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T-DNA-encoded auxin formation in crown-gall cells.
E Kemper1, S Wafenschmidt, E W Weiler
1Abt. Schell, Max-Planck-Institut für Züchtungsforschung, D-5000, Köln 30.
Crown gall tumor cells utilize Ti plasmid genes for auxin production. Gene 2 encodes an amidohydrolase that converts indole-3-acetamide to indole-3-acetic acid, crucial for plant growth.
Area of Science:
- Plant Molecular Biology
- Biochemistry
- Microbial Genetics
Background:
- Ti plasmids in Agrobacterium tumefaciens mediate crown gall formation.
- T-region genes 1 and 2 are essential for auxin production in crown gall cells.
- Gene 2 (IaaH) is known to encode an amidohydrolase converting indole-3-acetamide to indole-3-acetic acid.
Purpose of the Study:
- To develop a quantitative assay for the T-DNA gene 2-encoded amidohydrolase.
- To analyze the enzyme's properties in plant cells and Escherichia coli.
- To investigate the substrate specificity and potential broader function of the enzyme.
Main Methods:
- Quantitative enzyme assay development.
- Enzyme characterization in plant extracts and E. coli.
- Substrate specificity analysis using various indole derivatives and amides.
Main Results:
- A quantitative assay for the amidohydrolase was established.
- The enzyme exhibited similar properties (temperature/pH optima, Km) in plant cells and E. coli.
- The enzyme hydrolyzed indole-3-acetamide, indole-3-acetonitrile, IAA-glucose, IAA-myo-inositol, a-naphthaleneacetamide, and phenylacetamide.
Conclusions:
- T-DNA gene 2 product is a versatile amidohydrolase with broad substrate specificity.
- The enzyme likely plays a general role in converting auxin precursors to active auxins.
- Findings provide insights into auxin biosynthesis mechanisms in crown gall tumors and potential functions of T-DNA gene 1.
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