The Glucocorticoid-Inducible GVG System Causes Severe Growth Defects in Both Root and Shoot of the Model Legume <i>Lotus japonicus</i>

Andersen, Stig Uggerhøj; Cvitanich, Cristina; Hougaard, Birgit Kristine; Roussis, Andreas; Grønlund, Mette; Jensen, Dorthe Bødker; Frøkjær, Line Askou; Jensen, Erik Appel

doi:10.1094/mpmi.2003.16.12.1069

Cited by 44 publications

(37 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…While cre/lox-mediated excision of a genomic RBR fragment produced sectors with mutant phenotypes in root meristems (Wildwater et al, 2005) and disturbed cell proliferation in leaves (see Supplemental Figure 1 online), we found that the central zone of the shoot apical meristem was recalcitrant to loss of lox alleles and RBR function. We then established conditional RBR mutant lines using the dexamethasone (DEX)-inducible GVG transcription factor system (Aoyama and Chua, 1997) but found that the DEX system by itself may produce phenotypes that depend on the expression level of GVG, as was previously reported by others (Kang et al, 1999;Andersen et al, 2003;Amirsadeghi et al, 2007). Although RBR protein levels could be effectively manipulated in DEX-inducible plants (data not shown), the GVG-induced phenotypes may obscure or amplify phenotypes produced by deregulation of RBR expression.…”

Section: Construction Of Arabidopsis Lines With Inducible Rna Interfementioning

confidence: 84%

ArabidopsisRETINOBLASTOMA-RELATED Is Required for Stem Cell Maintenance, Cell Differentiation, and Lateral Organ Production

et al. 2010

View full text Add to dashboard Cite

Several genes involved in the regulation of postembryonic organ initiation and growth have been identified. However, it remains largely unclear how developmental cues connect to the cell cycle. RETINOBLASTOMA RELATED (RBR) is a plant homolog of the tumor suppressor Retinoblastoma (pRb), which is a key regulator of the cell cycle. Using inducible RNA interference (RNAi) against Arabidopsis thaliana RBR (RBRi), we reduced RBR expression levels at different stages of plant development. Conditional reduction or loss of RBR function disrupted cell division patterns, promoted context-dependent cell proliferation, and negatively influenced establishment of cell differentiation. Several lineages of toti-and pluripotent cells, including shoot apical meristem stem cells, meristemoid mother cells, and procambial cells, failed to produce appropriately differentiated cells. Meristem activity was altered, leading to a disruption of the CLAVATA-WUSCHEL feedback loop and inhibition of lateral organ formation. Release of RBR from RNAi downregulation restored meristem activity. Gene profiling analyses soon after RBRi induction revealed that a change in RBR homeostasis is perceived as a stress, even before genes regulated by RBR-E2F become deregulated. The results establish RBR as a key cell cycle regulator required for coordination of cell division, differentiation, and cell homeostasis. INTRODUCTIONRetinoblastoma (RB) was the first tumor suppressor gene identified in animals (Friend et al., 1986) and later was associated with cell cycle regulation, thus establishing pRb as a key regulator of cell proliferation. In animals, three members of the RB family of proteins (pRb, p107, and p130), also referred to as pocket proteins (Mulligan and Jacks, 1998), negatively regulate the G1-to-S phase transition during the cell cycle. It is now widely accepted that mitogenic signals activate several cyclin-dependent kinase (CDK)-cyclin complexes during progression through G1 to phosphorylate pRb, thereby releasing it from interactions with E2F/DP transcription factor complexes to facilitate S-phase entry (Weinberg, 1995). Subsequently, pRb was also found to regulate tissue-specific transcription factors that regulate cell differentiation, such as MyoD, which activates muscle-specific genes (De Falco et al., 2006). pRb also regulates germ cell proliferation, differentiation, and survival (Toppari et al., 2003) and is required for osteoblast, keratinocyte, and macrophage differentiation (Nead et al., 1998;Liu et al., 1999;Thomas et al., 2003). Together with the tumor suppressor p53, pRb regulates adipocyte differentiation and function (Hallenborg et al., 2009). The requirement for RB in cell differentiation therefore extends beyond regulation of cell cycle entry. For example, the loss of RB function in cardioblasts affects cell differentiation by modulating the activity of cardiogenic factors (Papadimou et al., 2005).RB-related genes, termed RETINOBLASTOMA-RELATED (RBR; Ach et al., 1997), are also found in monocots (Grafi et al., 1996;Xie et al...

