In Situ Hybridization to mRNA of Arabidopsis Tissue Sections

Engler, Janice de Almeida; Groodt, Ruth De; Montagu, Marc Van; Engler, Gilbert

doi:10.1006/meth.2000.1144

Cited by 64 publications

(48 citation statements)

References 9 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Nearly all steps of the in situ method were performed as described by de Almeida Engler et al (2001). Radish (Raphanus sativus) and Arabidopsis seedlings were germinated on K1 medium (Valvekens et al, 1988), and plant material was collected for fixation.…”

Section: Mrna In Situ Hybridizationmentioning

confidence: 99%

Auxin-Mediated Cell Cycle Activation during Early Lateral Root Initiation

et al. 2002

Self Cite

View full text Add to dashboard Cite

Lateral root formation can be divided into two major phases: pericycle activation and meristem establishment. In Arabidopsis, the first lateral root initiation event is spatially and temporally asynchronous and involves a limited number of cells in the xylem pericycle. To study the molecular regulation during pericycle activation, we developed a lateral rootinducible system. Successive treatments with an auxin transport inhibitor and exogenous auxin were used to prevent the first formative divisions and then to activate the entire pericycle. Our morphological and molecular data show that, in this inducible system, xylem pericycle activation was synchronized and enhanced to cover the entire length of the root. The results also indicate that the inducible system can be considered a novel in planta system for the study of synchronized cell cycle reactivation. In addition, the expression patterns of Kip-Related Protein2 ( KRP2 ) in the pericycle and its ectopic expression data revealed that the cyclin-dependent kinase inhibitor plays a significant role in the regulation of lateral root initiation. KRP2 appears to regulate early lateral root initiation by blocking the G1-to-S transition and to be regulated transcriptionally by auxin.

show abstract

Section: Mrna In Situ Hybridizationmentioning

confidence: 99%

Auxin-Mediated Cell Cycle Activation during Early Lateral Root Initiation

et al. 2002

Self Cite

View full text Add to dashboard Cite

show abstract

“…The whole-mount in situ RNA hybridization procedure has been described (de Almeida Engler et al, 1994). The RNA probe was designed to hybridize to both GL3 and EGL3 transcripts, so it included bp 550-1120 and bp 1400-1850 downstream from the start site of the EGL3-coding sequence corresponding to the most similar region of the GL3 and EGL3 proteins but excluding the bHLH signature region to eliminate the possibility of cross-hybridization to other bHLH proteins.…”

Section: In Situ Rna Hybridizationmentioning

confidence: 99%

The bHLH genesGL3andEGL3participate in an intercellular regulatory circuit that controls cell patterning in theArabidopsisroot epidermis

et al. 2005

View full text Add to dashboard Cite

“…Plant material was hybridized overnight at 42°C with the appropriate antisense and control 35 S-labeled mRNA probes (5 3 10 6 cpm per slide). After hybridization, the slides were washed in 2 3 SSC (1 3 SSC, 150 mM NaCl, NA 3 -citrate, pH 7.0) at room temperature for 1 h and in 0.1 3 SSC/50% (w/v) formamide at 42°C for 1 h. All posthybridization procedures, including RNase treatment and washes, were performed as described by de Almeida Engler et al (2001). Signal detection was achieved by autoradiography, using Kodak (Rochester, NY) NBT film emulsion.…”

Section: Cell Cycle Gene Promoter-gus Analysismentioning

confidence: 99%

The Role of the Cell Cycle Machinery in Resumption of Postembryonic Development

et al. 2005

Self Cite

View full text Add to dashboard Cite

Cell cycle activity is required for plant growth and development, but its involvement in the early events that initiate seedling development remains to be clarified. We performed experiments aimed at understanding when cell cycle progression is activated during seed germination, and what its contribution is for proper seedling establishment. To this end, the spatial and temporal expression profiles of a large set of cell cycle control genes in germinating seeds of Arabidopsis (Arabidopsis thaliana) and white cabbage (Brassica oleracea) were analyzed. The in vivo behavior of the microtubular cytoskeleton was monitored during Arabidopsis seed germination. Flow cytometry of Arabidopsis germinating seeds indicated that DNA replication was mainly initiated at the onset of root protrusion, when germination reached its end. Expression analysis of cell cycle genes with mRNA in situ localization, b-glucuronidase assays, and semiquantitative reverse transcription-polymerase chain reaction showed that transcription of most cell cycle genes was detected only after completion of germination. In vivo green fluorescent protein analysis of the microtubule cytoskeleton demonstrated that mitosis-specific microtubule arrays occurred only when the radicle had started to protrude, although the assembly of the microtubular cytoskeleton was promptly activated once germination was initiated. Thus, seed germination involves the synthesis and/or activation of a reduced number of core cell cycle proteins, which only trigger DNA replication, but is not sufficient to drive cells into mitosis. Mitotic divisions are observed only after the radicle has protruded and presumably rely on the de novo production of other cell cycle regulators.Seed germination is the process by which the plant embryo resumes growth after a period of quiescence. Under favorable conditions, rapid growth of the embryo culminates in rupture of the covering layers and emergence of the radicle, which is considered the completion of germination. At this stage, the decision of individual embryo cells to reenter the cell cycle or to remain arrested is crucial to determine seedling formation. The building of plant shape and function depends on the ability of the embryo cells to resume division and differentiate. Understanding how the cell cycle genes work at this particular phase of plant development might help to clarify the cellular and structural events that bring a quiescent embryo to a metabolically active plant.Cell cycle is a coordinated cyclic series of events, occurring between the end of subsequent cell divisions, by which cellular material is duplicated and divided between daughter cells. Thus, cell cycle consists of two major events, DNA replication (S phase) and mitosis (M phase) separated by two gap phases, G1 and G2 (for review, see Dewitte and Murray, 2003). The G1 phase is assumed to be the gate through which most cells resume cell cycle progression after a nonproliferative period. Additionally, G1-to-S transition is the central target of the cross-talk betwe...

show abstract

In Situ Hybridization to mRNA of Arabidopsis Tissue Sections

Cited by 64 publications

References 9 publications

Auxin-Mediated Cell Cycle Activation during Early Lateral Root Initiation

Auxin-Mediated Cell Cycle Activation during Early Lateral Root Initiation

The bHLH genesGL3andEGL3participate in an intercellular regulatory circuit that controls cell patterning in theArabidopsisroot epidermis

The Role of the Cell Cycle Machinery in Resumption of Postembryonic Development

Contact Info

Product

Resources

About