Changes in rocket salad phytochemicals within the commercial supply chain: Glucosinolates, isothiocyanates, amino acids and bacterial load increase significantly after processing.
ABSTRACT: Five cultivars of Eruca sativa and a commercial variety of Diplotaxis tenuifolia were grown in the UK (summer) and subjected to commercial growth, harvesting and processing, with subsequent shelf life storage. Glucosinolates (GSL), isothiocyanates (ITC), amino acids (AA), free sugars, and bacterial loads were analysed throughout the supply chain to determine the effects on phytochemical compositions. Bacterial load of leaves increased significantly over time and peaked during shelf life storage. Significant correlations were observed with GSL and AA concentrations, suggesting a previously unknown relationship between plants and endemic leaf bacteria. GSLs, ITCs and AAs increased significantly after processing and during shelf life. The supply chain did not significantly affect glucoraphanin concentrations, and its ITC sulforaphane significantly increased during shelf life in E. sativa cultivars. We hypothesise that commercial processing may increase the nutritional value of the crop, and have added health benefits for the consumer.
Project description:Seven accessions of Eruca sativa ("salad rocket") were subjected to a randomised consumer assessment. Liking of appearance and taste attributes were analysed, as well as perceptions of bitterness, hotness, pepperiness and sweetness. Consumers were genotyped for TAS2R38 status to determine if liking is influenced by perception of bitter compounds such as glucosinolates (GSLs) and isothiocyanates (ITCs). Responses were combined with previously published data relating to phytochemical content and sensory data in Principal Component Analysis to determine compounds influencing liking/perceptions. Hotness, not bitterness, is the main attribute on which consumers base their liking of rocket. Some consumers rejected rocket based on GSL/ITC concentrations, whereas some preferred hotness. Bitter perception did not significantly influence liking of accessions, despite PAV/PAV 'supertasters' scoring higher for this attribute. High sugar-GSL/ITC ratios significantly reduce perceptions of hotness and bitterness for some consumers. Importantly the GSL glucoraphanin does not impart significant influence on liking or perception traits.
Project description:Glucosinolates (GSLs) from <i>Lepidium graminifolium</i> L. were analyzed qualitatively and quantitatively by their desulfo-counterparts using UHPLC-DAD-MS/MS technique and by their volatile breakdown products-isothiocyanates (ITCs) using GC-MS analysis. Thirteen GSLs were identified with arylaliphatic as the major ones in the following order: 3-hydroxybenzyl GSL (glucolepigramin, <b>7</b>), benzyl GSL (glucotropaeolin, <b>9</b>), 3,4,5-trimethoxybenzyl GSL (<b>11</b>), 3-methoxybenzyl GSL (glucolimnanthin, <b>12</b>), 4-hydroxy-3,5-dimethoxybenzyl GSL (3,5-dimethoxysinalbin, <b>8</b>), 4-hydroxybenzyl GSL (glucosinalbin, <b>6</b>), 3,4-dimethoxybenzyl GSL (<b>10</b>) and 2-phenylethyl GSL (gluconasturtiin, <b>13</b>). GSL breakdown products obtained by hydrodistillation (HD) and CH<sub>2</sub>Cl<sub>2</sub> extraction after hydrolysis by myrosinase for 24 h (EXT) as well as benzyl ITC were tested for their cytotoxic activity using MTT assay. Generally, EXT showed noticeable antiproliferative activity against human bladder cancer cell line UM-UC-3 and human glioblastoma cell line LN229, and can be considered as moderately active, while IC<sub>50</sub> of benzyl ITC was 12.3 μg/mL, which can be considered as highly active.
Project description:Nutritional value and disease-preventive effects of cabbage are well-known. Levels of the antioxidant compounds ascorbic acid (AA) and glucosinolates (GSL) in new Czech cabbage cultivars were determined in the context of different production systems. The contents of AA and GSLs in cabbage biomass were determined by HPLC. Individual GSLs were identified according to their exact masses with sinigrin used as the external standard. Artificial infection with <i>A. brassicicola</i> generally raised the AA levels. The major GSLs (?10 mg kg<sup>-1</sup>) were glucobrassicin, sinigrin, and glucoiberin. Indole and aliphatic GSLs were present, but no aromatic ones were detected. Ecological growth conditions and the artificial fungal infection increased the total content of GSLs and, also, of the methoxylated indole GSLs. Sulforaphane, iberin, indole-3-carbinol, and ascorbigen resulting from the hydrolysis of GSLs were found in both cultivars. The amounts and profiles of GSLs present in the two Czech cultivars demonstrated their good nutritional value. The decomposition products sulforaphane, iberin, indole-3-carbinol, and ascorbigen detected improve its health-promoting qualities and represent a suitable component of the human diet.
