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Induction of potato steroidal glycoalkaloid biosynthetic pathway by overexpression of cDNA encoding primary metabolism HMG-CoA reductase and squalene synthase

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Abstract

Potato steroidal glycoalkaloids (SGAs) are toxic secondary metabolites whose total content in tubers must be regulated. SGAs are biosynthesized by the sterol branch of the mevalonic acid/isoprenoid pathway. In a previous study, we showed a correlation between SGA levels and the abundance of transcript coding for HMG-CoA reductase 1 (HMG1) and squalene synthase 1 (SQS1) in potato tissues and potato genotypes varying in SGA content. Here, Solanum tuberosum cv. Desirée (low SGA producer) was transformed with a gene construct containing the coding region of either HMG1 or SQS1 of Solanum chacoense Bitt. clone 8380-1, a high SGA producer. SGA levels in transgenic HMG-plants were either greater than (in eight of 14 plants) or no different from untransformed controls, whereas only four of 12 SQS-transgenics had greater SGA levels than control, as determined by HPLC. Quantitative real-time PCR was used to estimate relative steady-state transcript levels of isoprenoid-, steroid-, and SGA-related genes in leaves of the transgenic plants compared to nontransgenic controls. HMG-transgenic plants exhibited increased transcript accumulation of SQS1, sterol C24-methyltransferase type1 (SMT1), and solanidine glycosyltransferase 2 (SGT2), whereas SQS-transgenic plants, had consistently lower transcript levels of HMG1 and variable SMT1 and SGT2 transcript abundance among different transgenics. HMG-transgenic plants exhibited changes in transcript accumulation for some sterol biosynthetic genes as well. Taken together, the data suggest coordinated regulation of isoprenoid metabolism and SGA secondary metabolism.

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Abbreviations

HMGR:

3-Hydroxy-3-methylglutaryl coenzyme A reductase

CAS:

Cycloartenol synthase

Chc80-1:

S. chacoense clone 8380-1

CYP51G:

Obtusifoliol 14-α-demethylase

DWF:

DWARF

FK:

Fackel

HYD1:

C-8 sterol isomerase

LAS:

Lanosterol synthase

SGA:

Steroidal glycoalkaloids

SGT1:

Solanidine galactosyltransferase

SGT2:

Solanidine glucosyltransferase

SGT3:

Steroidal glycoalkaloid rhamnosyltransferase

SMO:

4- α methyl oxidase

SMT:

Sterol C24-methyltransferase

SQS:

Squalene synthase

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Acknowledgments

The authors would like to thank Pini Krits and Zechariah Tanami from the Volcani Center for technical assistance. This research was supported by Research Grant No. IS-4134-08 from BARD, The United States-Israel Binational Agricultural Research and Development Fund, and is a contribution No. 108/2011 from the ARO, The Volcani Center, Bet Dagan, Israel.

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  1. Muddarangappa Thippeswamy

    Present address: Department of Biochemistry, Indian Institute of Science, Bangalore, 560012, India

Authors and Affiliations

  1. Institute of Plant Sciences, ARO, the Volcani Center, 50250, Bet Dagan, Israel

    Idit Ginzberg, Muddarangappa Thippeswamy, Edna Fogelman & Ufuk Demirel

  2. Department of Horticulture, Virginia Tech Blacksburg, Virginia, USA

    Alice M. Mweetwa, James Tokuhisa & Richard E. Veilleux

  3. Department of Field Crops, Harran University, Sanliurfa, 63000, Turkey

    Ufuk Demirel

  4. University of Zambia, Lusaka, Zambia

    Alice M. Mweetwa

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  1. Idit Ginzberg
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Correspondence to Idit Ginzberg.

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Ginzberg, I., Thippeswamy, M., Fogelman, E. et al. Induction of potato steroidal glycoalkaloid biosynthetic pathway by overexpression of cDNA encoding primary metabolism HMG-CoA reductase and squalene synthase. Planta 235, 1341–1353 (2012). https://doi.org/10.1007/s00425-011-1578-6

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