Background: Previous trials have displayed augmented intracellular vitamin C concentrations in the leukocytes at 24 h after acute supplementation with 1000 mg calcium ascorbate (CA), compared to ascorbic acid (AA). Objective: The primary objective was to evaluate comparative leukocyte vitamin C accumulation kinetics over 32 h following acute 250 mg or 500 mg doses from the two sources. Secondary objectives were to evaluate neutrophil phagocytic function and lymphocyte differentiation between the two sources of vitamin C. Methods: Ninety-three healthy females (250 mg, n = 27; 500 mg, n = 24) and males (250 mg, n = 19; 500 mg, n = 23) were assigned to ingest a single dose of CA or AA providing 250 mg or 500 mg of vitamin C in two separate double-blind, randomized crossover trials. Results: There were no significant differences in the primary or secondary outcomes between the two treatments in the 250 mg low-dose study. Conversely, there was evidence that ingestion of 500 mg of CA increased DHA in plasma, increased neutrophil functionality during the first 8 h of the PK study, promoted increased natural killer cells, and altered weight-adjusted PK profiles, suggesting greater volume distribution and clearance from the blood. Conclusions: These findings indicate that 500 mg of CA may promote some immune benefits compared to 500 mg of AA ingestion.

Biodegradable and antimicrobial food packaging is preferred by consumers and has the potential to improve food safety. In this study, we develop and test films based on pelargonic acid (PA) and eugenol (EUG) entrapped in a zein (Z) emulsion (size 146.57 nm) dispersed in sodium alginate (SA) as wrap to improve the physiochemical, and antibacterial properties The moisture content, solubility, absorption, and biodegradation of the films were evaluated. Thermal properties of films were determined by differential scanning calorimetry and thermogravimetric analysis. Our results indicate that Ca2+ treatment enhanced the mechanical properties, imparted hydrophobicity, and produced a rough surface on films, as visualized with scanning electron microscopy. The untreated films demonstrated significant antimicrobial activity against L. monocytogenes V7, with a reduction of similar to 6.0 in log (colony-forming units/mL) after 24 h. These biodegradable films with enhanced antimicrobial activity and improved thermal and mechanical properties have potential applications in sustainable food packaging systems.

In this study, we used accelerated solvent extraction (ASE) to extract flavonoids and anthocyanins from dried red onion peels, producing red onion peel extract (ROPE). High-performance liquid chromatography analysis of ROPE obtained using ASE identified important flavonols and anthocyanins. Total flavonoid and anthocyanin contents measured from ROPE were 459.15 mg/g +/- 16.45 (DW) and 33.14 mg/g +/- 0.99 (DW), respectively. ROPE antimicrobial activity against L. monocytogenes showed strain-specific effects, where the minimum inhibitory concentration (MIC) for V7 was two-fold greater than for EGD and Scott A (i.e., 25.82 vs. 12.91 mg quercetin-3, 4 '-diglucoside equivalents/gram [mg QdGE/g] ROPE). ROPE significantly inhibited (P < 0.05) biofilm formation by all three strains at sub-inhibitory concentrations (0.40-6.45 mg QdGE/g) in a concentration-dependent manner. Pre-formed biofilms of mixed L. monocytogenes strains on stainless steel were inactivated after 48 h at 22 C only with higher concentrations of ROPE (25.82-56.63 mg QdGE/g). Sub -inhibitory concentrations of ROPE also inhibited L. monocytogenes motility and suppressed violacein pigment production in Chromobacterium violaceum ATCC 12472, suggesting that ROPE inhibited quorum sensing. Therefore, our data indicate that red onion peel waste has potential as a source of natural antibacterial com-pounds as an approach to food safety in the circular economy.

