Synchronous <i>Arabidopsis</i> suspension cultures for analysis of cell‐cycle gene activity

Menges, Margit; Murray, James A. H.

doi:10.1046/j.1365-313x.2002.01274.x

Cited by 306 publications

(318 citation statements)

References 63 publications

(109 reference statements)

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“…Arabidopsis seeds with T-DNA insertions in the AtNAP57 (SALK_031065) and AtKU70 (SALK_123114) genes were purchased from the Arabidopsis Biological Resource Center (Ohio State University, Columbus, OH), cold treated overnight at 4°C, and then placed in an environmental growth chamber and grown under a 16-h light/8-h dark photoperiod at 23°C. Arabidopsis suspension culture cells were maintained as described previously (31). Siliques from wild-type and AtNAP57 heterozygotes were dissected 10 days after fertilization and photographed using a Zeiss Axiocam digital camera coupled to a Zeiss microscope.…”

Section: Methodsmentioning

confidence: 99%

Dyskerin Is a Component of the Arabidopsis Telomerase RNP Required for Telomere Maintenance

Kannan

Nelson

Shippen

2008

Molecular and Cellular Biology

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Dyskerin binds the H/ACA box of human telomerase RNA and is a core telomerase subunit required for RNP biogenesis and enzyme function in vivo. Missense mutations in dyskerin result in dyskeratosis congenita, a complex syndrome characterized by bone marrow failure, telomerase enzyme deficiency, and progressive telomere shortening. Here we demonstrate that dyskerin also contributes to telomere maintenance in Arabidopsis thaliana. We report that both AtNAP57, the Arabidopsis dyskerin homolog, and AtTERT, the telomerase catalytic subunit, accumulate in the plant nucleolus, and AtNAP57 associates with active telomerase RNP particles in an RNA-dependent manner. Furthermore, AtNAP57 interacts in vitro with AtPOT1a, a novel component of Arabidopsis telomerase. Although a null mutation in AtNAP57 is lethal, AtNAP57, like AtTERT, is not haploinsufficient for telomere maintenance in Arabidopsis. However, introduction of an AtNAP57 allele containing a T66A mutation decreased telomerase activity in vitro, disrupted telomere length regulation on individual chromosome ends in vivo, and established a new, shorter telomere length set point. These results imply that T66A NAP57 behaves as a dominant-negative inhibitor of telomerase. We conclude that dyskerin is a conserved component of the telomerase RNP complex in higher eukaryotes that is required for maximal enzyme activity in vivo.

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Section: Methodsmentioning

confidence: 99%

Dyskerin Is a Component of the Arabidopsis Telomerase RNP Required for Telomere Maintenance

Kannan

Nelson

Shippen

2008

Molecular and Cellular Biology

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“…Particle bombardment was performed using the Biolistic PDS-1000/He system (Bio-Rad), and luciferase assays were performed using the DualLuciferase Reporter Assay System (Promega) as previously reported (Hiratsu et al, 2002). Arabidopsis MM2d cultured cells (Menges and Murray, 2002) were used as host cells and luciferase activities were quantified using the Mithras LB940 Microplate Luminometer (Berthold Technologies).…”

Section: Transient Expression Assaymentioning

confidence: 99%

WIND1 Promotes Shoot Regeneration through Transcriptional Activation of ENHANCER OF SHOOT REGENERATION1 in Arabidopsis

et al. 2016

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Many plant species display remarkable developmental plasticity and regenerate new organs after injury. Local signals produced by wounding are thought to trigger organ regeneration but molecular mechanisms underlying this control remain largely unknown. We previously identified an AP2/ERF transcription factor WOUND INDUCED DEDIFFERENTIATION1 (WIND1) as a central regulator of wound-induced cellular reprogramming in plants. In this study, we demonstrate that WIND1 promotes callus formation and shoot regeneration by upregulating the expression of the ENHANCER OF SHOOT REGENERATION1 (ESR1) gene, which encodes another AP2/ERF transcription factor in Arabidopsis thaliana. The esr1 mutants are defective in callus formation and shoot regeneration; conversely, its overexpression promotes both of these processes, indicating that ESR1 functions as a critical driver of cellular reprogramming. Our data show that WIND1 directly binds the vascular system-specific and wound-responsive cis-element-like motifs within the ESR1 promoter and activates its expression. The expression of ESR1 is strongly reduced in WIND1-SRDX dominant repressors, and ectopic overexpression of ESR1 bypasses defects in callus formation and shoot regeneration in WIND1-SRDX plants, supporting the notion that ESR1 acts downstream of WIND1. Together, our findings uncover a key molecular pathway that links wound signaling to shoot regeneration in plants.

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“…For high-throughput quantitative screening for plant immunepriming compounds, we used the Arabidopsis MM1 cell suspension cultures (Menges and Murray, 2002) challenged with Pst-avrRpm1 (Mackey et al, 2002). In this system, Arabidopsis cells induced immune-related cell death as a defense response 20 h after inoculation, as has been observed in planta (Mackey et al, 2002).…”

Section: Screening For Plant Immune-priming Compoundsmentioning

confidence: 99%

Novel Plant Immune-Priming Compounds Identified via High-Throughput Chemical Screening Target Salicylic Acid Glucosyltransferases in Arabidopsis

et al. 2012

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Plant activators are compounds, such as analogs of the defense hormone salicylic acid (SA), that protect plants from pathogens by activating the plant immune system. Although some plant activators have been widely used in agriculture, the molecular mechanisms of immune induction are largely unknown. Using a newly established high-throughput screening procedure that screens for compounds that specifically potentiate pathogen-activated cell death in Arabidopsis thaliana cultured suspension cells, we identified five compounds that prime the immune response. These compounds enhanced disease resistance against pathogenic Pseudomonas bacteria in Arabidopsis plants. Pretreatments increased the accumulation of endogenous SA, but reduced its metabolite, SA-O-β-d-glucoside. Inducing compounds inhibited two SA glucosyltransferases (SAGTs) in vitro. Double knockout plants that lack both SAGTs consistently exhibited enhanced disease resistance. Our results demonstrate that manipulation of the active free SA pool via SA-inactivating enzymes can be a useful strategy for fortifying plant disease resistance and may identify useful crop protectants.

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Synchronous Arabidopsis suspension cultures for analysis of cell‐cycle gene activity

Cited by 306 publications

References 63 publications

Dyskerin Is a Component of the Arabidopsis Telomerase RNP Required for Telomere Maintenance

Dyskerin Is a Component of the Arabidopsis Telomerase RNP Required for Telomere Maintenance

WIND1 Promotes Shoot Regeneration through Transcriptional Activation of ENHANCER OF SHOOT REGENERATION1 in Arabidopsis

Novel Plant Immune-Priming Compounds Identified via High-Throughput Chemical Screening Target Salicylic Acid Glucosyltransferases in Arabidopsis

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