Tom Shepherd

James Hutton Institute, Aberdeen, SCT, United Kingdom

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Publications (15)45.84 Total impact

  • Article: Utilisation of the MVA pathway to produce elevated levels of the sesquiterpene α-copaene in potato tubers.
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    ABSTRACT: Potato flavour is a complex trait resulting from the presence of a combination of volatile and non-volatile compounds. The aim of this work was to investigate the effect of specifically altering the volatile content of tubers and assess its impact on flavour. Tuber-specific over-expression of a potato α-copaene synthase gene resulted in enhanced levels (up to 15-fold higher than controls) of the sesquiterpene α-copaene. A positive correlation (R(2)=0.8) between transgene expression level and α-copaene abundance was observed. No significant changes in the levels of volatiles other than α-copaene were detected. Non-volatile flavour compounds (sugars, glycoalkaloids, major umami amino acids and 5'-ribonucleotides) were also determined. Relationships between flavour compounds and sensory evaluation data were investigated. Evaluators could not detect any aroma differences in the transgenic samples compared with controls and no significant differences in taste attributes were found. Thus although successful engineering of potato tubers to accumulate high levels of the flavour volatile α-copaene was achieved, sensory analysis suggests that α-copaene is not a major component of potato flavour.
    Phytochemistry 09/2011; 72(18):2288-93. · 3.35 Impact Factor
  • Article: Relationships between volatile and non-volatile metabolites and attributes of processed potato flavour.
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    ABSTRACT: Although the flavour of processed potatoes (Solanum tuberosum L.) is important to consumers, the blend of volatile and non-volatile metabolites that impact on flavour attributes is not well-defined. Additionally, it is important to understand how potato flavour changes during storage. In this study, quantitative descriptive analysis of potato samples by a trained taste panel was undertaken, comparing tubers from S. tuberosum group Phureja with those from S. tuberosum group Tuberosum, both at harvest and following storage. The cooked tuber volatile profile was analysed by solid phase micro extraction followed by gas chromatography-mass spectrometry analysis in sub-samples of the tubers that were assessed by taste panels. A range of non-volatile metabolites including the major umami compounds, glycoalkaloids and sugars was also measured in tuber sub-samples. Correlation and principal component analyses revealed differences between the potato cultivars and storage conditions and demonstrated associations of metabolites with the different sensory attributes.
    Phytochemistry 10/2010; 71(14-15):1765-73. · 3.35 Impact Factor
  • Article: A metabolomics study of cultivated potato (Solanum tuberosum) groups Andigena, Phureja, Stenotomum, and tuberosum using gas chromatography-mass spectrometry.
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    ABSTRACT: Phytochemical diversity was examined by gas chromatography-mass spectrometry in tubers of genotypes belonging to groups Andigena, Phureja, Stenotomum, and Tuberosum of the potato, Solanum tuberosum. Polar extracts (mainly amino acids, organic acids, sugars, and sugar alcohols) and nonpolar extracts (mainly fatty acids, fatty alcohols, and sterols) were examined. There was a large range in levels of metabolites, including those such as asparagine, fructose, and glucose, that are important to tuber quality, offering considerable scope for selecting germplasm for breeding programmes. There were significant differences in the levels of many metabolites among the groups. The metabolite profiles of genotypes belonging to Phureja and Stenotomum were similar and different from those of Tuberosum and the majority of Andigena genotypes. There was some agreement with the phylogeny of the groups in that Stenotomum is believed to be the ancestor of Phureja and they are both distinct from Tuberosum. Andigena genotypes could be partially distinguished according to geographical origin, Bolivian genotypes being particularly distinct from those from Ecuador. Biosynthetic links between metabolites were explored by performing pairwise correlations of all metabolites. The significance of some expected and unexpected strong correlations between many amino acids (e.g., between isoleucine, lysine, valine, and other amino acids) and between several nonpolar metabolites (e.g., between many fatty acids) is discussed. For polar metabolites, correlation analysis gave essentially similar results irrespective of whether the whole data set, only Andigena genotypes, or only Phureja genotypes were used. In contrast, for the nonpolar metabolites, Andigena only and Phureja only data sets resulted in weaker and stronger correlations, respectively, compared to the whole data set, and may suggest differences in the biochemistry of the two groups, although the interpretation should be viewed with some caution.
    Journal of Agricultural and Food Chemistry 12/2009; 58(2):1214-23. · 2.82 Impact Factor
  • Article: Transcriptional and metabolic profiles of Lolium perenne L. genotypes in response to a PEG-induced water stress.
