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Serpins constitute a family of proteins with a very wide distribution in nature. Serpins have a well-conserved tertiary structure enabling irreversible protease inhibition or other specific biochemical functions. We examined the 189 putative wheat serpin genes previously identified by Benbow et al. (2019) via analysis of gene annotations (RefSeq v1.0) and combined our previous examinations of wheat ESTs and the 454 genome assembly. We found that 81 of the 189 putative serpin genes, plus two manually annotated genes, encode full-length, structurally conserved serpins. Expression of these serpin genes during wheat development and disease/abiotic stress responses was analysed using a publicly available RNAseq database. Results showed that the wheat LR serpins, homologous to Arabidopsis AtSerpin1 and barley BSZx, are ubiquitously expressed across all tissues throughout the wheat lifecycle, whereas the expression of other wheat serpin genes is tissue-specific, including expression only in the grain, only in the root, and only in the anther and microspore. Nine serpin genes were upregulated in both biotic and abiotic responses. Two genes in particular were highly expressed during disease and abiotic challenges. Our findings provide valuable information for further functional study of the wheat serpins, which in turn may lead to their application as molecular markers in wheat breeding.

Application of the growth regulator salicylic acid (SA) and the polyamine spermidine (Spd) can be used to manage various plant abiotic stresses. We aimed to evaluate the sole and combined effects of SA and Spd on maize (Zea mays) under individual and combined drought and chromium (Cr) stress. Drought, Cr, and drought + Cr treatments caused oxidative stress by inducing higher production of reactive oxygen species (H2O2, O2−), enhanced malondialdehyde content and increased relative membrane permeability. Increased oxidative stress and higher Cr uptake in the host plant reduced the content of carotenoids, other photosynthetic pigments and protein, and changed carbohydrate metabolism. Combined drought + Cr stress was more damaging for the growth of maize plants than the individual stresses. Exogenous treatments of SA and Spd alleviated the adverse effects of drought and Cr toxicity, reflected by accumulations of osmolytes, antioxidants and endogenous polyamines. Single applications of Spd (0.1 mM) increased plant height, shoot fresh weight, leaf area, above-ground dry matter accumulation and polyamine content under drought, Cr, and drought + Cr stress conditions. However, the combined treatment SA + Spd (0.25 mM + 0.05 mM) was more effective in increasing protein and water contents, photosynthetic pigments, and carotenoids. The same treatment increased Cr tolerance in the maize plants by decreasing uptake of this heavy metal from root to shoot. The SA + Spd treatment also decreased oxidative stress by promoting antioxidant enzyme activities, and enhanced levels of proline, soluble sugars, and carbohydrate contents under individual and combined stress conditions. Results indicate that the combined half-dose application of SA + Spd may be utilized to boost the tolerance in maize under individual as well as combined drought and Cr stress conditions.

Rust fungi, which are responsible for some of the world's most destructive plant diseases, are named for the distinctive rusty colour of one or more of their spore forms. Rust spore pigments are thought to provide protection against UV radiation and oxidative stress, and may act as virulence factors. However, with the exception of daisy rust spores, the identity and relative abundance of the carotenoids in the rust spore cytoplasm have not been investigated using modern analytical methods, and little is known about the dependence of the carotenoid complement on species, pathotype, spore-colour mutations and season. We developed and validated a method to separate, identify and quantify rust carotenoids by reversed-phase high-performance liquid chromatography (RP-HPLC) combined with mass spectrometry. The carotenoids identified were lycopene, γ-carotene, β-carotene and phytoene. Rates of carotenoid degradation depended greatly on spore storage conditions, with freezing at −80 °C providing optimal stability. Carotenoid profiles of 103 isolates from 14 rust species were compared, showing that the ratio γ-carotene:β-carotene varied substantially among species. Total carotenoid content was generally lower in spring than in autumn (Sydney, Australia)—possibly due to differences in solar exposure—but the percentage of individual carotenoids was relatively stable. Among the colour mutants tested, chocolate mutants of Puccinia graminis f. sp. tritici (wheat stem rust) contained no carotenoid pigments, while albino mutants of P. striiformis f. sp. tritici (wheat stripe rust) contained only phytoene, a colourless carotenoid. We discuss our results in terms of the biogenesis and biological functions of carotenoids in rust fungi.

Thymoquinone (TQ) is a secondary metabolite found in abundance in very few plant species including Nigella sativa L., Monarda fistulosa L., Thymus vulgaris L. and Satureja montana L. Preclinical pharmacological studies have shown that TQ has many biological activities, such as anti-inflammatory, antioxidant and anticancer. Both in vivo and in vitro experiments have shown that TQ acts as an antitumor agent by altering cell cycle progression, inhibiting cell proliferation, stimulating apoptosis, inhibiting angiogenesis, reducing metastasis and affecting autophagy. In this comprehensive study, the evidence on the pharmacological potential of TQ on pancreatic cancer is reviewed. The positive results of preclinical studies support the view that TQ can be considered as an additional therapeutic agent against pancreatic cancer. The possibilities of success for this compound in human medicine should be further explored through clinical trials. © 2022 The Authors

Sorghum grain is a staple food for about 500 million people in 30 countries in Africa and Asia. Despite this contribution to global food production, most of the world's sorghum grain, and nearly all in Western countries, is used as animal feed. A combination of the increasingly important ability of sorghum crops to resist heat and drought, the limited history of the use of sorghum in Western foods, and the excellent functional properties of sorghum grain in healthy diets, suggests a greater focus on the development of new sorghum-based foods. An understanding of the structural and functional properties of sorghum grain to develop processes for production of new sorghum-based foods is required. In this review, we discuss the potential of sorghum in new food products, including sorghum grain composition, the functional properties of sorghum in foods, processing of sorghum-based products, the digestibility of sorghum protein and starch compared to other grains, and the health benefits of sorghum. In the potential for sorghum as a major ingredient in new foods, we suggest that the gluten-free status of sorghum is of relatively minor importance compared to the functionality of the slowly digested starch and the health benefits of the phenolic compounds present.