Understanding belowground chemical interactions between plant roots and plant-parasitic nematodes is immensely important for sustainable crop production and soilborne pest management. Due to metabolic diversity and ever-changing dynamics of root exudate composition, the impact of only certain molecules, such as nematode hatching factors, repellents, and attractants, has been examined in detail. Root exudates are a rich source of biologically active compounds, which plants use to shape their ecological interactions. However, the impact of these compounds on nematode parasitic behavior is poorly understood. In this study, we specifically address this knowledge gap in two cyst nematodes, Globodera pallida , a potato cyst nematode and the newly described species, Globodera ellingtonae . Globodera pallida is a devastating pest of potato ( Solanum tuberosum ) worldwide, whereas potato is a host for G. ellingtonae , but its pathogenicity remains to be determined. We compared the behavior of juveniles (J2s) hatched in response to root exudates from a susceptible potato cv. Desirée, a resistant potato cv. Innovator, and an immune trap crop Solanum sisymbriifolium (litchi tomato – a wild potato relative). Root secretions from S. sisymbriifolium greatly reduced the infection rate on a susceptible host for both Globodera spp. Juvenile motility was also significantly influenced in a host-dependent manner. However, reproduction on a susceptible host from juveniles hatched in S. sisymbriifolium root exudates was not affected, nor was the number of encysted eggs from progeny cysts. Transcriptome analysis by using RNA-sequencing (RNA-seq) revealed the molecular basis of root exudate-mediated modulation of nematode behavior. Differentially expressed genes are grouped into two major categories: genes showing characteristics of effectors and genes involved in stress responses and xenobiotic metabolism. To our knowledge, this is the first study that shows genome-wide root exudate-specific transcriptional changes in hatched preparasitic juveniles of plant-parasitic nematodes. This research provides a better understanding of the correlation between exudates from different plants and their impact on nematode behavior prior to the root invasion and supports the hypothesis that root exudates play an important role in plant-nematode interactions.

Accurate, rapid quantitation of the capsaicinoid and capsinoid compounds produced by peppers (Capsicum spp.) is essential to assess quality. Here, we developed a rapid ultra-high performance liquid chromatography method for the simultaneous separation of five major capsaicinoids and three major capsinoids from peppers. Optimal chromatographic separation was achieved using a phenyl-hexyl stationary phase with a mobile phase of acidified water and methanol with a flow rate of 0.5 ml/min at a column temperature of 55 degrees C over 5 min. The method was validated by testing linearity, precision, robustness, and limits of detection and quantification. The developed method was successfully employed to profile capsaicinoids and capsinoids from different pepper cultivars. Out of the 10 pepper cultivars analysed, all three major capsinoids were detected in two cultivars. To the best of our knowledge, this is the first report of successful separation of nordihydrocapsiate from capsiate and quantification of nordihydrocapsiate.

Solanum glycoalkaloids are gaining increased scientific attention due to their bioactive potential in the defense of plants against pests and pathogens. The comprehensive glycoalkaloid profiling from the leaves, stems, and roots of seven underexploited Solanum species (S. caripense, S. melanocerasum, S. muricatum, S. nigrum, S. quitoense, S. retroflexum, and S. sisymbriifolium) was conducted using high-performance liquid chromatography–time-of-flight mass spectrometry. A total of 51 glycoalkaloids were shared among the studied Solanum species, with concentrations ranging from 7 to 5.63 × 105 ng g−1. Based on the glycoalkaloid composition, plants were separated into two clusters, Cluster 1 (S. melanocerasum, S. nigrum, and S. retroflexum) and Cluster 2 (S. caripense, S. muricatum, S. quitoense, and S. sisymbriifolium). The inhibition activity of glycoalkaloid extracts on acetylcholinesterase showed a half-maximal inhibitory concentration (IC50), ranging from 0.4 (S. nigrum stems) to 344.9 µg mL−1 (S. sisymbriifolium leaves), that was not directly correlated to the total glycoalkaloid contents. This suggests that the composition of glycoalkaloids in the plant extract, rather than the total concentration, is a driver of biological activity. The study provides a framework for the bioprospecting of underexploited Solanum species for exploring bioactive glycoalkaloids and other compounds with potential pesticidal activities for the development of green bioformulation. This is the first comprehensive report on the glycoalkaloid profiles of S. retroflexum.

Steroidal glycoalkaloids (SGAs) are phytoanticipins found in solanaceous crops that act as the first line of chemical defense against pathogen attacks. Solanum sisymbriifolium, a trap crop for potato cyst nematodes, has been shown to effectively reduce populations of Globodera pallida. S. sisymbriifolium contains α-solamargine and other solasodine-type glycoalkaloids that may contribute to plant defenses. This study evaluated the influence of solanaceous SGAs on G. pallida hatch, development, and reproduction. Exposure to α-solamargine and α-solamarine reduced G. pallida hatch by 65 and 87%, respectively. Exposure of G. pallida cysts with the glycoalkaloids α-solamargine and solasodine significantly reduced infection in susceptible potato 'Russet Burbank' by 98 and 94% compared with the control. Exposure of cysts to either solasodine or solamargine significantly reduced reproduction of G. pallida on 'Russet Burbank' by 99% compared with the control. The study demonstrated the deleterious effect of SGAs on G. pallida hatch, infection, and reproduction.

