Terpene metabolic engineering via nuclear or chloroplast genomes profoundly and globally impacts off‐target pathways through metabolite signalling

Pasoreck, Elise K.; Su, Jin; Silverman, Ian M.; Gosai, Sager J.; Gregory, Brian D.; Yuan, Joshua S.; Daniell, Henry

doi:10.1111/pbi.12548

Title: Terpene metabolic engineering via nuclear or chloroplast genomes profoundly and globally impacts off‐target pathways through metabolite signalling

Journal Article · Tue Mar 08 00:00:00 EST 2016 · Plant Biotechnology Journal

DOI:https://doi.org/10.1111/pbi.12548· OSTI ID:1240755

Pasoreck, Elise K. ^[1]; Su, Jin ^[1]; Silverman, Ian M. ^[2]; Gosai, Sager J. ^[2]; Gregory, Brian D. ^[2]; Yuan, Joshua S. ^[3]; Daniell, Henry ^[1]

Department of Biochemistry School of Dental Medicine University of Pennsylvania Philadelphia PA USA
Department of Biology University of Pennsylvania Philadelphia PA USA
Department of Plant Pathology and Microbiology Texas A&,M University College Station TX USA

Summary The impact of metabolic engineering on nontarget pathways and outcomes of metabolic engineering from different genomes are poorly understood questions. Therefore, squalene biosynthesis genes FARNESYL DIPHOSPHATE SYNTHASE ( FPS ) and SQUALENE SYNTHASE ( SQS ) were engineered via the Nicotiana tabacum chloroplast (C), nuclear (N) or both ( CN ) genomes to promote squalene biosynthesis. SQS levels were ~4300‐fold higher in C and CN lines than in N, but all accumulated ~150‐fold higher squalene due to substrate or storage limitations. Abnormal leaf and flower phenotypes, including lower pollen production and reduced fertility, were observed regardless of the compartment or level of transgene expression. Substantial changes in metabolomes of all lines were observed: levels of 65–120 unrelated metabolites, including the toxic alkaloid nicotine, changed by as much as 32‐fold. Profound effects of transgenesis on nontarget gene expression included changes in the abundance of 19 076 transcripts by up to 2000‐fold in CN ; 7784 transcripts by up to 1400‐fold in N; and 5224 transcripts by as much as 2200‐fold in C. Transporter‐related transcripts were induced, and cell cycle‐associated transcripts were disproportionally repressed in all three lines. Transcriptome changes were validated by qRT ‐ PCR . The mechanism underlying these large changes likely involves metabolite‐mediated anterograde and/or retrograde signalling irrespective of the level of transgene expression or end product, due to imbalance of metabolic pools, offering new insight into both anticipated and unanticipated consequences of metabolic engineering.

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Research Organization:: Univ. of Pennsylvania, Philadelphia, PA (United States)

Sponsoring Organization:: USDOE Advanced Research Projects Agency - Energy (ARPA-E); National Institutes of Health (NIH)

Grant/Contract Number:: R01 HL107904; R01 HL109442

OSTI ID:: 1240755

Alternate ID(s):: OSTI ID: 1240756; OSTI ID: 1376055

Journal Information:: Plant Biotechnology Journal, Journal Name: Plant Biotechnology Journal Vol. 14 Journal Issue: 9; ISSN 1467-7644

Publisher:: Wiley-BlackwellCopyright Statement

Country of Publication:: United Kingdom

Language:: English

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Cited by: 23 works

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Title: Terpene metabolic engineering via nuclear or chloroplast genomes profoundly and globally impacts off‐target pathways through metabolite signalling

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