The peach-potato aphid (Myzus persicae Sulzer) is a major pest of potato (Solanum tuberosum L.) b... more The peach-potato aphid (Myzus persicae Sulzer) is a major pest of potato (Solanum tuberosum L.) but the molecular characterization of this interaction particularly with regard to oxidants and antioxidants remains to be undertaken. Aphid colonies reared on potato leaves containing high ascorbate were twice the size of those grown on leaves with low ascorbate. Infestation-dependent decreases in the abundance of key transcripts such as chloroplastic FeSOD, peroxisomal catalase 2, PR1 and JAZ1 preceded detectable leaf H2O2 or polyphenol accumulation. The leaf glutathione pool was increased 48 h after infestation, but the amount of ascorbate was unchanged. The ascorbate/ dehydroacorbate (DHA) ratio was lower at 48 h but the ratio of reduced glutathione (GSH) to oxidized glutathione (GSSG) was unchanged. While DHA reductase and GSSG reductase activities were unaffected by aphid feeding, nonspecific peroxidase activities were enhanced 48 h following aphid infestation. Brown ethanol-insoluble deposits were observed close to leaf veins following aphid infestation. Taken together, the results demonstrate that high ascorbate favours aphid colony expansion and that perturbations in the leaf antioxidant system are intrinsic to the potato leaf response to aphids. Moreover, these changes together with the induction of hormone-related transcripts precede the deposition of defence-associated oxidized polyphenols along the stylet track.
SIGNIFICANCE The concept that vitamin C (L-ascorbic acid) is at the heart of the peroxide-process... more SIGNIFICANCE The concept that vitamin C (L-ascorbic acid) is at the heart of the peroxide-processing and redox signalling hub in plants is well established but our knowledge of the precise mechanisms involved remains patchy at best. Recent Advances: Ascorbate participates in the multifaceted signaling pathways initiated by both reactive oxygen species (ROS) and reactive nitrogen species (RNS). Crucially, the apoplastic ascorbate/dehydroascorbate (DHA) ratio that is regulated by ascorbate oxidase (AO), sculpts the apoplastic ROS (apoROS) signal that controls polarised cell growth, biotic and abiotic defences and cell to cell signalling, as well as exerting control over the light-dependent regulation of photosynthesis. CRITICAL ISSUES Here we re-evaluate the roles of ascorbate in photosynthesis and other process, addressing the question of how much we really know about the regulation of ascorbate homeostasis and its functions in plants, or how AO is regulated to modulate apoROS signals. FUTURE DIRECTIONS The role of microRNAs in the regulation of AO activity in relation to stress perception and signaling must be resolved. Similarly, the molecular characterization of ascorbate transporters and mechanistic links between photosynthetic and respiratory electron transport and ascorbate synthesis/homeostasis is a prerequisite to understanding ascorbate homeostasis and function. Similarly, there is little in vivo evidence for ascorbate functions as an enzyme co-factor.
Plants co-evolved with an enormous variety of microbial pathogens and insect herbivores under dai... more Plants co-evolved with an enormous variety of microbial pathogens and insect herbivores under daily and seasonal variations in abiotic environmental conditions. Hence, plant cells display a high capacity to respond to diverse stresses through a flexible and finely balanced response network that involves components such as reduction-oxidation (redox) signalling pathways, stress hormones and growth regulators, as well as calcium and protein kinase cascades. Biotic and abiotic stress responses use common signals, pathways and triggers leading to cross-tolerance phenomena, whereby exposure to one type of stress can activate plant responses that facilitate tolerance to several different types of stress. While the acclimation mechanisms and adaptive responses that facilitate responses to single biotic and abiotic stresses have been extensively characterized, relatively little information is available on the dynamic aspects of combined biotic/abiotic stress response. In this review, we con...
