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Overexpression of MtABI3 splicing forms by ectopically expressing cDNA sequences in transgenic Medicago truncatula hairy roots generated from an abi3 mutant background (mutant NF6003)

ABSTRACT: Overexpression of MtABI3 splicing forms by ectopically expressing cDNA sequences in transgenic Medicago truncatula hairy roots generated from an abi3 mutant background (mutant NF6003)

PROVIDER: PRJNA750124 | ENA |

REPOSITORIES: ENA

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Overexpression of MtABI3 splicing forms by ectopically expressing cDNA sequences in transgenic Medicago truncatula hairy roots generated from an abi3 mutant background (mutant NF6003)

Project description:ABI3 is a B3-domain transcription factor that acts as a master regulator of seed maturation. To identify genes that are regulated specifically by different splicing forms in the model legume Medicago truncatula, Medicago hairy roots obtained from an abi3 mutant background (NF6003) were generated using Agrobacterium rhizogenes transformed with the three cDNA sequence of the ABI3 gene splicing forms of Medicago. Using the Medicago NimbleGen chip, a transcriptomic analysis was performed to identify differentially expressed genes compared to the GUS expressed control.

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Project description:Deciphering gene regulatory networks (GRNs) is a key for understanding gene expression regulations in living systems. Here, we describe the investigation of the ABSCISIC ACID INSENSITIVE 3 (ABI3) plant transcription factor GRN vicinity by a technique called Network Walking. The method involves transient transformation of protoplasts and inducible nuclear re-localization of transcription factors along with transcriptomic analysis. This genome-wide approach allowed the de novo recovery of i) direct and indirect ABI3 target genes, ii) cis-binding site preference, and iii) biological processes regulated by this canonical abscisic acid response factor. This work improves our knowledge of ABI3 action by inferring network motifs (such as Feed Forwar Loops) under its influence. The novel high-throughput-oriented technique will help accelerate GRN systems investigations in plants, as well as in other organisms. This work studies ABI3 direct and indirect targets by a technique named Network Walking. Root/protoplasts were treated with or without dexamethasone (DEX) and cycloheximide (CHX). 3 reps each.

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