Free proline accumulation in drought-stressed plants

Summary Free-proline accumulation was measured in leaves of intact wheat (Triticum vulgare L. cv. Kalyan Sona), plantago (Plantago ovata Forsk-Isabgool), papavar (Papaver somnifera L. Opium poppy) and mustard (Brassica juncea L. var. Varuna) grown in the field with low to high field water content and thus they were subjected to water stress. Leaf water deficit in percentage was used to determine the degree of stress at the time of proline anlysis.Free proline content was higher in mustard leaves as compared to wheat, plantago and papavar leaves. Water stress enhances the proline content but at same water deficit level the content differ in the leaves of the plants studied.     

Isolation and cryopreservation of O-methylthreonine-resistant Rosa cell lines altered in the feedback sensitivity of l-threonine deaminase

O-Methylthreonine (OMT) inhibits the growth of plated Rosa cells (ID₅₀6·10^-6M). Isoleucine is able to reverse efficiently and specifically this OMT toxicity. From OMT-resistant colonies occurring at a frequency of 1.58·10^-7 variants per cell plated at 10^-4M OMT, the variant strains OMT^R-1 and OMT^R-2 were isolated, cloned via protoplasts and characterized. Both variants were ten times more resistant to OMT than the wildtype and were cross-resistant to another isoleucine analog, dl-4-thiaisoleucine. The resistant variants retained their resistance after storage for three years in liquid nitrogen. Both resistant strains were stable for several months when subcultured in the absence of OMT although it was shown in a reconstitution experiment that wildtype cells overgrow OMT^R-2 variant cells if co-cultivated for many passages in drug-free medium. One case of instability was observed upon long-term subculturing in drug-free medium: the strain OMT^R-1D^* partially lost phenotypic properties. Resistance to OMT was followed qualitatively by a new method based on inhibition-zone formation in cell suspensions plated in agar medium. The OMT-resistant variants showed a reduction in sensitivity of the enzyme l-threonine deaminase to feedback inhibition by isoleucine, a decreased stability of l-threonine deaminase when stored at-18°C or incubated at +55°C and a two- to threefold increase of the free isoleucine pool within the cells. The genetical events and the biochemical mechanisms which might lead to the observed stable and biochemically defined character are discussed with particular reference to the high ploidy level of the Rosa cell line.Abbreviations OMT l-O-methylthreonine - TD l-threonine deaminase     

Availability of water controls Crassulacean acid metabolism in succulents of the Richtersveld (Namib desert,South Africa)

Features of Crassulacean acid metabolism (CAM) were studied in a variety of different succulents in response to climatic conditions between March 1977 and October 1983 in the southern Namib desert (Richtersveld). A screening in 1977 and 1978 revealed that nearly all investigated succulents performed a CAM, but overnight accumulation of malate declined gradually with decreasing soil water potential, tissue osmotic potential, and leaf water content. This was further substantiated by an extended period of insufficient rainfall in 1979 and 1980 which damaged the evergreen CAM succulents between 80 and 100%. In most of the species still living, neither CO₂-gas exchange nor diurnal acid fluctuation, indicative of CAM, could be detected unless an abundant rainfall restored both CAM features. Plants persisted in a stage of latent life.Water supply is one necessary prerequisite for CAM in the Richtersveld. But even well-watered plants with CAM were sensitive to short-term water stress caused by high water-vapour partialpressure deficit (VPD) in the night, which reduced or prevented CO₂ uptake and resulted in a linear relation between overnight accumulated malate and VPD. The results do not support the opinion that, for the Namib succulents, CAM is an adaptive mechanism to water stress since long-term and short-term water stress stopped nocturnal malate synthesis, but instead lead to the conclusion that nocuturnal CO₂ fixation is only performed when the water status of the plant can be improved simultaneously.Abbreviations CAM Crassulacean acid metabolism - VPD water vapour pressure deficit Dedicated to Professor H. Ziegler on the occasion of his 60th birthday     

Effect of host plant nitrogen fertilization on the biology of the two-spotted spider mite, Tetranychus urticae

Females of Tetranychus urticae Koch were reared on leaf discs of apple trees and bush beans grown at different N concentrations (0.6–75 mM NO _inf3 ^sup- ). N-deficiency increased pre-imaginal development time, preoviposition period, and decreased female weight, fecundity and oviposition rate of the mites. N, water, amino acid, and sugar content of the apple leaves were positively correlated with weight and egg production and negatively correlated with development time and pre-oviposition period. The reverse correlations were found with total phenol content of the leaves and above mite parameters. A reduction of leaf N by 50% was related with a tenfold decline in fecundity on apple leaves. The stress mainly affected the oviposition rate and to a lesser extent the oviposition period. On apple leaves the net reproductive rate (av. no. offspring per , R_o), mean length of a generation (T), and innate capacity for increase (r_m) were R_o=40.3, T=17.1, and r_m=0.22 for the standard N concentration, and 4.7, 25.0, and 0.06 for strong N-deficiency, respectively.

