Jiřina Švachulová

Repair of single-strand breaks and fate of N-7-methylguanine in DNA during the recovery from genetical damage induced by N-methyl-N-nitrosourea in barley seeds

Abstract The recovery from various types of N -methyl- N -nitrosourea (MNU)-induced genetical damage, observed after two weeks storage of alkylated seeds with 30% water content at 25°, was found to be connected with the repair of single-strand breaks and/or apurinic sites in DNA of the seed embryo. The amount of N -7-methylguanine (7-MeG) in DNA was not influenced by this type of seed storage, showing that the excision of this base is not a part of the repair mechanism. The changes in the alkylation of proteins, lipids and RNA observed after storage of the seed are considered to be irrelevant to the repair process. Single-strand breaks and/or apurinic sites in barley DNA are presumed to be the main lesions responsible for the reparable types of genetical damage.

Storage effect in barley changes in the amount of DNA lesions induced by methyl and ethyl methanesulphonates

Abstract Barley seeds were treated with methyl methanesulphonate (MMS) and ethyl methanesulphonate (EMS), stored at 15% water content and washed for 16–24 h. These treatments resulted in an increase of toxic and genetic effects. In teh DNA of embryos of such stored MMS- and EMS-treated seeds, a strong enhancement of the amount of single-strand breaks and/or alkali-labile sites took place. In contrast, the amount of alkylated sites, particularly of 7-methylguanine, was somewhat lower. It can be that the depurination and/or backbone breakage, which proceeds during the storage period, is responsible for the enhancement of toxic and genetic effects, whereas the influence of the alkylation of DNA during the storage period by the unreacted residual mutagen is negligible.

Immunochemical characterization of seed proteins of some species of the genusArachis L.

Relationships between seed protein patterns of 16 species ofArachis were studied by means of immunochemical methods (immunoelectrophoresis and double diffusion). The broad similarity of these patterns suggests that protein patterns are conservative in this genus from an evolutionary viewpoint. The differences which have evolved support recent schemes for classifying the genus, particularly the breakdown into sections. It also suggests thatA. villosa andA. correntina should probably be recognized as distinct species. The strong relationship indicated betweenA. hypogaea andA. batizocoi supports the hypothesis that the latter may be source of one of the genomes ofA. hypogaea. The possibility thatA. cardenasii might be the source of the other genome did not receive such strong positive support.

The comparison of seed protein patterns within the genusArachis by polyacrylamide gel electrophoresis

The seed protein patterns of 12Arachis species were compared by polyacrylamide gel electrophoresis (PAGE), similarities between patterns were measured by the Jaccard index. Results obtained confirm the close relationships established between members of the genus on morphological grounds and support the more recent classification schemes.A. villosa andA. correntina could well be regarded as distinct species on grounds of protein differences whileA. macedoi andA. villosulicarpa (although members of the same section, Extranervosae) show considerable differentiation of their protein patterns. Surprisingly, the formA. ×batizogaea showed less similarity in protein pattern to those of its parental species than might have been expected. The principle value of seed protein pattern data appears to be in distinguishing species within sections.

Use of esterase isoenzymes revealed by gel isoelectric focusing as an aid in chemotaxonomical study of the genusAllium

Eleven species of the genusAllium belonging to the sectionCepa, Phyllodolon, Rhizirideum, Melanocrommyum andAlliun ware investigated as to the presence of esterase isoenzymes by means of the gal isoelectric focusing which shows better resolution than polyaorylamide gel electrophoresis. In all examined species twenty six isoenzymes were found. The individual sections are well characterized by esterase isoenzymes revealed by this method, on the other hand, the differences, as shown by the Jaceard index, between the subganera are insignificant. Investigation of seven cultivars ofAllium cepa shows the isolated position of the cultivar Kaštická. These results are in full agreement with those found with use of the polyacrylamide gel electrophoresis and show the usefulness of gel isoelectric focusing for the solution of the chemotaxonomical problems.

Respiration in barley seeds treated with mutagenic methyl methanesulphonate and stored at different water contents

In water soaked barley seeds the respiration rate (QO2) at 25 °C is about 100 times higher at 30% than at 20% water content. The treatment of seeds with mutagenic methyl methanesulphonate led to the depression of the respiration rate in dependence on the applied mutagenic dose. Seed storage at 30% water content resulted in a recovery from the inhibition of seed respiration caused by methyl methanesulphonate. In contrast, at 20% seed water content the respiration rate either slightly decreased or remained unchanged in the course of storage. The results are discussed in relation to the previously described changes in the yield of induced mutagenic effects in barley seeds treated with monofunctional alkylating agents and stored at 30% and 20% seed water content.

Isolation from barley embryo of endonuclease specific for apurinic sites in DNA

The endonuclease activity specific for apurinic sites in DNA was detected in barley embryos. The enzyme was partially purified. It reveals high activity on partially depurinated DNA but low or nil activity on intact and alkylated DNA. The method used for the detection of enzyme activity was based on the changes in the sedimentation velocity of substrate DNA in neutral sucrose gradients with 80 % formamide.

Serological comparisons of seed proteins of someAllium (L.) species belonging to the subgenusRhizirideum (G.Don exKoch)Wendelbo

Similarity of seed protein spectra of 13 representatives of the genusAllium, subgenusRhizirideum, was estimated by means of immunochemical analysis; the species investigated were divided into four groups more or less corresponding to the contemporary systematic division of species into sections.

Response of “storage” effects in barley seeds to the dose of ethyl methanesulphonate and X-ray treatment

Abstract1. Ethyl methanesulphonate (BMS)-treated barley seeds stored at 30 % water content for 1 – 4 weeks are capable of recovering from the mutagen induced injury. The level and speed of the recovery depends on the EMS dose and storage period. Seeds treated with extremely severe EMS doses reducing the% of germination to 5 – 10 % cannot recover from the induced injury. 2. Due to a 1 to 6 weeks storage of EMS-treated seeds with 20 % water content the amount of induced injury increases in dependence on the applied EMS dose and storage period. Doses of EMS, reducing the seedling height in the M1 generation to 90 – 95 % of the control reveal no storage after-effects. 3. Storage of X-ray exposed seeds at 20 % or 30 % water content did not result in any significant changes in the yield of induced genetic damage.

Recovery from ethyl methanesulphonate-induced genetical damage in barley. Independence of sodium azide treatment

Barley seeds were treated for 3 h at 25°C with 240 mM ethyl methanesulphonate (EMS), washed for 18 h, treated with various concentrations of unbuffered sodium azide (pH 6.7–7.3) for 3 h at 25°C, re-dried to 30% water content and either sown immediately or stored at 25°C for 12 days and then sown. The synergistic action of sodium azide post-treatment has been demonstrated only for the EMS-induced M1 germination reduction, while the EMS-induced M1 sterility and the yield of M2 chlorophyll mutants were unaffected. The „storage” recovery from EMS-induced mutagenic effects was insensitive to sodium azide post-treatment. The 12 day-seed storage at 25°C brought about an improvement of M1 germination, M1 survival, M1 fertility and a decrease in the amount of M2 mutants, regardless of whether sodium azide post-treatment was applied or not.AbstractNa semena ječmene, ovlivněná etylmetansulfonátem (EMS), bylo působeno azidem sodným. Synergický účinek azidu sodného byl zjištěn jen pro EMS-indukovanou redukci M1 kličivosti a přežití, zatímeo EMS-indukovaná M1 sterilita a frekvence M2 chlorofylových mutant nebyly ovlivněny. Popůsobení azidem sodným také neovlivnilo zotavení z EMS-indukovaného mutačního poškození.