Phloem-feeding insects (PFIs), of which aphids are the largest group, are major agricultural pest... more Phloem-feeding insects (PFIs), of which aphids are the largest group, are major agricultural pests causing extensive damage to crop plants. In contrast to chewing insects, the nature of the plant response to PFIs remains poorly characterized. Scrutiny of the literature concerning transcriptional responses of model and crop plant species to PFIs reveals surprisingly little consensus with respect to the transcripts showing altered abundance following infestation. Nevertheless, core features of the transcriptional response to PFIs can be defined in Arabidopsis thaliana. This comparison of the PFI-associated transcriptional response observed in A. thaliana infested by the generalists Myzus persicae and Bemisia tabaci with the specialist Brevicoryne brassicae highlights the importance of calcium-dependent and receptor kinase-associated signalling. We discuss these findings within the context of the complex cross-talk between the different hormones regulating basal immune response mechani...
ABSTRACT Biochar is a carbon (C)-rich solid formed when biomass is used to produce bioenergy. Thi... more ABSTRACT Biochar is a carbon (C)-rich solid formed when biomass is used to produce bioenergy. This ‘black carbon’ has been suggested as a solution to climate change, potentially reducing global anthropogenic emissions of greenhouse gases by 12%, as well as promoting increased crop growth. How biochar application to soil leads to better crop yields remains open to speculation. Using the model plant Arabidopsis and the crop plant lettuce (Lactuca sativa L.), we found increased plant growth in both species following biochar application. Statistically significant increases for Arabidopsis in leaf area (130%), rosette diameter (61%) and root length (100%) were observed with similar findings in lettuce, where biochar application also increased leaf cell expansion. For the first time, global gene expression arrays were used on biochar-treated plants, enabling us to identify the growth-promoting plant hormones, brassinosteroid and auxin, and their signalling molecules, as key to this growth stimulation, with limited impacts on genes controlling photosynthesis. In addition, genes for cell wall loosening were promoted as were those for increased activity in membrane transporters for sugar, nutrients and aquaporins for better water and nutrient uptake and movement of sugars for metabolism in the plant. Positive growth effects were accompanied by down-regulation of a large suite of plant defence genes, including the jasmonic acid biosynthetic pathway, defensins and most categories of secondary metabolites. Such genes are critical for plant protection against insect and pathogen attack, as well as defence against stresses including drought. We propose a conceptual model to explain these effects in this biochar type, hypothesizing a role for additional K+ supply in biochar amended soils, leading to Ca2+ and Reactive Oxygen Species (ROS) –mediated signalling underpinning growth and defence signalling responses.
Over the past decade there has been increasing pressure to develop alternatives to the Reichstein... more Over the past decade there has been increasing pressure to develop alternatives to the Reichstein process, a largely chemical synthesis by which the vast majority of world vitamin C (L-ascorbic acid, L-AA) is produced. The pressures include increasing environmental concerns and legislation, and the need to increase process efficiency and reduce capital costs. The development of efficient fermentation processes in the past ten years has also represented a catalyst for change. Here, we describe the development of biotechnological alternatives for the synthesis of Reichstein intermediates by industrial microorganisms. The recent elucidation of the plant biosynthetic pathway represents new opportunities not only for the direct synthesis of L-AA by fermentation but also for the production of human crop plants and animal fodder with enhanced nutritional value. We discuss the potential for these developments in the light of recent findings concerning L-AA biosynthesis in plants.
