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Integrated metabolomic and transcriptomic analyses of high-tryptophan rice expressing a mutant anthranilate synthase alpha subunit.
J Exp Bot. 2007; 58(12):3309-21.JE

Abstract

Transgenic rice plants overexpressing a mutant rice gene for anthranilate synthase alpha subunit (OASA1D) accumulate large amounts of free tryptophan (Trp) with few adverse effects on the phenotype, except for poor germination and weak seedling growth. Metabolic profiling of 8-d-old seedlings of Nipponbare and two high-Trp lines, HW1 and HW5, by high performance liquid chromatography-photo diode array (HPLC-PDA) confirmed that, relative to Nipponbare, only the peak attributed to Trp was significantly changed in the profiles of the OASA1D lines. More detailed and targeted analysis using HPLC coupled with tandem mass spectrometry revealed that the OASA1D lines had higher levels of anthranilate, tryptamine, and serotonin than Nipponbare, but these metabolites were at much lower levels than free Trp. The levels of phenylalanine (Phe) and tyrosine (Tyr) were not affected by the overproduction of Trp. Transcriptomic analysis by microarray validated by quantitative Real-Time PCR (qRT-PCR) revealed that at least 12 out of 21 500 genes showed significant differential expression among genotypes. Except for the OASA1D transgene and a putative IAA beta-glucosyltransferase, these were not related to Trp metabolism. Most importantly, the overexpression of the OASA1D and the consequent accumulation of Trp in these lines had little effect on the overall transcriptome, consistent with the minimal effects on growth and the metabolome. Integrated analysis of the metabolome and transcriptome of these OASA1D transgenic lines indicates that the over-accumulation of free Trp may be partly due to the low activity of Trp decarboxylase or other metabolic genes that directly utilize Trp as a substrate.

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Authors+Show Affiliations

Dubouzet JG
CREST, Japan Science and Technology Agency, Tokyo 103-0027, Japan.
Ishihara A
No affiliation info available
Matsuda F
No affiliation info available
Miyagawa H
No affiliation info available
Iwata H
No affiliation info available
Wakasa K
No affiliation info available

MeSH

Anthranilate SynthaseBase SequenceChromatography, High Pressure LiquidDNA PrimersGene Expression Regulation, PlantOryzaRNA, MessengerReverse Transcriptase Polymerase Chain ReactionTandem Mass Spectrometry

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

17804429

Citation

Dubouzet, Joseph G., et al. "Integrated Metabolomic and Transcriptomic Analyses of High-tryptophan Rice Expressing a Mutant Anthranilate Synthase Alpha Subunit." Journal of Experimental Botany, vol. 58, no. 12, 2007, pp. 3309-21.
Dubouzet JG, Ishihara A, Matsuda F, et al. Integrated metabolomic and transcriptomic analyses of high-tryptophan rice expressing a mutant anthranilate synthase alpha subunit. J Exp Bot. 2007;58(12):3309-21.
Dubouzet, J. G., Ishihara, A., Matsuda, F., Miyagawa, H., Iwata, H., & Wakasa, K. (2007). Integrated metabolomic and transcriptomic analyses of high-tryptophan rice expressing a mutant anthranilate synthase alpha subunit. Journal of Experimental Botany, 58(12), 3309-21.
Dubouzet JG, et al. Integrated Metabolomic and Transcriptomic Analyses of High-tryptophan Rice Expressing a Mutant Anthranilate Synthase Alpha Subunit. J Exp Bot. 2007;58(12):3309-21. PubMed PMID: 17804429.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Integrated metabolomic and transcriptomic analyses of high-tryptophan rice expressing a mutant anthranilate synthase alpha subunit. AU - Dubouzet,Joseph G, AU - Ishihara,Atsushi, AU - Matsuda,Fumio, AU - Miyagawa,Hisashi, AU - Iwata,Hiroyoshi, AU - Wakasa,Kyo, Y1 - 2007/09/04/ PY - 2007/9/7/pubmed PY - 2008/2/6/medline PY - 2007/9/7/entrez SP - 3309 EP - 21 JF - Journal of experimental botany JO - J Exp Bot VL - 58 IS - 12 N2 - Transgenic rice plants overexpressing a mutant rice gene for anthranilate synthase alpha subunit (OASA1D) accumulate large amounts of free tryptophan (Trp) with few adverse effects on the phenotype, except for poor germination and weak seedling growth. Metabolic profiling of 8-d-old seedlings of Nipponbare and two high-Trp lines, HW1 and HW5, by high performance liquid chromatography-photo diode array (HPLC-PDA) confirmed that, relative to Nipponbare, only the peak attributed to Trp was significantly changed in the profiles of the OASA1D lines. More detailed and targeted analysis using HPLC coupled with tandem mass spectrometry revealed that the OASA1D lines had higher levels of anthranilate, tryptamine, and serotonin than Nipponbare, but these metabolites were at much lower levels than free Trp. The levels of phenylalanine (Phe) and tyrosine (Tyr) were not affected by the overproduction of Trp. Transcriptomic analysis by microarray validated by quantitative Real-Time PCR (qRT-PCR) revealed that at least 12 out of 21 500 genes showed significant differential expression among genotypes. Except for the OASA1D transgene and a putative IAA beta-glucosyltransferase, these were not related to Trp metabolism. Most importantly, the overexpression of the OASA1D and the consequent accumulation of Trp in these lines had little effect on the overall transcriptome, consistent with the minimal effects on growth and the metabolome. Integrated analysis of the metabolome and transcriptome of these OASA1D transgenic lines indicates that the over-accumulation of free Trp may be partly due to the low activity of Trp decarboxylase or other metabolic genes that directly utilize Trp as a substrate. SN - 0022-0957 UR - https://www.unboundmedicine.com/medline/citation/17804429/Integrated_metabolomic_and_transcriptomic_analyses_of_high_tryptophan_rice_expressing_a_mutant_anthranilate_synthase_alpha_subunit_ L2 - https://academic.oup.com/jxb/article-lookup/doi/10.1093/jxb/erm179 DB - PRIME DP - Unbound Medicine ER -