show abstract

Section: Construction Of Arabidopsis Lines With Inducible Rna Interfementioning

confidence: 84%

ArabidopsisRETINOBLASTOMA-RELATED Is Required for Stem Cell Maintenance, Cell Differentiation, and Lateral Organ Production

et al. 2010

View full text Add to dashboard Cite

show abstract

“…We have also to consider that some of these chemicals may be toxic for plant cells. For example, chemical agents can be at the origin of growth disturbance as it was observed for glucocorticoid-inducible system in the legume Lotus japonicus (Andersen et al, 2003). Another advantage of the heat-shock-induced gene silencing system is a more complete efficiency (i.e.…”

Section: Discussionmentioning

confidence: 99%

“…Their effects are reversible when the chemical treatment is stopped. However, it was reported that in some cases, the chemical application can causes growth disturbances as shown for the gluco-corticoid inducible system in the legume Lotus japonicus (Andersen et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

Gene silencing using a heat-inducible RNAi system in Arabidopsis

Masclaux

Charpenteau

Takahashi

et al. 2004

Biochemical and Biophysical Research Communications

View full text Add to dashboard Cite

Controlling gene expression during plant development is an efficient tool to explore gene function. In this paper, we describe a gene expression system driven by a heath shock gene promoter (HSP18.2), to trigger the expression of an intron-containing inverted repeat. RNA interference became a powerful way for gene functional analysis by reverse genetic approaches. However, constitutive gene silencing cannot be used with genes involved in fundamental processes such as embryo viability. Inducible promoters provide an alternative approach for temporal and spatial gene expression control. And we described here a new, system, complementary to those using chemical gene inducers. To evaluate the efficiency of this system, RNA corresponding to the phytoene desaturase gene of Arabidopsis thaliana was used as a reporter gene in transgenic plants and a comparative study was performed using either the CaMV35S constitutive promoter or the HSP18.2 inducible promoter.

show abstract

“…Position effects can alter the activity of the driver as well the reporter (2). High transcriptional levels of the driver can lead to posttranscriptional gene silencing (PTGS) (3) or nonspecific binding, thus promoting the inactivation of the system or causing unpredictable side effects on plant development, as shown in (4)(5)(6)(7). The choice of a binary system permits to select for functional driver and reporter lines without undesired side effects.…”

Section: Introductionmentioning

confidence: 99%

Inducible Gene Expression Systems for Plants

Borghi

2010

Plant Developmental Biology

View full text Add to dashboard Cite

Several systems for induction of transgene expression in plants have been described recently. Inducible systems were used mainly in tobacco, rice, Arabidopsis, tomato, and maize. Inducible systems offer researchers the possibility to deregulate gene expression levels at particular stages of plant development and in particular tissues of interest. The more precise temporal and spatial control, obtained by providing the transgenic plant with the appropriate chemical compound or treatment, permits to analyze also the function of those genes required for plant viability. In addition, inducible systems allow promoting local changes in gene expression levels without causing gross alterations to the whole plant development. Here, protocols will be presented to work with five different inducible systems: AlcR/AlcA (ethanol inducible); GR fusions, GVG, and pOp/LhGR (dexamethasone inducible); XVE/OlexA (beta-estradiol inducible); and heat shock induction.

show abstract

The Glucocorticoid-Inducible GVG System Causes Severe Growth Defects in Both Root and Shoot of the Model Legume Lotus japonicus

Cited by 44 publications

References 30 publications

ArabidopsisRETINOBLASTOMA-RELATED Is Required for Stem Cell Maintenance, Cell Differentiation, and Lateral Organ Production

ArabidopsisRETINOBLASTOMA-RELATED Is Required for Stem Cell Maintenance, Cell Differentiation, and Lateral Organ Production

Gene silencing using a heat-inducible RNAi system in Arabidopsis

Inducible Gene Expression Systems for Plants

Contact Info

Product

Resources

About