Background: Although plants are the main source of vitamin C in the human diet, we still have a l... more Background: Although plants are the main source of vitamin C in the human diet, we still have a limited understanding of how plants synthesise L-ascorbic acid (AsA) and what regulates its concentration in different plant tissues. In particular, the enormous variability in the vitamin C content of storage organs from different plants remains unexplained. Possible sources of AsA in plant storage organs include in situ synthesis and long-distance transport of AsA synthesised in other tissues via the phloem. In this paper we examine a third possibility, that of synthesis within the phloem. Results: We provide evidence for the presence of AsA in the phloem sap of a wide range of crop species using aphid stylectomy and histochemical approaches. The activity of almost all the enzymes of the primary AsA biosynthetic pathway were detected in phloem-rich vascular exudates from Cucurbita pepo fruits and AsA biosynthesis was demonstrated in isolated phloem strands from Apium graveolens petioles incubated with a range of precursors (D-glucose, D-mannose, L-galactose and L-galactono-1,4-lactone). Phloem uptake of D-[U-14 C]mannose and L-[1-14 C]galactose (intermediates of the AsA biosynthetic pathway) as well as L-[1-14 C]AsA and L-[1-14 C]DHA, was observed in Nicotiana benthamiana leaf discs. Conclusions: We present the novel finding that active AsA biosynthesis occurs in the phloem. This process must now be considered in the context of mechanisms implicated in whole plant AsA distribution. This work should provoke studies aimed at elucidation of the in vivo substrates for phloem AsA biosynthesis and its contribution to AsA accumulation in plant storage organs.
Background: Following on from recent advances in plant AsA biosynthesis there is increasing inter... more Background: Following on from recent advances in plant AsA biosynthesis there is increasing interest in elucidating the factors contributing to the L-ascorbic acid (AsA) content of edible crops. One main objective is to establish whether in sink organs such as fruits and tubers, AsA is synthesised in situ from imported photoassimilates or synthesised in source tissues and translocated via the phloem. In the current work we test the hypothesis that long-distance transport is involved in AsA accumulation within the potato tuber, the most significant source of AsA in the European diet. Results: Using the EDTA exudation technique we confirm the presence of AsA in the phloem of potato plants and demonstrate a correlation between changes in the AsA content of source leaves and that of phloem exudates. Comparison of carboxyflourescein and AgNO 3 staining is suggestive of symplastic unloading of AsA in developing tubers. This hypothesis was further supported by the changes in AsA distribution during tuber development which closely resembled those of imported photoassimilates. Manipulation of leaf AsA content by supply of precursors to source leaves resulted in increased AsA content of developing tubers. Conclusion: Our data provide strong support to the hypothesis that long-distance transport of AsA occurs in potato. We also show that phloem AsA content and AsA accumulation in sink organs can be directly increased via manipulation of AsA content in the foliage. We are now attempting to establish the quantitative contribution of imported AsA to overall AsA accumulation in developing potato tubers via transgenic approaches.
Sensory differentiation of blueberries from eight highbush cultivars grown in the UK was related ... more Sensory differentiation of blueberries from eight highbush cultivars grown in the UK was related to flavor volatile composition. A two-phase descriptive sensory analysis was used: initial FCP to understand the range of descriptors used by consumers to differentiate the fruit and a subsequent conventional profiling by trained assessors using a consensus vocabulary to generate a multivariate product space describing relationships between the cultivars. A generalized Procrustes analysis product was obtained for aroma volatiles, extracted by headspace solid-phase microextraction (HS-SPME), and analyzed by gas chromatography linked mass spectrometry (GC/MS). Sensory information on cultivar aroma characters could be correlated with differences in aroma volatiles.
To gain greater insight into the mechanism of dormancy release in the potato tuber, an investigat... more To gain greater insight into the mechanism of dormancy release in the potato tuber, an investigation into physiological and biochemical changes in tuber and bud tissues during the transition from bud dormancy (immediately after harvest) to active bud growth was undertaken. Within the tuber, a rapid shift from storage metabolism (starch synthesis) to reserve mobilization within days of detachment from the mother plant suggested transition from sink to source. Over the same period, a shift in the pattern of [U-(14)C]sucrose uptake by tuber discs from diffuse to punctate accumulation was consistent with a transition from phloem unloading to phloem loading within the tuber parenchyma. There were no gross differences in metabolic capacity between resting and actively growing tuber buds as determined by [U-(14)C]glucose labelling. However, marked differences in metabolite pools were observed with large increases in starch and sucrose, and the accumulation of several organic acids in growing buds. Carboxyfluorescein labelling of tubers clearly demonstrated strong symplastic connection in actively growing buds and symplastic isolation in resting buds. It is proposed that potato tubers rapidly undergo metabolic transitions consistent with bud outgrowth; however, growth is initially prevented by substrate limitation mediated via symplastic isolation.
