Unknown,Transcriptomics,Genomics,Proteomics

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Overexpression of MtABI3 in transgenic Medicago truncatula roots

ABSTRACT: ABI3 is a B3-domain transcription factor that acts as a master regulator of seed maturation. To identify genes that are regulated by this transcription factor in the model legume Medicago truncatula, Medicago hairy roots were generated using Agrobacterium rhizogenes transformed with the genomic sequence of the ABI3 gene of Medicago. Using the Medicago NimbleGen chip, a transciptomic analysis was performed to identify differentially expressed genes compared to the GUS expressed control. Two-condition experiment, GUS versus genomic. Biological replicates: 3 control (35S::GUS), 3 treatment (35S::Genomic ABI3 sequence), independently grown and harvested. One replicate per array.

ORGANISM(S): Medicago truncatula

SUBMITTER: Sandra Pelletier

PROVIDER: E-GEOD-44291 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Multigene families encoding diverse secreted peptide hormones play important roles in plant development. A need exists to efficiently elucidate the structures and post-translational-modifications of these difficult-to-isolate peptide hormones in planta so that their biological functions can be determined. A mass spectrometry and bioinformatics approach was developed to comprehensively analyse the secreted peptidome of Medicago hairy root cultures and xylem sap. We identified 759 spectra corresponding to the secreted products of twelve peptide hormones including four CEP (C-TERMINALLY ENCODED PEPTIDE), two CLE (CLV3/ENDOSPERM SURROUNDING REGION RELATED) and six XAP (XYLEM SAP ASSOCIATED PEPTIDE) peptides. The MtCEP1, MtCEP2, MtCEP5 and MtCEP8 peptides identified differed in post-translational-modifications. Most were hydroxylated at conserved proline residues but some MtCEP1 derivatives were tri-arabinosylated. In addition, many CEP peptides possessed unexpected N- and C-terminal extensions. The pattern of these extensions suggested roles for endo- and exoproteases in CEP peptide maturation. Longer -than -expected, hydroxylated and homogeneously modified mono- and tri-arabinosylated CEP peptides corresponding to their in vivo structures were chemically synthesised to probe the effect of these post-translational-modifications on function. The ability of CEP peptides to elevate root nodule number was increased by hydroxylation at key positions. MtCEP1 peptides with N-terminal extensions or with tri-arabinosylation modification, however, were unable to impart increased nodulation. The MtCLE5 and MtCLE17 peptides identified were of precise size, and inhibited main root growth and increased lateral root number. Six XAP peptides, each beginning with a conserved DY sulfation motif, were identified including MtXAP1a, MtXAP1b, MtXAP1c, MtXAP3, MtXAP5 and MtXAP7. MtXAP1a and MtXAP5 inhibited lateral root emergence. Transcriptional analyses demonstrated peptide hormone gene expression in the root vasculature and tip. Since hairy roots can be induced on many plants, their corresponding root cultures may represent ideal source materials to efficiently identify diverse peptide hormones in vivo in a broad range of species.

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Overexpression of MtABI3 in transgenic Medicago truncatula roots

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