Jacaranda mimosifolia trees are grown in frost-free regions globally. The aim of this study was to evaluate the methanol crude extract and various fractions of increasing polarity of J. mimosifolia leaves for bioactive metabolites, as well as antimicrobial, antioxidant and anticancer activities. The anti-inflammatory potential of the various fractions of J. mimosifolia leaf extract was studied via the lipoxygenase (LOX) inhibitory assay. Methanol crude extract (ME), derived fractions extracted with chloroform (CF) and ethyl acetate (EAF), and residual aqueous extract (AE) of dried J. mimosifolia leaves were assayed for polyphenolic compounds, their antioxidant, antimicrobial and lipoxygenase (LOX) inhibitory activities, and anticancer properties. Polyphenolic compounds were determined via HPLC while phytochemicals (total phenolics, flavonoids, tannins and ortho-diphenol contents), antioxidant activities (DPPH, hydrogen peroxideperoxide, hydroxyl and superoxide radical anions) and LOX were measured via spectrophotometry. Methanol extracts and various fractions were evaluated for antibacterial activities against Bacillus subtilis, Klebsiella pneumonia, Pseudomonas aeruginosa and Staphylococcus aureus. Antifungal potential of the fractions was tested against three species: Aspergillus flavus, Aspergillus fumigatus and Fusarium oxysporum. The highest values for total phenolic content (TPC), total flavonoid content (TFC), flavonols, tannins and ortho-diphenols were in the ME, followed by CF > EAF > AE. ME also had the highest antioxidant activity with EC50 values 48±1.3, 45±2.4, 42±1.3 and 46±1.3 μg/mL based on the DPPH, hydrogen peroxide, hydroxyl radical and superoxide radical assays, respectively. TPC and TFC showed a significant, strong and positive correlation with the values for each of these antioxidant activities. ME exhibited anti-inflammatory potential based on its LOX inhibitory activity (IC50 = 1.3 μg/mL). ME also had the maximum antibacterial and antifungal potential, followed by EAF > CF > AE. Furthermore, ME showed the strongest cytotoxic effect (EC50 = 10.7 and 17.3 μg/mL) against human hormone-dependent prostate carcinoma (LnCaP) and human lung carcinoma (LU-1) cell lines, respectively. Bioactive compounds present in leaf methanol extracts of J. mimosifolia were identified using gas chromatography-mass spectrometry (GC-MS). Fifteen compounds were identified including phenolic and alcoholic compounds, as well as fatty acids. Our results suggest that J. mimosifolia leaves are a good source of natural products with antioxidant, anti-inflammatory and anti-cancer properties for potential therapeutic, nutraceutical and functional food applications.

Fusarium verticillioides is an important fungal pathogen of maize, causing stalk rot and severely affecting crop production. The aim of this study was to characterize the protective effects of formulations based on Jacaranda mimosifolia leaf extracts against F. verticillioides in maize. We compared different seed treatments comprising J. mimosifolia extracts, chemical fungicide (mefenoxam) and salicylic acid to modulate the defense system of maize host plants. Both aqueous and methanolic leaf extracts of J. mimosifolia (1.2% w/v) resulted in 96–97% inhibition of mycelial growth of F. verticillioides. While a full-dose (1.2%) extract of J. mimosifolia provided significant protective effects on maize plants compared to the inoculated control, a half-dose (0.6% w/v) application of J. mimosifolia in combination with half-strength mefenoxam was the most effective treatment in reducing stalk rot disease in pot and field experiments. The same seed treatment significantly upregulated the expression of genes in the leaves encoding chitinase, glucanase, lipid transfer protein, and pathogenesis-related proteins PR-1, PR-5 and PR-10, 72 h after inoculation. This treatment also induced the activities of peroxidase, polyphenol oxidase, protease, acid invertase, chitinase and phenylalanine ammonia lyase. We conclude that seed pre-treatment with J. mimosifolia extract with half-strength chemical mefenoxam is a promising approach for the management of stalk rot in maize.

Diseases caused by rust fungi pose a significant threat to global plant production. Although carotenoid pigments are produced in spores of nearly all rust species, the corresponding biosynthesis pathway(s) have not been investigated. Here, candidate genes for carotenoid biosynthesis in Puccinia graminis f. sp. tritici (Pgt) were identified, cloned and functionally complemented using specifically engineered strains of Escherichia coli. A part of the carotenoid biosynthesis pathway in rust fungi was elucidated, with only two genes, CrtYB and CrtI, catalysing the reactions from geranyl–geranyl diphosphate (GGPP) to γ-carotene. The CrtYB gene encodes a bi-functional lycopene cyclase/phytoene synthase, which catalyses the condensation of two GGPP into phytoene, as well as the cyclisation of the ψ-end of lycopene to form γ-carotene. The CrtI gene encodes a phytoene desaturase that carries out four successive desaturations of phytoene, through the intermediates phytofluene and neurosporene to lycopene. The evolution of carotenoid pigmentation in rust fungi, including Pgt, P. graminis avenae, P. graminis secalis (Pgs), P. graminis lolli, P. striiformis f. sp. tritici, P. striiformis f. sp. pseudohordei, P. striiformis f. sp. hordei, the “scabrum” rust (putative hybrids between Pgt and Pgs), P. triticina, and P. hordei, was investigated by phylogenetic analysis. Both CrtYB and CrtI were found to be closely related among rust fungi, other pathogenic fungi, and some aphids. Our results provide a springboard to increase the understanding of the physiological role(s) of carotenoid pigments in rust fungi, to better understand evolution within the Pucciniales, and to develop robust molecular diagnostics for rust fungi.

Oryza sativa has many wild relatives that exhibit a wide range of canopy architectures. Two fast-growing ecotypes of an Australian wild rice (O. meridionalis) were compared with cultivated O. sativa to analyze the efficiency of light interception in mature vegetative canopies and how well light capture predicts biomass production. Canopies of individual plants were digitized and analyzed with YplantQMC. Plants were also grown at 700 ppm CO2 as a means of increasing the canopy density and assessing the relative impact of rising CO2 levels on photosynthesis, light interception and biomass. Simulations of light interception for these morphologically diverse canopies were combined with photosynthetic light response curves to estimate theoretical carbon gain. Both O. meridionalis genotypes had more tillers and leaves, and denser canopies, than O. sativa but one (‘Howard Springs’) intercepted more light than the other two genotypes, resulting in the highest simulated carbon gains. Photosynthesis per unit leaf area (Amax) was enhanced by 20–50% in 700 ppm CO2 in all genotypes but the proportion of total leaf area intercepting light decreased as a result of denser leaf canopies in these plants. Estimated total shoot carbon gain increased strongly at elevated CO2 despite these denser canopies and increased self-shading, reaching a peak in ‘Howard Springs’ at 700 ppm CO2. Thus, while high crown density and low leaf dispersion reduced the efficiency of light interception by the canopy, elevated CO2 still enhanced growth through consistently higher assimilation rates. Simulations showed that during the Summer Solstice, carbon gain was predicted to be greater than at the Equinox through superior light capture, particularly in ‘Howard Springs’. However, the relationship between light interception and carbon gain was broadly maintained, regardless of genotype, CO2 concentration and season.

