable. Truffles deteriorate rapidly showing undesirable changes within 10 days from harvest in aroma and visual appearance after harvest. The very short postharvest shelf life (about 7-10 days) limits the potential for export and domestic consumption all year round. Several preservation methods have been studied to prolong their shelf life without the loss of aroma. However, all traditional preservation techniques have their own shortcomings and remain challenging. The extraction of natural truffle aroma volatiles for food applications could be a potential alternative to replace the existing synthetic flavoring used for processed truffle products. Four commonly used extraction methods for recovering volatile compounds from plants, namely, supercritical carbon dioxide extraction, Soxhlet extraction, distillation, and cold pressing, are critically analyzed. Up to date, existing research about the extraction of aroma volatiles from truffles is limited in the literature but based on the volatility of the key truffle volatile compounds, supercritical carbon dioxide extraction may offer the best possibility so that a natural truffle-based product that can be used in food applications throughout the year can be made available.

"Wax On, Wax Off": In Vivo Imaging of Plant Physiology and Disease with Fourier Transform Infrared Reflectance Microspectroscopy.

Analysis of the epicuticular wax layer on the surface of plant leaves can provide a unique window into plant physiology and responses to environmental stimuli. Well-established analytical methodologies can quantify epicuticular wax composition, yet few methods are capable of imaging wax distribution in situ or in vivo. Here, the first report of Fourier transform infrared (FTIR) reflectance spectroscopic imaging as a non-destructive, in situ, method to investigate variation in epicuticular wax distribution at 25 µm spatial resolution is presented. The authors demonstrate in vivo imaging of alterations in epicuticular waxes during leaf development and in situ imaging during plant disease or exposure to environmental stressors. It is envisaged that this new analytical capability will enable in vivo studies of plants to provide insights into how the physiology of plants and crops respond to environmental stresses such as disease, soil contamination, drought, soil acidity, and climate change.

Synchrotron X-ray fluorescence microscopy-enabled elemental mapping illuminates the 'battle for nutrients' between plant and pathogen.

Metal homeostasis is integral to normal plant growth and development. During plant-pathogen interactions, the host and pathogen compete for the same nutrients, potentially impacting nutritional homeostasis. Our knowledge of outcome of the interaction in terms of metal homeostasis is still limited. Here, we employed the X-ray fluorescence microscopy (XFM) beamline at the Australian Synchrotron to visualize and analyse the fate of nutrients in wheat leaves infected with Pyrenophora tritici-repentis, a necrotrophic fungal pathogen. We sought to (i) evaluate the utility of XFM for sub-micron mapping of essential mineral nutrients and (ii) examine the spatiotemporal impact of a pathogen on nutrient distribution in leaves. XFM maps of K, Ca, Fe, Cu, Mn, and Zn revealed substantial hyperaccumulation within, and depletion around, the infected region relative to uninfected control samples. Fungal mycelia were visualized as thread-like structures in the Cu and Zn maps. The hyperaccumulation of Mn in the lesion and localized depletion in asymptomatic tissue surrounding the lesion was unexpected. Similarly, Ca accumulated at the periphery of the symptomatic region and as microaccumulations aligning with fungal mycelia. Collectively, our results highlight that XFM imaging provides the capability for high-resolution mapping of elements to probe nutrient distribution in hydrated diseased leaves in situ.

Crop-Zone Weed Mycobiomes of the South-Western Australian Grain Belt.

In the absence of a primary crop host, secondary plant hosts may act as a reservoir for fungal plant pathogens of agricultural crops. Secondary hosts may potentially harbor heteroecious biotrophs (e.g., the stripe rust fungus Puccinia striiformis) or other pathogens with broad host ranges. Agricultural grain production tends toward monoculture or a limited number of crop hosts over large regions, and local weeds are a major source of potential secondary hosts. In this study, the fungal phyllospheres of 12 weed species common in the agricultural regions of Western Australia (WA) were compared through high-throughput DNA sequencing. Amplicons of D2 and ITS were sequenced on an Illumina MiSeq system using previously published primers and BLAST outputs analyzed using MEGAN. A heatmap of cumulative presence-absence for fungal taxa was generated, and variance patterns were investigated using principal components analysis (PCA) and canonical correspondence analysis (CCA). We observed the presence of several major international crop pathogens, including basidiomycete rusts of the Puccinia spp., and ascomycete phytopathogens of the Leptosphaeria and Pyrenophora genera. Unrelated to crop production, several endemic pathogen species including those infecting Eucalyptus trees were also observed, which was consistent with local native flora. We also observed that differences in latitude or climate zones appeared to influence the geographic distributions of plant pathogenic species more than the presence of compatible host species, with the exception of Brassicaceae host family. There was an increased proportion of necrotrophic Ascomycete species in warmer and drier regions of central WA, compared to an increased proportion of biotrophic Basidiomycete species in cooler and wetter regions in southern WA.

