Enhanced transgene expression in sugarcane by co-expression of virus-encoded RNA silencing suppressors.

Post-transcriptional gene silencing is commonly observed in polyploid species and often poses a major limitation to plant improvement via biotechnology. Five plant viral suppressors of RNA silencing were evaluated for their ability to counteract gene silencing and enhance the expression of the Enhan...

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Autores principales:	San-Ji Gao, Mona B Damaj, Jong-Won Park, Getu Beyene, Marco T Buenrostro-Nava, Joe Molina, Xiaofeng Wang, Jessica J Ciomperlik, Shuga A Manabayeva, Veria Y Alvarado, Keerti S Rathore, Herman B Scholthof, T Erik Mirkov
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/2d96da64c3a34270b826f5e05a2356c6
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spelling	oai:doaj.org-article:2d96da64c3a34270b826f5e05a2356c62021-11-18T07:41:40ZEnhanced transgene expression in sugarcane by co-expression of virus-encoded RNA silencing suppressors.1932-620310.1371/journal.pone.0066046https://doaj.org/article/2d96da64c3a34270b826f5e05a2356c62013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23799071/?tool=EBIhttps://doaj.org/toc/1932-6203Post-transcriptional gene silencing is commonly observed in polyploid species and often poses a major limitation to plant improvement via biotechnology. Five plant viral suppressors of RNA silencing were evaluated for their ability to counteract gene silencing and enhance the expression of the Enhanced Yellow Fluorescent Protein (EYFP) or the β-glucuronidase (GUS) reporter gene in sugarcane, a major sugar and biomass producing polyploid. Functionality of these suppressors was first verified in Nicotiana benthamiana and onion epidermal cells, and later tested by transient expression in sugarcane young leaf segments and protoplasts. In young leaf segments co-expressing a suppressor, EYFP reached its maximum expression at 48-96 h post-DNA introduction and maintained its peak expression for a longer time compared with that in the absence of a suppressor. Among the five suppressors, Tomato bushy stunt virus-encoded P19 and Barley stripe mosaic virus-encoded γb were the most efficient. Co-expression with P19 and γb enhanced EYFP expression 4.6-fold and 3.6-fold in young leaf segments, and GUS activity 2.3-fold and 2.4-fold in protoplasts compared with those in the absence of a suppressor, respectively. In transgenic sugarcane, co-expression of GUS and P19 suppressor showed the highest accumulation of GUS levels with an average of 2.7-fold more than when GUS was expressed alone, with no detrimental phenotypic effects. The two established transient expression assays, based on young leaf segments and protoplasts, and confirmed by stable transgene expression, offer a rapid versatile system to verify the efficiency of RNA silencing suppressors that proved to be valuable in enhancing and stabilizing transgene expression in sugarcane.San-Ji GaoMona B DamajJong-Won ParkGetu BeyeneMarco T Buenrostro-NavaJoe MolinaXiaofeng WangJessica J CiomperlikShuga A ManabayevaVeria Y AlvaradoKeerti S RathoreHerman B ScholthofT Erik MirkovPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 6, p e66046 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q San-Ji Gao Mona B Damaj Jong-Won Park Getu Beyene Marco T Buenrostro-Nava Joe Molina Xiaofeng Wang Jessica J Ciomperlik Shuga A Manabayeva Veria Y Alvarado Keerti S Rathore Herman B Scholthof T Erik Mirkov Enhanced transgene expression in sugarcane by co-expression of virus-encoded RNA silencing suppressors.
description	Post-transcriptional gene silencing is commonly observed in polyploid species and often poses a major limitation to plant improvement via biotechnology. Five plant viral suppressors of RNA silencing were evaluated for their ability to counteract gene silencing and enhance the expression of the Enhanced Yellow Fluorescent Protein (EYFP) or the β-glucuronidase (GUS) reporter gene in sugarcane, a major sugar and biomass producing polyploid. Functionality of these suppressors was first verified in Nicotiana benthamiana and onion epidermal cells, and later tested by transient expression in sugarcane young leaf segments and protoplasts. In young leaf segments co-expressing a suppressor, EYFP reached its maximum expression at 48-96 h post-DNA introduction and maintained its peak expression for a longer time compared with that in the absence of a suppressor. Among the five suppressors, Tomato bushy stunt virus-encoded P19 and Barley stripe mosaic virus-encoded γb were the most efficient. Co-expression with P19 and γb enhanced EYFP expression 4.6-fold and 3.6-fold in young leaf segments, and GUS activity 2.3-fold and 2.4-fold in protoplasts compared with those in the absence of a suppressor, respectively. In transgenic sugarcane, co-expression of GUS and P19 suppressor showed the highest accumulation of GUS levels with an average of 2.7-fold more than when GUS was expressed alone, with no detrimental phenotypic effects. The two established transient expression assays, based on young leaf segments and protoplasts, and confirmed by stable transgene expression, offer a rapid versatile system to verify the efficiency of RNA silencing suppressors that proved to be valuable in enhancing and stabilizing transgene expression in sugarcane.
format	article
author	San-Ji Gao Mona B Damaj Jong-Won Park Getu Beyene Marco T Buenrostro-Nava Joe Molina Xiaofeng Wang Jessica J Ciomperlik Shuga A Manabayeva Veria Y Alvarado Keerti S Rathore Herman B Scholthof T Erik Mirkov
author_facet	San-Ji Gao Mona B Damaj Jong-Won Park Getu Beyene Marco T Buenrostro-Nava Joe Molina Xiaofeng Wang Jessica J Ciomperlik Shuga A Manabayeva Veria Y Alvarado Keerti S Rathore Herman B Scholthof T Erik Mirkov
author_sort	San-Ji Gao
title	Enhanced transgene expression in sugarcane by co-expression of virus-encoded RNA silencing suppressors.
title_short	Enhanced transgene expression in sugarcane by co-expression of virus-encoded RNA silencing suppressors.
title_full	Enhanced transgene expression in sugarcane by co-expression of virus-encoded RNA silencing suppressors.
title_fullStr	Enhanced transgene expression in sugarcane by co-expression of virus-encoded RNA silencing suppressors.
title_full_unstemmed	Enhanced transgene expression in sugarcane by co-expression of virus-encoded RNA silencing suppressors.
title_sort	enhanced transgene expression in sugarcane by co-expression of virus-encoded rna silencing suppressors.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/2d96da64c3a34270b826f5e05a2356c6
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Enhanced transgene expression in sugarcane by co-expression of virus-encoded RNA silencing suppressors.

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