Studies on vacuole regeneration in evacuolated tobacco mesophyll protoplasts. Protein analysis by one‐ and two‐dimensional microgel‐electrophoresis

Hoffmann, E.; Hampp, Rüdiger

doi:10.1111/j.1399-3054.1994.tb03024.x

Cited by 13 publications

(3 citation statements)

References 44 publications

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“…For isoelectric focussing, chitinases were dissolved in O'Farrell buer (O'Farrell 1975) and samples of 3 ll were focussed on ampholyte gradients ranging from pH 3 to 10, as described by Homann and Hampp (1994). After electrophoresis the gels were ®xed in 10% (w/v) sulfosalicylic acid and stained with Coomassie Brilliant Blue according to Neuho et al (1985).…”

Section: Methodsmentioning

confidence: 99%

Cleavage of chitinous elicitors from the ectomycorrhizal fungus Hebeloma crustuliniforme by host chitinases prevents induction of K + and Cl − release, extracellular alkalinization and H 2 O 2 synthesis of Picea abies cells

Salzer

Hebe

Hager

1997

Planta

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Rapid reactions comprising eux of K + and Cl A , phosphorylation of a 63-kDa protein (pp63), extracellular alkalinization and synthesis of H 2 O 2 are equally induced in cells of Picea abies (L.) Karst. by chitotetraose, colloidal chitin and cell wall elicitors from the ectomycorrhizal fungus Hebeloma crustuliniforme (Bull. ex Fries.) QueÂ l. an ectomycorrhizal partner of spruce. Cleavage of fungal cell wall elicitors and of arti®cial chitin elicitors to monomeric and dimeric fragments by apoplasmic spruce chitinases (36-kDa class I chitinase, pI 8.0, and 28-kDa chitinase, pI 8.7; EC 3.2.1.14) equally prevented induction of these rapid reactions. Also, N-acetylglucosamine oligomers and elicitors from the fungal cell walls showed a similar dependence of their activity on the degree of polymerisation. From these results it is suggested that, during ectomycorrhiza formation, only some of the chitin-derived elicitors reach their receptors at the plant plasma membrane, initiating reactions of the hypersensitive response in the host cells. The remaining fungal elicitors will be degraded to varying extents by wall-localized chitinases of the host root, reducing the defence reactions of the plant and allowing symbiotic interactions of both organisms. Abbreviations: DP = degree of polymerisation; HR = hypersensitive response; MS = mineral solution; Pi = inorganic phosphate; pp63 = phosphorylated protein of MW 63 kDa Correspondence to: P. Salzer; Fax

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Section: Methodsmentioning

confidence: 99%

Cleavage of chitinous elicitors from the ectomycorrhizal fungus Hebeloma crustuliniforme by host chitinases prevents induction of K + and Cl − release, extracellular alkalinization and H 2 O 2 synthesis of Picea abies cells

Salzer

Hebe

Hager

1997

Planta

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“…Electrophoretic separation of proteins and immunological detection of PEPCase protein was achieved by using a 7.5% polyacrylamide gel in a microgel system (Poehling and Neuhoff 1980; M. Guttenberger 1989, PhD thesis, Univ. of Tübingen, Germany) followed by a semidry western blot procedure (Hoffmann and Hampp 1994). The blots were developed with a 1:2 000 diluted antiserum raised against Sorghum C 4 PEPCase (Vidal et al 1981).…”

Section: Methodsmentioning

confidence: 99%

Phosphoenolpyruvate carboxylase in mistletoe leaves: Regulation of gene expression, protein content, and covalent modification