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    ABSTRACT: Metabolic profiling was carried out in the forage grass Lolium perenne L. (perennial ryegrass) to uncover mechanisms involved in the plants response to water stress. When leaf and root materials from two genotypes, with a contrasting water stress response, were analysed by GC-MS, a clear difference in the metabolic profiles of the leaf tissue under water stress was observed. Differences were principally due to a reduction in fatty acid levels in the more susceptible Cashel genotype and an increase in sugars and compatible solutes in the more tolerant PI 462336 genotype. Sugars with a significant increase included: raffinose, trehalose, glucose, fructose and maltose. Increasing the ability of perennial ryegrass to accumulate these sugars in response to a water deficit may lead to more tolerant varieties. The metabolomics approach was combined with a transcriptomics approach in the water stress tolerant genotype PI 462336, which has identified perennial ryegrass genes regulated under water stress.
    Plant Biotechnology Journal 09/2009; 7(8):719-32. · 5.44 Impact Factor
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    Article: Expression profiling of potato germplasm differentiated in quality traits leads to the identification of candidate flavour and texture genes.
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    ABSTRACT: Quality traits such as flavour and texture are assuming a greater importance in crop breeding programmes. This study takes advantage of potato germplasm differentiated in tuber flavour and texture traits. A recently developed 44,000-element potato microarray was used to identify tuber gene expression profiles that correspond to differences in tuber flavour and texture as well as carotenoid content and dormancy characteristics. Gene expression was compared in two Solanum tuberosum group Phureja cultivars and two S. tuberosum group Tuberosum cultivars; 309 genes were significantly and consistently up-regulated in Phureja, whereas 555 genes were down-regulated. Approximately 46% of the genes in these lists can be identified from their annotation and amongst these are candidates that may underpin the Phureja/Tuberosum trait differences. For example, a clear difference in the cooked tuber volatile profile is the higher level of the sesquiterpene alpha-copaene in Phureja compared with Tuberosum. A sesquiterpene synthase gene was identified as being more highly expressed in Phureja tubers and its corresponding full-length cDNA was demonstrated to encode alpha-copaene synthase. Other potential 'flavour genes', identified from their differential expression profiles, include those encoding branched-chain amino acid aminotransferase and a ribonuclease suggesting a mechanism for 5'-ribonucleotide formation in potato tubers on cooking. Major differences in the expression levels of genes involved in cell wall biosynthesis (and potentially texture) were also identified, including genes encoding pectin acetylesterase, xyloglucan endotransglycosylase and pectin methylesterase. Other gene expression differences that may impact tuber carotenoid content and tuber life-cycle phenotypes are discussed.
    Journal of Experimental Botany 12/2008; 59(15):4219-31. · 5.36 Impact Factor
  • Article: Phytochemical diversity in tubers of potato cultivars and landraces using a GC-MS metabolomics approach.
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    ABSTRACT: Phytochemical diversity with respect to a range of polar (including amino acids, organic acids, sugars, and sugar alcohols) and nonpolar (including fatty acids, alkanols, and sterols) metabolites was examined within tubers from a total of 29 genetically diverse potato cultivars and Chilean landraces using a metabolomics approach by gas chromatography-mass spectrometry. From principal component analysis of the polar and nonpolar metabolite data there was insufficient variation to differentiate the majority of cultivars and landraces. Analysis of all polar metabolite profiles revealed separation of two cultivars (Glenna and Morag) from the other cultivars and landraces and a separate cluster of one landrace line, largely due to higher levels of sugars. Pentland Javelin was distinct in containing high levels of many amino acids. The two Solanum tuberosum group phureja cultivars (Inca Sun and Mayan Gold) were not particularly similar and were not separated from the S. tuberosum group tuberosum cultivars. Analysis of the nonpolar metabolite data revealed partial separation of two landrace lines and, on the basis of some minor fatty acids, Mayan Gold was distinct. The differences in metabolite profiles are considered in terms of the taxonomy and breeding history of the cultivars and possible influences from other factors such as developmental stage of the tuber. With a view to exploring biosynthetic links between metabolites, a pairwise correlation analysis was performed on all metabolites. The significance of high correlations between many amino acids and between several nonpolar metabolites is discussed.
    Journal of Agricultural and Food Chemistry 11/2008; 56(21):10280-91. · 2.82 Impact Factor
  • Article: Comparison of the calystegine composition and content of potato sprouts and tubers from Solanum tuberosum Group Phureja and Solanum tuberosum Group Tuberosum.