Recent advances have defined some of the components of photoperiodic signalling that lead to tube... more Recent advances have defined some of the components of photoperiodic signalling that lead to tuberization in potato including orthologues of FLOWERING LOCUS T (StSP6A) and CYCLING DOF FACTOR (StCDF1). The aim of the current study is to investigate the molecular basis of permissive tuber initiation under long days in Solanum tuberosum Neo-Tuberosum by comparative analysis with an obligate short-day S. tuberosum ssp. Andigena accession. We show that the Neo-Tuberosum accession, but not the Andigena, contains alleles that encode StCDF1 proteins modified in the C-terminal region, likely to evade long day inhibition of StSP6A expression. We also identify an allele of StSP6A from the Neo-Tuberosum accession, absent in the Andigena, which is expressed under long days. Other leaf transcripts and metabolites that show different abundances in tuberizing and non-tuberizing samples were identified adding detail to tuberization-associated processes. Overall, the data presented in this study highlight the subtle interplay between components of the clock-CONSTANS-StSP6A axis which collectively may interact to fine-tune the timing of tuberization.
Although processed potato tuber texture is an important trait that influences consumer preference... more Although processed potato tuber texture is an important trait that influences consumer preference, a detailed understanding of tuber textural properties at the molecular level is lacking. Previous work has identified tuber pectin methyl esterase (PME) activity as a potential factor impacting on textural properties, and the expression of a gene encoding an isoform of PME (PEST1) was associated with cooked tuber textural properties. In this study, a transgenic approach was undertaken to investigate further the impact of the PEST1 gene. Antisense and over-expressing potato lines were generated. In over-expressing lines, tuber PME activity was enhanced by up to 2.3-fold; whereas in antisense lines, PME activity was decreased by up to 62%. PME isoform analysis indicated that the PEST1 gene encoded one isoform of PME. Analysis of cell walls from tubers from the over-expressing lines indicated that the changes in PME activity resulted in a decrease in pectin methylation. Analysis of processed tuber texture demonstrated that the reduced level of pectin methylation in the over-expressing transgenic lines was associated with a firmer processed texture. Thus, there is a clear link between PME activity, pectin methylation and processed tuber textural properties.
Analysis of phloem exudates from the fruit of Cucurbitaceae revealed the presence of several comp... more Analysis of phloem exudates from the fruit of Cucurbitaceae revealed the presence of several compounds with UV-visible absorption spectra identical to that of L-ascorbic acid. In Cucurbita pepo L. (zucchini), the compounds could be isolated from phloem exudates collected from aerial parts of the plant but were not detected in whole tissue homogenates. The compounds isolated from the phloem exudates of C. pepo fruit were eluted from strong anion exchange resin in the same fraction as L-ascorbic acid and were oxidised by ascorbate oxidase (E.C. 1.10.3.3). The major compound purified from C. pepo fruit exudates demonstrated similar redox properties to L-ascorbic acid and synthetic 6-O-glucosyl-L-ascorbic acid (6-GlcAsA) but differed from those of 2-O-glucosyl-L-ascorbic acid (2-GlcAsA) isolated from the fruit of Lycium barbarum L. Parent and fragment ion masses of the compound were consistent with hexosyl-ascorbate in which the hexose moiety was attached to C5 or C6 of AsA. Acid hydrolysis of the major C. pepo compound resulted in the formation of L-ascorbic acid and glucose. The purified compound yielded a proton NMR spectrum that was almost identical to that of synthetic 6-GlcAsA. A series of L-ascorbic acid conjugates have, therefore, been identified in the phloem of Cucurbitaceae and the most abundant conjugate has been identified as 6-GlcAsA. The potential role of such conjugates in the long-distance transport of Lascorbic acid is discussed.
