Molecular Characterization of the Plastidic Glucose-6-Phosphate Dehydrogenase from Potato in Comparison to its Cytosolic Counterpart

Schaewen, Antje von; Langenkämper, Georg; Graeve, K. G. de; Wenderoth, Irina; Scheibe, Renate

doi:10.1104/pp.109.4.1327

Cited by 85 publications

(88 citation statements)

References 28 publications

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“…1A). Immunoblot analyses with various G6PDH antisera (14,19) demonstrated that these defense-induced rises in G6PDH activity mainly result from up-regulated expression of cytosolic isoforms, encoded by G6PD (Fig. 1B), probably due to sensing elevated soluble sugar levels (7), known to accompany sourceto-sink transitions of infected tobacco source leaves (5,6).…”

Section: Resultsmentioning

confidence: 99%

“…Immunoblot analyses were conducted as described previously (14,19); blots were probed with various G6PDH antisera, developed by…”

Section: Methodsmentioning

confidence: 99%

“…S1) (22,23), and immunoblot analyses with isoform-specific G6PDH antisera (14,19). Although cP2-protein levels were reduced compared with those detected in the 2 parental lines (Fig.…”

Section: Expression Of a Plastidic G6pd Isoform In The Cytosol Of A Smentioning

confidence: 99%

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Isoenzyme replacement of glucose-6-phosphate dehydrogenase in the cytosol improves stress tolerance in plants

Scharte

Schön

Tjaden

et al. 2009

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

122

120

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In source leaves of resistant tobacco, oxidative burst and subsequent formation of hypersensitive lesions after infection with Phytophthora nicotianae was prevented by inhibition of glucose-6-phosphate dehydrogenase (G6PDH) or NADPH oxidases. This observation indicated that plant defense could benefit from improved NADPH availability due to increased G6PDH activity in the cytosol. A plastidic isoform of the G6PDH-encoding gene, G6PD, displaying high NADPH tolerance was engineered for cytosolic expression (cP2), and introduced into a susceptible cultivar. After infection, transgenic (previously susceptible) lines overexpressing cP2 showed early oxidative bursts, callose deposition, and changes in metabolic parameters. These responses resulted in timely formation of hypersensitive lesions similar to resistant plants, although their extent varied considerably between different transgenic lines. Additional RNAi suppression of endogenous cytosolic G6PD isoforms resulted in highly uniform defense responses and also enhanced drought tolerance and flowering. Cytosolic G6PDH seems to be a crucial factor for the outcome of plant defense responses; thus, representing an important target for modulation of stress resistance. Because isoenzyme replacement of G6PDH in the cytosol was beneficial under various kinds of cues, we propose this strategy as a tool to enhance stress tolerance in general.oxidative burst ͉ source-to-sink transition ͉ NADPH availability ͉ pathogen resistance ͉ tobacco P lants are under continuous threat of being challenged by pathogenic microorganisms, which try to exploit them as a source of carbohydrates and other assimilates. Timely recognition of invading microorganisms and rapid and effective induction of defense responses are presently considered the main difference between pathogen-resistant and susceptible plant lines (1). So-called compatible interactions between pathogens and plants result if the pathogen can overcome plant defense barriers and establish disease symptoms. During an incompatible interaction, a wealth of defense mechanisms [e.g., generation of reactive oxygen species (ROS), synthesis of pathogenesis-related (PR) proteins, cell-wall fortification, and the hypersensitive reaction (HR)] can efficiently limit pathogen growth. The HR, which includes ROS formation and programmed cell death (PCD), represents the most efficient mechanism of plant defense. ROS were previously proposed to orchestrate the establishment of plant defense responses (2, 3). NADPH oxidases at the plasma membrane are considered the main source of extracellular ROS formation during defense, the so-called oxidative burst. Down-regulation or elimination of NADPH oxidase leads to suppression of pathogen-induced oxidative bursts and HR (4).The oxidative burst, an evolutionarily conserved mechanism, is part of the innate immune response, and (besides triggering plant PCD) it is thought to contribute to direct killing of invading microorganisms. It also helps to mount an invasion barrier through radical-mediated polymer...

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Section: Resultsmentioning

confidence: 99%

“…Immunoblot analyses were conducted as described previously (14,19); blots were probed with various G6PDH antisera, developed by…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Isoenzyme replacement of glucose-6-phosphate dehydrogenase in the cytosol improves stress tolerance in plants

Scharte

Schön

Tjaden

et al. 2009

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

122

120

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“…RT of poly(A ϩ ) mRNA from total plant RNA was performed as by Graeve et al (1994), modified by von Schaewen et al (1995).…”

Section: Cdna Synthesis and Pcr Amplification (Rt-pcr)mentioning

confidence: 99%

“…For Southern and northern analyses and for screening of cDNA libraries, GntI-cDNA fragments were liberated by restriction digests, gel-purified, and radioactively labeled essentially as described in von Schaewen et al (1995), using 32 P(dCTP) and the HexaLabel DNA Labeling Kit (MBI Fermentas, St. Leon-Rot, Germany).…”

Section: Labeling Of Cdna Probesmentioning

confidence: 99%

Isolation and Characterization of Plant N-Acetyl Glucosaminyltransferase I (GntI) cDNA Sequences. Functional Analyses in the Arabidopsis cgl Mutant and in Antisense Plants

Wenderoth

Schaewen

2000

Plant Physiology

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We report on the isolation and characterization of full-length cDNA sequences coding for N-acetylglucosaminyltransferase I (GnTI) from potato (Solanum tuberosum L.), tobacco (Nicotiana tabacum L.), and Arabidopsis. The deduced polypeptide sequences show highest homology among the solanaceous species (93% identity between potato and tobacco compared with about 75% with Arabidopsis) but share only weak homology with human GnTI (35% identity). In contrast to the corresponding enzymes from animals, all plant GnTI sequences identified are characterized by a much shorter hydrophobic membrane anchor and contain one putative N-glycosylation site that is conserved in potato and tobacco, but differs in Arabidopsis. Southern-blot analyses revealed that GntI behaves as a single-copy gene. Northern-blot analyses showed that GntI-mRNA expression is largely constitutive. Arabidopsis cgl mutants deficient in GnTI activity also possess GntI mRNA, indicating that they result from point mutations. GntI-expression constructs were tested for the ability to relieve the GnTI block in protoplasts of the Arabidopsis cgl mutant and used to obtain transgenic potato and tobacco plants that display a substantial reduction of complex glycan patterns. The latter observation indicates that production of heterologous glycoproteins with little or no antigenic glycans can be achieved in whole plants, and not in just Arabidopsis, using antisense technology.

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Plastid Metabolic Pathways

Tetlow

Rawsthorne

Raines

et al. 2018

Annual Plant Reviews Online

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Molecular Characterization of the Plastidic Glucose-6-Phosphate Dehydrogenase from Potato in Comparison to its Cytosolic Counterpart

Cited by 85 publications

References 28 publications

Isoenzyme replacement of glucose-6-phosphate dehydrogenase in the cytosol improves stress tolerance in plants

Isoenzyme replacement of glucose-6-phosphate dehydrogenase in the cytosol improves stress tolerance in plants

Isolation and Characterization of Plant N-Acetyl Glucosaminyltransferase I (GntI) cDNA Sequences. Functional Analyses in the Arabidopsis cgl Mutant and in Antisense Plants

Plastid Metabolic Pathways

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