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Efficient Gene Silencing Mediated by Tobacco Rattle Virus in an Emerging Model Plant Physalis
The fruit of Physalis has a berry and a novelty called inflated calyx syndrome (ICS, also named the ‘Chinese lantern’). Elucidation of the underlying developmental mechanisms of fruit diversity demands an efficient gene functional inference platform. Here, we tested the application of the tobacco ra...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3891815/ https://www.ncbi.nlm.nih.gov/pubmed/24454885 http://dx.doi.org/10.1371/journal.pone.0085534 |
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author | Zhang, Ji-Si Zhao, Jing Zhang, Shaohua He, Chaoying |
author_facet | Zhang, Ji-Si Zhao, Jing Zhang, Shaohua He, Chaoying |
author_sort | Zhang, Ji-Si |
collection | PubMed |
description | The fruit of Physalis has a berry and a novelty called inflated calyx syndrome (ICS, also named the ‘Chinese lantern’). Elucidation of the underlying developmental mechanisms of fruit diversity demands an efficient gene functional inference platform. Here, we tested the application of the tobacco rattle virus (TRV)-mediated gene-silencing system in Physalis floridana. First, we characterized the putative gene of a phytoene desaturase in P. floridana (PfPDS). Infecting the leaves of the Physalis seedlings with the PfPDS-TRV vector resulted in a bleached plant, including the developing leaves, floral organs, ICS, berry, and seed. These results indicated that a local VIGS treatment can efficiently induce a systemic mutated phenotype. qRT-PCR analyses revealed that the bleaching extent correlated to the mRNA reduction of the endogenous PfPDS. Detailed comparisons of multiple infiltration and growth protocols allowed us to determine the optimal methodologies for VIGS manipulation in Physalis. We subsequently utilized this optimized VIGS methodology to downregulate the expression of two MADS-box genes, MPF2 and MPF3, and compared the resulting effects with gene-downregulation mediated by RNA interference (RNAi) methods. The VIGS-mediated gene knockdown plants were found to resemble the mutated phenotypes of floral calyx, fruiting calyx and pollen maturation of the RNAi transgenic plants for both MPF2 and MPF3. Moreover, the two MADS-box genes were appeared to have a novel role in the pedicel development in P. floridana. The major advantage of VIGS-based gene knockdown lies in practical aspects of saving time and easy manipulation as compared to the RNAi. Despite the lack of heritability and mosaic mutation phenotypes observed in some organs, the TRV-mediated gene silencing system provides an alternative efficient way to infer gene function in various developmental processes in Physalis, thus facilitating understanding of the genetic basis of the evolution and development of the morphological diversities within the Solanaceae. |
format | Online Article Text |
id | pubmed-3891815 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-38918152014-01-21 Efficient Gene Silencing Mediated by Tobacco Rattle Virus in an Emerging Model Plant Physalis Zhang, Ji-Si Zhao, Jing Zhang, Shaohua He, Chaoying PLoS One Research Article The fruit of Physalis has a berry and a novelty called inflated calyx syndrome (ICS, also named the ‘Chinese lantern’). Elucidation of the underlying developmental mechanisms of fruit diversity demands an efficient gene functional inference platform. Here, we tested the application of the tobacco rattle virus (TRV)-mediated gene-silencing system in Physalis floridana. First, we characterized the putative gene of a phytoene desaturase in P. floridana (PfPDS). Infecting the leaves of the Physalis seedlings with the PfPDS-TRV vector resulted in a bleached plant, including the developing leaves, floral organs, ICS, berry, and seed. These results indicated that a local VIGS treatment can efficiently induce a systemic mutated phenotype. qRT-PCR analyses revealed that the bleaching extent correlated to the mRNA reduction of the endogenous PfPDS. Detailed comparisons of multiple infiltration and growth protocols allowed us to determine the optimal methodologies for VIGS manipulation in Physalis. We subsequently utilized this optimized VIGS methodology to downregulate the expression of two MADS-box genes, MPF2 and MPF3, and compared the resulting effects with gene-downregulation mediated by RNA interference (RNAi) methods. The VIGS-mediated gene knockdown plants were found to resemble the mutated phenotypes of floral calyx, fruiting calyx and pollen maturation of the RNAi transgenic plants for both MPF2 and MPF3. Moreover, the two MADS-box genes were appeared to have a novel role in the pedicel development in P. floridana. The major advantage of VIGS-based gene knockdown lies in practical aspects of saving time and easy manipulation as compared to the RNAi. Despite the lack of heritability and mosaic mutation phenotypes observed in some organs, the TRV-mediated gene silencing system provides an alternative efficient way to infer gene function in various developmental processes in Physalis, thus facilitating understanding of the genetic basis of the evolution and development of the morphological diversities within the Solanaceae. Public Library of Science 2014-01-14 /pmc/articles/PMC3891815/ /pubmed/24454885 http://dx.doi.org/10.1371/journal.pone.0085534 Text en © 2014 Zhang et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Zhang, Ji-Si Zhao, Jing Zhang, Shaohua He, Chaoying Efficient Gene Silencing Mediated by Tobacco Rattle Virus in an Emerging Model Plant Physalis |
title | Efficient Gene Silencing Mediated by Tobacco Rattle Virus in an Emerging Model Plant Physalis
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title_full | Efficient Gene Silencing Mediated by Tobacco Rattle Virus in an Emerging Model Plant Physalis
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title_fullStr | Efficient Gene Silencing Mediated by Tobacco Rattle Virus in an Emerging Model Plant Physalis
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title_full_unstemmed | Efficient Gene Silencing Mediated by Tobacco Rattle Virus in an Emerging Model Plant Physalis
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title_short | Efficient Gene Silencing Mediated by Tobacco Rattle Virus in an Emerging Model Plant Physalis
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title_sort | efficient gene silencing mediated by tobacco rattle virus in an emerging model plant physalis |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3891815/ https://www.ncbi.nlm.nih.gov/pubmed/24454885 http://dx.doi.org/10.1371/journal.pone.0085534 |
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