The peach-potato aphid (Myzus persicae Sulzer) is a major pest of potato (Solanum tuberosum L.) b... more The peach-potato aphid (Myzus persicae Sulzer) is a major pest of potato (Solanum tuberosum L.) but the molecular characterization of this interaction particularly with regard to oxidants and antioxidants remains to be undertaken. Aphid colonies reared on potato leaves containing high ascorbate were twice the size of those grown on leaves with low ascorbate. Infestation-dependent decreases in the abundance of key transcripts such as chloroplastic FeSOD, peroxisomal catalase 2, PR1 and JAZ1 preceded detectable leaf H2O2 or polyphenol accumulation. The leaf glutathione pool was increased 48 h after infestation, but the amount of ascorbate was unchanged. The ascorbate/ dehydroacorbate (DHA) ratio was lower at 48 h but the ratio of reduced glutathione (GSH) to oxidized glutathione (GSSG) was unchanged. While DHA reductase and GSSG reductase activities were unaffected by aphid feeding, nonspecific peroxidase activities were enhanced 48 h following aphid infestation. Brown ethanol-insoluble deposits were observed close to leaf veins following aphid infestation. Taken together, the results demonstrate that high ascorbate favours aphid colony expansion and that perturbations in the leaf antioxidant system are intrinsic to the potato leaf response to aphids. Moreover, these changes together with the induction of hormone-related transcripts precede the deposition of defence-associated oxidized polyphenols along the stylet track.
SIGNIFICANCE The concept that vitamin C (L-ascorbic acid) is at the heart of the peroxide-process... more SIGNIFICANCE The concept that vitamin C (L-ascorbic acid) is at the heart of the peroxide-processing and redox signalling hub in plants is well established but our knowledge of the precise mechanisms involved remains patchy at best. Recent Advances: Ascorbate participates in the multifaceted signaling pathways initiated by both reactive oxygen species (ROS) and reactive nitrogen species (RNS). Crucially, the apoplastic ascorbate/dehydroascorbate (DHA) ratio that is regulated by ascorbate oxidase (AO), sculpts the apoplastic ROS (apoROS) signal that controls polarised cell growth, biotic and abiotic defences and cell to cell signalling, as well as exerting control over the light-dependent regulation of photosynthesis. CRITICAL ISSUES Here we re-evaluate the roles of ascorbate in photosynthesis and other process, addressing the question of how much we really know about the regulation of ascorbate homeostasis and its functions in plants, or how AO is regulated to modulate apoROS signals. FUTURE DIRECTIONS The role of microRNAs in the regulation of AO activity in relation to stress perception and signaling must be resolved. Similarly, the molecular characterization of ascorbate transporters and mechanistic links between photosynthetic and respiratory electron transport and ascorbate synthesis/homeostasis is a prerequisite to understanding ascorbate homeostasis and function. Similarly, there is little in vivo evidence for ascorbate functions as an enzyme co-factor.
Plants co-evolved with an enormous variety of microbial pathogens and insect herbivores under dai... more Plants co-evolved with an enormous variety of microbial pathogens and insect herbivores under daily and seasonal variations in abiotic environmental conditions. Hence, plant cells display a high capacity to respond to diverse stresses through a flexible and finely balanced response network that involves components such as reduction-oxidation (redox) signalling pathways, stress hormones and growth regulators, as well as calcium and protein kinase cascades. Biotic and abiotic stress responses use common signals, pathways and triggers leading to cross-tolerance phenomena, whereby exposure to one type of stress can activate plant responses that facilitate tolerance to several different types of stress. While the acclimation mechanisms and adaptive responses that facilitate responses to single biotic and abiotic stresses have been extensively characterized, relatively little information is available on the dynamic aspects of combined biotic/abiotic stress response. In this review, we con...
