Isolation of the genomic clone for pathogenesis-related protein 1a from Nicotiana tabacum cv. Xanthi-nc

Payne, George B.; Td, Parks; Burkhart, William; Dincher, Sandra; P, Ahl; Jp, Metraux; Ryals, John

doi:10.1007/bf00015662

Cited by 57 publications

(44 citation statements)

References 18 publications

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“…Samsun NN and Xanthi-nc (data not shown). Subsequently, two independent clones were sequenced in total both containing the same nucleotide sequence identical to the Samsun NN and Xanthi-nc PR-1 a sequences (Cornelissen et al, 1987;Ohshima et al, 1987;Payne et al, 1988). Taken together, these data suggest that there is no nucleotide sequence variation in the 902-bp upstream regions among PR-1 a genes from different tobacco cultivars.…”

Section: Isolation Of the 5'-flanking Region Of The Pr-la Gene From Nmentioning

confidence: 82%

“…1 A) was added to give plasmid -1533PRla [GUS]. The PR-1 a promoter/ GUS expression cassettes were excised from the respective pUC plasmids as HindIII-EcoRI fragments and ligated into (Cornelissen et al, 1987;Ohshima et al, 1987;Payne et al, 1988), the positions of the primers used in the PCR, the extents of the PCR products obtained, and the position of the previously isolated 1 clone W38/5 (Pfitzner et al, 1988) pBinl9 (Bevan, 1984). The mobilization of chimeric constructs into Agrobacterium and the transformation of tobacco plants have been described previously (Beilmann et al, 1991).…”

Section: Construction and Transformation Of Chimeric Reporter Genesmentioning

confidence: 99%

“…Four primers were designed based on the reported sequences of the PR-1 a genes from N. tabacum cvs. Samsun NN (Cornelissen et al, 1987;Ohshima et al, 1987) and Xanthi-nc (Payne et al, 1988), and the upstream region of the PR-la gene from N. tabacum cv. Wisconsin 38 was amplified from genomic DNA in independent reactions.…”

Section: Isolation Of the 5'-flanking Region Of The Pr-la Gene From Nmentioning

confidence: 99%

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The upstream region of the gene for the pathogenesis‐related protein 1 a from tobacco responds to environmental as well as to developmental signals in transgenic plants

Grüner

Pfitzner

1994

European Journal of Biochemistry

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Pathogenesis-related proteins (PR proteins) are a heterogeneous group of proteins which are induced in plants by diverse stimuli, e.g. PR proteins are elicited by pathogen attack in the course of the hypersensitive defense reaction of the plant. To examine the regulation of these genes, the 5'-flanking region of the tobacco (Nicotiana tabacum cv. Wisconsin 38) PR-1 a gene up to position -1533 was isolated from genomic DNA by the polymerase chain reaction. Two chimeric gene constructs containing 1533 bp and 906 bp, respectively, of the PR-1 a upstream region fused to the GUS reporter gene were stably integrated into the tobacco genome. All primary transformants exhibited induced expression of the reporter gene after infection of the plants with tobacco mosaic virus or treatment with acetylsalicylic acid. In addition, similar expression of the reporter gene was observed in leaves of adult transgenic plants-without any prior inductive treatments. To study this phenomenon in more detail, the F1 progeny of independent transgenic lines were monitored during the ontogeny of the plants. In normally developing tobacco plants, strong GUS activities were typically detected approximately 12 weeks after germination in the lowest leaves of vegetative plants. When successive leaves of individual plants were tested during the following weeks, a clear gradient of reporter gene activity had developed in the green leaves including the sepals from the bottom to the top of the plants. In all cases analyzed, this gradient of reporter gene expression was strictly parallelled by the expression of the endogenous PR-1 proteins. These results suggest that the acidic PR-1 proteins from tobacco fulfiIl a role during the later stages of plant development and that the PR-1 a upstream region -906 to -335 contains positive regulatory elements for both environmental and developmental signals.Infection of plants with pathogens usually results in a distinct host response depending on the genetic constitution of the plant and the pathogen. In an incompatible reaction, the plant succeeds in localizing the pathogen at the primary infection sites which often become visible as necrotic local lesions on the leaves (hypersensitive response). This local defense reaction is accompanied by a general resistance of the whole plant against further attack by pathogens, a phenomenon called systemic acquired resistance. During the defense reaction of the plant, several host-encoded proteins are induced which are known as pathogenesis-related proteins (PR proteins) and which are believed to play a role in restricting the pathogen. PR proteins were first described in 1970 in tobacco plants exhibiting the hypersensitive response after infection with toCorrespondence to U. M. Pfitzner, Botanisches Institut, MenFax: +49 89 1782 274.

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Section: Isolation Of the 5'-flanking Region Of The Pr-la Gene From Nmentioning

confidence: 82%

Section: Construction and Transformation Of Chimeric Reporter Genesmentioning

confidence: 99%

Section: Isolation Of the 5'-flanking Region Of The Pr-la Gene From Nmentioning

confidence: 99%

See 1 more Smart Citation

The upstream region of the gene for the pathogenesis‐related protein 1 a from tobacco responds to environmental as well as to developmental signals in transgenic plants

Grüner

Pfitzner

1994

European Journal of Biochemistry

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“…We reported the complete sequence of the PR-1a gene and two PR-1 pseudogenes , two active PR-S genes (Van Kan et al, 1989), and two genes encoding a glycine-rich protein (GRP) that does not show homology to any of the known PR proteins . In addition, the sequence of the PR-1a gene was reported by severa1 other groups (Ohshima et al, 1987;Payne et al, 1988;Pfitzner et al, 1988). The PR-1a and GRP genes are strongly induced by spraying tobacco with a 5 mM salicylate solution, whereas the PR-S genes are weakly induced by this treatment.…”

