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Gene Silencing Using Artificial miRNA in Sugarcane

Gene Silencing Using Artificial miRNA in Sugarcane One of the most common ways to study gene function is to analyze loss-of-function mutants. In polyploid species, although gene redundancy presents some advantages in nature, it offers an extra obstacle for genetic analyses due to the difficulty of obtaining loss-of-function mutants. Sugarcane presents several alleles at the same locus and the strong interaction between them contribute to the variation of phenotype and potential buffering effect, and this is one of the main challenges to study gene function in this species. Although RNA interference (RNAi) is largely used to produce loss-of-function mutants, due to the large fragments used to construct the hairpin structure, it may generate off-targets that can have confounding effect on the phenotype. The artificial micro RNA technique (amiRNA), an improved method of natural miRNA silencing, has been developed in the past years and is an useful tool for gene silencing in plants with high specificity and decreased off-target effects. In the present work we produced 26 transgenic events using amiRNA targeting the ScPDS gene, encoding a phytoene desaturase homolog. All events presented reduced expression levels of ScPDS. Although some events also showed distinguishable photobleached phenotypes, a high degree of expressivity was observed. Therefore, the use of amiRNA technique can be a good choice to silence genes in sugarcane with higher precision, and, consequently, is also a potential tool to be used in other polyploid species. However, a high number of transgenic events might be needed to achieve high levels of gene silencing. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Tropical Plant Biology Springer Journals

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Publisher
Springer Journals
Copyright
Copyright © The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature 2022
ISSN
1935-9756
eISSN
1935-9764
DOI
10.1007/s12042-022-09313-x
Publisher site
See Article on Publisher Site

Abstract

One of the most common ways to study gene function is to analyze loss-of-function mutants. In polyploid species, although gene redundancy presents some advantages in nature, it offers an extra obstacle for genetic analyses due to the difficulty of obtaining loss-of-function mutants. Sugarcane presents several alleles at the same locus and the strong interaction between them contribute to the variation of phenotype and potential buffering effect, and this is one of the main challenges to study gene function in this species. Although RNA interference (RNAi) is largely used to produce loss-of-function mutants, due to the large fragments used to construct the hairpin structure, it may generate off-targets that can have confounding effect on the phenotype. The artificial micro RNA technique (amiRNA), an improved method of natural miRNA silencing, has been developed in the past years and is an useful tool for gene silencing in plants with high specificity and decreased off-target effects. In the present work we produced 26 transgenic events using amiRNA targeting the ScPDS gene, encoding a phytoene desaturase homolog. All events presented reduced expression levels of ScPDS. Although some events also showed distinguishable photobleached phenotypes, a high degree of expressivity was observed. Therefore, the use of amiRNA technique can be a good choice to silence genes in sugarcane with higher precision, and, consequently, is also a potential tool to be used in other polyploid species. However, a high number of transgenic events might be needed to achieve high levels of gene silencing.

Journal

Tropical Plant BiologySpringer Journals

Published: Jun 1, 2022

Keywords: Bioenergy; PDS gene; Polyploid species; Gene expression; Chlorophyll content

References