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Living with high putrescine: expression of ornithine and arginine biosynthetic pathway genes in high and low putrescine producing poplar cells

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Abstract

Arginine (Arg) and ornithine (Orn), both derived from glutamate (Glu), are the primary substrates for polyamine (PA) biosynthesis, and also play important roles as substrates and intermediates of overall N metabolism in plants. Their cellular homeostasis is subject to multiple levels of regulation. Using reverse transcription quantitative PCR (RT-qPCR), we studied changes in the expression of all genes of the Orn/Arg biosynthetic pathway in response to up-regulation [via transgenic expression of mouse Orn decarboxylase (mODC)] of PA biosynthesis in poplar (Populus nigra × maximowiczii) cells grown in culture. Cloning and sequencing of poplar genes involved in the Orn/Arg biosynthetic pathway showed that they have high homology with similar genes in other plants. The expression of the genes of Orn, Arg and PA biosynthetic pathway fell into two hierarchical clusters; expression of one did not change in response to high putrescine, while members of the other cluster showed a shift in expression pattern during the 7-day culture cycle. Gene expression of branch point enzymes (N-acetyl-Glu synthase, Orn aminotransferase, Arg decarboxylase, and spermidine synthase) in the sub-pathways, constituted a separate cluster from those involved in intermediary reactions of the pathway (N-acetyl-Glu kinase, N-acetyl-Glu-5-P reductase, N-acetyl-Orn aminotransferase, N 2-acetylOrn:N-acetyl-Glu acetyltransferase, N 2-acetyl-Orn deacetylase, Orn transcarbamylase, argininosuccinate synthase, carbamoylphosphate synthetase, argininosuccinate lyase, S-adenosylmethionine decarboxylase, spermine synthase). We postulate that expression of all genes of the Glu-Orn-Arg pathway is constitutively coordinated and is not influenced by the increase in flux rate through this pathway in response to increased utilization of Orn by mODC; thus the pathway involves mostly biochemical regulation rather than changes in gene expression. We further suggest that Orn itself plays a major role in the regulation of this pathway.

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Acknowledgments

The authors are thankful to Dr. Karen Carlton for help in the RT-qPCR and to the UNH Hubbard Genome Center for sequencing. This work was partially supported by the USDA-NRI award # 2002-35318-12674, the NH Agricultural Experiment Station, and the US Forest Service, Northern Research Station.

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  1. Andrew F. Page

    Present address: Thermo Fisher Scientific, 3411 Silverside Rd, Bancroft Building, Wilmington, DE, 19810, USA

Authors and Affiliations

  1. Department of Biological Sciences, University of New Hampshire, Durham, NH, 03824, USA

    Andrew F. Page & Subhash C. Minocha

  2. US Forest Service, Northern Research Station, 271 Mast Road, Durham, NH, 03824, USA

    Rakesh Minocha

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  1. Andrew F. Page
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Correspondence to Subhash C. Minocha.

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Scientific contribution number 2374 from the NH Agricultural Experiment Station.

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Page, A.F., Minocha, R. & Minocha, S.C. Living with high putrescine: expression of ornithine and arginine biosynthetic pathway genes in high and low putrescine producing poplar cells. Amino Acids 42, 295–308 (2012). https://doi.org/10.1007/s00726-010-0807-9

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