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Abstract Detail

Metabolic Engineering

Collins-Hed, Andrew [1], Campbell, Michael [2], Wang, Yi-Hong [2].

Changes in Branched Chain Amino Acid Composition in the Fruit of Acetohydroxy Acid Synthase Silenced Tomato Mutants.

The branched chain amino acids (BCAAs) are essential for nutrition and can only be acquired through diet. The biosynthetic pathway resulting in the BCAAs involves a key enzyme acetohydroxy acid synthase (AHAS). The objective of this study is to determine if modification of AHAS expression can change BCAA composition in tomato (Solanum lycopersicum) fruit. To begin this study, amine profiles of developing wild type tomato fruits were characterized. Fruit development was broken into five stages from one, young and green, to five being red and ripe. Amines were extracted from the fruit and derivatized with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate. Profiles of fluorescently tagged amines were produced using high performance liquid chromatography. The fluorescent profiles were compared to profiles of single known amino acids and the common retention times were used to infer peak identities. All the fruit stages revealed peaks that correlated with glutamate and the BCAAs. A portion of the gene for AHAS was inserted into the ethanol inducible RNAi vector pMW4 creating the plasmid pMWAHAS. Plants were regenerated from leaf tissue transformed with pMWAHAS. Transformants were confirmed using PCR. Transformants were exposed to 1% ethanol over a period of several days to express the RNAi. Results of QT-PCR and HPLC analysis on extracts from the transformants will be presented.

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1 - Penn State Erie, The Behrend College, Biology, 4701 Station Road, Erie, Pennsylvania, 16563, United States of America
2 - Penn State Erie, The Behrend College, Biology

Branched-chain amino acid biosynthesis
Acetohydroxyacid synthase (AHAS)
Acetolactate synthase

Presentation Type: Plant Biology Abstract
Session: P
Location: Exhibit Hall (Northeast, Southwest & Southeast)/Hilton
Date: Sunday, July 8th, 2007
Time: 8:00 AM
Number: P43004
Abstract ID:422

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