Divide and conquer: development and cell cycle genes in plant transformation

Arias, Renée S.; Filichkin, Sergei A.; Strauss, Steven H.

doi:10.1016/j.tibtech.2006.04.007

Cited by 34 publications

(25 citation statements)

References 62 publications

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“…The process of making transgenic plant consist of various points including acquisition of totipotency, progress into the S-phase of the cell cycle, which makes cells competent for transformation, and differentiation of transformed cells into new plants (Arias et al 2006).…”

Section: Discussionmentioning

confidence: 99%

Stable transformation and actin visualization in callus cultures of dodder (Cuscuta europaea)

Švubová

Blehová

2013

Biologia

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Agrobacterium tumefaciens-mediated transformation of callus culture, combined with a visual selection of GFPtagged fimbrin actin binding domain (FABD2) expression is described for parasitic species (Cuscuta europaea). The conditions for callus induction from 1 cm-long explants from the basal part of 7-day-old dodder seedlings were defined. We obtained light-green calli, which were transformed with A. tumefaciens bacterial strain GV3101 carrying plasmid pCB302 (35S::ABD2:gfp) with neomycin phosphotransferase (nptII) gene. The limitations of selection procedures based on antibiotics were avoided using green fluorescent protein (GFP) detection, as a visual selection marker subcellularly targeted to the actin cytoskeleton. Fluorescence microscopy analyses demonstrated a network of nucleus-associated actin arrays and dense cortical actin arrangements in stably transformed Cuscuta callus cells. RT-PCR analyses confirmed gfp expression in transformed calli 7, 14 and 21 days after transformation. Although the GFP fluorescence associated with the actin cytoskeleton has retained for at least six months without silencing, no shoot regeneration was observed. It can be concluded that, C. europaea callus cells are competent for transformation, but under given conditions, these cells failed to realize their morphogenic and regeneration potentials.

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Section: Discussionmentioning

confidence: 99%

Stable transformation and actin visualization in callus cultures of dodder (Cuscuta europaea)

Švubová

Blehová

2013

Biologia

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“…The ability of plant cells to undergo transformation and regeneration is associated with cell cycle activity (29,32,33), and increasing evidence supports the idea of a critical role played by the RBR/E2F pathway (29,33,34). A compensatory mechanism between RBR1 and RBR3 could explain, at least in part, the well-known recalcitrance of maize and related grasses to genetic transformation (19,27,35).…”

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confidence: 96%

“…Ectopic expression of RepA provides an effective means to down-regulate RBR1 and release E2F activity in an inducible manner, and considerably stimulates cell transformation and callus growth in maize embryos (29). Inhibition of RBR3 expression by RBR1 suggests 2 possible alternative functions for RBR3 in cell cycle control: RBR3 could have a novel positive role or, similar to p107 in mammals, it could provide a fail-safe mechanism by restraining the cell cycle after loss or inactivation of RBR1 (15,19,27,31).The ability of plant cells to undergo transformation and regeneration is associated with cell cycle activity (29,32,33), and increasing evidence supports the idea of a critical role played by the RBR/E2F pathway (29,33,34). A compensatory mechanism between RBR1 and RBR3 could explain, at least in part, the well-known recalcitrance of maize and related grasses to genetic transformation (19,27,35).…”

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confidence: 99%

Positive regulation of minichromosome maintenance gene expression, DNA replication, and cell transformation by a plant retinoblastoma gene

