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Genome-wide analysis of gene expression in the soybean shoot apical meristem

ABSTRACT: The shoot apical meristem (SAM) contains undifferentiated stem cells that are responsible for the initiation of above-ground organs, and eventually the general architecture of the plant. To gain insight into the nature of genetic programs and the regulatory networks underlying SAM function in soybean, we have used Affymetrix soybean GeneChip® to investigate the transcript profiles associated with micro-dissected SAMs or axillary meristems (AMs). While the microarray data disclosed the conservation of transcriptional signature between the two types of meristems, subsequent comparison of SAM transcript profile with that of non-meristem (NM) tissue revealed a total of 1090 and 1523 transcripts that are significantly up- or down-regulated in the SAM. Further in situ hybridization analysis on selected transcripts has implicated their roles in SAM maintenance and the establishment of organ polarity. We also identified a gene that could potentially serve as a novel marker that distinguishes the differentiating cells in the meristem from the pluripotent stem cells. Along with many unknowns, transcripts with putative annotation have also been identified that has allowed us to infer SAM regulatory roles for various families of transcription factors as well as products associated with auxin-mediated responses, cell division and proliferation, epigenetic regulation, miRNA regulation and protein turnover. Computational analysis on the promoter regions of Arabidopsis orthologs of genes with high expression in the soybean SAM revealed a conserved over-representation of three cis-acting regulatory motifs. Our microarray data thus represents a rich source of target genes for further study into the meristem function and maintenance. Keywords: meristem transcript profile

ORGANISM(S): Glycine max

PROVIDER: GSE10607 | GEO | 2009/02/20

SECONDARY ACCESSION(S): PRJNA107777

REPOSITORIES: GEO

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Project description:The transition to flowering is characterized by a shift of the shoot apical meristem (SAM) from leaf production to the initiation of a floral meristem. In this study, we addressed the nature of SAM gene networks involved in the early floral initiation process in the crop legume soybean. Unique aspects (such as pod development and nitrogen fixation) of legume development make them appealing for plant development studies. Soybean, a major oilseed crop, possesses varied maturity groups; hence, understanding and unravelling initial transition control has implications in manipulating crop yield. To this end, we performed global gene expression analysis using Affymetrix® soybean GeneChip® with RNA isolated from micro-dissected soybean SAMs at various time points after plants were shifted from long-day to short-day growth conditions. Analysis of the resulting microarray data revealed a total of 331 transcripts that have differential expression profiles. Intriguingly, about 20% of the transcripts affected by the switch in the development program have orthologs reported to be responsive to abscisic acid (ABA), suggesting an increase in ABA levels in the SAM during this developmental change. A subsequent immunoassay verified this, thereby implicating its possible function as an endogenous signal during the floral evocation process. The striking occurrence of abiotic stress-related transcripts, including trehalose metabolism genes, in SAMs during the early transition to floral meristems points to an overlap of abiotic stress and floral signalling pathways in soybean. In addition, other hormones - auxin, jasmonic acid and brassinosteroids - and a number of candidate protein kinases may also act in the signalling process prior to or concurrently with the induction of the putative floral homeiotic transcripts. This indicates that molecular events mediated by multiple hormonal pathways are part of the mechanism employed by soybean to regulate the floral transition process. Keywords: transcript profiling floral transition soybean shoot apical meristem

Project description:Plant growth is the result of cell proliferation in the meristems, which requires a dynamic balance between the formation of new tissue and the maintenance of a set of undifferentiated stem cells. There is much that remains unknown about this vital developmental process. In this study, we have reported the generation of 2735 ESTs from three cDNA libraries derived from dissected garden pea (Pisum sativum cv Torsdag) shoot apical meristems. Clustering analysis of the resulting sequences gave rise to 1686 non-redundant ESTs. The unique sequences generated together with 500 other pea transcripts randomly selected from the GenBank pea protein database have been utilized to construct a 2K oligonucleotide array. Using this pea array, we have obtained the transcript profiles of pea shoot apical meristems in comparison to non-apical-meristem tissues. A total of 181 or 174 transcripts were identified to be significantly up- or down-regulated in the pea shoot apical meristem, respectively. As expected, close to 61% of the transcripts down-regulated in the shoot apical mersitem are those retrieved from the public database, whereas sequences from the same source only made up 12% of the genes that were expressed at higher levels in SAMs. This revelation highlights the under-representation of transcripts from the meristmatic tissues in the current public protein database. Manual inspection of the list of up-regulated transcripts reveals the presence of sequences predicted to encode products associated with cell division and proliferation, epigenetic regulation, auxin-mediated responses and miRNA regulation. Similar genes have been implicated in the regulation of plant stem cell activity. Taken together, our EST collection as well as the microarray data provides useful starting points for more in depth analysis of the meristem function and maintenance. Keywords: shoot apical meristem transcript profiling Total RNA was extracted from dissected SAM (approximately 80 SAMs per extraction) or other plant parts (primary stem, primary roots and mature leaves) using Qiagen RNeasy Mini Kit. Four independent tissue collections and RNA extractions (designated A, B, C and D) were performed for each of the microarray hybridization experiment.The Cy5- or Cy3-labelled cDNA was then hybridized to different sector of the chip according to a balanced block design dual label experiment scheme (Cochran & Cox, 1992): Sector 1: Cy3-SAM A vs Cy5-NM A Sector 2: Cy5-SAM B vs Cy3-NM B Sector 3: Cy3-SAM C vs Cy5-NM C Sector 4: Cy5-SAM D vs Cy3-NM D

Project description:SHOOTMERISTEMLESS (STM) is a well characterized key regulator of the shoot apical meristem (SAM) activity in diverse plant species. We used a transgenic approach to investigate how the altered expression of Brassica napus (Bn) STM affects the morphology and behavior of microspore-derived embryos (MDEs). Down-regulation of BnSTM repressed the formation of the SAM and reduced the number of MDEs able to regenerate viable plants at germination. This was in contrast to embryos over-expressing BnSTM which displayed enlarged SAMs with an improved regeneration frequency. To further investigate the regulatory mechanisms underpinning these different responses, global changes in transcript levels were analysed in laser micro dissected SAMs of the transformed MDEs. The induction of BnSTM up-regulated the expression of many transcription factors (TFs) some of which directly involved in the formation of the SAM, i.e. CUPSHAPED COTYLEDON1 and WUSCHEL, and regulatory components ofthe antioxidant response, hormone signalling, and cell wall synthesis and modification. Opposite expression patterns for some of these genes were observed in the SAMs of MDEs down-regulating BnSTM. These data were interpreted in relation to the ability of BnSTM to modulate the size and functionality of the SAM. Structural and expression studies were further conducted to confirm the role of BnSTM in mediating the regulatory network of cell wall and lignin synthesis. Besides lowering the transcript levels of early components of cell wall and lignin biosynthetic pathways in the SAM, BnSTM had a remarkable repressing effect on the expression of PHENYLALANINE AMMONIA LYASE2, CINNAMATE 4-4HYDROXYLASE, andCINNAMYL ALCOHOL DEHYDROGENASE. Since lignin formation is a feature of irreversible cell differentiation, these results suggest that one way in which BnSTM promotes indeterminate cell fate may be by preventing the expression of components of biochemical pathways involved in the accumulation of lignin in the meristematic cells. Overall these studies provide evidence for a novel function of BnSTM in enhancing the quality of in vitro produced meristems, and propose that this gene can be used as a potential target to improve regeneration of cultured embryos. 2 mutants and 1 wild type

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Genome-wide analysis of gene expression in the soybean shoot apical meristem

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