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Genes encoding nitrilase-like proteins from tobacco.

M Dohmoto1, H Tsunoda, G Isaji

  • 1Institute for Gene Research, Kanazawa University, Japan.

DNA Research : an International Journal for Rapid Publication of Reports on Genes and Genomes
|November 23, 2000
PubMed
Summary

Tobacco plants possess NIT4 homologs, unlike Arabidopsis thaliana which has NIT1-3 genes. Introducing Arabidopsis NIT2 into tobacco caused growth inhibition, while tobacco

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Area of Science:

  • Plant molecular biology
  • Biochemistry
  • Genetics

Background:

  • Nitrilases are enzymes catalyzing indole-3-acetonitrile (IAN) hydrolysis to indole-3-acetic acid (IAA), a key plant hormone.
  • The Arabidopsis thaliana genome contains four nitrilase genes (NIT1-4), with NIT1, NIT2, and NIT3 showing high similarity.

Purpose of the Study:

  • To investigate the presence and function of nitrilase homologs in tobacco (Nicotiana tabacum).
  • To compare the functional roles of Arabidopsis NIT2 and tobacco TNIT4A in response to IAN.

Main Methods:

  • Cloning of two NIT4 homologs (TNIT4A and TNIT4B) from tobacco.
  • Genomic Southern hybridization to assess the presence of nitrilase gene families in tobacco.
  • Introduction of Arabidopsis NIT2 and ectopic expression of TNIT4A in transgenic tobacco plants.

Related Experiment Videos

  • Assessment of IAN sensitivity in transgenic plants.

    • Main Results:

    • Genomic Southern hybridization indicated that tobacco possesses NIT4 homologs but lacks NIT1-3 homologs.
    • Transgenic tobacco expressing Arabidopsis NIT2 exhibited IAN-mediated growth inhibition.
    • Ectopic expression of tobacco TNIT4A in transgenic plants had minimal impact on IAN sensitivity.

    Conclusions:

    • Tobacco nitrilase gene composition differs from Arabidopsis, with a likely absence of NIT1-3 homologs.
    • Tobacco NIT4 homologs (e.g., TNIT4A) may have distinct functions or lower catalytic activity towards IAN compared to Arabidopsis NIT2.
    • The functional characterization of tobacco nitrilases provides insights into plant hormone metabolism and response diversity.