Diseases caused by rust fungi represent critical constraints to global plant production. A characteristic feature of rust pathogens is the striking pigments they produce in one or more spore forms, which give them a rusty appearance. Here, we review the literature published to date on the extraction, separation, quantification and characterisation of carotenoid pigments in rust fungi. These pigments are thought to protect rust fungi against UV radiation and oxidative stress, and possibly act as virulence factors. The yellow-orange colour of some rust species is due to carotenoid pigments. Four carotenoids have been found in rust fungi: phytoene, lycopene, γ-carotene and β-carotene, but their relative contributions to biological functions are largely unknown. Different pre-processes and storage of spore materials, as well as different extraction processes, have been applied in a wide range of investigations on rust spore pigments. We find that the value of the current literature on rust carotenoids for taxonomic diagnostics in understanding the evolution of pigment biosynthesis and in assessing their role in pathogenesis is limited. Re-investigation of rust carotenoid composition using modern analytical technologies is therefore critical to further these fields of research. Our review includes detailed guidance on choice of techniques for rust carotenoid experimental analyses.

RNA silencing is a powerful tool deployed by plants against viral infection and abnormal gene expression. Plant viruses have evolved a suite of silencing suppressors for counter-defense, which are also widely used to boost transcript and protein accumulation in transient assays. However, only wild type silencing suppressor proteins have been reported to date. Here we demonstrate that P0 of Potato leafroll virus (PLRV), PLP0, can be split into two proteins that only show silencing suppression activity upon co-expression. We cloned each of these proteins in two different constructs and transiently co-infiltrated them in N. benthamiana leaves. We expressed a fluorescent protein from one of the vectors and observed that cells expressing both halves of PLP0 suppressed gene silencing. Further, we showed that Q system of Neurospora crassa, based on co-expression of a transcription activator and inhibitor, is functional in agroinfiltrated leaves of N. benthamiana. Q system combined with the split PLP0 system showed very tight co-expression of Q system’s transcriptional activator and inhibitor. Altogether, our experiments demonstrate a functioning conditional silencing suppressor system and its potential as a powerful tool for transient expression in N. benthamiana leaves, as well as the application of the Q system in plants.

Wild rice species represent valuable germplasm for breeding of elite genotypes with enhanced resistance to biotic and abiotic stresses. However, differences in endosperm morphology between wild and cultivated rice, and how these are affected by enhanced levels of atmospheric CO2 (eCO2) predicted for later this century, are largely unknown. Accessions of native rice species from northern Australia, Oryza australiensis and O. meridionalis, and a local cultivar, O. sativa cv. Doongara, were grown to maturity in glasshouses at ambient (aCO2, 400 ppm) and eCO2 (700 ppm). Endosperm morphology was studied from transverse sections of mature grains using scanning electron microscopy. Aleurone cell length and width were higher in eCO2 than in aCO2 in the O. meridionalis accession Howard Springs, but lower in Doongara. The average starch granule size in Doongara was greater than in the wild genotypes in both CO2 treatments. The area and length of the starch granule images were larger in eCO2 than in aCO2 in the O. meridionalis accession Cape York but smaller in Doongara. Responses of rice endosperm morphology to eCO2 were dependent on genotype. We conclude that endosperm morphology responds differentially to eCO2, both between and within rice species. Contrasts in these responses can be exploited by breeders.

Susceptibility of plants to abiotic stresses, including extreme temperatures, salinity and drought, poses an increasing threat to crop productivity worldwide. Here the drought-induced response of maize was modulated by applications of methyl jasmonate (MeJA) and salicylic acid (SA) to seeds prior to sowing and to leaves prior to stress treatment. Pot experiments were conducted to ascertain the effects of exogenous applications of these hormones on maize growth, physiology and biochemistry under drought stress and well-watered (control) conditions. Maize plants were subjected to single as well as combined pre-treatments of MeJA and SA. Drought stress severely affected maize morphology and reduced relative water content, above and below-ground biomass, rates of photosynthesis, and protein content. The prolonged water deficit also led to increased relative membrane permeability and oxidative stress induced by the production of malondialdehyde (from lipid peroxidation), lipoxygenase activity (LOX) and the production of H2O2. The single applications of MeJA and SA were not found to be effective in maize for drought tolerance while the combined pre-treatments with exogenous MeJA+SA mitigated the adverse effects of drought-induced oxidative stress, as reflected in lower levels of lipid peroxidation, LOX activity and H2O2. The same pre-treatment also maintained adequate water status of the plants under drought stress by increasing osmolytes including proline, total carbohydrate content and total soluble sugars. Furthermore, exogenous applications of MeJA+SA approximately doubled the activities of the antioxidant enzymes catalase, peroxidase and superoxide dismutase. Pre-treatment with MeJA alone gave the highest increase in drought-induced production of endogenous abscisic acid (ABA). Pre-treatment with MeJA+SA partially prevented drought-induced oxidative stress by modulating levels of osmolytes and endogenous ABA, as well as the activities of antioxidant enzymes. Taken together, the results show that seed and foliar pre-treatments with exogenous MeJA and/or SA can have positive effects on the responses of maize seedlings to drought.