Wanek

Nehls

et al. 2001

Physiologia Plantarum

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Seasonal changes in the activity of phosphoenolpyruvate carboxylase (PEPCase, EC 4.1.1.31), a key enzyme in the interaction of carbohydrate and nitrogen metabolism, were studied in leaves of the C3 semiparasitic mistletoe, Viscum album, growing on different host trees. Maximum extractable PEPCase activities were higher in leaves of mistletoes growing on Betula pendula and Alnus glutinosa hosts compared with those on the conifers, Abies alba and Larix decidua. Independent of host, maximum extractable PEPCase activities were high in spring and autumn while low in summer. Samples with higher PEPCase activities showed higher amounts of PEPCase protein and higher PEPCase mRNA levels. A curvilinear correlation between leaf total nitrogen content and the maximum extractable PEPCase activity as well as PEPCase mRNA level suggested that nitrogen might affect the activity of PEPCase of mistletoe by up-regulating gene expression. In addition to extractable activity, seasonal changes of the PEPCase activation state, the ratio of activities resulting from limited:non-limited assays, were found, which was correlated to the variation of malate content in leaves of mistletoe. ATP-dependent activation of PEPCase was characterized by an increase in I0.5(L-malate), indicating that PEPCase of leaves of mistletoes is probably regulated via phosphorylation.

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“…For all samples, a protein extract of 12 pg root dry weight (equivalent to 18.4 [lg mycorrhizal dr\' weight at 35% fungal biomass) was applied per lane. Electrophoresis and blotting conditions were as described by Hoffmann and Hampp (1994). Polydonal rabbit antiserum against sorghum leaf PEPC was provided by J. Vidal, P,aris.…”

Section: Immunoblottlug Of Pepcmentioning

confidence: 99%

Mycorrhiza formation on Norway spruce (Picea abies) roots affects the pathway of anaplerotic CO2 fixation

Wingler¹,

Wallenda²,

Hampp³

1996

Physiol Plant

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R. 1996. Myeorrhiza formation on Norway spruce (Picea abies) roots affects the pathway of anaplerotic CO, Activities of carboxylaEion enzymes were analyzed in the mycelium of the mycorrhizal fungus Amanita muscaria (L. ex Fr.) Hooker, in non-myeorrhizal short roots of Norway spruce (Picea abies IL.] Karsl.) and in mycorrhizas of these two partnens. While pyruvate carboxylase (PC, EC 6.4.1.1) and phosphoenolpyruvate carboxykinase activities (PEPCK, EC 4.1.1.49) could be detected in the mycelium of A. muscaria, phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31) was only active in root tissue. In .4. muscaria, PC activity was generally low (around 10 nmol mg" protein min"') but PEPCK activity was above 250 nmol mg"' protein min . Mycorrhizal development on short roots decreased PEPC activity by more than 15%, although dilution by the funeal biomass in mycorrhizas was only 35%. This reduction in activity was paralleled by a decreased content of PEPC protein. By means of micro-analytical methods it was shown that PEPC activity was lowest in the central zones of the mycon-Mzas, whereas PEPC activity was highest in the corresponding central sections in non-mycorrhizal shon roots. '^c'Oj labelling, on Ihe other hand, revealed that in vivo CO, fixation was higher in mycontiizas compared to non-mycorrhizal short roots. It is concluded that fungal cai'boxylases (probably PEPCK) are important for anaplerotic CO; fixation during nitrogen assimilation in mycorrhizas of Noi-way spmce.Physiol. Plam. %. 19%

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Studies on vacuole regeneration in evacuolated tobacco mesophyll protoplasts. Protein analysis by one‐ and two‐dimensional microgel‐electrophoresis

Cited by 13 publications

References 44 publications

Cleavage of chitinous elicitors from the ectomycorrhizal fungus Hebeloma crustuliniforme by host chitinases prevents induction of K + and Cl − release, extracellular alkalinization and H 2 O 2 synthesis of Picea abies cells

Cleavage of chitinous elicitors from the ectomycorrhizal fungus Hebeloma crustuliniforme by host chitinases prevents induction of K + and Cl − release, extracellular alkalinization and H 2 O 2 synthesis of Picea abies cells

Phosphoenolpyruvate carboxylase in mistletoe leaves: Regulation of gene expression, protein content, and covalent modification

Mycorrhiza formation on Norway spruce (Picea abies) roots affects the pathway of anaplerotic CO2 fixation

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