Elicitation of ß-l,3-Glucanase and Chitinase Activities in Cell Suspension Cultures of Ascochyta rabiei Resistant and Susceptible Cultivars of Chickpea ( Cicer avietinum)

1990 ◽  
Vol 45 (3-4) ◽  
pp. 233-239 ◽  
Author(s):  
Ralph Vogelsang ◽  
Wolfgang Barz
Keyword(s):  
Cell Culture ◽  
Cell Suspension ◽  
Fungal Pathogen ◽  
Defense Mechanisms ◽  
Culture Medium ◽  
Cell Suspension Cultures ◽  
Chitinase Activity ◽  
Suspension Cultures ◽  
Ascochyta Rabiei ◽  
Incompatible Interaction

Abstract β-1,3-Glucanase and chitinase activities have been analyzed in cell suspension cultures of an A scochyta rabiei resistant ILC 3279 and a susceptible ILC 1929 cultivar of chickpea (Cicer arietinum) following treatment with an elicitor derived from this fungal pathogen. A facile method to determine both hydrolase activities in the cell culture medium was established. Significantly higher constitutive and elicitor-induced levels of both hydrolase activities were found in the medium of the ILC 3279 cell culture. The release of these enzyme activities is not due to cell lysis, but rather the consequence of a secretion mechanism. The cells of the resistant line contained a 5 times higher level of chitinase activity in comparison to the ILC 1929 cell culture, whereas the latter cells possessed a 3 times higher β -1,3-glucanase activity. The results are interpreted that accumulation of extracellular hydrolase activities may play an important role among the various plant defense mechanisms previously determined for the incompatible interaction between the resistant cultivar and its fungal pathogen.

Elicitation of Pterocarpan Phytoalexins in Cell Suspension Cultures of Different Chickpea (Cicer arietinum L.) Cultivars by an Elicitor from the Fungus Ascochyta rabiéi

1988 ◽  
Vol 43 (7-8) ◽  
pp. 529-535 ◽  
Author(s):  
Helmut Keßmann ◽  
Susanne Daniel ◽  
Wolfgang Barz
Keyword(s):  
Cell Culture ◽  
Cell Suspension ◽  
Cicer Arietinum ◽  
Time Course ◽  
Cell Suspension Cultures ◽  
Suspension Cultures ◽  
Ascochyta Rabiei ◽  
Cicer Arietinum L ◽  
Elicitor Treatment ◽  
Induced Changes

Cell suspension cultures of two chickpea (Cicer arietinum L.) cultivars, resistant and susceptible towards the chickpea pathogen Ascochyta rabiei, were compared with regard to elicitor-induced changes in phytoalexin and isoflavone accumulation. The elicitor was isolated from fermentergrown mycelium of A. rabiei and it mainly consisted of glucose, mannose and N-acetylgalactosamin. Time course and dose response studies on elicitor action demonstrated that the cell culture of the resistant cultivar ILC 3279 accumulated large amounts of the pterocarpan phytoalexins medicarpin and maackiain within 8 h. The cell culture of the susceptible cultivar ILC 1929 accumulated only small amounts of the phytoalexins some 12 h after elicitor treatment. Growth of the cell cultures and the accumulation of isoflavones and isoflavone conjugates were not altered by elicitor treatment except for a higher accumulation of formononetin 7-O-glucoside-6″-O-malonate in cell culture ILC 3279 subsequent to the maximum of phytoalexin accumulation.

A Comparison of Various Growth Parameters of Cell Suspension Cultures to Determine Phytotoxicity of Xenobiotics

Weed Science ◽  
1984 ◽  
Vol 32 (2) ◽  
pp. 235-242 ◽  
Author(s):  
David G. Davis ◽  
Rosa L. Stolzenberg ◽  
Joan A. Dusky
Keyword(s):  
Ethylene Oxide ◽  
Cell Suspension ◽  
Culture Medium ◽  
Growth Parameters ◽  
Cell Suspension Cultures ◽  
Dry Weight ◽  
Suspension Cultures ◽  
Weight Changes ◽  
Advantages And Disadvantages ◽  
Cell Volumes

