Transcription profiling of Medicago truncatula 2HA tissue culture with auxin treatment
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ABSTRACT: Leaf explants of the superembryogenic Medicago truncatula line 2HA were treated with auxin (1-naphthaleneacetic acid) for one week to induce the formation of roots (Imin et al J Exp Bot 58:439-451). Gene expression in the leaves and the NAA treated tissue cultures was compared to identify transcripts expressed during the commitment to root formation in tissue culture. We have used the Affymetrix Medicago Genome Array GeneChip to compared gene expression in Medicago truncatula leaves and leaf explants that have been cultured for one week on NAA, to identify genes expressed during the commitment to root formation in tissue culture. Experiment Overall Design: Medicago truncatula 2HA leaves and leaf explants treated with NAA for one week were collected; total RNA was extracted and used for hybridization to Affymetrix arrays
Project description:The Medicago truncatula line 2HA has a 500-fold greater capacity to regenerate plants in culture by somatic embryogenesis than wild-type Jemalong. We have compared transcriptomes of tissue cultures from leaf explants of these two lines. We have used the Affymetrix Medicago Genome Array GeneChip to compare leaf explants of Medicago truncatula Jemalong and the superembryogenic line 2HA after two weeks of tissue culture on 10 µM 1-naphthaleneacetic acid (NAA) and 4 µM 6-benzylaminopurine (BAP) (P4 10:4) Experiment Overall Design: Leaf explants from the two genotypes were treated with 10:4 for two weeks; total RNA was extracted and used for hybridization to Affymetrix arrays
Project description:Leaf explants of the superembryogenic Medicago truncatula line 2HA were treated with auxin (1-naphthaleneacetic acid) for one week to induce the formation of roots (Imin et al J Exp Bot 58:439-451). Gene expression in the leaves and the NAA treated tissue cultures was compared to identify transcripts expressed during the commitment to root formation in tissue culture. We have used the Affymetrix Medicago Genome Array GeneChip to compared gene expression in Medicago truncatula leaves and leaf explants that have been cultured for one week on NAA, to identify genes expressed during the commitment to root formation in tissue culture. Keywords: Cell type comparison
Project description:The Medicago truncatula line 2HA has a 500-fold greater capacity to regenerate plants in culture by somatic embryogenesis than wild-type Jemalong. We have compared transcriptomes of tissue cultures from leaf explants of these two lines. We have used the Affymetrix Medicago Genome Array GeneChip to compare leaf explants of Medicago truncatula Jemalong and the superembryogenic line 2HA after two weeks of tissue culture on 10 µM 1-naphthaleneacetic acid (NAA) and 4 µM 6-benzylaminopurine (BAP) (P4 10:4) Keywords: Genotype comparison
Project description:Background: The Medicago truncatula 2HA seed line is highly embryogenic while the parental line Jemalong rarely produces embryos. The 2HA line was developed from one of the rare Jemalong regenerates and this method for obtaining a highly regenerable genotype in M. truncatula is readily reproducible suggesting an epigenetic mechanism. Microarray transcriptomic analysis showed down regulation of an ETHYLENE INSENSITIVE 3-like gene in 2HA callus which provided an approach to investigating epigenetic regulation of genes related to ethylene signalling and the 2HA phenotype. Ethylene is involved in many developmental processes including somatic embryogenesis (SE) and is associated with stress responses. Results: Microarray transcriptomic analysis showed a significant number of up-regulated transcripts in 2HA tissue culture, including nodule and embryo specific genes and transposon-like genes, while only a few genes were down-regulated, including an EIN3-like gene we called MtEIL1. This reduced expression was associated with ethylene insensitivity of 2HA plants that was further investigated. The weak ethylene insensitivity affected root and nodule development. Sequencing of MtEIL1 found no difference between 2HA and wild-type plants. DNA methylation analysis of MtEIL1 revealed significant difference between 2HA and wild-type plants. Tiling arrays demonstrated an elevated level of miRNA in 2HA plants that hybridised to the antisense strand of the MtEIL1 gene. AFLP-like methylation profiling revealed more differences in DNA methylation between 2HA and wild-type. Segregation analysis demonstrated the recessive nature of the eil1 phenotype and the dominant nature of the SE trait. Conclusions: We have demonstrated that EIL1 of Medicago truncatula (MtEIL1) is epigenetically silenced in the 2HA seed line. The possible cause is an elevated level of miRNA that targets its 3M-bM-^@M-^YUTR leading to DNA methylation. Down regulation of MtEIL1 makes it possible to form nodules in the presence of ethylene and affects root growth under normal conditions. Segregation analysis showed no association between MtEIL1 expression and SE in culture but the role and mechanism of ethylene signalling in the process of plant regeneration through SE requires further investigation. Comparison of transcriptome of WT and highly embryogenic 2HA spontaneous mutant of Medicago truncatula in 4 weeks tissue culture
Project description:12plex_medicago-2012-06 - vegetative and reproductive leaves - Transcriptome in leaves of Medicago truncatula plants during the remobilization process . Effect of a nitrogen deficiency on this process.Note that lower leaves correspond to vegetative leaves (FV) and upper leaves correspond to leaves of the reproductive part (FR). - Analysis of expression in Medicago truncatula reproductive leaves in untreated and nitrogen deficient plants during remobilization process between beginning of flowering, pod filling and the end of pod filling.
Project description:We used an M. truncatula 16K oligonucleotide array to profile gene expression in the leaves of transgenic alfalfa plants expressing Medicago truncatula isoflavone synthase 1 (MtIFS1). RNA purified from leaves of MtIFS1-expressing lines C22 and B20 and vector control line VC11 was used to generate Cy5-labeled cDNA. RNA purified from a second vector control line, VB2, was used to generate Cy3-labeled reference cDNA. Three independently propagated cuttings of each line were used and a total of nine hybridizations were performed. Our results indicated that MtIFS1-expression does not significantly alter global gene expression in the leaves. Keywords = Medicago Keywords = isoflavone synthase Keywords = alfalfa Keywords: other
Project description:Genome-wide transcriptome analysis of Arabidopsis thaliana was performed to understand the role of auxin in the response of leaf growth to osmotic stress. We studied transcriptional changes in proliferating leaves of the seedlings grown in vitro on control medium, medium supplemented with 25mM mannitol, 0.1μM NAA and 0.1μM NAA + 25mM mannitol.
Project description:Hairy root lines over-expressing MtPAR using a 35S promoter compared with hairy lines over-expressing GUS gene. Hairy roots were generated in vitro using leaf explants from Medicago A17. The goal of this experiment is to prove that ectopic expression expression of MtPAR is sufficient to induce tannin biosynthesis