Phloem-feeding insects (PFIs), of which aphids are the largest group, are major agricultural pest... more Phloem-feeding insects (PFIs), of which aphids are the largest group, are major agricultural pests causing extensive damage to crop plants. In contrast to chewing insects, the nature of the plant response to PFIs remains poorly characterized. Scrutiny of the literature concerning transcriptional responses of model and crop plant species to PFIs reveals surprisingly little consensus with respect to the transcripts showing altered abundance following infestation. Nevertheless, core features of the transcriptional response to PFIs can be defined in Arabidopsis thaliana. This comparison of the PFI-associated transcriptional response observed in A. thaliana infested by the generalists Myzus persicae and Bemisia tabaci with the specialist Brevicoryne brassicae highlights the importance of calcium-dependent and receptor kinase-associated signalling. We discuss these findings within the context of the complex cross-talk between the different hormones regulating basal immune response mechani...
ABSTRACT Biochar is a carbon (C)-rich solid formed when biomass is used to produce bioenergy. Thi... more ABSTRACT Biochar is a carbon (C)-rich solid formed when biomass is used to produce bioenergy. This ‘black carbon’ has been suggested as a solution to climate change, potentially reducing global anthropogenic emissions of greenhouse gases by 12%, as well as promoting increased crop growth. How biochar application to soil leads to better crop yields remains open to speculation. Using the model plant Arabidopsis and the crop plant lettuce (Lactuca sativa L.), we found increased plant growth in both species following biochar application. Statistically significant increases for Arabidopsis in leaf area (130%), rosette diameter (61%) and root length (100%) were observed with similar findings in lettuce, where biochar application also increased leaf cell expansion. For the first time, global gene expression arrays were used on biochar-treated plants, enabling us to identify the growth-promoting plant hormones, brassinosteroid and auxin, and their signalling molecules, as key to this growth stimulation, with limited impacts on genes controlling photosynthesis. In addition, genes for cell wall loosening were promoted as were those for increased activity in membrane transporters for sugar, nutrients and aquaporins for better water and nutrient uptake and movement of sugars for metabolism in the plant. Positive growth effects were accompanied by down-regulation of a large suite of plant defence genes, including the jasmonic acid biosynthetic pathway, defensins and most categories of secondary metabolites. Such genes are critical for plant protection against insect and pathogen attack, as well as defence against stresses including drought. We propose a conceptual model to explain these effects in this biochar type, hypothesizing a role for additional K+ supply in biochar amended soils, leading to Ca2+ and Reactive Oxygen Species (ROS) –mediated signalling underpinning growth and defence signalling responses.
Over the past decade there has been increasing pressure to develop alternatives to the Reichstein... more Over the past decade there has been increasing pressure to develop alternatives to the Reichstein process, a largely chemical synthesis by which the vast majority of world vitamin C (L-ascorbic acid, L-AA) is produced. The pressures include increasing environmental concerns and legislation, and the need to increase process efficiency and reduce capital costs. The development of efficient fermentation processes in the past ten years has also represented a catalyst for change. Here, we describe the development of biotechnological alternatives for the synthesis of Reichstein intermediates by industrial microorganisms. The recent elucidation of the plant biosynthetic pathway represents new opportunities not only for the direct synthesis of L-AA by fermentation but also for the production of human crop plants and animal fodder with enhanced nutritional value. We discuss the potential for these developments in the light of recent findings concerning L-AA biosynthesis in plants.
