Methods for the Selection of Low-Temperature Tolerant Mutants of Chrysanthemum morifolium Ramat. by Using Irradiated Cell Suspension Cultures. I. Selection of Regenerants in vivo under Suboptimal Temperature Conditions

Changes in extracellular, cellular, and subcellular proteins during abscisic acid and low temperature induced cold hardening of alfalfa (Medicago sativa L. cv. Wisconsin 22C) cell suspension cultures were investigated by sodium dodecyl sulphate polyacrylamide gel electrophoresis. Extracellular proteins from 4- to 6-day-old ABA and low temperature grown alfalfa cells showed decreased electrophoretic mobilities, lacked a 190-kDa glycoprotein, and had reduced amounts of four other polypeptides. In total cell protein analyses, a 42-kDa protein was enriched in both ABA and low temperature treated alfalfa cells. Several proteins increased or induced by exogenous ABA treatment were identified in the extracellular (12.5 and 13 to 15 kDa), total cell and cell wall (24 kDa), and soluble (20, 37, and 41 kDa) fractions. However, no major protein changes were resolved by one-dimensional electrophoretic analyses of crude membrane proteins.

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Über den Abbau von Flavonolen in pflanzlichen Zellsuspensionskulturen / Degradation of Flavonols in Plant Suspension Cultures

Zeitschrift für Naturforschung B ◽

10.1515/znb-1972-0818 ◽

1972 ◽

Vol 27 (8) ◽

pp. 946-954 ◽

Cited By ~ 30

Author(s):

Wolfgang Hösel ◽

Paul D. Shaw ◽

Wolfgang Barz

Keyword(s):

Salicylic Acid ◽

Glycine Max ◽

Cell Suspension ◽

Cicer Arietinum ◽

Cell Suspension Cultures ◽

Suspension Cultures ◽

Enzyme Preparations ◽

Cicer Arietinum L

The flavonols kaempferol, quercetin and isorhamnetin were labelled with 14C by keeping seven day old Cicer arietinum L. plants in an atmosphere of 14CO2 for five days. The purified (U-14C) flavonols were applied to cell suspension cultures of Cicer arietinum L., Phaseolus aureus Roxb., Glycine max and Petroselinum hortense. Based on the rates of 14CO2 formation and distribution of radioactivity after fractionation of the cells, the flavonols were shown to be catabolized to a very high extent.All four cell suspension cultures possess the enzymatic activity transforming flavonols to the recently discovered 2,3-dihydroxyflavanones. Upon incubation of the flavonols datiscetin and kaempferol with enzyme preparations from Cicer arietinum L. cell suspension cultures, it was demonstrated that the enzymatically formed 2,3-dihydroxyflavanones are further transformed in an enzyme catalyzed reaction. Salicylic acid was found as a degradation fragment of ring B of the 2,3,5,7,2′-pentahydroxyflavanone derived from datiscetin. Neither phloroglucinol nor phloroglucinol carboxylic acid were observed as metabolites of ring A. These in vitro findings were further substantiated by in vivo data because the flavonols kaempferol, quercetin and datiscetin when applied to cell suspension cultures of Cicer arietinum L. and Glycine max gave rise to para-hydroxybenzoic acid, protocatechuic acid and salicylic acid, respectively. It was thus concluded that flavonols are catabolized via 2,3-dihydroxyflavanones with the B-ring liberated as the respective benzoic acid. The data are discussed in connection with earlier findings on the catabolism of chalcones, cinnamic and benzoic acids.

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