Salinity is a major constraint on rice productivity worldwide. However, mechanisms of salt tolerance in wild rice relatives are unknown. Root microsomal proteins are extracted from two Oryza australiensis accessions contrasting in salt tolerance. Whole roots of 2-week-old seedlings are treated with 80 mM NaCl for 30 days to induce salt stress. Proteins are quantified by tandem mass tags (TMT) and triple-stage Mass Spectrometry. More than 200 differentially expressed proteins between the salt-treated and control samples in the two accessions (p-value

Interspecific germplasm from wild species of Oryza will be increasingly applied to breeding programs over the coming decades. However, the concurrent impact of rising atmospheric CO2 concentrations (atm-CO2) on grain properties is unknown. Wild accessions of Australian native rice species (Oryza australiensis and O. meridionalis) and the local cultivar (O. sativa cv. Doongara) were grown to maturity in glasshouses at ambient (400 ppm; aCO2) and elevated (700 ppm; eCO2) atm-CO2 and physicochemical properties of the grains were compared. Seed length (Sl), seed width (Sw) and 1000-seed weight (TSW) were higher at eCO2 than aCO2, while nitrogen content (N) and apparent amylose content (AC) were not significantly changed, irrespective of genotype. Rapid viscosity analyser (RVA) profiles revealed increased peak viscosity (PV) for rice under eCO2, indicating differences in starch gelatinization. However, final viscosity (FV) increased in the wild rices in eCO2, whereas it decreased in Doongara. Increases in PV were accompanied by lower estimates of protein contents (PC) at eCO2 in all species. When AgNO3 was excluded from the RVA, it became apparent that its presence had enhanced the effects of eCO2 on flour pasting properties through inhibition of α-amylase activity of the flour. Our results suggest that grain morphology and pasting properties are differently affected by eCO2 in the wild rices compared with cultivated rice. These data will inform the breeding criteria for rice grain properties and inform the introgression of wild germplasm.

1. The two red grain sorghums were extensively characterised. Kafirin, polyphenolic compounds, free, conjugated and bound phenolic acids, phytate concentrations and starch pasting profiles were determined. 2. The experiment consisted of a 2 × 4 factorial array of dietary treatments comprising two red sorghum varieties (Tiger and Block I) ground through 4 hammer-mill screen sizes (2.0, 3.2, 4.8 6.0 mm) prior to incorporation into nutritionally equivalent diets. Eight steam-pelleted dietary treatments were each offered to 7 replicates (6 male Ross 308 birds per cage) from 7 to 28 d post-hatch. 3. Effects of dietary treatments on growth performance, relative gizzard and pancreas weights, nutrient utilisation, apparent starch and protein (N) digestibility coefficients and disappearance rates from 4 small intestinal segments were determined. 4. The 2.0-mm hammer-mill screen generated an average geometric mean particle size of 794 μm and the 6.0-mm screen a mean particle size of 1405 μm. However, hammer-mill screen size did not influence weight gain or FCR. The 6.0-mm screen size generated significantly higher starch and protein (N) digestibility coefficients in the distal jejunum and distal ileum than the 2.0-mm hammer-mill screen. 5. Tiger sorghum was superior to Block I sorghum, as significant advantages were observed for feed conversion ratios (3.25%), AME (0.37 MJ), ME:GE ratios (4.15%), AMEn (0.53 MJ), distal ileal starch digestibility coefficients (2.46%) and protein (N) digestibility coefficients in the distal jejunum (4.66%), proximal ileum (1.96%) and distal ileum (2.16%). The inferior Block I sorghum contained more kafirin (67.1 versus 51.3 g/kg), phytate (9.79 versus 8.40 g/kg), total phenolic compounds (4.68 versus 4.12 mg GAE/g), flavan-4-ols (7.98 versus 5.04 ABS/ml/g), total phenolic acids (554 versus 402 μg/g) and total ferulic acid (375 versus 281 μg/g) in comparison to Tiger sorghum.

The use of aleurone-layer protoplasts as a model system for barley germination research offers several advantages over working with the whole tissue, including the ability to more precisely control the level of hormone exposure during experiments, and to more easily perform cell biology studies using microscopy. However, protoplasts from the cultivar Himalaya have dominated these studies, probably owing to the availability of a suitable isolation protocol tailored to these hull-less grains. Here the development of a protocol to isolate aleurone-layer protoplasts from commercial malting barley cultivars is described. By implementing the protocol, protoplasts were isolated from three commercial Australian varieties and their responses to incubation with gibberellic acid evaluated. Key properties of the protoplasts that make them a good model for the aleurone layer are demonstrated. The soluble proteome of the isolated protoplasts was shown to be highly similar to that of the intact tissue. The cell death response of the protoplasts to treatment with gibberellic acid, which involves increased vacuolization, was visualized as increasingly transparent protoplasts under microscopy. The use of isolated protoplasts offers several applications of great potential to further understand the behaviour of the aleurone layer during germination and thus during the malting process. Copyright © 2016 The Institute of Brewing & Distilling

Germination of barley grain is central to the malting industry and is a valuable model for cereal grain germination. Our current understanding of the complexity of germination at the molecular level is facilitated by access to genomic, transcriptomic, proteomic and metabolomic data. Here we review recent progress in barley germination research and discuss the factors to be considered when designing ‘omics’ experiments and interpreting the results. These factors include the structural and functional relationships between the various tissues of the barley caryopsis and the timing of the events of germination in the context of industrial malting. For transcriptomics, recent advances in sequencing the barley genome allow next-generation sequencing approaches to reveal novel effects of variety and environment on germination. For proteomics, selection of the source tissue(s) and the protein extraction conditions continue to be key to discovering the roles of individual protein forms and posttranslational modifications, such as glycosylation. Activity-based proteomics, particularly in combination with new gene editing technologies, has great potential to elucidate the network of enzymes in barley germination. Lastly, the application of metabolomics to barley grain germination provides essential data on biochemical processes, including insights into the formation of compounds that contribute to malt quality. To maximize the benefits of the ‘omics’ revolution to the malting industry, there is a need to integrate these data, taking into account barley variety, time, tissue, and specific physiological processes.

