Reverse genetics in ecological research.

By precisely manipulating the expression of individual genetic elements thought to be important for ecological performance, reverse genetics has the potential to revolutionize plant ecology. However, untested concerns about possible side-effects of the transformation technique, caused by Agrobacteri...

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Autores principales:	Jens Schwachtje, Susan Kutschbach, Ian T Baldwin
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2008
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/61883011122c49e59da2a0841aed9116
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spelling	oai:doaj.org-article:61883011122c49e59da2a0841aed91162021-11-25T06:13:29ZReverse genetics in ecological research.1932-620310.1371/journal.pone.0001543https://doaj.org/article/61883011122c49e59da2a0841aed91162008-02-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/18253491/?tool=EBIhttps://doaj.org/toc/1932-6203By precisely manipulating the expression of individual genetic elements thought to be important for ecological performance, reverse genetics has the potential to revolutionize plant ecology. However, untested concerns about possible side-effects of the transformation technique, caused by Agrobacterium infection and tissue culture, on plant performance have stymied research by requiring onerous sample sizes. We compare 5 independently transformed Nicotiana attenuata lines harboring empty vector control (EVC) T-DNA lacking silencing information with isogenic wild types (WT), and measured a battery of ecologically relevant traits, known to be important in plant-herbivore interactions: phytohormones, secondary metabolites, growth and fitness parameters under stringent competitive conditions, and transcriptional regulation with microarrays. As a positive control, we included a line silenced in trypsin proteinase inhibitor gene (TPI) expression, a potent anti-herbivore defense known to exact fitness costs in its expression, in the analysis. The experiment was conducted twice, with 10 and 20 biological replicates per genotype. For all parameters, we detected no difference between any EVC and WT lines, but could readily detect a fitness benefit of silencing TPI production. A statistical power analyses revealed that the minimum sample sizes required for detecting significant fitness differences between EVC and WT was 2-3 orders of magnitude larger than the 10 replicates required to detect a fitness effect of TPI silencing. We conclude that possible side-effects of transformation are far too low to obfuscate the study of ecologically relevant phenotypes.Jens SchwachtjeSusan KutschbachIan T BaldwinPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 3, Iss 2, p e1543 (2008)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Jens Schwachtje Susan Kutschbach Ian T Baldwin Reverse genetics in ecological research.
description	By precisely manipulating the expression of individual genetic elements thought to be important for ecological performance, reverse genetics has the potential to revolutionize plant ecology. However, untested concerns about possible side-effects of the transformation technique, caused by Agrobacterium infection and tissue culture, on plant performance have stymied research by requiring onerous sample sizes. We compare 5 independently transformed Nicotiana attenuata lines harboring empty vector control (EVC) T-DNA lacking silencing information with isogenic wild types (WT), and measured a battery of ecologically relevant traits, known to be important in plant-herbivore interactions: phytohormones, secondary metabolites, growth and fitness parameters under stringent competitive conditions, and transcriptional regulation with microarrays. As a positive control, we included a line silenced in trypsin proteinase inhibitor gene (TPI) expression, a potent anti-herbivore defense known to exact fitness costs in its expression, in the analysis. The experiment was conducted twice, with 10 and 20 biological replicates per genotype. For all parameters, we detected no difference between any EVC and WT lines, but could readily detect a fitness benefit of silencing TPI production. A statistical power analyses revealed that the minimum sample sizes required for detecting significant fitness differences between EVC and WT was 2-3 orders of magnitude larger than the 10 replicates required to detect a fitness effect of TPI silencing. We conclude that possible side-effects of transformation are far too low to obfuscate the study of ecologically relevant phenotypes.
format	article
author	Jens Schwachtje Susan Kutschbach Ian T Baldwin
author_facet	Jens Schwachtje Susan Kutschbach Ian T Baldwin
author_sort	Jens Schwachtje
title	Reverse genetics in ecological research.
title_short	Reverse genetics in ecological research.
title_full	Reverse genetics in ecological research.
title_fullStr	Reverse genetics in ecological research.
title_full_unstemmed	Reverse genetics in ecological research.
title_sort	reverse genetics in ecological research.
publisher	Public Library of Science (PLoS)
publishDate	2008
url	https://doaj.org/article/61883011122c49e59da2a0841aed9116
work_keys_str_mv	AT jensschwachtje reversegeneticsinecologicalresearch AT susankutschbach reversegeneticsinecologicalresearch AT iantbaldwin reversegeneticsinecologicalresearch
_version_	1718413986363342848

Reverse genetics in ecological research.

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