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Germline transformation of Artemisia annuaL. plant via in planta transformation technology “Floral dip”
The therapeutic efficacy of Artemisia annua L. is governed by artemisinin (ART), prevalently produced by A. annua extraction. Due to the modest amount of ART (0.01-1 %dw) in this plant, commercialization of ACTs is difficult. In this study, the floral-dip based transformation protocol for A. annua w...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9489500/ https://www.ncbi.nlm.nih.gov/pubmed/36159743 http://dx.doi.org/10.1016/j.btre.2022.e00761 |
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author | Qamar, Firdaus Khan, Shazia Ashrafi, Kudsiya Iqrar, Sadia Quadri, Syed Naved Saifi, Monica Abdin, M.Z. |
author_facet | Qamar, Firdaus Khan, Shazia Ashrafi, Kudsiya Iqrar, Sadia Quadri, Syed Naved Saifi, Monica Abdin, M.Z. |
author_sort | Qamar, Firdaus |
collection | PubMed |
description | The therapeutic efficacy of Artemisia annua L. is governed by artemisinin (ART), prevalently produced by A. annua extraction. Due to the modest amount of ART (0.01-1 %dw) in this plant, commercialization of ACTs is difficult. In this study, the floral-dip based transformation protocol for A. annua was developed to enhance expression of artemisinin biosynthesis genes and ART content. For dipping, the effective infiltration media components were optimized, and to obtain high transformation (26.9%) partially open bud stage capitulum of floral development was used. Hygromycin phospho-transferase (hptII) selection marker was used to validate the transformed T(1) progenies. The copy numbers of the transgene (hptII) in T(1) progenies were determined using a sensitive, high-throughput SYBR Green based quantitative RT-PCR. The results of the hptII transgene were compared with those of the low copy number, internal standard (hmgr). Using optimised PCR conditions, one, two and three transgene copies in T(1) transformants were achieved. |
format | Online Article Text |
id | pubmed-9489500 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-94895002022-09-22 Germline transformation of Artemisia annuaL. plant via in planta transformation technology “Floral dip” Qamar, Firdaus Khan, Shazia Ashrafi, Kudsiya Iqrar, Sadia Quadri, Syed Naved Saifi, Monica Abdin, M.Z. Biotechnol Rep (Amst) Research Article The therapeutic efficacy of Artemisia annua L. is governed by artemisinin (ART), prevalently produced by A. annua extraction. Due to the modest amount of ART (0.01-1 %dw) in this plant, commercialization of ACTs is difficult. In this study, the floral-dip based transformation protocol for A. annua was developed to enhance expression of artemisinin biosynthesis genes and ART content. For dipping, the effective infiltration media components were optimized, and to obtain high transformation (26.9%) partially open bud stage capitulum of floral development was used. Hygromycin phospho-transferase (hptII) selection marker was used to validate the transformed T(1) progenies. The copy numbers of the transgene (hptII) in T(1) progenies were determined using a sensitive, high-throughput SYBR Green based quantitative RT-PCR. The results of the hptII transgene were compared with those of the low copy number, internal standard (hmgr). Using optimised PCR conditions, one, two and three transgene copies in T(1) transformants were achieved. Elsevier 2022-08-30 /pmc/articles/PMC9489500/ /pubmed/36159743 http://dx.doi.org/10.1016/j.btre.2022.e00761 Text en © 2022 The Authors. Published by Elsevier B.V. https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Research Article Qamar, Firdaus Khan, Shazia Ashrafi, Kudsiya Iqrar, Sadia Quadri, Syed Naved Saifi, Monica Abdin, M.Z. Germline transformation of Artemisia annuaL. plant via in planta transformation technology “Floral dip” |
title | Germline transformation of Artemisia annuaL. plant via in planta transformation technology “Floral dip” |
title_full | Germline transformation of Artemisia annuaL. plant via in planta transformation technology “Floral dip” |
title_fullStr | Germline transformation of Artemisia annuaL. plant via in planta transformation technology “Floral dip” |
title_full_unstemmed | Germline transformation of Artemisia annuaL. plant via in planta transformation technology “Floral dip” |
title_short | Germline transformation of Artemisia annuaL. plant via in planta transformation technology “Floral dip” |
title_sort | germline transformation of artemisia annual. plant via in planta transformation technology “floral dip” |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9489500/ https://www.ncbi.nlm.nih.gov/pubmed/36159743 http://dx.doi.org/10.1016/j.btre.2022.e00761 |
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