An assessment was made of various parameters to measure growth of soybean [Glycine max (L.) Merr. ‘Wilkin’] and einkorn (Triticum monococcum L.) cell suspension cultures to establish convenient methods of screening the effects of chemicals. Methods assessed were settled cell volumes, packed cell volumes, absorbance at 525 nm of sonicated aliquots, dry weights (of aliquots or entire flask contents), and electrical conductivity and pH of the culture medium. Settled cell volumes, conductivity, and dry-weight changes were the most useful of the methods tested for determining the phytotoxicity of a nonionic linear alcohol ethylene oxide detergent (an adduct of 1-dodecanol containing eight ethylene oxide units) and the methyl ester of diclofop {2-[4-(2,4-dichlorophenoxy)phenoxy] propanoic acid}. Because 3 to 4 weeks were required to assess whether the cultures could grow out of the initial inhibition by the detergent or herbicide, none of the methods was rapid. Advantages and disadvantages of the various methods and their relative values for screening compounds are described.

Indole Alkaloids from Cell Suspension Cultures of Stemmadenia tomentosa and Voacanga africana

1982 ◽  
Vol 37 (10) ◽  
pp. 857-860 ◽  
Author(s):  
Joachim Stöckigt ◽  
Karl-Heinz Pawelka ◽  
Ana Rother ◽  
Brigitte Deus
Keyword(s):  
Cell Culture ◽  
Cell Suspension ◽  
Growth Condition ◽  
Indole Alkaloids ◽  
Normal Growth ◽  
Cell Suspension Cultures ◽  
Suspension Cultures ◽  
Normal Growth Condition ◽  
Different Types

Abstract Cell suspension cultures of Stemmadenia tomentosa synthesized under normal growth condition the eight major indole alkaloids: (-)-tabersonine, (-)-minovincinine, (+)-conoflorine (voaphyl-line), condylocarpine, (+)-tubotaiwine (dihydrocondylocarpine), (-)-norfluorocurarine (vin-canine), (-)-vinervine, and (-)-coronaridine. These alkaloids consist of the three different types, Aspidosperma, Strychnos and Iboga. In contrast, cultures o f Voacanga africana produced mainly one alkaloid group (Aspidosperma-type) represented by (-)-tabersonine, lochnericine and (-)-minovincinine. Therefore this cell culture seems to be qualified for investigation concerning the biosynthesis of Aspidosperma alkaloids.

Elicitor-induced Changes of Enzyme Activities Related to Isoflavone and Pterocarpan Accumulation in Chickpea (Cicer arietinum L.) Cell Suspension Cultures

1988 ◽  
Vol 43 (7-8) ◽  
pp. 536-544 ◽  
Author(s):  
Susanne Daniel ◽  
Walter Hinderer ◽  
Wolfgang Barz
Keyword(s):  
Cell Culture ◽  
Cell Suspension ◽  
Cicer Arietinum ◽  
Enzyme Activities ◽  
Cell Suspension Cultures ◽  
Suspension Cultures ◽  
Biochanin A ◽  
Primary Metabolism ◽  
Cicer Arietinum L ◽  
L Cell

The extractable activities of thirteen enzymes of primary and secondary metabolism have been measured in chickpea (Cicer arietinum L.) cell suspension cultures after treatment with an elicitor from the fungus Ascochyta rabiei (Pass.) Lab. The cell culture, derived from the A. rabiei resistant cultivar ILC 3279, constitutively accumulated the isoflavones biochanin A and formononetin together with their 7-O-glucosides and the 7-O-glucoside-6″-malonates. After elicitor application the cells rapidly form the pterocarpan phytoalexins medicarpin and maackiain. Among the enzymes of primary metabolism only the glucose 6-phosphate dehydrogenase exhibited a significant increase in activity with a maximum four hours after application of the elicitor. In phenylpropane metabolism the activities of phenylalanine ammonia lyase and chalcone synthase were enhanced by the elicitor and exhibited highest levels after four hours. In contrast the chalcone isomerase activity was not influenced by the elicitor. A substantial enhancement occurred with the isoflavone 7-O-glucosyltransferase activity eight hours after elicitor application. The results suggest that in this cell culture the elicitor-induced biosynthesis of pterocarpan phytoalexins was accompanied with a rapid and transient increase of those enzyme activities which are located at branching points of related pathways, i.e. pentose phosphate cycle, general phenylpropane metabolism, flavonoid formation and isoflavone conjugation.

scholarly journals Demonstration of lactase activity in culture medium of melon cells

