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  Metabolic engineering of tomato fruit organic Acid content guided by biochemical analysis of an introgression line

Morgan, M. J., Osorio, S., Gehl, B., Baxter, C. J., Kruger, N. J., Ratcliffe, R. G., et al. (2013). Metabolic engineering of tomato fruit organic Acid content guided by biochemical analysis of an introgression line. Plant Physiology, 161(1), 397-407. doi:10.1104/pp.112.209619.

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Morgan, M. J.1, Author
Osorio, S.2, Author              
Gehl, B.1, Author
Baxter, C. J.1, Author
Kruger, N. J.1, Author
Ratcliffe, R. G.1, Author
Fernie, A. R.2, Author              
Sweetlove, L. J.1, Author
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1External Organizations, ou_persistent22              
2Central Metabolism, Department Willmitzer, Max Planck Institute of Molecular Plant Physiology, Max Planck Society, ou_1753339              

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 Abstract: Organic acid content is regarded as one of the most important quality traits of fresh tomato (Solanum lycopersicum). However, the complexity of carboxylic acid metabolism and storage means that it is difficult to predict the best way to engineer altered carboxylic acid levels. Here, we used a biochemical analysis of a tomato introgression line with increased levels of fruit citrate and malate at breaker stage to identify a metabolic engineering target that was subsequently tested in transgenic plants. Increased carboxylic acid levels in introgression line 2-5 were not accompanied by changes in the pattern of carbohydrate oxidation by pericarp discs or the catalytic capacity of tricarboxylic acid cycle enzymes measured in isolated mitochondria. However, there was a significant decrease in the maximum catalytic activity of aconitase in total tissue extracts, suggesting that a cytosolic isoform of aconitase was affected. To test the role of cytosolic aconitase in controlling fruit citrate levels, we analyzed fruit of transgenic lines expressing an antisense construct against SlAco3b, one of the two tomato genes encoding aconitase. A green fluorescent protein fusion of SlAco3b was dual targeted to cytosol and mitochondria, while the other aconitase, SlAco3a, was exclusively mitochondrial when transiently expressed in tobacco (Nicotiana tabacum) leaves. Both aconitase transcripts were decreased in fruit from transgenic lines, and aconitase activity was reduced by about 30% in the transgenic lines. Other measured enzymes of carboxylic acid metabolism were not significantly altered. Both citrate and malate levels were increased in ripe fruit of the transgenic plants, and as a consequence, total carboxylic acid content was increased by 50% at maturity.

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Language(s): eng - English
 Dates: 2012-11-212013
 Publication Status: Published in print
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 Identifiers: Other: 23166354
DOI: 10.1104/pp.112.209619
ISSN: 1532-2548 (Electronic)0032-0889 (Linking)
URI: http://www.ncbi.nlm.nih.gov/pubmed/23166354http://www.plantphysiol.org/content/161/1/397.full.pdf
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Title: Plant Physiology
  Other : Plant Physiol.
Source Genre: Journal
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Publ. Info: Bethesda, Md. : American Society of Plant Biologists
Pages: - Volume / Issue: 161 (1) Sequence Number: - Start / End Page: 397 - 407 Identifier: ISSN: 0032-0889
CoNE: https://pure.mpg.de/cone/journals/resource/991042744294438