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.     

Ontogenetic development of antibody formation in response to different doses of gram-negative microorganisms in young rabbits

Rabbits at different stages of development were immunized with different doses of heatinactivated suspension ofEscherichia coli 086 andSalmonella paratyphi B. The dynamics and the site of formation of bactericidal and haemolytic antibodies during the primary reaction was investigated. An increase and an acceleration of antibody formation after increasing the dose of antigen was found in the serum and at the cellular level. The magnitude of the response and the rate of the reaction were directly proportional to the age of the experimental rabbits. The site of antibody formation depends on the character, route of administration, antigen dose and age of rabbits. After intraperitoneal and also after intravenous immunization withEscherichia coli andSalmonella paratyphi B antigens the site of antibody production in 15-day-old rabbits was the lymphatic tissue of the intestine, the appendix, and mesenteric lymph nodes. As the antigen dose was increased and the age of rabbits rose, i.e. in correlation with the increase of the antibody response, antibody formation shifted to the spleen which is the chief site of antibody production following immunization by these bacterial antigens from the first month of life of rabbits. In contrast with this type of antigen, after intraperitoneal or intravenous immunization with sheep erythrocytes of new-born or older rabbits antibody formation was concentrated in the spleen. The development of the immunological competence and the significance of intestinal lymphatic tissue as one of peripheral type is discussed.     

Die wirkung der β-(3-Pyridyl)-propion-und der β-(3-Pyridyl)-acrylsäure auf die Atmung der Wurzeln vonSinapis alba L.

V práci byly studovány rozdíly v ú?innosti kyseliny β-(3-pyridyl)-propionové (VIII) a β-(3-pyridyl)-akrylové (IX) v závislosti na stupni nenasycení pobo?ného ?etězce. Obě kyseliny inhibují r?st ko?ene i hypokotyluSinapis alba L. a zp?sobují inhibici sukcinátdehydrogenázového enzymového systému. Kyselina IX inhibuje zmíněný systém p?ibli?ně na dvojnásobek inhibice kyseliny VIII. Cytochromoxydázový systém z?stává témě? neovlivněn. Na základě experimen tálních výsledk? byla zp?esněna strukturní podmínka fytotoxické aktivity modelových slou?enin typu I v tom smyslu, ?e v?echny slou?eniny této struktury inhibují kromě r?stu také aktivitu sukcinátdehydrogenázového systému.     

Chromosome aberrations caused by the effect of ultrasound in the meristematic cells ofvicia faba

In our present work the formation of chromosome aberrations has been studied in dependence on the tima interval between sonication and fixation of the primary root tips of Vicia faba. Maximum occurrence of aberrations was recorded immediately after sonication. The results of our experiments pointed to the fact that the frequency of the induced changes was independent on the sonic waves intensity within the range of 0-2—3-0 W/cm² and on ultrasond treatment duration within the range of 1—20 min. Studies of the distribution of chromosome abnormalities caused by ultrasound between the large and small chromosomes of theVicia faba meristematic cells in various time intervals showed that the frequency of the aberrations in both chromosome groups was proportional to its total metaphase lengths. Analysis of the type of aberrations observed in various time intervals after sonication indicated the simultaneous formation of chromosome and chromatide abnormalities.     

Spores of microorganisms

6-Azauracil at a concentration of 1 μmole/ml inhibits by 50% the outgrowth of germinated spores of a strain ofBacillus cereus, concentration of 1.5 μmole/ml resulting in 100% inhibition. Two distinct maxima of sensitivity to 6-azauracil are observed during postgerminative development of spores. The first occurs during early stages of development (immediately after depolymerization period) and the second after about 60 min of cultivation (late stage of swelling). Uracil reverses the inhibition of the outgrowth of spores caused by 6-azauracil when added during 0–30 min of the spore development. The addition of uracil after 30 min of the germination does not bring about the reversion of the effect of 6-azauracil. An important role of pyrimidine pathway via orotidine 5′-phosphate in germinating spores was proved, suggesting a possible use of 6-azauracil in synchronization of the postgerminative development of spores.