Flowering phenology in a Eucalyptus loxophleba seed orchard, heritability and genetic correlation with biomass production and cineole: breeding strategy implications.

Reproductive synchronicity within a seed orchard facilitates gene exchange and reduces self-fertilisation. Here we assessed key flowering traits, biomass and foliar 1,8-cineole concentrations of Eucalyptus loxophleba (subsp. lissophloia and gratiae) in an open-pollinated seed orchard. Monthly flowering observations were made on 1142 trees from 60 families and nine provenances across 2 years. The percentage of trees flowering in both years was similar at 87%. There were differences between provenances and families within provenances for flowering traits, biomass and 1,8-cineole and interactions between provenances and year for flowering traits. Heritability of start and end flowering, and 1,8-cineole were high to moderate ([Formula: see text] = 0.75-0.45) and duration of flowering, propensity to flower and biomass estimates were moderate to low ([Formula: see text] = 0.31-0.10). Genetic and phenotypic correlations between flowering traits were high (rg = 0.96-0.63 and rp = 0.93-0.34) except between duration and end of flowering. The correlations were weaker between flowering traits and biomass or 1,8-cineole. 'Dual flowering', when trees underwent two reproductive cycles in a year, was responsible for out-of-phase flowering and those with low biomass and 1,8-cineole concentration should be removed from the breeding programme to hasten selection for desirable traits.

Real-Time PCR for Diagnosing and Quantifying Co-infection by Two Globally Distributed Fungal Pathogens of Wheat.

Co-infections - invasions of a host-plant by multiple pathogen species or strains - are common, and are thought to have consequences for pathogen ecology and evolution. Despite their apparent significance, co-infections have received limited attention; in part due to lack of suitable quantitative tools for monitoring of co-infecting pathogens. Here, we report on a duplex real-time PCR assay that simultaneously distinguishes and quantifies co-infections by two globally important fungal pathogens of wheat: Pyrenophora tritici-repentis and Parastagonospora nodorum. These fungi share common characteristics and host species, creating a challenge for conventional disease diagnosis and subsequent management strategies. The assay uses uniquely assigned fluorogenic probes to quantify fungal biomass as nucleic acid equivalents. The probes provide highly specific target quantification with accurate discrimination against non-target closely related fungal species and host genes. Quantification of the fungal targets is linear over a wide range (5000-0.5 pg DNA µl-1) with high reproducibility (RSD ≤ 10%). In the presence of host DNA in the assay matrix, fungal biomass can be quantified up to a fungal to wheat DNA ratio of 1 to 200. The utility of the method was demonstrated using field samples of a cultivar sensitive to both pathogens. While visual and culture diagnosis suggested the presence of only one of the pathogen species, the assay revealed not only presence of both co-infecting pathogens (hence enabling asymptomatic detection) but also allowed quantification of relative abundances of the pathogens as a function of disease severity. Thus, the assay provides for accurate diagnosis; it is suitable for high-throughput screening of co-infections in epidemiological studies, and for exploring pathogen-pathogen interactions and dynamics, none of which would be possible with conventional approaches.

Analysis of the nexus between population, water resources and Global Food Security highlights significance of governance and research investments and policy priorities.

BACKGROUND: Analyses of sensitivity of Global Food Security (FS) score to a key set of supply or demand factors often suggest population and water supply as being the most critical and on which policies tend to focus. To explore other policy options, we characterized the nexus between GFS and a set of supply or demand factors including population, agricultural and industrial water uses, agricultural publications (as a surrogate for investment in agricultural research and development (R&D)) and corruption perception index (CPI), to reveal opportunities for attaining enduring GFS. RESULTS: We found that despite being the primary driver of demand for food, population showed no significant correlation with FS scores. Similarly, agricultural water use was poorly correlated with GFS scores, except in countries where evaporation exceeds precipitation and irrigation is significant. However, FS had a strong positive association with industrial water use as a surrogate for overall industrialization. Recent expansions in cultivated land area failed to yield concomitant improvements in FS score since such expansions have been mostly into marginal lands with low productivity and thus barely compensated for lands retired from cropping in several developed economies. However, FS was positively associated with agricultural R&D investments, as it was with the CPI scores. The apparent and relative strengths of these drivers on FS outcome amongst countries were in the order: industrial water-use ≈ publication rate ≈ corruption perception ≫ agricultural water use > population. CONCLUSIONS: We suggest that to enshrine enduring food security, policies should prioritize (1) increased R&D investments that address farmer needs and (2) governance mechanisms that promote accountability in both research and production value chains. © 2018 Society of Chemical Industry.

