Genetic manipulation of 6-phosphofructokinase in potato tubers

Burrell, M. M.; Mooney, Peter; Blundy, Margaret; Carter, Dawn; Wilson, Frank J.; Green, John; Blundy, Keith S.; Rees, Tom ap

doi:10.1007/bf00201039

Cited by 98 publications

(83 citation statements)

References 29 publications

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“…These measurements were confirmed by recovery experiments, have low standard errors, and the wild-type values are comparable to those previously reported for potato tubers (e.g. Jelitto et al, 1992;Burrell et al, 1994). In U-IN2-30 there is a significant three-to fourfold increase in the hexose phosphate pools relative to the wild-type.…”

Section: Determination Of Hexose Phosphatessupporting

confidence: 87%

“…The levels of 3-phosphoglycerate (3PGA), phosphoenolpyruvate (PEP) and pyruvate were comparable to those measured previously in potato tubers (e.g. Burrell et al, 1994;Jellito et al, 1992), and the measurements were authenticated by recovery experiments. The measurements were made on the same extracts that were used for the hexose phosphate measurements.…”

Section: Determination Of Glycolytic Intermediatessupporting

confidence: 83%

“…To ascertain whether changes in glycolytic enzymes played a role in the increase in glycolytic intermediates, the maximum catalytic activities of all the enzymes in the glycolytic sequence were determined using established protocols for potato tubers (Burrell et al, 1994). These measurements were made on the same tissue used for the analysis of metabolic intermediates (Table 3).…”

Section: Measurement Of the Activities Of The Glycolytic Enzymesmentioning

confidence: 99%

“…Extracts and supernatants were prepared and maintained at 4°C until assayed. Phosphoglucoisomerase (EC 5.3.1.9), 6-phosphofructokinase (EC 2.1.7.11), pyrophosphate: fructose-6-phosphate 1-phosphotransferase (PFP; EC 2.7.1.90), fructose 1,6-bisphosphate aldolase (EC 4.1.2.13), triose phosphate isomerase (EC 5.3.1.1), glyceraldehyde-3-phosphate dehydrogenase (GAP-DH, EC 1.2.1.12), phosphoglycerate kinase (EC 2.7.2.3), phosphoglycerate mutase (EC 2.7.5.3), enolase (EC 4.2.1.11), and pyruvate kinase (EC 2.7.1.40) were assayed as described by Burrell et al (1994). Acid invertase activity was determined in a stopped assay as described by Zrenner et al (1994).…”

Section: Determination Of Enzyme Activitiesmentioning

confidence: 99%

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Combined expression of glucokinase and invertase in potato tubers leads to a dramatic reduction in starch accumulation and a stimulation of glycolysis

et al. 1998

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SummaryThe original aim of this work was to increase starch accumulation in potato tubers by enhancing their capacity to metabolise sucrose. We previously reported that specific expression of a yeast invertase in the cytosol of tubers led to a 95% reduction in sucrose content, but that this was accompanied by a larger accumulation of glucose and a reduction in starch. In the present paper we introduced a bacterial glucokinase from Zymomonas mobilis into an invertase-expressing transgenic line, with the intention of bringing the glucose into metabolism. Transgenic lines were obtained with up to threefold more glucokinase activity than in the parent invertase line and which did not accumulate glucose. Unexpectedly, there was a further dramatic reduction in starch content, down to 35% of wild-type levels. Biochemical analysis of growing tuber tissue revealed large increases in the metabolic intermediates of glycolysis, organic acids and amino acids, two-to threefold increases in the maximum catalytic activities of key enzymes in the respiratory pathways, and three-to fivefold increases in carbon dioxide production. These changes occur in the lines expressing invertase, and are accentuated following introduction of the second transgene, glucokinase. We conclude that the expression of invertase in potato tubers leads to an increased flux through the glycolytic pathway at the expense of starch synthesis and that heterologous overexpression of glucokinase enhances this change in partitioning.

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Section: Determination Of Hexose Phosphatessupporting

confidence: 87%

Section: Determination Of Glycolytic Intermediatessupporting

confidence: 83%

Section: Measurement Of the Activities Of The Glycolytic Enzymesmentioning

confidence: 99%

Section: Determination Of Enzyme Activitiesmentioning

confidence: 99%

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Combined expression of glucokinase and invertase in potato tubers leads to a dramatic reduction in starch accumulation and a stimulation of glycolysis

et al. 1998

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“…The contents of Glc, Fru, Suc, starch, and adenylates were determined as in Mustroph et al (2006a) and phosphorylated metabolites as in Burrell et al, (1994). The content of Pi was determined as described by Itaya and Ui (1966).…”

Section: Metabolite Extraction and Analysismentioning

confidence: 99%

A Shoot-Specific Hypoxic Response of Arabidopsis Sheds Light on the Role of the Phosphate-Responsive Transcription Factor PHOSPHATE STARVATION RESPONSE1

et al. 2014

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Plant responses to biotic and abiotic stresses are often very specific, but signal transduction pathways can partially or completely overlap. Here, we demonstrate that in Arabidopsis (Arabidopsis thaliana), the transcriptional responses to phosphate starvation and oxygen deficiency stress comprise a set of commonly induced genes. While the phosphate deficiency response is systemic, under oxygen deficiency, most of the commonly induced genes are found only in illuminated shoots. This jointly induced response to the two stresses is under control of the transcription factor PHOSPHATE STARVATION RESPONSE1 (PHR1), but not of the oxygen-sensing N-end rule pathway, and includes genes encoding proteins for the synthesis of galactolipids, which replace phospholipids in plant membranes under phosphate starvation. Despite the induction of galactolipid synthesis genes, total galactolipid content and plant survival are not severely affected by the up-regulation of galactolipid gene expression in illuminated leaves during hypoxia. However, changes in galactolipid molecular species composition point to an adaptation of lipid fluxes through the endoplasmic reticulum and chloroplast pathways during hypoxia. PHR1-mediated signaling of phosphate deprivation was also light dependent. Because a photoreceptor-mediated PHR1 activation was not detectable under hypoxia, our data suggest that a chloroplast-derived retrograde signal, potentially arising from metabolic changes, regulates PHR1 activity under both oxygen and phosphate deficiency.

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Simultaneous analysis of a series of phosphorylated sugars in small tissue samples by anion exchange chromatography and pulsed amperometric detection

1999

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Genetic manipulation of 6-phosphofructokinase in potato tubers

Cited by 98 publications

References 29 publications

Combined expression of glucokinase and invertase in potato tubers leads to a dramatic reduction in starch accumulation and a stimulation of glycolysis

Combined expression of glucokinase and invertase in potato tubers leads to a dramatic reduction in starch accumulation and a stimulation of glycolysis

A Shoot-Specific Hypoxic Response of Arabidopsis Sheds Light on the Role of the Phosphate-Responsive Transcription Factor PHOSPHATE STARVATION RESPONSE1

Simultaneous analysis of a series of phosphorylated sugars in small tissue samples by anion exchange chromatography and pulsed amperometric detection

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