Section: Introductionmentioning

confidence: 79%

Analysis of regulatory elements involved in the induction of two tobacco genes by salicylate treatment and virus infection.

et al. 1990

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Tobacco genes encoding the PR-la protein and a glycine-rich protein are expressed after treatment of plants with salicylate or infection with tobacco mosaic virus. Upstream sequences of these genes were fused to reporter genes, and these constructs were used to transform tobacco. Upstream sequences of the PR-la gene of 689 base pairs or longer were sufficient for induction of the reporter gene in tobacco mosaic virus-inoculated leaves, systemically induced leaves from infected plants, and leaves treated with salicylate. N o such induction was found with upstream sequences of 643 base pairs or shorter of the PR-la gene. When the PR-la upstream sequence from nucleotides -625 to -902 was fused to the cauliflower mosaic virus 35s core promoter, a construct was obtained that conferred tobacco mosaic virus and salicylate inducibility to the reporter gene in transgenic plants. This confirmed the localization of tobacco mosaic vírus-and salicylate-responsive elements between positions -643 and -689 in the PR-la promoter. With the glycine-rich protein gene, an upstream sequence of 645 base pairs was sufficient for tobacco mosaic virus and salicylate inducibility of the reporter gene, whereas constructs containing 400 base pairs or fewer of the glycine-rich protein promoter were largely inactive.

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“…The PR-la messenger transcribed from this gene will contain 28 5'-noncoding nucleotides in front of the start codon and possibly terminate somewhere on one of several polyadenylation signals in the 322-nucleotide-long 3'-noncoding region. Payne et al (1988) found that PR-la mRNA from Xanthi-nc tobacco is polyadenylated at at least two sites in this nontranslated region. If transcription termination of the transgenic messenger takes place at these internal polyadenylation sites, the resulting messenger will be only one nucleotide shorter than the endogenous PR-la messengers.…”

mentioning

confidence: 97%

Constitutive expression of pathogenesis-related proteins PR-1, GRP, and PR-S in tobacco has no effect on virus infection.

et al. 1989

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Samsun NN tobacco cells were transformed with chimeric genes for pathogenesis-related (PR) proteins derived from genomic (PR-la, GRP) or cDNA (PR-S) clones under the transcriptional control of the cauliflower mosaic virus 35S promoter. Regenerated plants were assayed by RNA and protein gel blotting, and plants showing high specific expression of the inserted genes were selected for self-pollination and seed formation. Inspection of second generation transformants showed that constitutive expression of PR-la, GRP, and PR-S in tobacco in general does not have an effect on the phenotypic appearance of the plants or the expression of other endogenous PR genes. Furthermore, constitutive expression of the above genes does not affect the susceptibility of the plants to infection with tobacco mosaic virus or alfalfa mosaic virus. INTRODUCTIONSamsun NN tobacco responds to infection with tobacco mosaic virus (TMV) in a hypersensitive way. Small necrotic lesions are formed within 2 to 3 days after inoculation on the inoculated leaves, and the infection usually does not spread to other parts of the plant. However, these noninfected parts appear to have acquired a resistance to the virus when subsequently challenged with TMV. In addition, infection with other pathogens to which the plant reacts in a hypersensitive way with the formation of local necrosis induces resistance to further infection with either virus, fungus, or bacterium (Gianninazzi, 1983). This phenomenon of acquired resistance is paralleled with the appearance of a set of proteins collectively known as pathogenesis-related (PR) proteins (Van Loon, 1982). These proteins appear to have acidic pl values, to be highly proteaseresistant, and to be excreted to the intercellular fluid (Van Loon, 1988).Recently, characterization of cDNA clones and serologic studies have revealed that several of these acidic PR proteins have homologous basic counterparts (Cornelissen et al., 1987;Kauffman et al., 1987;Legrand et al., 1987;Shinshi et al., 1987). These basic proteins are also induced upon TMV infection and are possibly transported to the vacuole (Singh et al., 1987).In vitro studies have indicated the possible functions of ~To whom correspondence should be addressed. Current address: Gorlaeus Laboratories, State University of Leiden, P.O. Box 9502, 2300 RA Leiden, The Netherlands.several of these induced proteins. PR proteins P and Q have chitinase activity , whereas PR-2, N and O are/3-1,3-glucanases (Kauffman et al., 1987), suggesting that these hydrolyzing enzymes could be involved in the degradation of bacterial and fungal cell walls or insect exoskeletons. Indeed, in vitro experiments have shown the inhibitory effect on fungal growth of mixtures of purified chitinase and glucanase from pea (Mauch et al., 1988). PR-S has extensive sequence homology with a maize protein inhibiting the activity of proteinase and insect e-amylase (Richardson et al., 1987), which suggests a possible function in the resistance to insect attack. The function of the three almost identical ...

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Isolation of the genomic clone for pathogenesis-related protein 1a from Nicotiana tabacum cv. Xanthi-nc

Cited by 57 publications

References 18 publications

The upstream region of the gene for the pathogenesis‐related protein 1 a from tobacco responds to environmental as well as to developmental signals in transgenic plants

The upstream region of the gene for the pathogenesis‐related protein 1 a from tobacco responds to environmental as well as to developmental signals in transgenic plants

Analysis of regulatory elements involved in the induction of two tobacco genes by salicylate treatment and virus infection.

Constitutive expression of pathogenesis-related proteins PR-1, GRP, and PR-S in tobacco has no effect on virus infection.

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