Potato cyst nematode (PCN) cysts consist of heterogenous populations of eggs, juveniles, and eggshells that make manual sorting of individual life stages cumbersome. The number of viable PCN eggs is a major determinant of crop damage. An accurate high-throughput PCN egg viability assay is useful for developing effective management and eradication plans. In this study, we present a method for rapid and precise enumeration and sorting of PCN eggs and juveniles, along with an egg viability assessment by staining eggs with the fluorescent stain, acridine orange, and sorting with the Complex Object Parametric Analyzer and Sorter (COPAS) system, a large particle flow cytometer. Both size sorting and fluorescent sorting capabilities of the COPAS were explored. By using the COPAS, sorting efficiency for eggs and preparasitic second-stage juveniles (J2s) was 97.6 and 97.2%, respectively, with 99% recovery at a flow rate of 15 events/s. Purity of sorted live and dead eggs was 95.5 and 94.1%, respectively. Sorting of J2s by size indicated that 15 to 16.4% of Globodera ellingtonae or G. pallida had an average body length of 436.1 ± 3.4 µm compared with an average size of 512.9 ± 4.4 µm for the majority of the J2 population for both species. A red autofluorescing J2 population was also identified through sorting. Sorting of eggs by flow cytometry did not significantly affect hatching (55.1 ± 1.2 and 53.9 ± 1.6%, respectively, for sorted or nonsorted eggs) or juvenile motility (91.3 ± 1.0 or 90.1 ± 1.1%, respectively), thus confirming that the method does not impair the biological activity of the nematode.

Potato cyst nematodes (PCNs), Globodera pallida and Globodera rostochiensis, are quarantine pests of potato which cause significant damage to production and farm gate revenue worldwide. Accurately assessing viability of PCN eggs is important for eradication and management programs. The goal of this study was to develop a quick and reliable fluorescent staining method to evaluate viability of G. pallida and Globodera ellingtonae eggs. The staining efficiency of eight fluorescent stains was evaluated using G. pallida eggs compared with the conventional Meldola’s Blue (MB) staining method. The staining efficiency of the fluorescent stains ranged from 80.33 ± 2.99 (Sytox Green) to 100% (Acridine Orange) for non-viable eggs. Two stains were further evaluated for their efficiency in assessing viability of encysted eggs from five different greenhouse-reared G. pallida cyst sources which contained both viable and non-viable eggs. For the G. pallida cyst sources, viability ofencysted eggs were estimated to be 41.02 ± 3.81 to 62.66% ± 3.12% when stained with Acridine Orange (AO) and 79.52% ± 1.54% viability for G. ellingtonae. Both staining time and stain concentration were significant for staining efficiency of released and encysted eggs. Staining time and concentration were optimized for released eggs at 4 h at 10 μg/ml and for encysted eggs at 16 h at 25 μg/ml respectively for AO. Fluorescent stains accurately and rapidly assessed percent egg viability and were determined to be as sensitive as a seven-day incubation with the Meldola’s Blue staining method.

Seven high yielding, new, promising lines of black pepper (Piper nigrum) were characterized using molecular markers (randomly amplified polymorphic DNA) and morphological features. Out of the 14 random decatner primers studied, 9 could generate unique bands in 6 lines. Maximum unique bands of the primers were observed in the line OPKrn followed by HP-1411 and HP-105. HP-780 could not be discriminated by any of the primers. Sixteen unique bands were prod uced by the nine primers making an average of 1.7 bands per primer. The lines OPKm, HP-1411 and HP-105 exhibited distinct morphological features also.

Sixty-five Solanum tuberosum group Andigena, Phureja and Stenotomum genotypes from an initial population of 1,500 were analyzed for phenylpropanoids, carotenoids, and antioxidant capacity. Total phenolic content ranged from 3 to 49 mg g−1 DW, total carotenoids from 4.1 to 154 μg/g DW, anthocyanins from 0.27 to 34 mg g−1 DW and antioxidant capacity from 60 to 1,767 μmol TE/g DW. HPLC analysis of phenolic extracts revealed that 5-O-chlorogenic acid (5CGA) was the most abundant polyphenol in all genotypes. Ten genotypes were independently grown out for more in-depth phytonutrient analysis. The Phureja genotypes RN 27.01 had the highest polyphenol, anthocyanin and antioxidant content, while RN 39.05 had the highest carotenoid content. The tuber percentage dry matter varied markedly among the ten genotypes, influencing the phytonutrient values when expressed on a dry weight basis. Chlorogenic acid concentrations ranged from 1.7 to 29.4 mg g−1 DW and kaempferol-3-rutinose was present up to 3 mg g−1 DW. Petunidin-3-O-coum-rutinoside-5-O-glu or pelargonidin-3-O-coum-rutinoside-5-O-glu were the most abundant anthocyanins. The principal carotenoids were lutein, zeaxanthin, violaxanthin, and antheraxanthin, but no one carotenoid was predominant in all genotypes. These findings further support utilization of Phureja group germplasm for phytonutrient enhancement efforts.