Background: Although plants are the main source of vitamin C in the human diet, we still have a l... more Background: Although plants are the main source of vitamin C in the human diet, we still have a limited understanding of how plants synthesise L-ascorbic acid (AsA) and what regulates its concentration in different plant tissues. In particular, the enormous variability in the vitamin C content of storage organs from different plants remains unexplained. Possible sources of AsA in plant storage organs include in situ synthesis and long-distance transport of AsA synthesised in other tissues via the phloem. In this paper we examine a third possibility, that of synthesis within the phloem. Results: We provide evidence for the presence of AsA in the phloem sap of a wide range of crop species using aphid stylectomy and histochemical approaches. The activity of almost all the enzymes of the primary AsA biosynthetic pathway were detected in phloem-rich vascular exudates from Cucurbita pepo fruits and AsA biosynthesis was demonstrated in isolated phloem strands from Apium graveolens petioles incubated with a range of precursors (D-glucose, D-mannose, L-galactose and L-galactono-1,4-lactone). Phloem uptake of D-[U-14 C]mannose and L-[1-14 C]galactose (intermediates of the AsA biosynthetic pathway) as well as L-[1-14 C]AsA and L-[1-14 C]DHA, was observed in Nicotiana benthamiana leaf discs. Conclusions: We present the novel finding that active AsA biosynthesis occurs in the phloem. This process must now be considered in the context of mechanisms implicated in whole plant AsA distribution. This work should provoke studies aimed at elucidation of the in vivo substrates for phloem AsA biosynthesis and its contribution to AsA accumulation in plant storage organs.
Background: Following on from recent advances in plant AsA biosynthesis there is increasing inter... more Background: Following on from recent advances in plant AsA biosynthesis there is increasing interest in elucidating the factors contributing to the L-ascorbic acid (AsA) content of edible crops. One main objective is to establish whether in sink organs such as fruits and tubers, AsA is synthesised in situ from imported photoassimilates or synthesised in source tissues and translocated via the phloem. In the current work we test the hypothesis that long-distance transport is involved in AsA accumulation within the potato tuber, the most significant source of AsA in the European diet. Results: Using the EDTA exudation technique we confirm the presence of AsA in the phloem of potato plants and demonstrate a correlation between changes in the AsA content of source leaves and that of phloem exudates. Comparison of carboxyflourescein and AgNO 3 staining is suggestive of symplastic unloading of AsA in developing tubers. This hypothesis was further supported by the changes in AsA distribution during tuber development which closely resembled those of imported photoassimilates. Manipulation of leaf AsA content by supply of precursors to source leaves resulted in increased AsA content of developing tubers. Conclusion: Our data provide strong support to the hypothesis that long-distance transport of AsA occurs in potato. We also show that phloem AsA content and AsA accumulation in sink organs can be directly increased via manipulation of AsA content in the foliage. We are now attempting to establish the quantitative contribution of imported AsA to overall AsA accumulation in developing potato tubers via transgenic approaches.
Sensory differentiation of blueberries from eight highbush cultivars grown in the UK was related ... more Sensory differentiation of blueberries from eight highbush cultivars grown in the UK was related to flavor volatile composition. A two-phase descriptive sensory analysis was used: initial FCP to understand the range of descriptors used by consumers to differentiate the fruit and a subsequent conventional profiling by trained assessors using a consensus vocabulary to generate a multivariate product space describing relationships between the cultivars. A generalized Procrustes analysis product was obtained for aroma volatiles, extracted by headspace solid-phase microextraction (HS-SPME), and analyzed by gas chromatography linked mass spectrometry (GC/MS). Sensory information on cultivar aroma characters could be correlated with differences in aroma volatiles.
To gain greater insight into the mechanism of dormancy release in the potato tuber, an investigat... more To gain greater insight into the mechanism of dormancy release in the potato tuber, an investigation into physiological and biochemical changes in tuber and bud tissues during the transition from bud dormancy (immediately after harvest) to active bud growth was undertaken. Within the tuber, a rapid shift from storage metabolism (starch synthesis) to reserve mobilization within days of detachment from the mother plant suggested transition from sink to source. Over the same period, a shift in the pattern of [U-(14)C]sucrose uptake by tuber discs from diffuse to punctate accumulation was consistent with a transition from phloem unloading to phloem loading within the tuber parenchyma. There were no gross differences in metabolic capacity between resting and actively growing tuber buds as determined by [U-(14)C]glucose labelling. However, marked differences in metabolite pools were observed with large increases in starch and sucrose, and the accumulation of several organic acids in growing buds. Carboxyfluorescein labelling of tubers clearly demonstrated strong symplastic connection in actively growing buds and symplastic isolation in resting buds. It is proposed that potato tubers rapidly undergo metabolic transitions consistent with bud outgrowth; however, growth is initially prevented by substrate limitation mediated via symplastic isolation.