Cocoa butter is a key ingredient in many chocolate products but its partial substitution with mango (Mangifera indica L.) seed kernel fat (MSKF) has the potential to reduce chocolate production costs and improve shelf-life. Here, MSKF was extracted from three cultivars of mango grown in Pakistan: Lal Badshah, Anwar Retual, and Chaunsa. Physicochemical and antioxidant properties of the MSKF samples were studied at 0, 30, and 60 days of storage at 30 °C, a temperature reflecting typical storage conditions in the tropics. Overall, the Lal Badshah MSKF had the most favorable physicochemical properties, including the highest DPPH antioxidant activity among the three cultivars. Thus, Lal Badshah MSKF was used to formulate cocoa butter substitute chocolate (CBSC), substituting the cocoa butter at 20 to 80 g/100 g. CBSC had a lower value for hardness (3.80 N) compared with the control chocolate (4.42 N). Color values L* , a* , and b* were not significantly affected by the different rates of substitution or by length of storage. Oxidative stability and antioxidant potential of CBSC increased with both higher substitution levels of MSKF and length of storage. The results suggest that MSKF can be utilized as a cocoa butter substitute at levels up to 60 g/100 g. This potential for substitution is particularly valuable for tropical regions where refrigerated storage may not be available or financially viable. PRACTICAL APPLICATION: Mango seed kernel fat (MSKF) has potential to be used as a cocoa butter substitute in confectionery products, particularly chocolate. The mango industry could utilize fat extraction from mango seeds, which are normally a waste product, for value adding.; © 2019 Institute of Food Technologists®.

Durum wheat semolina (DWS) can be enriched with legume flours to produce more nutritious but high-quality pasta. DWS was substituted with detoxified matri (Lathyrus sativus) flour (DMF) at 5–25%, which in spaghetti increased the levels of protein, lipid, fibre and ash but decreased nitrogen-free extract. Water absorption, arrival time and dough development time increased from 63.1 to 69.2%, 1.7 to 2.4 and 2.3 to 3.3 min, respectively, while dough stability, consistency and tolerance index decreased. DMF addition increased cooking loss (4.8–5.8%) and hardness (13.2–16.5 N) but decreased percent rehydration. Based on farinographic (departure time), cooking quality (adhesiveness) and cooking loss thresholds for DMF at 15%, the effects of xanthan gum (XG) addition on the cooking qualities of the corresponding spaghetti were determined. XG up to 3% limited cooking loss (4.97 vs 5.4%) and improved hardness, compared to samples lacking XG. Considering functional, cooking and nutritional properties of spaghetti, incorporation of 15% DMF and 3% XG appeared optimal.

Concentrations of polyphenolic compounds and free, conjugated and bound phenolic acids were determined in six red grain sorghum varieties harvested on the Liverpool Pains in 2009. Categories of polyphenolic compounds quantified included total phenolics, anthocyanins, flavan-4ols, luteolinidin, apigeninidin, 5-methoxy-luteolinidin, 7-methoxy-apigeninidin, apigenin, luteolin, eriodictyol and naringenin. Phenolic acids that were quantified included p-hydroxybenzoic, vanillic, caffeic, p-coumaric, ferulic, syringic and sinapic acids. These sorghums were also extensively characterised for additional properties including rapid visco-analysis (RVA) of starch pasting profiles, protein solubility and quantification of kafirin and phytate. A pigmented testa was not detected in the six varieties by the quantal Clorox bleach test, which indicated that they were Type I sorghums that do not possess condensed tannin. This was confirmed by vanillin assays. The six varieties were incorporated into unprocessed sorghum–casein mash diets and offered to broiler chickens to compare nutrient utilisation parameters and protein (N) digestibility coefficients. One objective of this study was to assess if non-tannin phenolic compounds influence energy utilisation in broiler chickens. Negative correlations between certain conjugated phenolic acids in sorghum and parameters of energy utilisation (AME, ME:GE ratios, AMEn) in birds were observed. For example, p-coumaric (r = −0.826), ferulic (r = −0.831) and total conjugated phenolic acids (r = −0.832) were negatively correlated with ME:GE ratios to significant (P

The Liverpool Plains is a fertile agricultural region in New South Wales, Australia. Two sorghums from the 2009 Liverpool Plains harvest, sorghums #3 and #5, were extensively characterised which included concentrations of kafirin and phenolic compounds plus rapid visco-analysis (RVA) starch pasting profiles. Diets based on these two sorghums were formulated to be iso-nitrogenous and iso-energetic and were offered to male Ross 308 broiler chicks from 7 to 28 days post--hatch as either intact pellets or reground mash following steam-pelleting at conditioning temperatures of either 65 or 97°C. Thus the feeding study consisted of a 2 × 2 × 2 factorial array of dietary treatments: two sorghum varieties, two feed forms and two conditioning temperatures. Each of the eight treatments was replicated six times with six birds per replicate cage. Assessed parameters included growth performance, nutrient utilisation, apparent starch and protein (N) digestibility coefficients and disappearance rates from the distal jejunum and distal ileum. Intact pellets supported higher (P < 0.001) feed intakes and weight gains by 9.83 and 9.08%, respectively, than reground mash diets. Feed conversion ratios of broilers offered diets steam-conditioned at 97°C were 2.46% inferior (P < 0.001) in comparison to 65°C diets and both apparent metabolizable energy (AME) and N-corrected AME (AMEn) were compromised. Broilers offered sorghum #3-based diets significantly (P < 0.001) outperformed their sorghum #5 counterparts in terms of weight gain by 3.75% (1,334 versus 1,223 g/bird), FCR by 4.81% (1.524 versus 1.601), AME by 1.06 MJ (13.61 versus 12.55 MJ/kg), ME:GE ratio (ME:GE) by 4.81% (0.806 versus 0.769) and AMEn by 1.03 MJ (12.38 versus 11.35 MJ/kg). The inferiority of sorghum #5 appeared to be associated with higher concentrations of kafirin (61.5 versus 50.7 g/kg) and conjugated phenolic acids, including ferulic acid (31.1 versus 25.6 µg/g). There were no significant differences in jejunal and ileal starch and protein (N) digestibility coefficients between the two sorghums. However, starch to protein (N) disappearance rate ratios from the distal jejunum were significantly (P < 0.001) correlated with ME:GE and AME. The multiple linear regression equations indicated that energy utilisation was enhanced by coupling rapidly digestible protein with slowly digestible starch, which suggests that bilateral bioavailability of starch and protein is pivotal to efficient energy utilisation. Keywords: Broiler chickens, Ferulic acid, Kafirin, Protein, Sorghum, Starch

