Plastidial Expression of 3β-Hydroxysteroid Dehydrogenase and Progesterone 5β-Reductase Genes Confer Enhanced Salt Tolerance in Tobacco

The short-chain dehydrogenase/reductase (SDR) gene family is widely distributed in all kingdoms of life. The SDR genes, 3β-hydroxysteroid dehydrogenase (3β-HSD) and progesterone 5-β-reductases (P5βR1, P5βR2) play a crucial r...

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Autores principales:	Muhammad Sameeullah, Muhammet Yildirim, Noreen Aslam, Mehmet Cengiz Baloğlu, Buhara Yucesan, Andreas G. Lössl, Kiran Saba, Mohammad Tahir Waheed, Ekrem Gurel
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	short chain dehydrogenase/reductase (SDR) genes transplastomic plants salt tolerance 3β-HSD NMR sucrose Biology (General) QH301-705.5 Chemistry QD1-999
Acceso en línea:	https://doaj.org/article/e1942547d50f4b03af832f185363bf4b
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spelling	oai:doaj.org-article:e1942547d50f4b03af832f185363bf4b2021-11-11T17:11:38ZPlastidial Expression of 3β-Hydroxysteroid Dehydrogenase and Progesterone 5β-Reductase Genes Confer Enhanced Salt Tolerance in Tobacco10.3390/ijms2221117361422-00671661-6596https://doaj.org/article/e1942547d50f4b03af832f185363bf4b2021-10-01T00:00:00Zhttps://www.mdpi.com/1422-0067/22/21/11736https://doaj.org/toc/1661-6596https://doaj.org/toc/1422-0067The short-chain dehydrogenase/reductase (SDR) gene family is widely distributed in all kingdoms of life. The <i>SDR</i> genes, 3β-hydroxysteroid dehydrogenase (<i>3β-HSD</i>) and progesterone 5-β-reductases (<i>P5βR1</i>, <i>P5βR2</i>) play a crucial role in cardenolide biosynthesis pathway in the <i>Digitalis</i> species. However, their role in plant stress, especially in salinity stress management, remains unexplored. In the present study, transplastomic tobacco plants were developed by inserting the <i>3β-HSD</i>, <i>P5βR1</i> and <i>P5βR2</i> genes. The integration of transgenes in plastomes, copy number and transgene expression at transcript and protein level in transplastomic plants were confirmed by PCR, end-to-end PCR, qRT-PCR and Western blot analysis, respectively. Subcellular localization analysis showed that 3β-HSD and P5βR1 are cytoplasmic, and P5βR2 is tonoplast-localized. Transplastomic lines showed enhanced growth in terms of biomass and chlorophyll content compared to wild type (WT) under 300 mM salt stress. Under salt stress, transplastomic lines remained greener without negative impact on shoot or root growth compared to the WT. The salt-tolerant transplastomic lines exhibited enhanced levels of a series of metabolites (sucrose, glutamate, glutamine and proline) under control and NaCl stress. Furthermore, a lower Na<sup>+</sup>/K<sup>+</sup> ratio in transplastomic lines was also observed. The salt tolerance, mediated by plastidial expression of the <i>3β-HSD</i>, <i>P5βR1</i> and <i>P5βR2</i> genes, could be due to the involvement in the upregulation of nitrogen assimilation, osmolytes as well as lower Na<sup>+</sup>/K<sup>+</sup> ratio. Taken together, the plastid-based expression of the <i>SDR</i> genes leading to enhanced salt tolerance, which opens a window for developing saline-tolerant plants via plastid genetic engineering.Muhammad SameeullahMuhammet YildirimNoreen AslamMehmet Cengiz BaloğluBuhara YucesanAndreas G. LösslKiran SabaMohammad Tahir WaheedEkrem GurelMDPI AGarticleshort chain dehydrogenase/reductase (SDR) genestransplastomic plantssalt tolerance3β-HSDNMRsucroseBiology (General)QH301-705.5ChemistryQD1-999ENInternational Journal of Molecular Sciences, Vol 22, Iss 11736, p 11736 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	short chain dehydrogenase/reductase (SDR) genes transplastomic plants salt tolerance 3β-HSD NMR sucrose Biology (General) QH301-705.5 Chemistry QD1-999