Recent advances have defined some of the components of photoperiodic signalling that lead to tube... more Recent advances have defined some of the components of photoperiodic signalling that lead to tuberization in potato including orthologues of FLOWERING LOCUS T (StSP6A) and CYCLING DOF FACTOR (StCDF1). The aim of the current study is to investigate the molecular basis of permissive tuber initiation under long days in Solanum tuberosum Neo-Tuberosum by comparative analysis with an obligate short-day S. tuberosum ssp. Andigena accession. We show that the Neo-Tuberosum accession, but not the Andigena, contains alleles that encode StCDF1 proteins modified in the C-terminal region, likely to evade long day inhibition of StSP6A expression. We also identify an allele of StSP6A from the Neo-Tuberosum accession, absent in the Andigena, which is expressed under long days. Other leaf transcripts and metabolites that show different abundances in tuberizing and non-tuberizing samples were identified adding detail to tuberization-associated processes. Overall, the data presented in this study highlight the subtle interplay between components of the clock-CONSTANS-StSP6A axis which collectively may interact to fine-tune the timing of tuberization.
Although processed potato tuber texture is an important trait that influences consumer preference... more Although processed potato tuber texture is an important trait that influences consumer preference, a detailed understanding of tuber textural properties at the molecular level is lacking. Previous work has identified tuber pectin methyl esterase (PME) activity as a potential factor impacting on textural properties, and the expression of a gene encoding an isoform of PME (PEST1) was associated with cooked tuber textural properties. In this study, a transgenic approach was undertaken to investigate further the impact of the PEST1 gene. Antisense and over-expressing potato lines were generated. In over-expressing lines, tuber PME activity was enhanced by up to 2.3-fold; whereas in antisense lines, PME activity was decreased by up to 62%. PME isoform analysis indicated that the PEST1 gene encoded one isoform of PME. Analysis of cell walls from tubers from the over-expressing lines indicated that the changes in PME activity resulted in a decrease in pectin methylation. Analysis of processed tuber texture demonstrated that the reduced level of pectin methylation in the over-expressing transgenic lines was associated with a firmer processed texture. Thus, there is a clear link between PME activity, pectin methylation and processed tuber textural properties.
Analysis of phloem exudates from the fruit of Cucurbitaceae revealed the presence of several comp... more Analysis of phloem exudates from the fruit of Cucurbitaceae revealed the presence of several compounds with UV-visible absorption spectra identical to that of L-ascorbic acid. In Cucurbita pepo L. (zucchini), the compounds could be isolated from phloem exudates collected from aerial parts of the plant but were not detected in whole tissue homogenates. The compounds isolated from the phloem exudates of C. pepo fruit were eluted from strong anion exchange resin in the same fraction as L-ascorbic acid and were oxidised by ascorbate oxidase (E.C. 1.10.3.3). The major compound purified from C. pepo fruit exudates demonstrated similar redox properties to L-ascorbic acid and synthetic 6-O-glucosyl-L-ascorbic acid (6-GlcAsA) but differed from those of 2-O-glucosyl-L-ascorbic acid (2-GlcAsA) isolated from the fruit of Lycium barbarum L. Parent and fragment ion masses of the compound were consistent with hexosyl-ascorbate in which the hexose moiety was attached to C5 or C6 of AsA. Acid hydrolysis of the major C. pepo compound resulted in the formation of L-ascorbic acid and glucose. The purified compound yielded a proton NMR spectrum that was almost identical to that of synthetic 6-GlcAsA. A series of L-ascorbic acid conjugates have, therefore, been identified in the phloem of Cucurbitaceae and the most abundant conjugate has been identified as 6-GlcAsA. The potential role of such conjugates in the long-distance transport of Lascorbic acid is discussed.
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Papers by Robert Hancock