Coleoptiles of rice (Oryza sativa) seedlings grown under water commonly elongate by up to 1 mm h(-1) to reach the atmosphere. We initially analysed this highly specialized phenomenon by measuring epidermal cell lengths along the coleoptile axis to determine elongation rates. This revealed a cohort of cells in the basal zone that elongated rapidly following emergence from the embryo, reaching 200 μm within 12 h. After filming coleoptiles in vivo for a day, kinematic analysis was applied. Eight time-sliced 'segments' were defined by their emergence from the embryo at four-hourly intervals, revealing a mathematically simple growth model. Each segment entering the coleoptile from the embryo elongated at a constant velocity, resulting in accelerating growth for the entire organ. Consistent with the epidermal cell lengths, relative rates of elongation (mm mm(-1) h(-1)) were tenfold greater in the small, newly emerged basal segments than the older distal tip segments. This steep axial gradient defined two contrasting growth zones (bases versus tips) in which we measured ATP production and protein, RNA and DNA content, and analysed the global transcriptome under steady-state normoxia, hypoxia (3% O2) and anoxia. Determination of the transcriptome revealed tip-specific induction of genes encoding TCP [Teosinte Branched1 (Tb1) of maize, Cycloidea (Cyc), and Proliferating Cell Factor (Pcf)] transcription factors, RNA helicases, ribosomal proteins and proteins involved in protein folding, whilst expression of F-box domain-containing proteins in the ubiquitin E3-SCF complex (Skp, Cullin, F-box containing complex) was induced specifically in bases under low oxygen conditions. We ascribed the sustained elongation under hypoxia to hypoxia-specific responses such as controlled suppression of photosystem components and induction of RNA binding/splicing functions, indicating preferential allocation of energy to cell extension.

Plants belonging to the genus Taraxacum have been used in traditional healthcare to treat infectious diseases including food-borne infections. This review aims to summarize the available information on Taraxacum spp., focusing on plant cultivation, ethnomedicinal uses, bioactive phytochemicals, and antimicrobial properties. Phytochemicals present in Taraxacum spp. include sesquiterpene lactones, such as taraxacin, mongolicumin B, and taraxinic acid derivatives; triterpenoids, such as taraxasterol, taraxerol, and officinatrione; and phenolic derivatives, such as hydroxycinnamic acids (chlorogenic, chicoric, and caffeoyltartaric acids), coumarins (aesculin and cichoriin), lignans (mongolicumin A), and taraxacosides. Aqueous and organic extracts of different plant parts exhibit promising in vitro antimicrobial activity relevant for controlling fungi and Gram-positive and Gram-negative bacteria. Therefore, this genus represents a potential source of bioactive phytochemicals with broad-spectrum antimicrobial activity. However, so far, preclinical evidence for these activities has not been fully substantiated by clinical studies. Indeed, clinical evidence for the activity of Taraxacum bioactive compounds is still scant, at least for infectious diseases, and there is limited information on oral bioavailability, pharmacological activities, and safety of Taraxacum products in humans, though their traditional uses would suggest that these plants are safe.

The effects of phytase supplementation on growth performance, nutrient utilisation, starch and protein digestive dynamics in broiler chickens offered maize-, sorghum- and wheat-based diets were determined in a previous study (Liu et al., 2014). Responses to phytase were most pronounced in maize-based diets, which suggest that more phytate was degraded in these diets. Relevant retained samples of grain, diets and digesta from four small intestinal segments were retrospectively analysed for concentrations of phytate, sodium and starch pasting properties to investigate the hypothesis that phytate in maize-based diets was more completely degraded by exogenous phytase. Exogenous phytase significantly (P

Seven sorghum-based broiler diets containing the reducing agent, sodium metabisulphite (SMBS) at levels of 0, 1.50, 2.25, 3.00, 3.75, 4.50 and 5.25 g/kg were prepared. The in vitro starch pasting properties of the sorghum-based diets and starch extracted from the diets were assessed by rapid viscosity analysis (RVA). The seven dietary treatments were offered to 294 male Ross 308 chicks from 10 to 24 days post-hatch to determine SMBS effects on starch and nitrogen digestion dynamics. Digesta samples were collected, freeze-dried and weighed for determination of digestibility coefficients and mean retention times in the proximal jejunum, distal jejunum, proximal ileum and distal ileum. The digestion kinetics of starch and nitrogen (N) were determined using an exponential mathematical model to relate digestibility coefficients to mean retention times in each small intestinal segment. SMBS inclusions linearly reduced apparent starch digestibility coefficients in the proximal jejunum (r = −0.444, P

Pomegranate (Punica granatum L.) seeds are normally waste products from pomegranate fruit processing but they are a valuable source of certain pharmaceutical and nutraceutical compounds. This study was performed to determine and compare the physico-chemical properties of oils of pomegranate seed extracted by cold pressing from the variety Torshe Malas Iran (TMOI) and two commercial oils from Iran (COI) and Turkey (COT). Fatty acid analysis indicated that all the oil samples had a dominant fatty acid, punicic acid, with the other prominent fatty acids being linoleic and oleic acids. The thermal behavior of the three oils was determined using differential scanning calorimetry, which gave melting points of TMOI (−12.70°C), COI (1.87°C), and COT (2.25°C). TMOI was found to have the most favorable chemical quality attributes, including a low peroxide value (4.67 meq/kg), low free fatty acid content (0.65% as punicic acid), and high total phenolic content (10.44 mg GAE/g sample), followed by COT. Analysis of flavor compounds was conducted using fast gas chromatography-surface acoustic wave technology, which allowed detection of 13 distinct aromas among the three oils. Overall, the physico-chemical properties of the three oils were superior to those of pomegranate seed oils extracted using organic solvents reported previously. Practical application: Medical applications of pomegranate seed oil are gaining increasing attention, particularly due to the health benefits of punicic acid. Pomegranate seed oil is also highly suited to specific industrial applications, such as tanning. Despite its importance, little research has been conducted on the physico-chemical properties of the oil extracted by cold press compared to conventional extraction methods and super critical CO2 extraction. This study shows that superior quality pomegranate seed oil can be extracted simply and without using organic solvents. Extraction of the highly nutritious cold-pressed oil from pomegranate seeds could enable the generation of substantial revenue for pomegranate processing factories without associated environmental impacts.

