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Transcription profiling of Arabidopsis wild type or mp mutant seedlings treated with auxin

ABSTRACT: Comparison of mp mutant seedlings with transgenics carrying a dexamethasone-inducible GR-bodenlos gene. Seedlings were pretreated with control medium or medium containing dexamethasone, followed by a 1 hour auxin treatment, and expression profiles were compared using Affymetrix ATH1 arrays.

ORGANISM(S): Arabidopsis thaliana

SUBMITTER: Dolf Weijers

PROVIDER: E-MEXP-1948 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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MONOPTEROS controls embryonic root initiation by regulating a mobile transcription factor.

Schlereth Alexandra A Möller Barbara B Liu Weilin W Kientz Marika M Flipse Jacky J Rademacher Eike H EH Schmid Markus M Jürgens Gerd G Weijers Dolf D

Nature 20100310 7290

Acquisition of cell identity in plants relies strongly on positional information, hence cell-cell communication and inductive signalling are instrumental for developmental patterning. During Arabidopsis embryogenesis, an extra-embryonic cell is specified to become the founder cell of the primary root meristem, hypophysis, in response to signals from adjacent embryonic cells. The auxin-dependent transcription factor MONOPTEROS (MP) drives hypophysis specification by promoting transport of the hor ...[more]

PMID: 20220754

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Transcriptional profiles between mp mutant seedlings and transgenics carrying the dexamethasone-inducible GR-bdl protein

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Project description:In order to identify targets of the transcription factor AUXIN RESPONSE FACTOR5 / MONOPTEROS (ARF5/MP), we compared transcriptomes of mp-B4149 mutant seedlings (9 day-old) and seedlings carrying the dexamethasone-inducible version of the MP inhibitor protein BODENLOS (GR-bdl). Without dexamethasone (DEX) treatment, this line is identical to the wild-type, while DEX treatment leads to strong inhibition of ARF-dependent transcription. To remove all endogenous MP-inhibiting Aux/IAA proteins, we treated mp or GR-bdl seedlings during 1 hour with auxin (50 micromolar Indole-3-Acetic Acid), either with or without a pretreatment with 10 micromolar DEX for 1 hour. Genes that are activated by MP are expected to br downregulated in mp seedlings and in the GR-bdl line afer DEX treatment. We used biological duplicates for each of the three treatments. Keywords: Transcriptional profiling of transcription factor targets Mp mutant and GR-bdl seeds were germinated on solid 1/2MS media, and transferred to liquid medium after 9 days of horizontal growth. Duplicate cultures were generated for mp mutants, and quadruplicates for the GR-bdl line. 10 micromolar DEX was added to both mp cultures and to two of the GR-bdl cultures, and seedlings were gently shaken during 1 hour. Then, 50 micromolar IAA was added to all 6 cultures and seedlings were incubated with gentle shaking for another hour. Total RNA was isolated from the seedlings using a QIAgen RNeasy kit, and reverse transcription, in-vitro transcription, cRNA labling and fragemntation was done according to established methods (Schmid et al., Nature Genetics [2005] 37, 501-506). Labeled cRNA was hybridized to Arabidopsis ATH1 arrays.

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