Contribution to the study on the reparative effect of exogenous DNA in the irradiated meristem ofVicia faba

The present report is focused on the study of participation of exogenous DNA in the process of postirradiation reparation of meristematic cells ofVicia faba primary roots. It is aimed at comparison of the positive reparative effect of isologous DNA with postirradiation action of heterologous DNA in its native, thermally denatured and DNAase-degraded forms, or DNA degraded by ultrasound, and with the effect of other biologically important macromolecules (RNA, histone, heparin, and dextran sulphate). For this purpose, the roots ofVicia faba irradiated by 150 r exposure were cultivated in solutions containing the above substances for an appropriate time interval. In squash slides both mitotic activity of the investigated cell population and frequency of postmetaphase chromosomal aberrations induced by radiation were evaluated. It was shown that a stimulation of cell division and reparation of chromosome damages were supported exclusively by isologous DNA. On the contrary, exogenously applied heterologous DNA increased postirradiation frequency of aberrations; maintenance of native structure of applied DNA was an essential condition for the above effect. Other macromolecules investigated on the course of postirradiation reparation ofVicia faba meristematic cells were without effect.     

Localization of L-asparaginase inEscherichia coli

The localization ofl-asparaginase (l-asparagine amidohydrolase, EC 3.5.1.1) EC-2 isoenzyme was studied inEscherichia coli ATCC 9637 grown under conditions of moderate aeration. The enzyme was determined in cell fractions obtained by fraction centrifugation of lysed spheroplasts. When the synthesis of the enzyme was induced byl-asparagine, its amount in the cytoplasmic fraction at the beginning of the induction exceeded as much as five times that in uninduced cells, attaining up to 20% of the total activity. In the course of growth of the culture this activity decreased gradually to zero. The membrane fraction of induced cells contained considerable amount of EC-2l-asparaginase which, at the beginning of the induction, reached up to 6% ot the total enzymic activity; in membrane fraction of control cells the activity was close to zero. The results indicate a relationship of cell structures to thel-asparagine-induced synthesis of the enzyme.     

Oxidative phosphorylation in a yeast mutant lacking α-ketoglutarate dehydrogenase activity

The effect of α-ketoglutarate deficiency on the oxidative phosphorylation in yeast mitochondria was studied. By determining the properties of electron transport and energy transduction systems of mutant mitochondria it was found that the lack of α-ketoglutarate dehydrogenase activity in mitochondria does not result in any functional defect in the oxidative phosphorylation system.     

Effects of the new antifungal antibiotic mucidin

The antibiotic activity of the antifungal substance mucidin was compared with the activity of nystatin and pimaricin. The antibiotics were tested by the plate method using 19 fungal species, mainly phytopathogenic ones. Toward 14 species, mucidin was ten times more active than nystatin and pimaricin, toward 5 species the activities were roughly the same. The antibiotics differed also in the sharpness of the inhibition zone boundaries.     

On the media improving freeze-sectioning of plant material

Using a semiconductor freezing microtome, there is no difficulty in getting an ample supply of quite suitable sections of Ca formol fixed root tips ofVicia faba, applying Holt’s sirup or plain 0.88 M sucrose as media for keeping the material, getting the sections and treating them after sectioning, but the protoplasts shrink and often fall out from the cells. No improvement in this respect was revealed using — at different concentrations — sucrose and arabic gum alone. The changes in the proportions of the ingredients of the sirup were also ineffective. A series of experiments were therefore performed with plain sucrose, but none of the factors tested was efficient enough to improve the results as compared with the standard procedure, the gradual transfer being the only exception. Since this makes the procedure more laborious and time consuming, certain other media were tested, 0.88 M ethanol and 0.88 M dimethylsulf-oxide yielding the best results. Attempts are being made to understand some points of the present experience in connection with the results of other authors and on the basis of general theory respectively.     

On the interaction of growth retardants with IAA and kinetin

In the pea test a highly positive response to the treatment with IAA reversed to a negative one or became 5 to 6 times weaker when CCC was applied together with IAA. In cultivating pea seedlings, following their decapitation, for two days in a 0.25 per cent CCC solution and then in water, growth of their cotyledonous axillaries (cotylaries) were inhibited. This inhibitive action of CCC could be made ineffective when the seedlings, following two-days’ cultivation in the CCC solution, were grown further in kinetin solutions (0.37–3 mg per 1). Cotylaries of decapitated pea seedlings, when grown in kinetin solutions were inhibited. With kinetin solutions of 6–12 mg/l a strong inhibition also occured in the growth of roots at the apical parts of which spherical swellings were developing. The CCC supplied to the roots of intact etiolated pea seedlings is translocated acropetally into the stem at a rate of about 5 cm per hour. Decapitation of the plant causes retardation of this transport, yet a coat of 0.00001–1% IAA or kinetin paste produces acceleration of the stream. Existence of an antagonism between CCC and IAA, demonstrated earlier, was found holding true also for B-9 (N, N-dimethyl-aminesuccinamic acid) and IAA, as the inhibitive action of B-9, 0.06% solution on the growth of lettuce hypocotyls was reduced to a highly significant degree when the plants were supplied with B-9 together with IAA at a concentration of 10 mg/l.     

Influence of exogenous DNA on Ypenyl-treated chromosomes ofVicia faba L.

This paper is a study of the effect of exogenous DNA of different genetic origins on the repair of meristematic cells of primary roots ofVicia faba, damaged by 24 hour treatment with 0·01mm solution of Ypenyl. Both kinds of DNA,i.e. isologous and heterologous, stimulated cell proliferation which was decreased by the action of the radiomimetic and influenced both dynamics of production of chromosome aberrations and the interchromosomal distribution of induced damage. While heterologous DNA increased the frequency of aberrations after all recovery periods studied, isologous DNA significantly decreased the number of chromosomal aberrations. Heterologous DNA increased at the same time the relative number of breaks in the group of small chromosomes, while by the action of isologous DNA the number of aberrations related to this group of chromosomes was relatively decreased.     

Serological analysis of eleven strains ofRhizobium japonicum

The present communication reports a serological analysis of eleven strains ofRhizobium japonicum. The slow-diffusing thermostable antigens were found to be suitable for the basic differentiation of the somatic serogroups inRhizobium japonicum. One to three precipitation bands of the slow-diffusing thermostable antigens, one to two bands of the fast-diffusing thermostable antigens and one to three bands of the thermolabile antigens were detectable in the whole cell cultures ofR. japonicum by means of the immunodiffusion technique. Two basic somatic serogroups were differentiated on the basis of the slow-diffusing thermostable antigens. The thermolabile antigens were identical in most of the strains.The author is greatly indebted to Mrs. M. Kabelovà for technical assistance.This investigation forms part of a contribution prepared by the Czechoslovak National Committee for the International Biological Programme (Section PP: Production Processes).     

Antibacterially active substances

Synthesis of two hydroxy-derivatives of nalidixic acid as a result of microbial transformation was demonstrated in certain species of the genusAspergillus. Aspergillus alliaceus produced 7-hydroxy-nalidixic acid andAspergillus niger 6-hydroxy-nalidixic acid. It was demonstrated that the antibacterial activity of both hydroxy-derivatives (tested inEscherichia coli) was lower than that of the initial nalidixis acid.     

The inactivation ofBacillus subtilis transforming DNA by mutagenic agents

The inactivation of three genetic markers inBacillus subtilis transforming DNA by UV rays, nitrous acid and hydroxylamine was studied. The inactivation of markers by UV and HNO₂ was marker specific, whereas the hydroxylamine inactivates the markers approximately to the same degree.