Project description:It is proposed that post-harvest longevity and appearance of salad crops is closely linked to pre-harvest leaf morphology (cell and leaf size) and biophysical structure (leaf strength). Transgenic lettuce plants (Lactuca sativa cv. Valeria) were produced in which the production of the cell wall-modifying enzyme xyloglucan endotransglucosylase/hydrolase (XTH) was down-regulated by antisense inhibition. Independently transformed lines were shown to have multiple members of the LsXTH gene family down-regulated in mature leaves of 6-week-old plants and during the course of shelf life. Consequently, xyloglucan endotransglucosylase (XET) enzyme activity and action were down-regulated in the cell walls of these leaves and it was established that leaf area and fresh weight were decreased while leaf strength was increased in the transgenic lines. Membrane permeability was reduced towards the end of shelf life in the transgenic lines relative to the controls and bacteria were evident inside the leaves of control plants only. Most importantly, an extended shelf-life of transgenic lines was observed relative to the non-transgenic control plants. These data illustrate the potential for engineering cell wall traits for improving quality and longevity of salad crops using either genetic modification directly, or by using markers associated with XTH genes to inform a commercial breeding programme.
Project description:Sweet cherry fruits (Prunus avium cvs. 'Canada Giant', 'Ferrovia') were harvested at commercial maturity stage and analyzed at harvest and after maintenance at room temperature (storage at ?20°C, shelf life) for 1, 2, 4, 6, and 8 days, respectively. Fruit were initially analyzed for respiration rate, qualitative attributes and textural properties: 'Canada Giant' fruit were characterized by higher weight losses and stem browning index, being more intense over the late stages of shelf life period; meanwhile 'Ferrovia' possessed appreciably better performance even after extended shelf life period. A gradual decrease of respiration rate was monitored in both cultivars, culminated after 8 days at 20°C. The sweet cherry fruit nutraceutical profile was monitored using an array of instrumental techniques (spectrophotometric assays, HPLC, (1)H-NMR). Fruit antioxidant capacity was enhanced with the progress of shelf life period, concomitant with the increased levels of total anthocyanin and of phenolic compounds. 'Ferrovia' fruit presented higher contents of neochlorogenic acid and p-coumaroylquinic acid throughout the shelf life period. We further developed an (1)H-NMR method that allows the study of primary and secondary metabolites in a single running, without previous separation and isolation procedures. Diagnostic peaks were located in the aliphatic region for sugars and organic acids, in the aromatic region for phenolic compounds and at 8.2-8.6 ppm for anthocyanins. This NMR-based methodology provides a unifying tool for quantitative and qualitative characterization of metabolite changes of sweet cherry fruits; it is also expected to be further exploited for monitoring temporal changes in other fleshy fruits.
Project description:This study was conducted to investigate doubled haploid (DH) lines produced between high GSL (HGSL) <i>B</i><i>rassica rapa</i> ssp. <i>trilocularis</i> (yellow sarson) and low GSL (LGSL) <i>B. rapa</i> ssp. <i>chinensis</i> (pak choi) parents. In total, 161 DH lines were generated. GSL content of HGSL DH lines ranged from 44.12 to 57.04 μmol·g<sup>-1</sup>·dry weight (dw), which is within the level of high GSL <i>B. rapa</i> ssp. <i>trilocularis</i> (47.46 to 59.56 μmol g<sup>-1</sup> dw). We resequenced five of the HGSL DH lines and three of the LGSL DH lines. Recombination blocks were formed between the parental and DH lines with 108,328 single-nucleotide polymorphisms in all chromosomes. In the measured GSL, gluconapin occurred as the major substrate in HGSL DH lines. Among the HGSL DH lines, BrYSP_DH005 had glucoraphanin levels approximately 12-fold higher than those of the HGSL mother plant. The hydrolysis capacity of GSL was analyzed in HGSL DH lines with a Korean pak choi cultivar as a control. Bioactive compounds, such as 3-butenyl isothiocyanate, 4-pentenyl isothiocyanate, 2-phenethyl isothiocyanate, and sulforaphane, were present in the HGSL DH lines at 3-fold to 6.3-fold higher levels compared to the commercial cultivar. The selected HGSL DH lines, resequencing data, and SNP identification were utilized for genome-assisted selection to develop elite GSL-enriched cultivars and the industrial production of potential anti-cancerous metabolites such as gluconapin and glucoraphanin.
