Robust Agrobacterium-Mediated Transient Expression in Two Duckweed Species (Lemnaceae) Directed by Non-replicating, Replicating, and Cell-to-Cell Spreading Vectors

Plant-based transient expression systems have recognized potential for use as rapid and cost-effective alternatives to expression systems based on bacteria, yeast, insect, or mammalian cells. The free-floating aquatic plants of the Lemnaceae family (duckweed) have compact architecture and can be ve...

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Autores principales:	Anton Peterson, Olena Kishchenko, Yuzhen Zhou, Maksym Vasylenko, Anatoli Giritch, Jian Sun, Nikolai Borisjuk, Mykola Kuchuk
Formato:	article
Lenguaje:	EN
Publicado:	Frontiers Media S.A. 2021
Materias:	transient expression recombinant proteins expression vector duckweed plant expression system Biotechnology TP248.13-248.65
Acceso en línea:	https://doaj.org/article/c0c91b7d5b1e4b44b5a05cb921f18598
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spelling	oai:doaj.org-article:c0c91b7d5b1e4b44b5a05cb921f185982021-11-04T07:18:47ZRobust Agrobacterium-Mediated Transient Expression in Two Duckweed Species (Lemnaceae) Directed by Non-replicating, Replicating, and Cell-to-Cell Spreading Vectors2296-418510.3389/fbioe.2021.761073https://doaj.org/article/c0c91b7d5b1e4b44b5a05cb921f185982021-11-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fbioe.2021.761073/fullhttps://doaj.org/toc/2296-4185 Plant-based transient expression systems have recognized potential for use as rapid and cost-effective alternatives to expression systems based on bacteria, yeast, insect, or mammalian cells. The free-floating aquatic plants of the Lemnaceae family (duckweed) have compact architecture and can be vegetatively propagated on low-cost nutrient solutions in aseptic conditions. These features provide an economically feasible opportunity for duckweed-based production of high-value products via transient expression of recombinant products in fully contained, controlled, aseptic and bio-safe conditions in accordance with the requirements for pharmaceutical manufacturing and environmental biosafety. Here, we demonstrated Agrobacterium-mediated high-yield transient expression of a reporter green fluorescent protein using deconstructed vectors based on potato virus X and sweet potato leaf curl virus, as well as conventional binary vectors, in two representatives of the Lemnaceae (Spirodela polyrhiza and Landoltia punctata). Aseptically cultivated duckweed populations yielded reporter protein accumulation of >1 mg/g fresh biomass, when the protein was expressed from a deconstructed potato virus X-based vector, which is capable of replication and cell-to-cell movement of the replicons in duckweed. The expression efficiency demonstrated here places duckweed among the most efficient host organisms for plant-based transient expression systems, with the additional benefits of easy scale-up and full containment.Anton PetersonAnton PetersonOlena KishchenkoOlena KishchenkoYuzhen ZhouMaksym VasylenkoAnatoli GiritchJian SunNikolai BorisjukMykola KuchukFrontiers Media S.A.articletransient expressionrecombinant proteinsexpression vectorduckweedplant expression systemBiotechnologyTP248.13-248.65ENFrontiers in Bioengineering and Biotechnology, Vol 9 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	transient expression recombinant proteins expression vector duckweed plant expression system Biotechnology TP248.13-248.65
spellingShingle	transient expression recombinant proteins expression vector duckweed plant expression system Biotechnology TP248.13-248.65 Anton Peterson Anton Peterson Olena Kishchenko Olena Kishchenko Yuzhen Zhou Maksym Vasylenko Anatoli Giritch Jian Sun Nikolai Borisjuk Mykola Kuchuk Robust Agrobacterium-Mediated Transient Expression in Two Duckweed Species (Lemnaceae) Directed by Non-replicating, Replicating, and Cell-to-Cell Spreading Vectors
description	Plant-based transient expression systems have recognized potential for use as rapid and cost-effective alternatives to expression systems based on bacteria, yeast, insect, or mammalian cells. The free-floating aquatic plants of the Lemnaceae family (duckweed) have compact architecture and can be vegetatively propagated on low-cost nutrient solutions in aseptic conditions. These features provide an economically feasible opportunity for duckweed-based production of high-value products via transient expression of recombinant products in fully contained, controlled, aseptic and bio-safe conditions in accordance with the requirements for pharmaceutical manufacturing and environmental biosafety. Here, we demonstrated Agrobacterium-mediated high-yield transient expression of a reporter green fluorescent protein using deconstructed vectors based on potato virus X and sweet potato leaf curl virus, as well as conventional binary vectors, in two representatives of the Lemnaceae (Spirodela polyrhiza and Landoltia punctata). Aseptically cultivated duckweed populations yielded reporter protein accumulation of >1 mg/g fresh biomass, when the protein was expressed from a deconstructed potato virus X-based vector, which is capable of replication and cell-to-cell movement of the replicons in duckweed. The expression efficiency demonstrated here places duckweed among the most efficient host organisms for plant-based transient expression systems, with the additional benefits of easy scale-up and full containment.
format	article
author	Anton Peterson Anton Peterson Olena Kishchenko Olena Kishchenko Yuzhen Zhou Maksym Vasylenko Anatoli Giritch Jian Sun Nikolai Borisjuk Mykola Kuchuk
author_facet	Anton Peterson Anton Peterson Olena Kishchenko Olena Kishchenko Yuzhen Zhou Maksym Vasylenko Anatoli Giritch Jian Sun Nikolai Borisjuk Mykola Kuchuk
author_sort	Anton Peterson
title	Robust Agrobacterium-Mediated Transient Expression in Two Duckweed Species (Lemnaceae) Directed by Non-replicating, Replicating, and Cell-to-Cell Spreading Vectors
title_short	Robust Agrobacterium-Mediated Transient Expression in Two Duckweed Species (Lemnaceae) Directed by Non-replicating, Replicating, and Cell-to-Cell Spreading Vectors
title_full	Robust Agrobacterium-Mediated Transient Expression in Two Duckweed Species (Lemnaceae) Directed by Non-replicating, Replicating, and Cell-to-Cell Spreading Vectors
title_fullStr	Robust Agrobacterium-Mediated Transient Expression in Two Duckweed Species (Lemnaceae) Directed by Non-replicating, Replicating, and Cell-to-Cell Spreading Vectors
title_full_unstemmed	Robust Agrobacterium-Mediated Transient Expression in Two Duckweed Species (Lemnaceae) Directed by Non-replicating, Replicating, and Cell-to-Cell Spreading Vectors
title_sort	robust agrobacterium-mediated transient expression in two duckweed species (lemnaceae) directed by non-replicating, replicating, and cell-to-cell spreading vectors
publisher	Frontiers Media S.A.
publishDate	2021
url	https://doaj.org/article/c0c91b7d5b1e4b44b5a05cb921f18598
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Robust Agrobacterium-Mediated Transient Expression in Two Duckweed Species (Lemnaceae) Directed by Non-replicating, Replicating, and Cell-to-Cell Spreading Vectors

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