The effects of plant growth conditions on concentrations of proteins, including allergens, in peanut ( Arachis hypogaea L.) kernels are largely unknown. Peanuts (cv. Walter) were grown at five sites (Taabinga, Redvale, Childers, Bundaberg, and Kairi) covering three commercial growing regions in Queensland, Australia. Differences in temperature, rainfall, and solar radiation during the growing season were evaluated. Kernel yield varied from 2.3 t/ha (Kairi) to 3.9 t/ha (Childers), probably due to differences in solar radiation. Crude protein appeared to vary only between Kairi and Childers, whereas Ara h 1 and 2 concentrations were similar in all locations. 2D-DIGE revealed significant differences in spot volumes for only two minor protein spots from peanuts grown in the five locations. Western blotting using peanut-allergic serum revealed no qualitative differences in recognition of antigens. It was concluded that peanuts grown in different growing regions in Queensland, Australia, had similar protein compositions and therefore were unlikely to show differences in allergenicity.

Phenolic compounds are well-known phytochemicals found in all plants. They consist of simple phenols, benzoic and cinnamic acid, coumarins, tannins, lignins, lignans and flavonoids. Substantial developments in research focused on the extraction, identification and quantification of phenolic compounds as medicinal and/or dietary molecules have occurred over the last 25 years. Organic solvent extraction is the main method used to extract phenolics. Chemical procedures are used to detect the presence of total phenolics, while spectrophotometric and chromatographic techniques are utilized to identify and quantify individual phenolic compounds. This review addresses the application of different methodologies utilized in the analysis of phenolic compounds in plant-based products, including recent technical developments in the quantification of phenolics.

Background The plant species Aristolochia indica (AI), Melilotus indicus (MI), Tribulus terrestris (TT) and Cuscuta pedicellata (CP) are widely used in folk medicine in the villages around Chowk Azam, South Punjab, Pakistan. The aim of this study was to evaluate the antioxidant activity, phytochemical composition, and the antibacterial, antifungal, cytotoxic and anti-inflammatory potential of the four medicinal plants listed above. For CP stem, this study represents (to the best of our knowledge) the first time phytochemicals have been identified and the antioxidant and anti-inflammatory potential determined. Methods Phytochemicals were analyzed through chemical tests, thin layer chromatography (TLC) and spectrophotometric methods. Antioxidant activities (DPPH and H2O2) were also determined through spectrophotometric methods. Extracts were evaluated for antibacterial potential via the agar well diffusion method against Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumonia and Acinetobacter baumannii. The minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) were determined by the microdilution method. Antifungal activities were tested using the agar tube dilution method against three species: Aspergillus fumigatus, Aspergillus flavus and Rhizopus oryzae. The cytotoxic potential of the plant extracts was checked using the brine shrimp assay. In vitro anti-inflammatory activity of the selected plant extracts was evaluated using albumin denaturation, membrane stabilization and proteinase inhibitory assays. Results Of all the methanolic extracts tested, those from CP (stem) and TTF (T. terrestris fruit) had the highest phenolic, flavonoid and flavonol contents (497±4 mg GAE/g, 385±8 mg QE/g and 139±4 mg QE/g; 426±5 mg GAE/g, 371±8 mg QE/g and 138±6 mg QE/g, respectively) and also exhibited strong antioxidant potential in scavenging DPPH and hydrogen peroxide (IC50 values; 20±1 and 18±0.7 μg/mL; 92±2 and 26±2 μg/mL, respectively). CP, TTF and TTL (T. terrestris leaf) extracts substantially inhibited the growth of the bacteria A. baumannii, S. aureus, and K. pneumonia and also exhibited the highest antifungal potential. The ranking of the plant extracts for cytotoxicity was TTF > TTL > AI > CP > MI, while the ranking for in vitro anti-inflammatory potential at a concentration of 200 μg/mL of the selected plant extracts was CP > TTL, TTF > AI > MI. The lowest IC50 (28 μg/mL) observed in the albumin denaturation assay was for CP. Positive correlations were observed between total phenolics, antioxidants, antibacterial, antifungal and anti-inflammatory potential of the selected plant extracts, indicating a significant contribution of phenolic compounds in the plant extracts to these activities. Conclusions This study revealed the strong antimicrobial, antioxidant, cytotoxic and anti-inflammatory potential of the plant species CP and TT used in folk medicine.

During barley germination, the aleurone layer secretes most of the enzymes required to degrade the endosperm, many of which are yet to be characterized. We used activity-based protein profiling (ABPP) to detect a range of active enzymes extracted from aleurone layers isolated from grains of a commercial malting barley variety incubated with or with out gibberellic acid (GA). Enzymes found to be induced by GA were putative aleurains, cathepsin-B-like proteases and serine hydrolases. By using an inhibitory sugar panel, a specific active retaining β-glycosidase in the barley aleurone was identified as a xylanase. Our results show that ABPP can be used rapidly to identify a variety of active enzyme isoforms in cereal aleurone without the need for enzyme purification.

The clinical importance of peanut (Arachis hypogaea) allergies demands standardized allergen extraction protocols. We determined the effectiveness of common extraction conditions (20 buffers, defatting reagents, extraction time/temperatures, processing, extraction repeats) on crude protein and Ara h 1 and 2 yields. Despite similar 1D-gel profiles, defatting with n-hexane resulted in significantly higher yields of crude protein, Ara h 1, and Ara h 2 than with diethyl ether. The yields were affected by the composition and pH of the extraction buffers and other conditions, but crude protein yield did not always correlate with Ara h 1 and 2 yields. Denaturants, reducing agents, acidic buffers, and thermal processing of peanuts perturbed allergen quantification in ELISAs, probably via exposure of additional epitopes. Allergen detection in 2D-Western blots with PBS resulted in greater sensitivity than with TBS or Tris. We recommend that allergen extraction conditions be selected based on the research question being investigated.