Phenylpropanoid metabolite and transcript expression during different developmental stages were examined in field grown potatoes. Carbohydrate and shikimic acid metabolism was assessed to determine how tuber primary metabolism influences phenylpropanoid metabolism. Phenylpropanoid concentrations were highest in immature tubers, as were some transcript levels and enzyme activities including phenylalanine ammonia lyase (PAL). Phenylpropanoid concentration differences between mature and immature tubers varied by genotype, but in some cases were approximately three-fold. The most abundant phenylpropanoid was chlorogenic acid (5CGA), which decreased during tuber maturation. Hydroxycinnamoyl-CoA:quinate hydroxycinnamoyl transferase (HQT) transcripts were highly expressed relative to other phenylpropanoid genes, but were not well correlated with 5CGA concentrations (r = -0.16), whereas HQT enzyme activity was. In contrast to 5CGA, less abundant chlorogenic isomers increased during development. Concentrations of hydroxycinnamic acid amides were higher in immature tubers, as was expression of arginine- and ornithine decarboxylases. Expression of several genes involved in carbohydrate or shikimate metabolism, including sucrose synthase and DAHP, showed similar developmental patterns to phenylpropanoid pools, as did shikimate dehydrogenase enzyme activity. Sucrose, glucose and fructose concentrations were highest in immature tubers. Exogenous treatment of potatoes with sugars stimulated phenylpropanoid biosynthesis, suggesting sugars contribute to the higher phenylpropanoid concentrations in immature tubers. These changes in phenylpropanoid expression suggest the nutritional value of potatoes varies during development. Published by Elsevier Masson SAS.

BACKGROUND:Plant secondary metabolites, including phenylpropanoids and carotenoids, are stress inducible, have important roles in potato physiology and influence the nutritional value of potatoes. The type and magnitude of environmental effects on tuber phytonutrients is unclear, especially under modern agricultural management that minimizes stress. Understanding factors that influence tuber secondary metabolism could facilitate production of more nutritious crops. Metabolite pools of over forty tuber phenylpropanoids and carotenoids, along with the expression of twenty structural genes, were measured in high-phenylpropanoid purple potatoes grown in environmentally diverse locations in North America (Alaska, Texas and Florida).RESULTS:Phenylpropanoids, including chlorogenic acid (CGA), were higher in samples from the northern latitudes, as was the expression of phenylpropanoid genes including phenylalanine ammonia lyase (PAL), which had over a ten-fold difference in relative abundance. Phenylpropanoid gene expression appeared coordinately regulated and was well correlated with metabolite pools, except for hydroxycinnamoyl-CoA:quinatehydroxcinnamoyl transferase (HQT; r = -0.24). In silico promoter analysis identified two cis-acting elements in the HQT promoter not found in the other phenylpropanoid genes. Anthocyanins were more abundant in Alaskan samples and correlated with flavonoid genes including DFR (r = 0.91), UFGT (r = 0.94) and F3H (r = 0.77). The most abundant anthocyanin was petunidin-3-coum-rutinoside-5-glu, which ranged from 4.7 mg g-1 in Alaska to 2.3 mg g-1 in Texas. Positive correlations between tuber sucrose and anthocyanins (r = 0.85), suggested a stimulatory effect of sucrose. Smaller variation was observed in total carotenoids, but marked differences occurred in individual carotenoids, which had over a ten-fold range. Violaxanthin, lutein or zeaxanthin were the predominant carotenoids in tubers from Alaska, Texas and Florida respectively. Unlike in the phenylpropanoid pathway, poor correlations occurred between carotenoid transcripts and metabolites.CONCLUSION:Analysis of tuber secondary metabolism showed interesting relationships among different metabolites in response to collective environmental influences, even under conditions that minimize stress. The variation in metabolites shows the considerable phenotypical plasticity possible with tuber secondary metabolism and raises questions about to what extent these pathways can be stimulated by environmental cues in a manner that optimizes tuber phytonutrient content while protecting yields. The differences in secondary metabolites may be sufficient to affect nutritional quality.