Project description:It is important for the poultry industry to maximize product safety and quality by understanding the connection between bacterial diversity on chicken carcasses throughout poultry processing to the end of shelf life and the impact of the local processing environment. Enumeration of total aerobic bacteria, <i>Campylobacter</i> and <i>Pseudomonas</i>, and 16S rRNA gene amplicon sequencing were used to evaluate the processing line by collecting 10 carcasses from five processing steps: prescald, postplucker, pre- and post-immersion chill, and post-air chill. The diversity throughout a 12-day shelf life was also determined by examining 30 packaged carcasses. To identify the sources of possible contamination, scald water tank, immersion chilling water tank, air samples, and wall surfaces in the air-chill room were analyzed. Despite bacterial reductions on carcasses (>5 log<sub>10</sub> CFU/ml) throughout the process, each step altered the bacterial diversity. <i>Campylobacter</i> was a minor but persistent component in the bacterial community on carcasses. The combination of scalding, defeathering, and plucking distributed thermophilic spore-forming <i>Anoxybacillus</i> to carcasses, which remained at a high abundance on carcasses throughout subsequent processes. <i>Pseudomonas</i> was not isolated from carcasses after air chilling but was abundant on the wall of the air-chill room and became the predominant taxon at the end of shelf life, suggesting possible contamination through air movement. The results suggest that attention is needed at each processing step, regardless of bacterial reductions on carcasses. Changing scalding water regularly, maintaining good hygiene practices during processing, and thorough disinfection at the end of each processing day are important to minimize bacterial transmission.<b>IMPORTANCE</b> Culture-based and culture-independent approaches were utilized to reveal bacterial community changes on chicken carcasses at different processing steps and potential routes from the local processing environment. Current commercial processing effectively reduced bacterial loads on carcasses. Poultry processes have similar processes across facilities, but various processing arrangements and operating parameters could impact the bacterial transmission and persistence on carcasses differently. This study showed the use of a single tunnel incorporating scalding, defeathering and plucking may undesirably distribute the thermoduric bacteria, e.g., <i>Campylobacter</i> and <i>Anoxybacillus</i>, between the local environment and carcasses, whereas this does not occur when these steps are separated. The length of immersion and air chilling also impacted bacterial diversity on carcasses. Air chilling can transfer <i>Pseudomonas</i> from wall surfaces onto carcasses; this may subsequently influence chicken product shelf life. This study helps poultry processors understand the impact of current commercial processing and improve the chicken product quality and safety.
Project description:Cucumis melo fruit is highly valued for its sweet and refreshing flesh, however the flavour and value are also highly influenced by aroma as dictated by volatile organic compounds (VOCs). A simple and robust method of sampling VOCs on polydimethylsiloxane (PDMS) has been developed. Contrasting cultivars of C. melo subspecies melo were investigated at commercial maturity: three cultivars of var. Cantalupensis group Charentais (cv. Cézanne, Escrito, and Dalton) known to exhibit differences in ripening behaviour and shelf-life, as well as one cultivar of var. Cantalupensis group Ha'Ogan (cv. Noy Yisre'el) and one non-climacteric cultivar of var. Inodorus (cv. Tam Dew). The melon cultivar selection was based upon fruits exhibiting clear differences (cv. Noy Yisre'el and Tam Dew) and similarities (cv. Cézanne, Escrito, and Dalton) in flavour. In total, 58 VOCs were detected by thermal desorption (TD)-GC-MS which permitted the discrimination of each cultivar via Principal component analysis (PCA). PCA indicated a reduction in VOCs in the non-climacteric cv. Tam Dew compared to the four Cantalupensis cultivars. Within the group Charentais melons, the differences between the short, mid and long shelf-life cultivars were considerable. ¹H NMR analysis led to the quantification of 12 core amino acids, their levels were 3-10-fold greater in the Charentais melons, although they were reduced in the highly fragrant cv. Cézanne, indicating their role as VOC precursors. This study along with comparisons to more traditional labour intensive solid phase micro-extraction (SPME) GC-MS VOC profiling data has indicated that the high-throughput PDMS method is of great potential for the assessment of melon aroma and quality.