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    ABSTRACT: The calystegines detected in tubers from 17 Phureja ( S. tuberosum Group Phureja) lines and five Tuberosum ( S. tuberosum Group Tuberosum) cultivars were identified as the A 3 and B 2 structural types. Their concentration in whole tubers was of a similar order of magnitude in both species, as was the variability in the ratio of B 2 to A 3. On average, calystegine concentrations in the peel were about 13 times that found in the flesh for the five Tuberosum cultivars, and 4 times higher for four Phureja lines. Removal of the peel reduced the calystegine content by an average of over 50% in Tuberosum but by only 30% in Phureja, despite the latter having the greater proportion of peel. The calystegine content of sprouts was also determined for five Tuberosum cultivars and four Phureja lines and was found to include small amounts of four additional types, B 3, B 4, N 1, and X 2, in addition to the more abundant A 3 and B 2. Concentrations in the sprouts of Tuberosum were on average 100 times higher than that in the tuber flesh and 8 times higher than in the peel, whereas for Phureja, the equivalent values were 30 and 7 times higher, respectively. No correlation was found between sprout concentration and either flesh or peel calystegine concentration.
    Journal of Agricultural and Food Chemistry 08/2008; 56(13):5197-204. · 2.82 Impact Factor
  • Article: Potato metabolomics by GC–MS: what are the limiting factors?
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    ABSTRACT: Metabolic profiling methods are not ideally suited to the simultaneous analysis of all metabolite classes within a biological sample and must be optimized for maximum applicability. Several factors related to the optimization, validation and limitations of a GC–MS-based metabolic profiling method for potato were examined. A key step is conversion of reducing sugars to methyloximes, and optimum reaction conditions were 50°C for 4h. Shorter times or lower temperatures resulted in incomplete oximation whereas longer times and higher temperatures caused hydrolysis of sucrose, the major tuber dissacharide. Metabolite concentration gradients were observed in tuber sections. Glucose, fructose, alanine, methionine, threonine and tyrosine were more concentrated in the interior, whereas asparagine, putrescine, and caffeic and chlorogenic acids were higher in the skin and citrate was concentrated at the tuber’s bud end. These results impact upon choice of sampling strategy, consequently the use of freeze-dried (FD) material from a sampling protocol developed to avoid gradient-induced bias was examined. Using FD material, the method was highly linear and there was little qualitative or quantitative difference in the metabolite composition between fresh and FD material. The short- and long-term repeatability of the method was studied, and the use of reference materials to monitor and to improve data quality is discussed. Ascorbate is an important tuber metabolite that is readily measured by targeted approaches, but can be a problem in metabolic profiling. It was shown for standards and FD potato that ascorbate was largely degraded during oximation, although some survived in FD material.
    Metabolomics 11/2007; 3(4):475-488. · 4.51 Impact Factor
  • Chapter: Profiling of Metabolites and Volatile Flavour Compounds from Solanum Species Using Gas Chromatograph-Mass Spectrometry
    03/2007: pages 209-219;
  • Article: Application of metabolite and flavour volatile profiling to studies of biodiversity in solanum species
    Concepts in Plant Metabolomics. 01/2007;
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    Article: The effects of stress on plant cuticular waxes.
    Tom Shepherd, D Wynne Griffiths
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    ABSTRACT: Plants are subject to a wide range of abiotic stresses, and their cuticular wax layer provides a protective barrier, which consists predominantly of long-chain hydrocarbon compounds, including alkanes, primary alcohols, aldehydes, secondary alcohols, ketones, esters and other derived compounds. This article discusses current knowledge relating to the effects of stress on cuticular waxes and the ways in which the wax provides protection against the deleterious effects of light, temperature, osmotic stress, physical damage, altitude and pollution. Topics covered here include biosynthesis, morphology, composition and function of cuticular waxes in relation to the effects of stress, and some recent findings concerning the effects of stress on regulation of wax biosynthesis are described.
    New Phytologist 02/2006; 171(3):469-99. · 6.64 Impact Factor
  • Article: Metabolic engineering of high carotenoid potato tubers containing enhanced levels of beta-carotene and lutein.
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    ABSTRACT: In order to enhance the carotenoid content of potato tubers, transgenic potato plants have been produced expressing an Erwinia uredovora crtB gene encoding phytoene synthase, specifically in the tuber of Solanum tuberosum L. cultivar Desiree which normally produces tubers containing c. 5.6 microg carotenoid g(-1) DW and also in Solanum phureja L. cv. Mayan Gold which has a tuber carotenoid content of typically 20 microg carotenoid g(-1) DW. In developing tubers of transgenic crtB Desiree lines, carotenoid levels reached 35 microg carotenoid g(-1) DW and the balance of carotenoids changed radically compared with controls: beta-carotene levels in the transgenic tubers reached c. 11 microg g(-1) DW, whereas control tubers contained negligible amounts and lutein accumulated to a level 19-fold higher than empty-vector transformed controls. The crtB gene was also transformed into S. phureja (cv. Mayan Gold), again resulting in an increase in total carotenoid content to 78 microg carotenoid g(-1) DW in the most affected transgenic line. In these tubers, the major carotenoids were violaxanthin, lutein, antheraxanthin, and beta-carotene. No increases in expression levels of the major carotenoid biosynthetic genes could be detected in the transgenic tubers, despite the large increase in carotenoid accumulation. Microarray analysis was used to identify a number of genes that were consistently up- or down-regulated in transgenic crtB tubers compared with empty vector controls. The implications of these data from a nutritional standpoint and for further modifications of tuber carotenoid content are discussed.
