Ameliorating the effects of multiple stresses on agronomic traits in crops: modern biotechnological and omics approaches.

While global climate change poses a significant environmental threat to agriculture, the increasing population is another big challenge to food security. To address this, developing crop varieties with increased productivity and tolerance to biotic and abiotic stresses is crucial. Breeders must identify traits to ensure higher and consistent yields under inconsistent environmental challenges, possess resilience against emerging biotic and abiotic stresses and satisfy customer demands for safer and more nutritious meals. With the advent of omics-based technologies, molecular tools are now integrated with breeding to understand the molecular genetics of genotype-based traits and develop better climate-smart crops. The rapid development of omics technologies offers an opportunity to generate novel datasets for crop species. Identifying genes and pathways responsible for significant agronomic traits has been made possible by integrating omics data with genetic and phenotypic information. This paper discusses the importance and use of omics-based strategies, including genomics, transcriptomics, proteomics and phenomics, for agricultural and horticultural crop improvement, which aligns with developing better adaptability in these crop species to the changing climate conditions.

Indigenous Australian grass seeds as grains: macrostructure, microstructure and histochemistry.

Utilization of grains of local grasses by Australia's First Nations people for food and connection to Country has largely been lost due to colonization. Native Australian grain production has the potential to deliver environmental, economic, nutritional and cultural benefits to First Nations people and the wider community. Revitalization of the native grain food system can only be achieved if relevant properties of the grains are elucidated. This study aimed to characterize the grain structure and histochemistry of four Australian native grasses: Dactyloctenium radulans (Button Grass), Astrebla lappacea (Curly Mitchell Grass), Panicum decompositum (Native Millet) and Microlaena stipoides (Weeping Grass). For these species, as well as wheat and sorghum, whole-grain images were obtained via stereo microscopy, starch and the embryo were visualized, and sections of fixed grains were imaged via bright-field and fluorescence microscopy. The shape, size and colour of the whole native grains varied between the species. The aleurone layer was one-cell thick in the native species, as in the domesticated grains, except for Weeping Grass, which had a two-cell-thick aleurone. In the native grains, endosperm cell walls appeared thinner than in wheat and sorghum. Starch granules in Button Grass, Curly Mitchell Grass and Native Millet were found mainly in the central region of the starchy endosperm, with very few granules in the sub-aleurone layer, whereas Weeping Grass had abundant starch in the sub-aleurone. Protein appeared most abundant in the aleurone and sub-aleurone layers of the native grains, although in Button Grass, the starchy endosperm was observed to be rich in protein, as in wheat and sorghum. As a proportion of the whole grain, the embryo was larger in the native species than in wheat. The differences found in the grain properties among the four native Australian species have important implications for the agri-food industry in a changing climate.

Genes Encoding Structurally Conserved Serpins in the Wheat Genome: Identification and Expression Profiles during Plant Development and Abiotic and Biotic Stress.

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.

Sorghum in foods: Functionality and potential in innovative products.

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.

Rhizosphere bacteria associated with Chenopodium quinoa promote resistance to Alternaria alternata in tomato.

Microorganisms can interact with plants to promote plant growth and act as biocontrol agents. Associations with plant growth-promoting rhizobacteria (PGPR) enhance agricultural productivity by improving plant nutrition and enhancing protection from pathogens. Microbial applications can be an ideal substitute for pesticides or fungicides, which can pollute the environment and reduce biological diversity. In this study, we isolated 68 bacterial strains from the root-adhering soil of quinoa (Chenopodium quinoa) seedlings. Bacterial strains exhibited several PGPR activities in vitro, including nutrient solubilization, production of lytic enzymes (cellulase, pectinase and amylase) and siderophore synthesis. These bacteria were further found to suppress the mycelial growth of the fungal pathogen Alternaria alternata. Nine bacterial strains were selected with substantial antagonistic activity and plant growth-promotion potential. These strains were identified based on their 16S rRNA gene sequences and selected for in planta experiments with tomato (Solanum lycopersicum) to estimate their growth-promotion and disease-suppression activity. Among the selected strains, B. licheniformis and B. pumilus most effectively promoted tomato plant growth, decreased disease severity caused by A. alternata infection by enhancing the activities of antioxidant defense enzymes and contributed to induced systemic resistance. This investigation provides evidence for the effectiveness and viability of PGPR application, particularly of B. licheniformis and B. pumilus in tomato, to promote plant growth and induce systemic resistance, making these bacteria promising candidates for biofertilizers and biocontrol agents.

The effects of thymoquinone on pancreatic cancer: Evidence from preclinical studies.

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.

Carotenoid biosynthesis and the evolution of carotenogenesis genes in rust fungi.

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.

Combined ability of salicylic acid and spermidine to mitigate the individual and interactive effects of drought and chromium stress in maize (Zea mays L.).

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.

Induction of defense-related enzymes and enhanced disease resistance in maize against Fusarium verticillioides by seed treatment with Jacaranda mimosifolia formulations.

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.

GC-MS analysis, antimicrobial, antioxidant, antilipoxygenase and cytotoxic activities of Jacaranda mimosifolia methanol leaf extracts and fractions.

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.

Combined seed and foliar pre-treatments with exogenous methyl jasmonate and salicylic acid mitigate drought-induced stress in maize.

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.