Project description:The incipient legalization and commercialization of Cannabis sativa in Canada have promulgated research into characterizing the plant's microbiome as it promotes many facets of plant growth and health. The emblematic production of commercially important secondary metabolites, namely tetrahydrocannabinol (THC), cannabidiol (CBD) and terpenes, has warranted investigating the modulating capacity of these molecules on the plant microbiome. C. sativa cultivars can be classified into chemotypes depending on the relative levels of THC and CBD they produce; their biosynthesis also varies spatially and temporally during the life cycle of the plant. To study the differential microbiome structure and diversity between cultivars in a spatio-temporal manner, we extracted microbial DNA from the rhizosphere, endorhizosphere, and phyllosphere during the entire life cycle of three different chemotypes; CBD Yummy (<1% THC/13% CBD), CBD shark (6% THC/10% CBD) and Hash (14% THC/ < 1% CBD). Illumina marker gene sequencing of bacterial (16S) and fungal (ITS) communities were coupled to the QIIME2, PICRUSt, and LEfSe pipelines for analysis. Our study describes spatio-temporal and cultivar-dependent variations in the fungal and bacterial microbiome of C. sativa, and details strong cultivar-dependent variance in the belowground microbiome. Furthermore, the predicted pathway abundance of the bacterial microbiome is concomitantly subject to spatio-temporal variations; pathways related to lipid, amino acid, glucose and pentose metabolism were noteworthy. These results describe, for the first time, spatio-temporal and cultivar-dependent variations in the microbiome of C. sativa produced under strict commercial settings. Describing the microbiome is the first step in discoveries that could help in engineering a plant growth and health promoting microbiome in future works.
Project description:The present investigation was carried out to study the response of two commercial pomegranate cultivars to individual shrink wrapping in extending the storage life and quality maintenance. Pomegranate fruits ('Mridula' and 'Bhagwa') were individually shrink wrapped using three semi-permeable films (Cryovac® BDF-2001, D-955 and normal LDPE) and stored at ambient (25-32 °C and 49-67% RH) and low temperature (8 °C and 75-80% RH). Shrink wrapping greatly reduced weight loss in both cultivars irrespective of the film used and storage temperature. Weight loss in shrink wrapped (D-955 film) 'Mridula' and 'Bhagwa' after 1 month storage at ambient temperature was respectively 1.40 and 1.05%, when compared to 22.92 and 22.53% in non-wrapped fruits. After 3 months at 8 °C, shrink wrapped 'Mridula' and 'Bhagwa' fruits lost only 0.43 and 0.68% weight respectively, compared to 17.23 and 21.67% in non-wrapped ones. Shrink wrapping significantly reduced the respiration rate at ambient temperature and the response varied with variety and film used. Shrink wrapped fruits of both cultivars retained the original peel colour (Hunter h? and C* values) to a maximum extent during 3 months storage at 8 °C and shelf-life period at ambient temperature. Irrespective of variety and film, shrink wrapping maintained the peel thickness and peel moisture content, significantly much higher than non-wrapped fruits at both temperatures. Compared to 'Mridula' cultivar, 'Bhagwa' responded well to shrink wrapping during prolonged storage at both temperatures with better maintenance of quality in terms of appearance, colour, juice content, TSS, acidity, sugars and sensory attributes. At ambient temperature, shrink wrapping with D-955 or LDPE film extended the storage life of 'Mridula' and 'Bhagwa' for 3 weeks and 1 month respectively, whereas at 8 °C both could be stored for 3 months with 3 days of shelf life.