Quantitation of gibberellins and indoleacetic acid in Begonia leaves: Relationship with environment, regeneration and flowering

Indole-3-acetic acid (IAA) was purified by high performance liquid chromatography (HPLC) and identified by gas chromatography - mass spectrometry (GC-MS) in leaf extracts of Begonia × cheimantha Everett cv. Nova. The content of IAA and of gibberellins A4, A9, A19 and A20 (GAs) previously identified in this material, were quantified by GC-MS in leaves of Begonia plants grown under different temperature and daylength conditions, using deuterated compounds as internal standards. GA1, which was also identified, was present in too low quantities for reliable quantitation. Rapid and significant decreases (within 2–4 days) occurred in the content of both IAA and GAs when the plants were transferred from conditions which are non-inductive for adventitious bud formation and flowering (24°C/long day) to inductive conditions (24°C/short day, 15°C/long day or 15°C/short day). GA4 and GA9 were affected by photoperiod only, whereas IAA, GA19 and GA20 were affected by both photoperiod and temperature. The data suggest that biosynthesis of GA9 and GA4 are blocked in short days at a step located before GA9. Conversion of GA19 to GA20 seemed to be blocked by both short days and low temperature, while an additional block located before GA19 seemed to be imposed in 15°C/short day. The results confirm earlier results and support the hypothesis that photoperiod and temperature effects in Begonia are mediated by endogenous hormones.     

Cytokinin Activity in Begonia and Bryophyllum

Cytokinin activity has been obtained in ethanol extracts of Begonia and Bryophyllum plants. Begonia x cheimantha Everett yielded 30 to 300 μg kinetin equivalents per kg of fresh leaves in the tobacco callus bioassay. Short day conditions appeared to increase the extractable cytokinin content in the tissue. Purification by fractionation on Dowex 50 H⁺ columns followed by organic solvent extraction, silver precipitation, and repeated paper chromatography yielded an apparently homogeneous product which accounted for most of the activity in the Begonia extracts. It was indistinguishable from zeatin in chromatograms developed with six solvent systems. Other cytokinin active fractions were also obtained from both Begonia and Bryophyllum. Crystalline picrate preparations of active products were insufficient for identification by mass spectrometry.     

Stratification in the filling phase of the man-made lake Brokopondo in Surinam (S. America)

Summary Temperature and dissolved oxygen content measured weekly at various depths during 3 1/2 years in the filling phase of a tropical man-made lake, show that at the dam-site fluctuations of epilimnion values occurred, whereas the hypolimnion remained almost constant in temperature and permanently devoid of oxygen. This confirms previous reports concerning another mid-lake station. Apart from rather steep superficial temperature gradients, classical thermoclines did not occur. It is proposed that the lake could not become fully mixed by wind action because of the great vertical density differences at the prevailing temperatures, and because of the drowning forest still giving shelter by the extending crowns. On the other hand, the stratification was not static. Fluctuations between high and low epilimnion values for temperature, oxygen content and transparency oocurred yearly, but could not be related satisfactorily to the prevailing seasons. The depth of the epilimnion increased each year. It is suggested that the observed kind of stratification was effective though not absolute. Vertical exchange may have been reduced by increasing density differences, but it was not fully eliminated. This explains the varying degree of incorporation of upper hypolimnetic layers in the epilimnion in relation with epilimnetic temperature. Comparison with large African reservoirs accentuates the exceptional limnological development of Lake Brokopondo during ifs filling phase.     

Synthesis of guanosine 5'-diphosphate, 3'-diphosphate in spot mutants of Escherichia coli.

The synthesis of ppGpp in spoT- mutants of Escherichia coli has been invesitgated. In these mutants the first-order rate constant for ppGpp breakdown is low, and pppGpp is barely detectable. It is shown that the rate of pppGpp, and hence ppGpp, synthesis is strongly reduced compared with that observed in spot+ strains. The low rate of magic spot synthesis satisfactorily explains the low levels of pppGpp in spoT- mutants. The pentaphosphate very probably is the precursor of ppGpp as it is in wild-type, i.e. spoT+, strains.     

A Sensitive and Reliable Assay for Dopamine (β-Hydroxylase in Tissue

A new assay procedure for dopamine β-hydroxylase (DBH) in tissue extracts is described. Solubilized DBH was adsorbed from crude extracts on Concanavalin A-Sepharose (Con A-Sepharose), resulting in enrichment of the enzyme as well as removal of endogenous catecholamines and inhibitory substances. The enzymatic assay was carried out with DBH still adsorbed to Con A-Sepharose. The adsorption of the DBH to Con A-Sepharose offers three advantages over previous assay procedures. (1) Because of removal of the endogenous inhibitory substances, a single Cu²⁺ concentration can be used for the determination of DBH activity, regardless of the tissue dilution or inhibitor content of the analysed sample. Using this procedure, the optimal Cu²⁺ concentration for DBH of bovine adrenal gland extracts was 3 μM and for rat brain 10 μM. (2) Because of removal of endogenous catecholamines, dopamine, the main physiological substrate of DBH in noradrenergic neurons, can be used for the assay. The enzymatic reaction product, noradrenaline, was determined by high performance liquid chromatography and electrochemical detection (hplc-ec). This procedure resulted in an approx. 10-fold increase in sensitivity of the assay compared with other procedures, e.g., the radioenzymatic assay. (3) Direct determination of the immediate product of the enzymatic reaction (noradrenaline) permits kinetic analysis. It was found that the Michaelis constants for the substrate (dopamine) and co-factor (ascorbic acid) (2 mM and 0.65 mM, respectively) determined in bovine adrenal tissue extracts by the described procedure were identical with the values for the purified DBH preparation.