Aminopeptidase, carboxypeptidase and proteinase activities were measured in endosperms from unripe and ripe seeds of ×Haynaldoticum sardoum. Aminopeptidase and proteinase activities were high during the early maturation stages and then decreased. In contrast, carboxypeptidase activity increased with maturation.
Localization studies demonstrated that aminopeptidase and carboxypeptidase activities were present in the three tissues examined (pericarp, green layer plus aleurone, and starchy endosperm). Proteinase activity against gliadin was located in the pericarp and in the green layer plus aleurone, but was absent in the starchy endosperm.
The presence of proteolytic activities in the outer kernel layers might be correlated to the hydrolysis of transitory protein reserves during the senescence of the seed coatś.
Aminopeptidase and carboxypeptidase activities located in the starchy endosperm could participate in the breakdown of protein reserves during the early phases of seed germination.

The equilibrium density of chloroplasts from barley (Hordeum vulgare L. cv. Hassan) was analyzed by sucrose gradient centrifugation. Natural and detachment-induced leaf senescence were associated with a decrease in density and an increase in heterogeneity of the chloroplast population. Treatments (with growth regulators and light) which retarded or accelerated senescence, respectively, retarded or accelerated chloroplast density decrease. Accelerators as well as retardants of senescence decreased the heterogeneity of the chloroplast population.

Influx and efflux of inorganic phosphate was determined in freshly cut and "aged" slices of sweet potato tubers, in the presence or absence of abscisic acid. Slices aged in the phytohormone took up considerably less phosphate than did the control slices. No difference in the rates of uptake of phosphate by the fresh tissue was observed. Kinetics of uptake of the aged untreated slices, proved to be bi-phasic. This was not the case for the treated slices. Neither the treated nor the untreated uptake curves for the fresh slices were bi-phasic. Efflux of phosphate from the slices aged in abscisic acid was greater than from the untreated slices. The results strongly suggest an effect of abscisic acid on the cell membrane.

Regeneration abilities of buds on shoot segment explants isolated from adult trees of oak (Quercus robur), aspen (Populus tremula), black locust (Robinia pseudacacia), Japan pagoda tree (Sophora japonica), and English walnut (Ailanthus glandulosa) were studied during the growing season. Optimum BAP concentrations for the regeneration of oak bud meristems were dependent on the date of sampling. Axillary shoots could be induced from winter and summer buds of oak and aspen on Dustan and Short media supplemented with activated charcoal and BAP at concentrations from 0.5 to 2 mg 1^-1. More intensive rooting of segments of newly formed shoots was observed on MS medium diluted to one half and supplemented with 2 % sucrose and 0.2 mg 1^-1 of IBA.Populus tremula formed longer axillary shoots on DS media supplemented with 0.5 mg 1^-1 of BAP and 1 mg 1^-1 of GA₃.
Regeneration capacities of black locust, Japan pagoda tree, and English walnut were higher. In addition to the induction of multiple shoots from buds, shoots could also be obtained from calluses formed on basal segment parts. Asparagine and glutamine at a concentration of 25 mg 1^-1 stimulated the percentage of differentiated stems on callus surface. Inhibitory effects of substances which accumulated in buds in the second half of the growing season could be reduced by means of pulse treatments in 50 mg 1^-1 BAP solutions or using short-term dipping into 0.1 % AgNO₃ solution.

Organogenic callus cultures ofPapaver somniferum L. were studied with the aim of describing the morphology of the callus and observing the changes occurring during differentiation and induction of organogenesis. The morphology of the cells and the formation of meristemoid areas were studied on a light microscopic level. Histological evidence showed that from the inner meristematic centres, protracheal elements are differentiated, from the surface meristematic centres meristemoids are formed and therefrom green leaf-like organoids.

Phaseolus coccineus storage globulins were extracted from mature cotyledons, purified and characterized. Three major proteins were separated. A component showing erythroagglutinating activity was thoroughly purified by thyroglobulin-Sepharose chromatography. The relative molecular masses of the three fractions are Mr = 330, 178, and 500 kDa as determined by polyacrylamide gel electrophoresis (PAGE). They correspond to the proteins found in other systems and classified as phytohaemagglutinin (PHA), vicilin and legumin, respectively. Electrophoretic analyses under denaturating conditions (SDS-PAGE) evidenced the major subunits for the three proteins. Isoelectrofocusing of the isolated proteins indicated a large heterogeneity for vicilin.

The study deals with the effect of indol-3-ylacetic acid (IAA) on endogenous rhythm of flowering inChenopodium rubrum L. ecotype 374. Phase setting of the rhythm and length of its period were not affected, its amplitude decreased or changed insignificantly depending on the time and the site of IAA application. The intervals of significant inhibitory effect of IAA applied to apical buds terminated later than after IAA application to photoreceptive organs. The inhibitory effect did not correlate with the level of IAA uptake. Results obtained support the hypothesis that both photoreceptive organs and apical buds are the sites of inhibitory effect of applied IAA.

In the long-term cultivated callus cultures ofMatricaria recutita L. the identical concentration changes in the biosynthesis of glutathione, glutamate, aspartate, total thiols and proteins were detected within the subculture. The level of oxidized glutathione during the growth of callus culture was low with the highest value 10.66 nmol g^-1 on the 13th day of subculture. The ratio GSH/GSSG which significantly influences the redox processes in a cell, and the activity of glutathione reductase increased from the 8th day. Ascorbate formation was detected on the 17th day, although no relation between the ascorbate synthesis and the concentration of glutathione and glutathione reductase was found.

