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Fruit-specific over-expression of human S-adenosylmethionine decarboxylase gene results in polyamine accumulation and affects diverse aspects of tomato fruit development and quality

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Abstract

Polyamines (PAs) have been implicated in fruit ripening where they antagonize the action of ethylene: a ripening inducing phytohormone. S-adenosylmethionine decarboxylase (SAMDC) is a key enzyme involved biosynthesis of higher PAs- spermidine and spermine. Here, we report the genetic modification of tomato fruit ripening and quality by over-expressing human-SAMDC driven by fruit-specific promoter (2A11). The PA analysis of ripening fruits from these transgenics showed elevated PA levels in comparison to wild-type (WT). The increased levels of higher PAs are correlated with the accumulation of heterologous SAMDC transcripts in such fruits. Transgenic fruits exhibited reduced levels of ethylene (~50 %) production, ~10 days delay in on-vine ripening and extended post-harvest storage of ~11 days as compared to the WT fruits. As a result, these fruits showed improvement in various ripening traits like enhanced lycopene, vitamin C and total soluble solid. In Lesam fruits, an up-regulated expression of SlySAMDC, SlyEXP1, SlyTBG4, SlyDXS 1 and SlyPSY 1 was observed, while ethylene biosynthesis genes were down-regulated. Here, we have demonstrated the important role of PAs in altering the molecular and biochemical processes underlying fruit ripening by interfering with the ethylene biosynthesis.

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Abbreviations

Put:

Putrescine

SAMDC:

S-adenosylmethionine decarboxylase

Spd:

Spermidine

Spm:

Spermine

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Acknowledgments

This work was generously supported by grants from the Department of Biotechnology (Govt. of India), New Delhi (Grant Nos. BT/PR/2990/Agr/16/232/2002 and BT/PR8657/PBD/16/738/2007), University Grants Commission–Special Assistance Programme and Department of Science and Technology–FIST programme. Research fellowships to P. Madhulatha and Aarti Gupta by the Council of Scientific and Industrial Research are acknowledged.

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Authors and Affiliations

  1. Plant Polyamine, Transgenic and RNAi Research Laboratory, Department of Genetics, University of Delhi–South Campus, New Delhi, 110021, India

    P. Madhulatha, Aarti Gupta, Saaraj Gupta & M. V. Rajam

  2. Division of Post-Harvest Technology, Indian Agricultural Research Institute, New Delhi, 110012, India

    Anuj Kumar & R. K. Pal

Authors
  1. P. Madhulatha
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  2. Aarti Gupta
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  3. Saaraj Gupta
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  4. Anuj Kumar
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  5. R. K. Pal
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  6. M. V. Rajam
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Corresponding author

Correspondence to M. V. Rajam.

Additional information

P. Madhulatha and Aarti Gupta contributed equally

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Supplementary Fig. 1

Molecular characterization of tomato primary transformants. (a) PCR with primers specific to NPT II gene: L-1 kb ladder; PC-Plasmid DNA; NC-DNA from untransformed control; Lesam 1–70-DNA from different transgenic tomato lines, (b) PCR with SAMDC gene-specific primers: L-1 kb ladder; PC-Plasmid DNA, NC-DNA from untransformed control; Lesam 1–56-DNA from different tomato transgenic lines, (c) Southern blot analysis of Lesam transgenics for copy number using NPT II gene probe. NC-DNA from untransformed control; Lesam 1–56- DNA from different transgenic tomato lines digested with XbaI enzyme. (d) PCR analysis of Lesam T3 progenies. L-Ladder; PC-Plasmid DNA; NC-DNA from untransformed control; D.1) PCR of Lesam16 D.2) PCR of Lesam24 D.3) PCR of Lesam56. 1–12 DNA from Lesam T3 progenies. (JPEG 85 kb)

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Table S1

List of primer sets used in the study (DOC 50 kb)

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Madhulatha, P., Gupta, A., Gupta, S. et al. Fruit-specific over-expression of human S-adenosylmethionine decarboxylase gene results in polyamine accumulation and affects diverse aspects of tomato fruit development and quality. J. Plant Biochem. Biotechnol. 23, 151–160 (2014). https://doi.org/10.1007/s13562-013-0194-x

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