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. 1985 Apr;4(4):853–859. doi: 10.1002/j.1460-2075.1985.tb03710.x

Tumour genes in plants: T-DNA encoded cytokinin biosynthesis

Ilka Buchmann 1, Franz-Josef Marner 1, Gudrun Schröder 1, Sabine Waffenschmidt 1, Joachim Schröder 1
PMCID: PMC554271  PMID: 15938044

Abstract

Gene 4 from the T-region of Ti plasmids is responsible for cytokinin effects in crown gall cells; we investigated whether it codes for an enzyme of hormone biosynthesis. In a first set of experiments, gene 4 from octopine plasmid pTiAch5 and nopaline plasmid pTiC58 was expressed in Escherichia coli, and the gene products were identified by reaction with antiserum raised against a decapeptide derived from the DNA sequence of the gene. Extracts from cells expressing the gene contained high isopentenyl-transferase activity catalyzing the formation of N6-(△2-isopentenyl)adenosine from 5'-AMP and △2-isopentenylpyrophosphate. The cytokinin was identified by sequential h.p.l.c. chromatography and mass spectrometry. In a second set of experiments it was shown that crown gall cells contained isopentenyltransferase activity and a protein of mol. wt. 27 000 which was identified as the product of gene 4 by reaction with the antiserum. Isopentenyltransferase activity was specifically inhibited by the antiserum. No comparable enzyme activity or immunoreactive protein was detected in cytokinin-autotrophic, T-DNA free tobacco cells. The results establish that gene 4 from the T-region of octopine and nopaline Ti plasmids codes for an enzyme of cytokinin biosynthesis.

Keywords: cytokinin biosynthesis, isopentenyltransferase, peptide antiserum, crown gall, Ti plasmids

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Selected References

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