    Journal of Experimental Botany 02/2005; 56(409):81-9. · 5.36 Impact Factor
  • Article: A comparison of the composition of epicuticular wax from red raspberry (Rubus idaeus L.) and hawthorn (Crataegus monogyna Jacq.) flowers
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    ABSTRACT: Epicuticular waxes have been characterised from the flowers of raspberry and hawthorn, on both of which adult raspberry beetles (Byturus tomentosus) can feed. The flower wax from both species had similar alkane profiles and also contained long-chain alcohols, aldehydes and fatty acids. The range of the carbon numbers detected for these classes of compounds was broadly similar in both but the relative amounts of each differed between species. Raspberry flower wax also contained fatty acid methyl esters, a group of compounds that has rarely been detected in plant epicuticular waxes, however, these were not observed in hawthorn flower wax. Long-chain alcohol-fatty acid esters with carbon numbers ranging from C36 to C48 were also detected in both plant species. However, an examination of their constituent acids indicated that in hawthorn the esters based on the C16 fatty acid predominated, whilst in raspberry flower wax, esters based on the C20 fatty acid were most abundant. Both species also contained pentacyclic triterpenoids, which accounted for, on average, over 16 and 48% of the total wax extracted from raspberry and hawthorn flowers respectively. In the former, ursolic and oleanolic acids accounted for over 90% of the pentacyclic triterpenes, whilst hawthorn flower wax, in addition to containing these acids, also contained high relative concentrations of both free and esterified α- and β-amyrins.
    Phytochemistry.
  • Article: Epicuticular wax composition in relation to aphid infestation and resistance in red raspberry (Rubus idaeus L.)
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    ABSTRACT: Epicuticular waxes from the aphid-resistant red raspberry (Rubus idaeus) cultivar Autumn Bliss and the aphid-susceptible cultivar Malling Jewel were collected from the newly emerging crown leaves, and also from the group of four more mature leaves immediately below the crown. Resistance and susceptibility status of the leaves to infestation by the large raspberry aphid, Amphorophora idaei, were determined by bioassay with the insect just prior to collection of the wax. Analysis showed the waxes to consist of a complex mixture of free fatty acids; free primary alcohols and their acetates; secondary alcohols; ketones; terpenoids including squalene, phytosterols, tocopherol and amyrins; alkanes and long chain alkyl and terpenyl esters. Compositional differences which may relate to A. idaei-resistance status were noticeably higher levels of sterols, particularly cycloartenol, together with the presence of branched alkanes, and an absence of C29 ketones and the symmetrical C29 secondary alcohol in wax from the resistant cultivar Bliss. There were also differences between the cultivars in the distribution of individual amyrins and tocopherols and in the chain length distribution for homologues of fatty acids, primary alcohols and alkanes, and these may also be related to resistance to A. idaei. Emerging leaves had lower levels of primary alcohols and terpenes, but higher levels of long-chain alkyl esters, and in general, more compounds of shorter chain-length than the more mature leaves. During bioassay A. idaei displayed a preference to settle on the more mature leaves. This may be due to greater wax coverage and higher levels of the compounds of shorter chain length found in the newly emerged younger leaves at the crown of the plant.
    Phytochemistry 52(7):1239-1254. · 3.35 Impact Factor
  • Article: Epicuticular waxes and volatiles from faba bean (Vicia faba) flowers
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    ABSTRACT: The floral bouquet of faba bean flowers was shown to be a complex mixture of some 27 identifiable compounds. In addition to the ubiquitous mono and sesquiterpenes, the porous-polymer entrained volatiles included a diverse range of phenylpropenoids which together accounted for over 7% of the total. Cinnamyl alcohol was also found to be the most abundant free alcohol in the epicuticular wax of faba bean flowers. Two new classes of epicuticular wax esters consisting of saturated C16, C18, C20, C22 and C24 fatty acids esterified with the phenylpropenoid, cinnamyl alcohol and with the diterpene, phytol have been identified.
    Phytochemistry.