Sabelli

Hoerster

Lizarraga

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

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Retinoblastoma-related (RBR) genes inhibit the cell cycle primarily by repressing adenovirus E2 promoter binding factor (E2F) transcription factors, which drive the expression of numerous genes required for DNA synthesis and cell cycle progression. The RBR-E2F pathway is conserved in plants, but cereals such as maize are characterized by having a complex RBR gene family with at least 2 functionally distinct members, RBR1 and RBR3. Although RBR1 has a clear cell cycle inhibitory function, it is not known whether RBR3 has a positive or negative role. By uncoupling RBR3 from the negative regulation of RBR1 in cultured maize embryos through a combination of approaches, we demonstrate that RBR3 has a positive and critical role in the expression of E2F targets required for the initiation of DNA synthesis, DNA replication, and the efficiency with which transformed plants can be obtained. Titration of endogenous RBR3 activity through expression of a dominant-negative allele with a compromised pocket domain suggests that these RBR3 functions require an activity distinct from its pocket domain. Our results indicate a cell cycle pathway in maize, in which 2 RBR genes have specific and opposing functions. Thus, the paradigm that RBR genes are negative cell cycle regulators cannot be considered universal.cell cycle ͉ DNA synthesis ͉ E2F ͉ RepA R etinoblastoma-related (RBR) proteins are negative cell cycle regulators conserved in animals and plants (1-5). They are known as pocket proteins by virtue of a conserved region located in their C-terminal half (i.e., the pocket domain), which is largely responsible for protein-protein interactions and biological function. It comprises 2 highly conserved A and B domains separated by a nonconserved spacer of varying length, as well as a less-conserved C-terminal region. In mammals, these proteins are well characterized and comprise RB, p107 and p130. RBR proteins repress cell cycle progression primarily through the inhibition of E2 promoter binding factor (E2F)-dependent transcription, which is required to express many genes involved in S-phase and cell cycle progression, including the minichromosome maintenance 2-7(MCM2-7) family of DNA replication factors (4, 6-11). Thus, repression or expression of E2F targets, such as MCM2-7 genes, is a good indicator of the activity of RBR and E2F proteins.In overexpression studies, all 3 mammalian pocket proteins inhibit E2F-dependent gene expression, recruit chromatin remodeling complexes, actively repress transcription, and arrest cell growth (3, 12, 13). However, gene knockouts in mice have shown that, in addition to having overlapping functions, individual pocket proteins have unique properties (14). In particular, although only RB is considered a bona fide tumor suppressor, both p107 and p130 can functionally compensate for RB inactivation or loss in certain contexts and restrain cell proliferation (15).Inactivation of RBRs can occur by various mechanisms. For example, it is well documented that several oncoviral proteins, such as SV40 large...

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“…Breakthroughs are needed that employ genes, such as the rol gene cassette from Agrobacterium rhizogenes, which promote the regeneration of transgenic cells (Arias et al, 2006). These genes could then be removed via methods such as recombinase mediated excision to produce normal plant phenotypes.…”

Section: Future Prospects and Challengesmentioning

confidence: 99%

Transformation as a Tool for Genetic Analysis in Populus

Busov

Strauss

Pilate³

2009

Genetics and Genomics of Populus

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We summarize the outlook for using transformation as a genetic tool in Populus. Transformation approaches avoid the major obstacle to performing genetics experiments in trees -namely long generation cycles and the difficulty of inbreeding to reveal loss of function alleles. Dominant transgenic alleles allow modifications in gene function to be readily observed in primary transformants. Although transformation has been mainly used for reverse genetics (where the gene sequence of interest is known), transgenic mutagenesis approaches such as activation tagging and gene/enhancer traps have also been shown to enable forward genetics (where the phenotype, not the gene, is known). We outline challenges and needs for more efficient use of transformation tools. These include expansion of the transformation toolbox (e.g., promoters, vectors, targeting), and improved ability to conduct field trials to study gene function in native and plantation environments (in spite of regulatory obstacles). Because of the power of transformation, it will remain a major genetic research tool for dissection of gene function in Populus for many years to come. It is the key biological attribute that makes poplar the most powerful model organism for genetic analysis of woody plant growth, adaptation, and development.

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Divide and conquer: development and cell cycle genes in plant transformation

Cited by 34 publications

References 62 publications

Stable transformation and actin visualization in callus cultures of dodder (Cuscuta europaea)

Stable transformation and actin visualization in callus cultures of dodder (Cuscuta europaea)

Positive regulation of minichromosome maintenance gene expression, DNA replication, and cell transformation by a plant retinoblastoma gene

Transformation as a Tool for Genetic Analysis in Populus

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