Potatoes from over fifty genotypes representing cultivars, breeding lines, primitive germplasm and wild species were analysed for phenolic content and hydrophilic antioxidant capacity. Genotypes with markedly higher amounts than the most commonly consumed potatoes were identified. Chlorogenic acid was the most abundant phenolic and ranged from 22 to 473 mg/100 g dry weight. Rutin and kaempferol-3-rutinose were the most abundant flavonols. Total phenolics ranged from 1.8 to 11 mg/g DW and antioxidant capacity ranged from 27 to 219 μmol TE/g DW. Total phenolics and antioxidants in these high-phytonutrient potatoes compared favourably to 15 other analysed vegetables. With the high per capita consumption of potatoes, widespread adoption of high-phytonutrient cultivars could significantly increase dietary intake of phytonutrients.

PurposeThis study aims to treat the development and application of sequence characterised amplified region (SCAR) markers for the detection of plant based adulterants (dried red beet pulp and powdered Ziziphus nummularia fruits) in traded ground chilli.Design/methodology/approachAdulterant‐specific DNA fragments (red beet pulp specific – “Beet 01” and Z. nummularia specific – “Ziz 01”) identified by random amplified polymorphic DNA polymerase chain reaction (RAPD‐PCR) analysis were cloned and sequenced for SCAR marker development. Red beet pulp specific SCAR primer pair, B1, and Z. nummularia specific SCAR primer pair, Z1, were designed from the corresponding RAPD marker sequences to amplify SCAR markers of 320 bp and 389 bp, respectively. The utility of the SCAR markers for adulterant detection was verified in model blends of chilli powder with the adulterants. Six commercial samples of ground chilli powder were analysed using the SCAR markers.FindingsSCAR markers could detect the adulterants at a concentration as low as 10 g adulterant kg−1 of blended sample. The Z. nummularia SCAR marker could detect the presence of Z. nummularia fruit adulteration in one of the commercial samples. All the market samples tested were free from red beet pulp adulteration.Practical implicationsThe PCR‐based method developed in the study is simple, rapid, and sensitive and has the potential to be developed into a quantitative analytical method and for commercial PCR kits for the large‐scale screening of ground chilli to detect and prevent plant‐based adulterants. The work has public health significance too, as ground chilli is one of the major spices consumed worldwide.Originality/valueThe study is the first report on the development of SCAR markers for adulterant detection in ground chilli. This work has relevance, as adulteration is a major concern of the sanitary and phytosanitary issues of the World Trade Organization (WTO) agreement.

Turmeric powder (Curcuma longa L.), an important medicinal spice product traded internationally, is subjected to adulteration by design or default with powders of related curcumin containing wild species like Curcuma zedoaria and Curcuma malabarica leading to toxicity and poor quality of the produce. The present study aims at development of specific, sensitive and reproducible Sequence Characterized Amplified Region (SCAR) markers to detect these adulterants in traded turmeric powder. Two putative RAPD markers, ‘Cur 01’ and ‘Cur 02’, generated by random primers OPA 01 and OPE 18 were identified as C. zedoaria/C. malabarica specific by comparative RAPD analysis of genuine turmeric and market samples of turmeric powder, C. zedoaria and C. malabarica. These specific RAPD markers were cloned and sequenced. Two pairs of SCAR primers were designed from the RAPD markers ‘Cur 01’ and ‘Cur 02’, respectively. Six market samples of turmeric powder and four simulated standards besides the genuine samples were analyzed using the specific SCAR markers. Both the SCAR markers detected the presence of C. zedoaria/C. malabarica adulteration in four market samples and all the simulated standards prepared in different concentrations. The two SCAR markers developed in the study would be potentially useful for the regulatory agencies to detect C. zedoaria/C. malabarica adulteration in traded turmeric powder. The analytical strategy being very simple could be used for large scale screening of turmeric powder samples intended for export and domestic uses.

Development of a robust set of 18 genomic microsatellite markers from turmeric (Curcuma longa L.) and its effective utilization in estimating the genetic diversity of 20 turmeric accessions are described. A total of 103 alleles were detected with an average of 5.7 alleles per locus. These markers displayed varied levels of polymorphism as evident from its discriminating power ranging from 0.19 to 0.70. The UPGMA cluster analysis of genetic distance values resolved the 20 turmeric accessions into five main groups. Three sets of genetically identical accessions were detected within the analyzed accessions, suggesting a revisit of the germplasm collection strategy based on vernacular identity. The entire grouping pattern of the entities was loose and independent of their geographical origins. These polymorphic SSR markers would be useful for the population genetic studies and germplasm management of turmeric.