Plant Physiol. Journal of Pharmacology and Experimental Therapeutics
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Published on April 13, 2007; 10.1104/pp.107.096511

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Received January 26, 2007
Accepted April 9, 2007

Genetic Dissection of Histidine Biosynthesis in Arabidopsis

Rosanna Muralla , Colleen Sweeney , Asya Stepansky , Thomas Leustek , and David Meinke *

Department of Botany, Oklahoma State University, Stillwater, Oklahoma, 74078; Biotechnology Center for Agriculture and the Environment, Department of Plant Biology and; Pathology, Rutgers University, New Brunswick, New Jersey, 08901

* Corresponding author; email: meinke{at}okstate.edu.

The biosynthesis of histidine in microorganisms, long studied through the isolation and characterization of auxotrophic mutants, has emerged as a paradigm for the regulation of metabolism and gene expression. Much less is known about histidine biosynthesis in flowering plants. One limiting factor has been the absence of large collections of informative auxotrophs. We describe here the results of a systematic screen for histidine auxotrophs of Arabidopsis thaliana. Ten insertion mutants disrupted in four different biosynthetic genes (HISN2, HISN3, HISN4, HISN6A) were identified through a combination of forward and reverse genetics and were shown to exhibit an embryo-defective (emb) phenotype that could be rescued by watering heterozygous plants with histidine. Male transmission of the mutant allele was in several cases reduced. Knockouts of two redundant genes (HISN1B, HISN5A) had no visible phenotype. Another mutant blocked in the final step of histidine biosynthesis (hisn8) and a double mutant altered in the redundant first step of the pathway (hisn1a hisn1b) exhibited a combination of gametophytic and embryonic lethality in heterozygotes. Homozygous mutant seedlings and callus tissue produced from rescued seeds appeared normal when grown in the presence of histidine but typically senesced after continued growth in the absence of histidine. These knockout mutants document the importance of histidine biosynthesis for plant growth and development, provide valuable insights into amino acid transport and source-sink relationships during seed development, and represent a significant addition to the limited collection of well-characterized auxotrophs in flowering plants.




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