J Integr Plant Biol. ›› 2011, Vol. 53 ›› Issue (12): 942-950.DOI: 10.1111/j.1744-7909.2011.01083.x

• Metabolism and Biochemistry • Previous Articles     Next Articles

Characterization of Three Homoeologous cDNAs Encoding Chloroplast-targeted Aminolevulinic Acid Dehydratase in Common Wheat

Yu Takenouchi1, Haruka Nakajima2, Kengo Kanamaru2 and Shigeo Takumi1*   

  1. 1Laboratory of Plant Genetics, Graduate School of Agricultural Science, Kobe University, Rokkodai 1-1, Nada, Kobe 657-8501, Japan
    2Laboratory of Biological Chemistry, Graduate School of Agricultural Science, Kobe University, Rokkodai 1-1, Nada, Kobe 657-8501, Japan
  • Received:2011-08-18 Accepted:2011-10-23 Published:2011-12-01
  • About author:*Corresponding author. Tel(Fax): +81 78 803 5860, E-mail: takumi@kobe-u.ac.jp

Abstract:

In the tetrapyrrole biosynthetic pathway of higher plants, 5-aminolevulinic acid (ALA) is metabolized by ALA dehydratase (ALAD). Here, we isolated ALAD1 cDNA from common wheat (Triticum aestivum L.) and its diploid progenitors, and produced transgenic tobacco plants expressing the wheat ALAD1 gene. The ALAD1 genes were highly conserved among wheat relatives, and three homoeologous loci of wheat ALAD1 (TaALAD1) were equally transcribed in common wheat. A transient expression assay of a TaALAD1-GFP (green fluorescent protein) fusion protein suggested that TaALAD1 is localized in chloroplasts. Overexpression of TaALAD1 in transgenic tobacco resulted in a significant increase in ALAD activity in leaves. Moreover, the transgenic tobacco showed vigorous growth and increased survival rate on medium containing ALA at herbicidal concentrations. These results indicate that wheat ALAD1 has catalytic activity in metabolizing ALA in plastids, and that ectopic expression of TaALAD1 in transgenic plants increases their tolerance to ALA application at high concentrations.

Takenouchi Y, Nakajima H, Kanamaru K, Takumi S (2011) Characterization of three homoeologous cDNAs encoding chloroplast-targeted aminolevulinic acid dehydratase in common wheat. J. Integr. Plant Biol. 53(12), 942–950.

Key words: 5-aminolevulinic acid, chloroplast targeting, herbicide resistance, transgenic tobacco, Triticum aestivum, 5-aminolevulinic acid dehydratase, transcriptional control

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