Different radiosensitivities of Nicotiana plumbaginifolia leaves and regenerating protoplasts

Magnien, E.; Dalschaert, X.; Devreux, M.

doi:10.1016/0304-4211(80)90077-2

Cited by 11 publications

(7 citation statements)

References 29 publications

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“…D~-sirre. The protoplast isolation procedure and culture media are as described by Magnien [27], except for the enzymatic solution which contained 2 ~o w/v cellulysin and 0.5 ~o w/v macerase. Electroporation of protoplasts was conducted with a home-made capacitor discharge system, using the disposable electroporation chambers (0.4 cm) of the Cell-Porator System of Gibco-BRL (Gaithersburg, MD).…”

Section: Protoplast Isolation and Electroporationmentioning

confidence: 99%

Identification of cis-acting elements involved in the regulation of the pathogenesis-related gene STH-2 in potato

et al. 1993

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We have characterized a genomic clone containing the potato pathogenesis-related genes STH-2 and STH-21. The two genes are found 4 kb apart on the same chromosome and their sequences are highly similar. They present the same transcriptional orientation and are both interrupted by a single intron. A chimaeric gene consisting of 1015 bp of 5'-flanking sequence and part of the first exon of STH-2 fused to the bacterial beta-glucuronidase gene was highly-expressed in tubers of transgenic potato plants after wounding and elicitor treatments. The levels of activity observed in these transgenic plants parallel those observed for the accumulation of STH-2 mRNAs under similar conditions. This indicates that cis-acting elements necessary for the proper activation of the gene are present within 1 kb of 5'-flanking sequences. Functional analysis of 5' deletions of the STH-2/GUS constructs by transient expression in leaf protoplasts revealed the presence of an upstream regulatory sequence between -135 and -52 which contains a TGAC motif, and a possible negative regulatory region between -52 and -28. A factor present in nuclear extracts of wounded potato tubers was found to bind specifically to nucleotides located between -135 to -105, suggesting that this region contains important cis-regulatory elements.

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Section: Protoplast Isolation and Electroporationmentioning

confidence: 99%

Identification of cis-acting elements involved in the regulation of the pathogenesis-related gene STH-2 in potato

et al. 1993

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“…Tubers were stored in the dark at 4ЊC and brought to room temperature 24 hr before use. Leaf mesophyll protoplasts were isolated from 7-to 8-week-old potato plants (cv Kennebec) grown in growth chambers as described by Magnien et al (1980). …”

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confidence: 99%

PBF-2 Is a Novel Single-Stranded DNA Binding Factor Implicated in PR-10a Gene Activation in Potato

et al. 2000

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Elicitor-induced activation of the potato pathogenesis-related gene PR-10a requires a 30-bp promoter sequence termed the ERE (elicitor response element) that is bound by the nuclear factor PBF-2 ( PR-10a binding factor 2). In this study, PBF-2 has been purified to near homogeneity from elicited tubers through a combination of anion-exchange and DNA affinity chromatography. Evidence demonstrates that inactive PBF-2 is stored in the nuclei of fresh tubers and becomes available for binding to the ERE upon elicitation. A protein with an apparent molecular mass of 24 kD (p24) is a DNA binding component of PBF-2. A cDNA encoding p24 has been cloned and encodes a novel protein with a potential transcriptional activation domain that could also act as a single-stranded DNA binding domain. Both PBF-2 and the cDNA-encoded protein bind with high affinity to the single-stranded form of the ERE in a sequence-specific manner. The inverted repeat sequence of the ERE, TGACAnnnnTGTCA, is critical for binding of this factor in vitro and for PR-10a expression in vivo, supporting the role of PBF-2 as a transcriptional regulator. INTRODUCTIONPlants defend themselves against fungal pathogens by a variety of mechanisms, including preexisting physical barriers and inducible defenses (Lamb et al., 1989). Attack by an avirulent strain of pathogen results in a rapid localized necrosis at the site of infection (termed the hypersensitive response), which contributes to pathogen limitation (Keen, 1992). With a few exceptions, deployment of the inducible defenses requires massive gene induction (Lamb et al., 1989). Despite the importance of transcriptional activation during the plant defense response, very little is known about the players involved and the exact mechanisms that lead to defense gene induction.PR (pathogenesis-related) genes are among the best characterized genes induced by pathogens. Heterogeneous in structure and function, PR genes are subdivided into 11 groups (Van Loon et al., 1994). Although the function of certain PR proteins is unknown, some display in vitro antifungal properties (Schlumbaum et al., 1986;Vigers et al., 1991;Ponstein et al., 1994;Niderman et al., 1995). Genes of the PR-10 group are present in numerous dicots (Somssich et al., 1988;Breiteneder et al., 1989;Matton and Brisson, 1989;Walter et al., 1990) and monocots (Warner et al., 1992;Moons et al., 1997). Evidence is accumulating that some PR-10 proteins might possess ribonuclease activity (Moiseyev et al., 1994;Bufe et al., 1996;Swoboda et al., 1996). More recently, structural and sequential homology between the PR-10 proteins and a group of latex proteins has been described (Osmark et al., 1998). Genes of the PR-10 group encode small, primarily acidic intracellular proteins with molecular masses ranging from 15 to 18 kD and have been shown to be transcriptionally regulated (Linthorst, 1991).In only two cases have cis elements and their trans -acting factors been characterized in the promoters of PR-10 genes. These studies revealed that the processes of tr...

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“…This stage of development corresponds in this particular species to a good cytological homogeneity, with >90% of Grphase diploid nuclei [21 ]. The isolation procedure implied carborundum brushing, enzymatic digestion and stationary density gradient purification, as in [19].…”

Section: Plantlet Culture and Protoplast Isolationmentioning

confidence: 89%

“…The synthetic substrate was an agar medium [19] supplemented with the vitamins recommended in [20], 200 mg glutamine/1, 5 g sucrose/1 and 6 g activated charcoal/l. Controlled growth conditions were 16 h photo-periods at 3000 lux at 24-28°C.…”

Section: Plantlet Culture and Protoplast Isolationmentioning

confidence: 99%

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DNA repair synthesis in plant protoplasts is aphidicolin‐resistant

Sala

Magnien

Galli³

et al. 1982

FEBS Letters

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Different radiosensitivities of Nicotiana plumbaginifolia leaves and regenerating protoplasts

Cited by 11 publications

References 29 publications

Identification of cis-acting elements involved in the regulation of the pathogenesis-related gene STH-2 in potato

Identification of cis-acting elements involved in the regulation of the pathogenesis-related gene STH-2 in potato

PBF-2 Is a Novel Single-Stranded DNA Binding Factor Implicated in PR-10a Gene Activation in Potato

DNA repair synthesis in plant protoplasts is aphidicolin‐resistant

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