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Rational design and testing of abiotic stress‐inducible synthetic promoters from poplar cis‐regulatory elements

Abiotic stress resistance traits may be especially crucial for sustainable production of bioenergy tree crops. Here, we show the performance of a set of rationally designed osmotic‐related and salt stress‐inducible synthetic promoters for use in hybrid poplar. De novo motif‐detecting algorithms yiel...

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Autores principales: Yang, Yongil, Lee, Jun Hyung, Poindexter, Magen R., Shao, Yuanhua, Liu, Wusheng, Lenaghan, Scott C., Ahkami, Amir H., Blumwald, Eduardo, Stewart, Charles Neal
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8313130/
https://www.ncbi.nlm.nih.gov/pubmed/33471413
http://dx.doi.org/10.1111/pbi.13550
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author Yang, Yongil
Lee, Jun Hyung
Poindexter, Magen R.
Shao, Yuanhua
Liu, Wusheng
Lenaghan, Scott C.
Ahkami, Amir H.
Blumwald, Eduardo
Stewart, Charles Neal
author_facet Yang, Yongil
Lee, Jun Hyung
Poindexter, Magen R.
Shao, Yuanhua
Liu, Wusheng
Lenaghan, Scott C.
Ahkami, Amir H.
Blumwald, Eduardo
Stewart, Charles Neal
author_sort Yang, Yongil
collection PubMed
description Abiotic stress resistance traits may be especially crucial for sustainable production of bioenergy tree crops. Here, we show the performance of a set of rationally designed osmotic‐related and salt stress‐inducible synthetic promoters for use in hybrid poplar. De novo motif‐detecting algorithms yielded 30 water‐deficit (SD) and 34 salt stress (SS) candidate DNA motifs from relevant poplar transcriptomes. We selected three conserved water‐deficit stress motifs (SD18, SD13 and SD9) found in 16 co‐expressed gene promoters, and we discovered a well‐conserved motif for salt response (SS16). We characterized several native poplar stress‐inducible promoters to enable comparisons with our synthetic promoters. Fifteen synthetic promoters were designed using various SD and SS subdomains, in which heptameric repeats of five‐to‐eight subdomain bases were fused to a common core promoter downstream, which, in turn, drove a green fluorescent protein (GFP) gene for reporter assays. These 15 synthetic promoters were screened by transient expression assays in poplar leaf mesophyll protoplasts and agroinfiltrated Nicotiana benthamiana leaves under osmotic stress conditions. Twelve synthetic promoters were induced in transient expression assays with a GFP readout. Of these, five promoters (SD18‐1, SD9‐2, SS16‐1, SS16‐2 and SS16‐3) endowed higher inducibility under osmotic stress conditions than native promoters. These five synthetic promoters were stably transformed into Arabidopsis thaliana to study inducibility in whole plants. Herein, SD18‐1 and SD9‐2 were induced by water‐deficit stress, whereas SS16‐1, SS16‐2 and SS16‐3 were induced by salt stress. The synthetic biology design pipeline resulted in five synthetic promoters that outperformed endogenous promoters in transgenic plants.
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spelling pubmed-83131302021-07-31 Rational design and testing of abiotic stress‐inducible synthetic promoters from poplar cis‐regulatory elements Yang, Yongil Lee, Jun Hyung Poindexter, Magen R. Shao, Yuanhua Liu, Wusheng Lenaghan, Scott C. Ahkami, Amir H. Blumwald, Eduardo Stewart, Charles Neal Plant Biotechnol J Research Articles Abiotic stress resistance traits may be especially crucial for sustainable production of bioenergy tree crops. Here, we show the performance of a set of rationally designed osmotic‐related and salt stress‐inducible synthetic promoters for use in hybrid poplar. De novo motif‐detecting algorithms yielded 30 water‐deficit (SD) and 34 salt stress (SS) candidate DNA motifs from relevant poplar transcriptomes. We selected three conserved water‐deficit stress motifs (SD18, SD13 and SD9) found in 16 co‐expressed gene promoters, and we discovered a well‐conserved motif for salt response (SS16). We characterized several native poplar stress‐inducible promoters to enable comparisons with our synthetic promoters. Fifteen synthetic promoters were designed using various SD and SS subdomains, in which heptameric repeats of five‐to‐eight subdomain bases were fused to a common core promoter downstream, which, in turn, drove a green fluorescent protein (GFP) gene for reporter assays. These 15 synthetic promoters were screened by transient expression assays in poplar leaf mesophyll protoplasts and agroinfiltrated Nicotiana benthamiana leaves under osmotic stress conditions. Twelve synthetic promoters were induced in transient expression assays with a GFP readout. Of these, five promoters (SD18‐1, SD9‐2, SS16‐1, SS16‐2 and SS16‐3) endowed higher inducibility under osmotic stress conditions than native promoters. These five synthetic promoters were stably transformed into Arabidopsis thaliana to study inducibility in whole plants. Herein, SD18‐1 and SD9‐2 were induced by water‐deficit stress, whereas SS16‐1, SS16‐2 and SS16‐3 were induced by salt stress. The synthetic biology design pipeline resulted in five synthetic promoters that outperformed endogenous promoters in transgenic plants. John Wiley and Sons Inc. 2021-02-08 2021-07 /pmc/articles/PMC8313130/ /pubmed/33471413 http://dx.doi.org/10.1111/pbi.13550 Text en © 2021 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd. https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ (https://creativecommons.org/licenses/by-nc-nd/4.0/) License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.
spellingShingle Research Articles
Yang, Yongil
Lee, Jun Hyung
Poindexter, Magen R.
Shao, Yuanhua
Liu, Wusheng
Lenaghan, Scott C.
Ahkami, Amir H.
Blumwald, Eduardo
Stewart, Charles Neal
Rational design and testing of abiotic stress‐inducible synthetic promoters from poplar cis‐regulatory elements
title Rational design and testing of abiotic stress‐inducible synthetic promoters from poplar cis‐regulatory elements
title_full Rational design and testing of abiotic stress‐inducible synthetic promoters from poplar cis‐regulatory elements
title_fullStr Rational design and testing of abiotic stress‐inducible synthetic promoters from poplar cis‐regulatory elements
title_full_unstemmed Rational design and testing of abiotic stress‐inducible synthetic promoters from poplar cis‐regulatory elements
title_short Rational design and testing of abiotic stress‐inducible synthetic promoters from poplar cis‐regulatory elements
title_sort rational design and testing of abiotic stress‐inducible synthetic promoters from poplar cis‐regulatory elements
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8313130/
https://www.ncbi.nlm.nih.gov/pubmed/33471413
http://dx.doi.org/10.1111/pbi.13550
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