2011 ◽  
Vol 31 (No. 4) ◽  
pp. 132-135
Author(s):  
J. Stano ◽  
K. Mičieta ◽  
E. Tokhtaeva ◽  
M. Valšíková ◽  
M. Koreňová ◽  
...  
Keyword(s):  
Cell Suspension ◽  
Culture Medium ◽  
Agar Plate ◽  
Cell Suspension Cultures ◽  
Callus Cultures ◽  
Suspension Cultures ◽  
Lactase Activity ◽  
Seedling Roots ◽  
Intracellular Activities

Lactase activity was detected in a culture medium of the cell suspension culture of watermelon (Citrullus vulgaris L.). A simple, rapid and reproducible procedure for identification of extracellular lactase is described using callus cultures of seedlings from the tested plant, hairy roots of 2.5 days old seedlings of watermelon germinating on agar plates as well as cell suspension cultures derived from callus cultures. For the determination of intracellular activities of lactase, 6-bromo-2-naphthyl-β-D-galactopyranoside and p-nitrophenyl-β-D-galactopyranoside were used as synthetic substrates. The extracellular lactase activity was determined by evaluating the day-zone in agar medium. The enzyme from watermelon callus cultures and seedling roots, cultivated on agar plates supplemented with 6-bromo-2-naphthyl-2-bromo-β-D-galactopyranoside, hydrolyzed this substrate releasing 6-bromo-naphthyl. By simultaneous coupling with hexazonium p-rosaniline or Fast Blue BB the corresponding azo dye was formed. The parallel extracellular and intracellular activities were determined in cell suspension cultures derived from callus cultures. The results show a 43.8% intracellular and 54.2% extracellular distribution of lactase activity. The described agar plate method enables a rapid, simple and specific detection of plant processes of extracellular lactase.  

Changes in culture medium pH by cell suspension cultures of Dioscorea deltoidea

1984 ◽  
Vol 35 (3) ◽  
pp. 207-212 ◽  
Author(s):  
R.G. Butenko ◽  
A.Kh. Lipsky ◽  
N.D. Chernyak ◽  
H.C. Arya
Keyword(s):  
Cell Suspension ◽  
Culture Medium ◽  
Cell Suspension Cultures ◽  
Suspension Cultures ◽  
Medium Ph ◽  
Dioscorea Deltoidea

Elicitor Induction of Cytochrome P-450 Monooxygenases in Cell Suspension Cultures of Chickpea (Cicer arietinum L.) and Their Involvement in Pterocarpan Phytoalexin Biosynthesis

1991 ◽  
Vol 46 (1-2) ◽  
pp. 58-66 ◽  
Author(s):  
Werner Gunia ◽  
Walter Hinderer ◽  
Uta Wittkampf ◽  
Wolfgang Barz
Keyword(s):  
Cell Culture ◽  
Cell Suspension ◽  
Cicer Arietinum ◽  
Cell Suspension Cultures ◽  
Suspension Cultures ◽  
Resistant Cultivar ◽  
Susceptible Cultivar ◽  
A Cell ◽  
Cell Culture Line ◽  
Cytochrome P 450

Abstract A yeast glucan elicitor causes the accumulation of the pterocarpan phytoalexins medicarpin and maackiain in chickpea (Cicer arietinum) cell suspension cultures established from seeds. A cell culture line from a chickpea cultivar resistant against its main fungal pathogen Ascochyta rabiei accumulates large amounts (944 nm ol/g fr. wt.) whereas a cell culture line from a susceptible cultivar accumulates only low amounts (38 nm ol/g fr. wt.) of the phytoalexins. This is consistent with differential accumulation of pterocarpan phytoalexins in intact plants [1], The first reactions in the pterocarpan-specific branch of biosynthesis are hydroxylation of the isoflavone intermediate form ononetin in position 2′ or 3′, catalyzed by microsomal cytochrome P-450 monooxygenases. Upon elicitation form ononetin 2′-hydroxylase undergoes a strong transient induction in the cell suspension culture of the resistant cultivar, whereas in the cell culture from the susceptible cultivar it is only slightly induced. In both cell suspension cul­tures the induction of cinnamic acid 4-hydroxylase and of form ononetin 3′-hydroxylase does not show a clear correlation with phytoalexin accumulation. Experiments with different elici­tor concentrations confirm that formononetin 2′-hydroxylase is much more induced in cell cul­tures from the resistant cultivar than from the susceptible one. It is concluded that the massive difference in phytoalexin accumulation between cell suspension cultures from the resistant and susceptible cultivar is determined mainly by the differential induction of form ononetin 2′-hydroxylase activity.

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