Functional Analysis of Root microRNAs by a Constitutive Overexpression Approach in a Composite Plant System
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Agrobacterium-mediated transformation is a fast and efficient method for genome modification in plants. In this protocol, we apply this technique for the analysis of root microRNA functionality. The induction of hairy roots constitutively overexpressing a given microRNA precursor allows us, in a simple way, to modify the accumulation of specific mature microRNA and analyze the consequence of this alteration on a phenotype of interest. This method generates ready-to-phenotype “composite plants” with untransformed aerial part and microRNA-overexpressing root system, in about 20 days.
Key wordsmicroRNA Overexpression Agrobacterium rhizogenes T-DNA Hairy roots
The work in our group has been supported by Dirección General de Asuntos del Personal Académico (DGAPA)—UNAM (grants no. PAPIIT IN200816 and IA203218).