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Zusammenfassung Die polyphage Gemeine Spinnmilbe ist ein wirtschaftlich bedeutender Schädling. Die Rolle einzelner Nährstoffe bei der Milbenvermehrung wurde schon von einigen Autoren mit unterschiedlichen Ergebnissen untersucht. Der Stickstoff scheint aber einer der Hauptfaktoren zu sein.Ueber Gewebekulturen klonierte Apfelbäume sowie aus Samen gezogene Buschbohnen wurden in Nährlösungen mit verschiedenen N-Angeboten kultiviert. Aus solchen Pflanzen wurden Blattscheiben ausgestanzt und mit Weibchen von T. urticae besetzt. Gemessen wurden die tägliche Eiablage bis zum natürlichen Tod, die Entwicklungsdauer und das Weibchengewicht. Mit dem bei N-Mangel abnehmenden N, Aminosäuren- und Zuckergehalt (v.a. Sorbit) im Blatt korrelierend nahmen auch Gewicht, Ablagerate und Fekundität ab, bzw. die Entwicklungsdauer und Praeovipositionsperiode zu. Die umgekehrte Wirkung auf diese Milbenparameter hatte der Gesamtphenolgehalt. Eine Reduktion des N-Gehalts der Blätter auf die Hälfte (1.5% N) bewirkte auf Apfelblättern eine Abnahme von Fekundität und Ovipositionsrate um das zehnfache, bzw. eine Zunahme der Präovipositionsperiode um mehr als das dreifache. Der Stress beeinflusste v.a. die Maxima der Ablageverläufe und weniger die Lebensdauer. Der Wassergehalt war ebenfalls mit der Fekundität positiv korreliert.Auf Bohnenblättern legten die Tiere mehr Eier und erreichten ein höheres Gewicht als auf Apfel, dies bei gleichem N-Gehalt beider Pflanzen. Es wirken somit noch andere Faktoren auf die Reproduktion der Spinnmilben.Die Populationsparameter wurden ebenfalls sehr stark beeinflusst. Auf Apfelblättern ergab sich bei der Kontrolle eine Nettoreproduktionsrate R_o von 40.3 und bei starker N-Defizienz 4.7. Die mittlere Generationsdauer T sowie die spezifische natürliche Wachstumsrate r_m betrugen für die Kontrolle 17.1 Tage, bzw. 0.22 und für starken N-Mangel 25.0 Tage, bzw. 0.06.Die Ergebnisse zeigen, dass sich bereits kleinere Unterschiede im Stickstoffgehalt und damit zusammenhängend im Zucker- und Phenolgehalt stark auf die Populationsdynamik der Gemeinen Spinnmilbe auswirken können.

     

Control of l-amino acid oxidase in Neurospora crassa by different regulatory circuits

l-Amino acid oxidase is synthesized in Neurospora crassa in response to three different physiological stimuli: (i) starvation in phosphate buffer, (ii) mating, and (iii) nitrogen derepression in the presence of amino acids. During starvation in phosphate buffer, or after mating, l-amino acid oxidase synthesis occurred in parallel with that of tyrosinase. Exogenous sulfate repressed the formation of the two enzymes in starved cultures, but not in mated cultures. Sulfate repression was relieved by protein synthesis inhibitors, suggesting that the effect of sulfate required the synthesis of a metabolically unstable protein repressor. With amino acids as the sole nitrogen source only l-amino acid oxidase was produced. Under these conditions enzyme synthesis was repressed by ammonium and was insensitive to sulfate. Biochemical evidence suggested that the l-amino acid oxidase formed under the three different conditions was the same protein. Therefore, the expression of l-amino acid oxidase appeared to be under the control of least two regulatory circuits. One, also controlling tyrosinase, seems to respond to developmental signals related to sexual morphogenesis. The other, controlling other enzymes of the nitrogen catabolic system, is used by the organism to obtain nitrogen from alternative sources such as proteins and amino acids.     