spellingShingle	short chain dehydrogenase/reductase (SDR) genes transplastomic plants salt tolerance 3β-HSD NMR sucrose Biology (General) QH301-705.5 Chemistry QD1-999 Muhammad Sameeullah Muhammet Yildirim Noreen Aslam Mehmet Cengiz Baloğlu Buhara Yucesan Andreas G. Lössl Kiran Saba Mohammad Tahir Waheed Ekrem Gurel Plastidial Expression of 3β-Hydroxysteroid Dehydrogenase and Progesterone 5β-Reductase Genes Confer Enhanced Salt Tolerance in Tobacco
description	The short-chain dehydrogenase/reductase (SDR) gene family is widely distributed in all kingdoms of life. The <i>SDR</i> genes, 3β-hydroxysteroid dehydrogenase (<i>3β-HSD</i>) and progesterone 5-β-reductases (<i>P5βR1</i>, <i>P5βR2</i>) play a crucial role in cardenolide biosynthesis pathway in the <i>Digitalis</i> species. However, their role in plant stress, especially in salinity stress management, remains unexplored. In the present study, transplastomic tobacco plants were developed by inserting the <i>3β-HSD</i>, <i>P5βR1</i> and <i>P5βR2</i> genes. The integration of transgenes in plastomes, copy number and transgene expression at transcript and protein level in transplastomic plants were confirmed by PCR, end-to-end PCR, qRT-PCR and Western blot analysis, respectively. Subcellular localization analysis showed that 3β-HSD and P5βR1 are cytoplasmic, and P5βR2 is tonoplast-localized. Transplastomic lines showed enhanced growth in terms of biomass and chlorophyll content compared to wild type (WT) under 300 mM salt stress. Under salt stress, transplastomic lines remained greener without negative impact on shoot or root growth compared to the WT. The salt-tolerant transplastomic lines exhibited enhanced levels of a series of metabolites (sucrose, glutamate, glutamine and proline) under control and NaCl stress. Furthermore, a lower Na<sup>+</sup>/K<sup>+</sup> ratio in transplastomic lines was also observed. The salt tolerance, mediated by plastidial expression of the <i>3β-HSD</i>, <i>P5βR1</i> and <i>P5βR2</i> genes, could be due to the involvement in the upregulation of nitrogen assimilation, osmolytes as well as lower Na<sup>+</sup>/K<sup>+</sup> ratio. Taken together, the plastid-based expression of the <i>SDR</i> genes leading to enhanced salt tolerance, which opens a window for developing saline-tolerant plants via plastid genetic engineering.
format	article
author	Muhammad Sameeullah Muhammet Yildirim Noreen Aslam Mehmet Cengiz Baloğlu Buhara Yucesan Andreas G. Lössl Kiran Saba Mohammad Tahir Waheed Ekrem Gurel
author_facet	Muhammad Sameeullah Muhammet Yildirim Noreen Aslam Mehmet Cengiz Baloğlu Buhara Yucesan Andreas G. Lössl Kiran Saba Mohammad Tahir Waheed Ekrem Gurel
author_sort	Muhammad Sameeullah
title	Plastidial Expression of 3β-Hydroxysteroid Dehydrogenase and Progesterone 5β-Reductase Genes Confer Enhanced Salt Tolerance in Tobacco
title_short	Plastidial Expression of 3β-Hydroxysteroid Dehydrogenase and Progesterone 5β-Reductase Genes Confer Enhanced Salt Tolerance in Tobacco
title_full	Plastidial Expression of 3β-Hydroxysteroid Dehydrogenase and Progesterone 5β-Reductase Genes Confer Enhanced Salt Tolerance in Tobacco
title_fullStr	Plastidial Expression of 3β-Hydroxysteroid Dehydrogenase and Progesterone 5β-Reductase Genes Confer Enhanced Salt Tolerance in Tobacco
title_full_unstemmed	Plastidial Expression of 3β-Hydroxysteroid Dehydrogenase and Progesterone 5β-Reductase Genes Confer Enhanced Salt Tolerance in Tobacco
title_sort	plastidial expression of 3β-hydroxysteroid dehydrogenase and progesterone 5β-reductase genes confer enhanced salt tolerance in tobacco
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/e1942547d50f4b03af832f185363bf4b
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Plastidial Expression of 3β-Hydroxysteroid Dehydrogenase and Progesterone 5β-Reductase Genes Confer Enhanced Salt Tolerance in Tobacco

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