Salt-Treated Roots of Oryza australiensis Seedlings are Enriched with Proteins Involved in Energetics and Transport.

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 <0.05) are found. Gene Ontology (GO) analysis shows that proteins categorized as "metabolic process," "transport," and "transmembrane transporter" are highly responsive to salt treatment. In particular, mitochondrial ATPases and SNARE proteins are more abundant in roots of the salt-tolerant accession and responded strongly when roots are exposed to salinity. mRNA quantification validated the elevated protein abundances of a monosaccharide transporter and an antiporter observed in the salt-tolerant genotype. The importance of the upregulated monosaccharide transporter and a VAMP-like protein by measuring salinity responses of two yeast knockout mutants for genes homologous to those encoding these proteins in rice are confirmed. Potential new mechanisms of salt tolerance in rice, with implications for breeding of elite cultivars are also discussed.

Mango Seed Kernel Fat as a Cocoa Butter Substitute Suitable for the Tropics.

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.

Carotenoid complement of rust spores: Variation among species and pathotype.

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, ß-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:ß-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.

Salinity tolerance in Australian wild Oryza species varies widely and matches that observed in O. sativa.

BACKGROUND: Soil salinity is widespread in rice-producing areas globally, restricting both vegetative growth and grain yield. Attempts to improve the salt tolerance of Asian rice, Oryza sativa-the most salt sensitive of the major cereal crops-have met with limited success, due to the complexity of the trait and finite variation in salt responses among O. sativa lines. Naturally occurring variation among the more than 20 wild species of the Oryza genus has great potential to provide breeders with novel genes to improve resistance to salt. Here, through two distinct screening experiments, we investigated variation in salinity tolerance among accessions of two wild rice species endemic to Australia, O. meridionalis and O. australiensis, with O. sativa cultivars Pokkali and IR29 providing salt-tolerant and sensitive controls, respectively. RESULTS: Rice plants were grown on soil supplemented with field-relevant concentrations of NaCl (0, 40, 80, and 100 mM) for 30 d, a period sufficient to reveal differences in growth and physiological traits. Two complementary screening approaches were used: destructive phenotyping and high-throughput image-based phenotyping. All genotypes displayed clear responses to salt treatment. In the first experiment, both salt-tolerant Pokkali and an O. australiensis accession (Oa-VR) showed the least reduction in biomass accumulation, SES score and chlorophyll content in response to salinity. Average shoot Na+/K+ values of these plants were the lowest among the genotypes tested. In the second experiment, plant responses to different levels of salt stress were quantified over time based on projected shoot area calculated from visible red-green-blue (RGB) and fluorescence images. Pokkali grew significantly faster than the other genotypes. Pokkali and Oa-VR plants displayed the same absolute growth rate under 80 and 100 mM, while Oa-D grew significantly slower with the same treatments. Oa-VR showed substantially less inhibition of growth in response to salinity when compared with Oa-D. Senescence was seen in Oa-D after 30 d treatment with 40 mM NaCl, while the putatively salt-tolerant Oa-VR had only minor leaf damage, even at higher salt treatments, with less than a 40% increase in relative senescence at 100 mM NaCl compared to 120% for Oa-VR. CONCLUSION: The combination of our two screening experiments uncovered striking levels of salt tolerance diversity among the Australian wild rice accessions tested and enabled analysis of their growth responses to a range of salt levels. Our results validate image-based phenotyping as a valuable tool for quantitative measurement of plant responses to abiotic stresses. They also highlight the potential of exotic germplasm to provide new genetic variation for salinity tolerance in rice.

Ethnobotany of the genus Taraxacum-Phytochemicals and antimicrobial activity.

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.

A conditional silencing suppression system for transient expression.

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.

Fortification of durum wheat semolina with detoxified matri (Lathyrus sativus) flour to improve the nutritional properties of pasta.

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.

Effects of Sorghum Malting on Colour, Major Classes of Phenolics and Individual Anthocyanins.

Sorghum (Sorghum bicolor) grain contains many health-promoting phytochemicals, including a broad range of phenolic compounds. Malting of cereal grains is known to increase the bioavailability of macro- and micronutrients. However, the detailed effects of malting on sorghum grain anthocyanins, a major class of phenolics that influence the taste and colour of sorghum-based foods, requires further investigation. Eight commercial sorghum hybrids harvested from three regions in eastern Australia were malted and analysed for colour, tannin content, total phenolic content (TPC), flavan-4-ols, total flavonoids, total anthocyanins and 3-deoxyanthocyanins. Grains of all the sorghums were found to be tannin-free. Malting decreased the TPC of all samples. For TPC, the grand means among all the sorghum cultivars for raw and malted grain were 2.77 and 2.48 mg gallic acid equivalents (GAE)/g, respectively. For flavan-4-ols, the grand means for raw and malted sorghum grains were 2.98 and 2.23 abs/mL/g, respectively. Remarkably, total anthocyanin levels more than doubled upon malting whereas total flavonoid levels decreased by 12%. The average abundance of 3-deoxyanthocyanins in raw sorghum grains increased for about 8-fold upon malting. Our results will be valuable for sorghum breeders in the selection of lines for specific end uses and for food scientists developing sorghum-based products.

Antimicrobial activity, toxicity and anti-inflammatory potential of methanolic extracts of four ethnomedicinal plant species from Punjab, Pakistan.

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.