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Gene Silencing Using Artificial miRNA in Sugarcane

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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.

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Acknowledgements

This work was funded by São Paulo Research Foundation (FAPESP), research grants 2013/15576-5 and 2014/50884-5 (MM) and National Council for Scientific and Technological Development (CNPq, Brazil; Grants 333270/2018-7 and 465319/2014-9). ALGLP received a fellowship from the Brazilian Federal Agency for Support and Evaluation of Graduate Education (CAPES; Grant 88882.329508/2019-01).

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Authors and Affiliations

  1. Departamento de Genética, Evolução, Microbiologia e Imunologia, Instituto de Biologia, Universidade Estadual de Campinas, Campinas, SP, Brazil

    Ana Laura Garcia Leme Peres, José Sérgio Soares & Marcelo Menossi

  2. Department of Agronomy and Plant Genetics, University of Minnesota, Saint Paul, MN, USA

    Rafael Della Coletta

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  1. Ana Laura Garcia Leme Peres
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  2. Rafael Della Coletta
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  3. José Sérgio Soares
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  4. Marcelo Menossi
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Correspondence to Marcelo Menossi.

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Communicated by: Graham Bonnett

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José Sérgio Soares: In memoriam

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Peres, A.L.G.L., Coletta, R.D., Soares, J.S. et al. Gene Silencing Using Artificial miRNA in Sugarcane. Tropical Plant Biol. 15, 111–120 (2022). https://doi.org/10.1007/s12042-022-09313-x

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  • DOI: https://doi.org/10.1007/s12042-022-09313-x

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