Gene Silencing Using Artificial miRNA in Sugarcane

Author(s)

Garcia Leme Peres, Ana Laura ; Della Coletta, Rafael ; Soares, Jose Sergio ; Menossi, Marcelo

• Document type: Journal article
• Source: TROPICAL PLANT BIOLOGY; v. 15, n. 2, p. 10-pg., 2022-03-25.

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. (AU)

• FAPESP's process: 13/15576-5 - Molecular approaches to study the relationship between ethylene and sugarcane ripening
• Grantee: Marcelo Menossi Teixeira
• Support Opportunities: Program for Research on Bioenergy (BIOEN) - Thematic Grants
• FAPESP's process: 14/50884-5 - INCT 2014: National Institute of Science and Technology of Bioethanol
• Grantee: Marcos Silveira Buckeridge
• Support Opportunities: Research Projects - Thematic Grants