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Enhanced flux of substrates into polyamine biosynthesis but not ethylene in tomato fruit engineered with yeast S-adenosylmethionine decarboxylase gene

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Abstract

S-adenosylmethionine (SAM), a major substrate in 1-C metabolism is a common precursor in the biosynthetic pathways of polyamines and ethylene, two important plant growth regulators, which exhibit opposing developmental effects, especially during fruit ripening. However, the flux of various substrates including SAM into the two competing pathways in plants has not yet been characterized. We used radiolabeled 14C-Arg, 14C-Orn, L-[U-14C]Met, 14C-SAM and 14C-Put to quantify flux through these pathways in tomato fruit and evaluate the effects of perturbing these pathways via transgenic expression of a yeast SAM decarboxylase (ySAMDC) gene using the fruit ripening-specific promoter E8. We show that polyamines in tomato fruit are synthesized both from Arg and Orn; however, the relative contribution of Orn pathway declines in the later stages of ripening. Expression of ySAMDC reversed the ripening associated decline in spermidine (Spd) and spermine (Spm) levels observed in the azygous control fruit. About 2- to 3-fold higher levels of labeled-Spd in transgenic fruit (556HO and 579HO lines) expressing ySAMDC confirmed the enzymatic function of the introduced gene. The incorporation of L-[U-14C]Met into Spd, Spm, ethylene and 1-aminocyclopropane-1-carboxylic acid (ACC) was used to determine Met-flux into these metabolites. The incorporation of 14C-Met into Spd/Spm declined during ripening of the control azygous fruit but this was reversed in fruits expressing ySAMDC. However, incorporation of 14C-Met into ethylene or ACC during ripening was not altered by the expression of ySAMDC in the fruit. Taken together these results show that: (1) There is an inverse relationship between the production of higher polyamines and ethylene during fruit ripening, (2) the inverse relationship between higher polyamines and ethylene is modulated by ySAMDC expression in that the decline in Spd/Spm during fruit ripening can be reversed without significantly altering ethylene biosynthesis, and (3) cellular flux of SAM in plants is homeostatically regulated based on its demand for competing pathways.

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Abbreviations

SAM:

S-adenosylmethionine

ACS:

1-aminocyclopropane-1-carboxylate (ACC) synthase

ACO:

ACC oxidase

ADC:

Arginine decarboxylase

dcSAM:

Decarboxylated SAM

MTA:

Methylthioadenosine

ODC:

Ornithine decarboxylase

SAMDC:

SAM decarboxylase

Put:

Putrescine

Spd:

Spermidine

SPDS:

Spd synthase

Spm:

Spermine

SPMS:

Spm synthase

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Acknowledgments

The authors would like to thank Stephanie Long and Ken Dudzik for their technical assistance. The New Hampshire Agricultural Experiment Station (NHAES) provided partial funding for this work at UNH; this is Scientific Contribution Number 2508 from the NHAES. Mention of trade names or commercial products in this article is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the U.S. Department of Agriculture.

Conflict of interest

The authors declare no conflicts of interest.

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Author notes

  1. Yi Lasanajak

    Present address: Emory University School of Medicine, Atlanta, GA, 30322, USA

  2. Ravinder Goyal

    Present address: Ag Fd Canada, Lethbridge, AB, Canada

  3. Tahira Fatima

    Present address: University of Western Ontario, London, ON, N6a 5b7, Canada

Authors and Affiliations

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

    Yi Lasanajak & Subhash C. Minocha

  2. USDA Forest Service, Durham, NH, 03824, USA

    Rakesh Minocha

  3. Henry A. Wallace Beltsville Agricultural Research Center, USDA-ARS, Sustainable Agricultural Systems Laboratory, Building 001, Beltsville, MD, 20705, USA

    Ravinder Goyal, Tahira Fatima & Autar K. Mattoo

  4. Department of Horticulture and Landscape Architecture, Purdue University, West Lafayette, IN, 47907, USA

    Avtar K. Handa

Authors
  1. Yi Lasanajak
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Correspondence to Autar K. Mattoo.

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Lasanajak, Y., Minocha, R., Minocha, S.C. et al. Enhanced flux of substrates into polyamine biosynthesis but not ethylene in tomato fruit engineered with yeast S-adenosylmethionine decarboxylase gene. Amino Acids 46, 729–742 (2014). https://doi.org/10.1007/s00726-013-1624-8

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