Both developmental and metabolic signals activate the promoter of a class I patatin gene

Rocha-Sosa, Mario; Sonnewald, Uwe; Frommer, Wolf B; Stratmann, M.; Schell, Jeff; Willmitzer, Lothar

doi:10.1002/j.1460-2075.1989.tb03344.x

Cited by 393 publications

(316 citation statements)

References 24 publications

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“…Potato leaves of U-IN2-30 (Sonnewald et al, 1997) were transformed with this construct using an Agrobacteriummediated protocol (Rocha-Sosa et al, 1989), and 80 transgenic lines were transferred to the greenhouse for screening. Tubers were formed in small 0.2 litre pots, and samples were taken for activity tests.…”

Section: Preparation and Selection Of Transgenic Plantsmentioning

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: Preparation and Selection Of Transgenic Plantsmentioning

confidence: 99%

“…This construct was introduced into line U-IN2-30 of Solanum tuberosum L. cv. Desiree (Sonnewald et al, 1997) by Agrobacterium-mediated transformation (Rocha-Sosa et al, 1989). Transgenic plants were selected on hygromycin containing medium (Dietze et al, 1995).…”

Section: Construction Of Glucokinase Construct For Potato Transformationmentioning

confidence: 99%

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 patatin promoter region was isolated by PCR using oligonucleotide primers based on the published sequence by Rocha-Sosa et al (1989). The PCR primers, containing an Ncol site at the initiation codon, produced a 1-0 Kb fragment.…”

Section: Construction Of Chimeric Sacb Genesmentioning

confidence: 99%

“…1 b) cassette was transformed into potato leaf disks. Tuber-specific expression of the gene was accomplished by replacing the promoter in pSSU-Sac with a 1-0 Kb patatin promoter region (Rocha-Sosa et al, 1989). Transformed lines were selected by growth on kanamycin media and confirmed by PCR using SacB coding sequence primers (data not shown).…”

Section: Tissue-specific Expression Of Sacb Protein In Potato Tubersmentioning

confidence: 99%

Cytosolic expression of the Bacillus amyloliquefaciens SacB protein inhibits tissue development in transgenic tobacco and potato

Caimi¹,

McCOLE²,

KLEIN³

et al. 1997

New Phytologist

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SUMMARYFructans are linear or brancbed polymers containing a single sucrose and repeating fructose residues. An early model for fructan biosyntbesis in bigber plants suggested that partial synthesis of tbe polymer occurred in tbe cell cytosol. Tbe current model suggests tbat synthesis requires the interaction of two separate fructosyltransferases located in the vacuole. Tobacco lines containing a chemically induced promoter, directing expression of the Bacillus amyloliquefaciens SacB gene in tbe present study, provided an opportunity to regulate and target fructan synthesis to tbe cytosol of transgenic plants. Induced expression of tbe gene led to rapid destruction of leaf tissue. Amino acid substitution at a bigbly conserved site (Arg^^^) in tbe SacB gene reduced tbe fructosyltransferase efficiency witbout reducing tbe invertase activity of tbe enzyme. Expression of tbe mutant gene in transgenic tobacco also resulted in leaf damage. Hov^^ever, the appearance of necrotic tissue was greatly delayed. The results suggest tbat tbe pbenotype is due to accumulation of fructan in tbe cytosol. Fructan metabolism in tbe cytosol of potato tubers was also detrimental to development. Tuber size and starcb synthesis was significantly reduced in lines containing tbe untargeted gene. Transgenic tobacco and potato containing tbe SacB gene offer an opportunity to study tbe metabolism of fructan and the effect of accumulation on plant cell development.

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“…Previous experiments have already shown that both develo pmental and metabolic signals activate the promoter of cert~Lin tuber storage protein genes [24][25][26]. Recent studies on patatin promoter have identified tuber-specific and sucroseiiLducible cis elements [27,28].…”

Section: Discussionmentioning

confidence: 99%

Starch synthesis‐, and tuber storage protein genes are differently expressed in Solanum tuberosum and in Solanum brevidens

Bánfalvi¹,

Molnár²,

Molnár³

et al. 1996

FEBS Letters

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Studying in vitro stem cuttings of Solanum tuberosum induced for tuberization and those of a non-tuberizing Solanum species, differences both in morphology and in gene expression were detected. Stolon formation essentially depended on light while tuberization was triggered by the elevated level of sucrose in the medium. Genes involved in starch synthesis were induced b) sucrose in both species, however, starch granules were detected only in potato. A new tuber specific cDNA clone, GM7, encoding a putative metallocarboxypeptidase inhibitor and the cDNA of a proline rich cell wall protein with S. brevidens specific expression were isolated by differential screening. Sucrose mediated transcription of the tuber storage proteins like patatin and proteinase inhibitors (Kunitz-type, winI, GM7) failed in S. brevidens. K~ y words."Patatin; Proteinase inhibitor; Solanum brevidens; S~lanum tuberosurn; Starch synthesis; Tuberization synthase (GBSS) branching enzyme (BE) and plastidic starch synthase (STP) expressions and activities [4]. Starch is synthesized from sucrose that is assimilated in leaves and transported to the tuber. Translocation of sucrose from mesophyll into the phloem is done via the sucrose transporter (SUT) [5]. Transgenic plants with antisense-RNA inhibition of SUT show a dramatic reduction in tuber yield [6]. In the absence of Susy or AGPase sugar-storing tubers with decreased amount of soluble proteins are formed [4,7].An alternative approach toward understanding the molecular mechanism of tuber morphogenesis might be to study the evolution of tuberization. Solanum brevidens is a close relative of potato (S. tuberosum), however, with no tuber-forming ability. Here we report that by isolating single-node stem segments and growing them under various conditions, in vitro, differences both in morphology and in gene expression could be detected between the two species.

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Both developmental and metabolic signals activate the promoter of a class I patatin gene

Cited by 393 publications

References 24 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

Cytosolic expression of the Bacillus amyloliquefaciens SacB protein inhibits tissue development in transgenic tobacco and potato

Starch synthesis‐, and tuber storage protein genes are differently expressed in Solanum tuberosum and in Solanum brevidens

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