Oxygen deprivation limits the energy available for cellular processes and yet no comprehensive ATP budget has been reported for any plant species under O(2) deprivation, including Oryza sativa. Using 3-d-old coleoptiles of a cultivar of O. sativa tolerant to flooding at germination, (i) rates of ATP regeneration in coleoptiles grown under normoxia (aerated solution), hypoxia (3% O(2)), and anoxia (N(2)) and (ii) rates of synthesis of proteins, lipids, nucleic acids, and cell walls, as well as K(+) transport, were determined. Based on published bioenergetics data, the cost of synthesizing each class of polymer and the proportion of available ATP allocated to each process were then compared. Protein synthesis consumed the largest proportion of ATP synthesized under all three oxygen regimes, with the proportion of ATP allocated to protein synthesis in anoxia (52%) more than double that in normoxic coleoptiles (19%). Energy allocation to cell wall synthesis was undiminished in hypoxia, consistent with preferential elongation typical of submerged coleoptiles. Lipid synthesis was also conserved strongly in O(2) deficits, suggesting that membrane integrity was maintained under anoxia, thus allowing K(+) to be retained within coleoptile cells. Rates of protein synthesis in coleoptiles from rice cultivars with contrasting tolerance to oxygen deficits (including mutants deficient in fermentative enzymes) confirmed that synthesis and turnover of proteins always accounted for most of the ATP consumed under anoxia. It is concluded that successful establishment of rice seedlings under water is largely due to the capacity of coleoptiles to allocate energy to vital processes, particularly protein synthesis.

Pomegranate oil is the unique edible oil produced from the seed of the pomegranate fruit (Punica granatum L.), which can be extracted by various methods including supercritical CO2, superheated hexane, cold pressing, microwaves and ultrasound. The oil composition of pomegranate seed has received particular attention recently due to its content of antioxidant and anticancer agents, which may be important components for the production of new medicines and cosmetics.

Protease inhibitors of the serpin family are ubiquitous in the plant kingdom but relatively little is known about their biological functions in comparison with their counterparts in animals. X-ray crystal structures have provided crucial insights into animal serpin functions. The recently solved structure of AtSerpin1 from Arabidopsis thaliana, which has the highly conserved reactive center P2-P1' Leu-Arg-Xaa (Xaa = small residue), displays both conserved and plant-specific serpin features. Sequence homology suggests that AtSerpin1 belongs to serpin Clade B, composed of intracellular mammalian serpins, which is consistent with the lack of strong evidence for secretion of serpins from plant cells. The major in vivo target protease for AtSerpin1 is the papain-like cysteine RD21 protease, a match reminiscent of the inhibition of cathepsins K, L and S by the Clade-B mammalian serpin, SCCA-1 (SERPINB3). The function of AtSerpin1 and other serpins that contain P2-P1' Leu-Arg-Xaa (the 'LR' serpins) in plants remains unknown. However, based on its homology and interactive partners, AtSerpin1 and perhaps other serpins are likely to be involved in regulating programmed cell death or associated processes such as senescence. Abundant accumulation of serpins in seeds and their presence in phloem sap suggest additional functions in plant defense by irreversible inhibition of digestive proteases from pests or pathogens. Here we review the most recent findings in plant serpin biology, focusing on advances in describing the structure and inhibitory specificity of the LR serpins.

In animals, protease inhibitors of the serpin family are associated with many physiological processes, including blood coagulation and innate immunity. Serpins feature a reactive center loop (RCL), which displays a protease target sequence as a bait. RCL cleavage results in an irreversible, covalent serpin-protease complex. AtSerpin1 is an Arabidopsis protease inhibitor that is expressed ubiquitously throughout the plant. The x-ray crystal structure of recombinant AtSerpin1 in its native stressed conformation was determined at 2.2 A. The electrostatic surface potential below the RCL was found to be highly positive, whereas the breach region critical for RCL insertion is an unusually open structure. AtSerpin1 accumulates in plants as a full-length and a cleaved form. Fractionation of seedling extracts by nonreducing SDS-PAGE revealed the presence of an additional slower migrating complex that was absent when leaves were treated with the specific cysteine protease inhibitor L-trans-epoxysuccinyl-L-leucylamido (4-guanidino)butane. Significantly, RESPONSIVE TO DESICCATION-21 (RD21) was the major protease labeled with the L-trans-epoxysuccinyl-L-leucylamido (4-guanidino)butane derivative DCG-04 in wild type extracts but not in extracts of mutant plants constitutively overexpressing AtSerpin1, indicating competition. Fractionation by nonreducing SDS-PAGE followed by immunoblotting with RD21-specific antibody revealed that the protease accumulated both as a free enzyme and in a complex with AtSerpin1. Importantly, both RD21 and AtSerpin1 knock-out mutants lacked the serpin-protease complex. The results establish that the major Arabidopsis plant serpin interacts with RD21. This is the first report of the structure and in vivo interaction of a plant serpin with its target protease.

Germination in the soft wheat (Triticum aestivum L.) cultivar Rosella was followed for three days after imbibition by proteomic analysis of the germ tissue. Two-dimensional electrophoresis was performed in triplicate for proteins extracted from embryos dissected from mature grain and from grains germinated for 1, 2, and 3 days. For this period, 63 proteins (in 86 spots) in the germ were identified as decreasing in abundance, 35 proteins (in 60 spots) as increasing in abundance, and 28 proteins (in 39 spots) as exhibiting no significant abundance change. Proteins with significant abundance changes are discussed in relation to physiology; these include proteases, amylases and amylase inhibitors, enzymes in lipid metabolism, proteins related to water stress, cell wall hydrolases, oxalate oxidase, and H+-ATPases. Functions associated with proteins synthesized during the germination period are inconsistent with the embryo of mature grain being fully primed for germination.

Aims Identification of fungi isolated from koala faeces and screening for their enzyme activities of biotechnological interest. Methods and results Thirty-seven fungal strains were isolated from koala faeces and identified by the amplification and direct sequencing of the internal transcribed spacer (ITS) region of the ribosomal DNA. The fungi were screened for selected enzyme activities using agar plates containing a single substrate for each target class of enzyme. For xylanase, endoglucanase, ligninase (ligninolytic phenoloxidase) and protease over two-thirds of the isolates produced a clearing halo at 25 degrees C, indicating the secretion of active enzyme by the fungus, and one-third produced a halo indicating amylase, mannanase and tannase activity. Some isolates were also able to degrade crystalline cellulose and others displayed lipase activity. Many of the fungal isolates also produced active enzymes at 15 degrees C and some at 39 degrees C. Conclusions Koala faeces, consisting of highly lignified fibre, undigested cellulose and phenolics, are a novel source of fungi with high and diverse enzyme activities capable of breaking down recalcitrant substrates. Significance and impact of the study To our knowledge, this is the first time fungi from koala faeces have been identified using ITS sequencing and screened for their enzyme activities.