Terpene metabolic engineering via nuclear or chloroplast genomes profoundly and globally impacts off‐target pathways through metabolite signalling
Abstract
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 largemore »
- Authors:
-
- 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
- Publication Date:
- Research Org.:
- Univ. of Pennsylvania, Philadelphia, PA (United States)
- Sponsoring Org.:
- USDOE Advanced Research Projects Agency - Energy (ARPA-E); National Institutes of Health (NIH)
- OSTI Identifier:
- 1240755
- Alternate Identifier(s):
- OSTI ID: 1240756; OSTI ID: 1376055
- Grant/Contract Number:
- R01 HL107904; R01 HL109442
- Resource Type:
- Published Article
- Journal Name:
- Plant Biotechnology Journal
- Additional Journal Information:
- Journal Name: Plant Biotechnology Journal Journal Volume: 14 Journal Issue: 9; Journal ID: ISSN 1467-7644
- Publisher:
- Wiley-Blackwell
- Country of Publication:
- United Kingdom
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; Secondary metabolism; retrograde/anterograde signalling; transgenic/transplastomic plants; squalene
Citation Formats
Pasoreck, Elise K., Su, Jin, Silverman, Ian M., Gosai, Sager J., Gregory, Brian D., Yuan, Joshua S., and Daniell, Henry. Terpene metabolic engineering via nuclear or chloroplast genomes profoundly and globally impacts off‐target pathways through metabolite signalling. United Kingdom: N. p., 2016.
Web. doi:10.1111/pbi.12548.
Pasoreck, Elise K., Su, Jin, Silverman, Ian M., Gosai, Sager J., Gregory, Brian D., Yuan, Joshua S., & Daniell, Henry. Terpene metabolic engineering via nuclear or chloroplast genomes profoundly and globally impacts off‐target pathways through metabolite signalling. United Kingdom. https://doi.org/10.1111/pbi.12548
Pasoreck, Elise K., Su, Jin, Silverman, Ian M., Gosai, Sager J., Gregory, Brian D., Yuan, Joshua S., and Daniell, Henry. Tue .
"Terpene metabolic engineering via nuclear or chloroplast genomes profoundly and globally impacts off‐target pathways through metabolite signalling". United Kingdom. https://doi.org/10.1111/pbi.12548.
@article{osti_1240755,
title = {Terpene metabolic engineering via nuclear or chloroplast genomes profoundly and globally impacts off‐target pathways through metabolite signalling},
author = {Pasoreck, Elise K. and Su, Jin and Silverman, Ian M. and Gosai, Sager J. and Gregory, Brian D. and Yuan, Joshua S. and Daniell, Henry},
abstractNote = {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.},
doi = {10.1111/pbi.12548},
journal = {Plant Biotechnology Journal},
number = 9,
volume = 14,
place = {United Kingdom},
year = {Tue Mar 08 00:00:00 EST 2016},
month = {Tue Mar 08 00:00:00 EST 2016}
}
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