Host-Multi-Pathogen Warfare: Pathogen Interactions in Co-infected Plants.

Studies of plant-pathogen interactions have historically focused on simple models of infection involving single host-single disease systems. However, plant infections often involve multiple species and/or genotypes and exhibit complexities not captured in single host-single disease systems. Here, we review recent insights into co-infection systems focusing on the dynamics of host-multi-pathogen interactions and the implications for host susceptibility/resistance. In co-infection systems, pathogen interactions include: (i) Competition, in which competing pathogens develop physical barriers or utilize toxins to exclude competitors from resource-dense niches; (ii) Cooperation, whereby pathogens beneficially interact, by providing mutual biochemical signals essential for pathogenesis, or through functional complementation via the exchange of resources necessary for survival; (iii) Coexistence, whereby pathogens can stably coexist through niche specialization. Furthermore, hosts are also able to, actively or passively, modulate niche competition through defense responses that target at least one pathogen. Typically, however, virulent pathogens subvert host defenses to facilitate infection, and responses elicited by one pathogen may be modified in the presence of another pathogen. Evidence also exists, albeit rare, of pathogens incorporating foreign genes that broaden niche adaptation and improve virulence. Throughout this review, we draw upon examples of co-infection systems from a range of pathogen types and identify outstanding questions for future innovation in disease control strategies.

Deconvoluting effects of vine and soil properties on grape berry composition.

BACKGROUND: Grape berry composition is influenced by several factors including grapevine and soil properties and their interactions. Understanding how these factors interact to determine berry composition is integral to producing berries with desired composition. Here we used extensive spatio-temporal data to identify significant vine and soil features that influence Shiraz berry composition. RESULTS: The concentrations of berry flavonoids (anthocyanins, tannin and total phenolics), total soluble solids and pH were typically negatively associated with canopy, crop and berry size factors whereas titratable acidity was positively associated. The strengths of the associations, however, were generally greater with the crop and berry size factors than with the canopy size factor. The analyses also resolved separate influences of berry and crop size on berry composition. Soil properties had significant influences on berry composition; however, when influences of soil factors on vine-attributes were accounted for, the apparent effects of soil factors on berry composition were largely non-existent. CONCLUSION: At each site, variations in berry composition were more strongly associated with crop and berry size than with canopy size factors. Apparent influences of soil properties on berry composition are indirect, being mediated via their effects on vine attributes (canopy, crop and berry sizes).

Berry composition and climate: responses and empirical models.

Climate is a strong modulator of berry composition. Accordingly, the projected change in climate is expected to impact on the composition of berries and of the resultant wines. However, the direction and extent of climate change impact on fruit composition of winegrape cultivars are not fully known. This study utilised a climate gradient along a 700 km transect, covering all wine regions of Western Australia, to explore and empirically describe influences of climate on anthocyanins, pH and titratable acidity (TA) levels in two or three cultivars of Vitis vinifera (Cabernet Sauvignon, Chardonnay and Shiraz). The results showed that, at a common maturity of 22° Brix total soluble solids, berries from the warmer regions had low levels of anthocyanins and TA as well as high pH compared to berries from the cooler regions. Most of these regional variations in berry composition reflected the prevailing climatic conditions of the regions. Thus, depending on cultivar, 82-87 % of TA, 83 % of anthocyanins and about half of the pH variations across the gradient were explained by climate-variable-based empirical models. Some of the variables that were relevant in describing the variations in berry attributes included: diurnal ranges and ripening period temperature (TA), vapour pressure deficit in October and growing degree days (pH), and ripening period temperatures (anthocyanins). Further, the rates of change in these berry attributes in response to climate variables were cultivar dependent. Based on the observed patterns along the climate gradient, it is concluded that: (1) in a warming climate, all other things being equal, berry anthocyanins and TA levels will decline whereas pH levels will rise; and (2) despite variations in non-climatic factors (e.g. soil type and management) along the sampling transect, variations in TA and anthocyanins were satisfactorily described using climate-variable-based empirical models, indicating the overriding impact of climate on berry composition. The models presented here are useful tools for assessing likely changes in berry TA and anthocyanins in response to changing climate for the wine regions and cultivars covered in this study.