Effects of fructose on human fibroblast metabolism: the application of DNA measurements as a basis for interpretation

Summary A fluorometric procedure for measuring DNA was used to study growth and metabolic responses of eight cell strains of human foreskin fibroblasts. In preliminary studies this procedure gave more precise specific activity changes inN-acetyl-β-d-glucosaminidase (NAG) than did a protein activity basis, when changes in this enzyme's specific activity were investigated as a function of experimental cell manipulation. When fibroblast growth in eight cell strains was compared in 134 mM d-fructose vs. 13.4 mM glucose-supplemented minimum essential media, a significant increase in cellular DNA (50%) and protein (45%) occurred over an 11-d period. No significant differences in media pH change, lactate production, or carbohydrate uptake occurred on a DNA basis when cell metabolism was compared over the last 24 h of culture in the two media. Cells grown in fructose-containing media tended to show a reduction in NAG specific activity when compared with those grown in glucose-containing media.     

Patterns of protein synthesis during the germination of pea axes,and the effects of an interrupting desiccation period

As germination of axes of Pisum sativum L. seeds progressed, profound quantitative and qualitative changes occurred in the patterns of protein synthesis. This was shown by fluorography of gels following two-dimensional polyacrylamide gel electrophoresis separation of [³⁵S]methioninelabelled proteins. The effects of desiccation during germination on these in-vivo protein-synthesis patterns were followed. Desiccation differentially affected the synthesis of proteins. Usually, however, upon rehydration following desiccation the types of proteins being synthesized were recognizable as those synthesized earlier during imbibition of control, once-imbibed axes: seeds imbibed for 8 h, and then dried, did not recommence synthesis of proteins typical of 8-h-imbibed control seeds, but rather of 4-h-imbibed control seeds. Seeds imbibed for 12 h, and then dried and rehydrated, synthesized proteins typical of 4-h-and 8-h-control seeds. Thus drying of germinating pea axes caused the proteinsynthesizing mechanism to revert to producing proteins typical of earlier stages of imbibition. Drying during germination never caused the seed to revert to the metabolic status of the initial mature dry state, however.Abbreviation DR dried and rehydrated     

The effect of drought on levels of abscisic acid,cytokinins, gibberellins and ethylene in aeroponically-grown sunflower plants

Abscisic acid (ABA), cytokinins and gibberellin-like substances (GAs) were extracted from the roots and shoots of 17-day-old sunflower seedlings which had been droughted or were unstressed. Plants were grown in an aeroponic chamber which allowed for good control over degree of water stress and easy access to roots. Following methanolic extraction of lyophilized material, cytokinins were separated from the acidic growth-regulators on a cellulose PO₄ cationic exchange column. The cytokinins were analysed by paper chromatography and HPLC and the soybean hypocotyl section assay. Semipurified acidic regulators were chromatographed on SiO₂ columns and HPLC and aliquots assayed with the dwarf rice cv. Tan-ginbozu bioassay for GAs. Fractions known to contain ABA were purified by sequential reverse-phase HPLC of the acid and then of the methyl ester forms followed by quantitation as Me-ABA on GLC-EC. ABA losses were measured by using an internal standard [³H]-ABA). Ethylene production was also monitored in stressed and unstressed seedlings.The effect of drought on GAs and ethylene was minimal. The ABA levels were markedly higher in droughted plants. Stressed roots had 32 times more ABA than controls. The levels of cytokinins in the shoots of droughted plants were about half those in unstressed shoots, and qualitative differences occurred in the roots. Under stress a large peak of activity was present similar to zeatin glucoside which was not present in the unstressed condition. The results are discussed in relation to drought-effects on metabolism.     

The possible involvement of gibberellins and calcium in tipburn of Chinese cabbage: Study of intact plants and detached leaves

Spraying Chinese cabbage seedlings [Brassica pekinensis (Lour.) Rupr.] with the growth retardant daminozide (succinic acid-2,2-dimethylhydrazide) reduced tipburn of the mature plants. As the concentration of daminozide increased, the reduction in tipburn damage was correlated with increased calcium content in young susceptible leaves. This effect was much more pronounced in plants that were misted once a day during the head formation period.Incubation of detached Chinese cabbage leaves for 48 h (in the dark) in solutions which contained either EDTA or EGTA caused characteristic lesions at the leaf tips. The extent of the damage was reduced by including CaCl₂ in the solutions. Leaves which were incubated in a solution of EDTA+GA₃ or EGTA+GA₃ were severely affected, with the latter solution being the more harmful. GA₃ alone did not enhance tipburn. CaCl₂ greatly reduced the effect of a complex of chelating agents and GA₃. Leaves derived from daminozide-treated plants which were incubated in EDTA+GA₃ were less affected with tipburn lesions than leaves of control plants treated with the same solutions. When detached leaves were water-stressed for 24 h prior to incubation in these solutions, the severity of tipburn symptoms increased. The possible interactions between GA, calcium chelation and tipburn development are discussed.Contribution no. 1171-E, 1984 series, from the ARO, The Volcani Center Bet Dagan, Israel.