Continuous treatment with spermidine or 1-aminocyclopropane-1-carboxylic acid stimulated ethylene production and ethylene-forming enzyme activity and accelerated chlorophyll breakdown in detached tobacco leaves. The treatments also induced the production of eleven major acidic pathogenesis-related proteins, which were also produced during the hypersensitive reaction to tobacco necrosis virus. A delay between the onset of the stimulated ethylene increase and the detection of PR-proteins was found; ethylene production was stimulated after a few hours of treatment, whereas one, three and all the eleven PR-proteins were detected by polyacrylamide gel electrophoresis of fluid extracts after 2, 4 and 6 days of treatments, respectively. The possible causal relationship between stimulation of ethylene production and PR-protein accumulation is discussed.

The growth of primordia of marginal buds (marginals) which differentiate on leaf margins is correlatively inhibited on intactBryophyllum crenatum plants. Following leaf isolation, the marginals are released from this correlative inhibition, which process is accompanied within 2 to 10 h after leaf isolation with a decrease in the content of endogenous IAA in the leaf blade. This decrease can be enhanced by transversal cutting of the leaf blade into three parts which also results in enhanced subsequent growth of the marginals. The growth which follows after the release of the marginals from correlative inhibition is accompanied in cut leaf blades with an increased content of endogenous IAA in the period from 12 h to 7 d after leaf isolation when compared with uncut leaf blades. The highest content of endogenous IAA was recorded in the middle section, and the lowest IAA content in the basal section of the leaf blade.

Lead was detected and located in definite regions of thePopulus adventitious roots with the light microscope. Incubations of 24, 48, and 72 h in solutions of Pb(NO₃)₂ resulted in clear sequences of microscopic images. Interpretation of these images allows the construction of a scheme for lead transport pathways in roots.

Changes in macromolecule syntheses, especially RNA synthesis, and the energy providing system were investigated in seeds ofAgrostemma githago aged for different periods. In embryos of aged seeds all macromolecule syntheses start later and reach a lower level than young ones. It was found that the synthesis of rRNA in embryos of aged seeds is reduced whereas the synthesis of poly (A⁺) RNA in relation to the total RNA synthesis is highly increased as well as the amount of this RNA species with long poly (A) chains. The results are discussed in connection with the decreased protein synthesis and the reduced ATP content and ATP formation ability in embryos during the long time storage of seeds.

The inheritance of the gliadin spectra in F₂ hybrids between cultivars hexaploid wheat Bezostaya 1 and Bisel has been investigated. The results from the electrophoretic examination and analysis of the gliadin spectra reveal that the components 1,2,3,11 and 15 from the Bezostaya 1 spectrum and 1,2, 3, 7,11 and 15 from the Bisel spectrum are controlled by allelic variants of one gliadin coding locus in 1D chromosome.

Protein complex patterns of cloves and esterase isoenzyme patterns of apical buds of cloves were studied with Czechoslovak virus-free cultivars ofAllium sativum L. and the wild speciesA. longicuspis Regel, Similarly, four clones-regenerants obtained using explant culture techniques from A.sativum L. cv. Bzenecký paličák (two somaclones and two clones derived from plants regenerated from meristem cultures treatedin vitro with colchicine) differing in their ploidy, morphology, and yields were studied. Immunophoreograms of protein complexes of theA. sativum L. cultivars under investigation differed from one another in the number and mobility of protein fractions in both the cathodic and anodic regions and thus these cultivars can be distinguished. On the basis of esterase isoenzyme patterns, the Czechoslovak cultivars of A. sativum L. can be arranged into three groups - bolting winter cultivars with broad leaves, non-bolting winter cultivars with broad leaves, and non-bolting spring cultivars with narrow leaves. All the clones-regenerants showed the same protein complex and esterase isoenzyme patterns as their original cultivar.A. longicuspis Regel markedly differed in its protein complex and esterase isoenzyme patterns from all the other genotypes studied.Received May 17, 1989: accepted January 5,1990

Embryogenic callus which has maintained its embryogenic ability on media without growth regulators for three years has been induced at the base of shoots of a genotype with CMS propagated for a long timein vitro by transferring the shoots onto media richer in inorganic and organic components. The effect of two basal media (MS and PG₀) on the intensity and completeness of the proliferation of somatic embryos was examined with different combinations of growth regulators. Pollen fertility was evaluated in 87 plants regenerated from somatic embryos. Cytoplasmic male sterility was conserved in all of them.

Phenolic content and IAA-oxidase (IAA-o) activity have been assayed in cells and medium of tobacco crown gall suspension culture in several stages of culture cycle. The highest content of total phenolics in the cells were found prior to cell division and in the middle stage of intensive growth. The beginning of intensive growth is accompanied by temporary reduction in phenolic level in the cells as well as their intensive secretion to the medium. In the second part of the culture cycle, when the phenolic production was weaker, the majority of these compounds were maintained in the cells.
The highest activity of IAA-o in the cells was detected in the middle stage of intensive growth, simultaneously with high phenolic content; following it a considerable decrease of IAA-o activity is correlated with maximum of chlorogenic acid (ChA) content (at reduced amount of total phenolics). IAA-o activity increased again at the end of the stage of intensive growth when the level of phenolics was low including ChA. These data suggest that IAA-o in relation to phenolic level determines cell growth in the culture.
In the culture medium - fairly distinct negative correlation between IAA-o activity and phenolic content suggests that the latter participates in enzyme activity regulation. During intensive growth IAA-o activity is strongly inhibited.
The results prove that phenolic level. IAA-o activity and auxin level are closely correlated and may constitute essential elements of a mechanism of regulation crown gall cell growth in culture.

A simple equipment was developed to cultivate young cereal plants under enhanced CO₂ concentration. Cultivation system permits growing of about 40 barley seedlings for about 2 to 3 weeks. The system consists of two identical growth chambers (volume about 30 dm³), gas conditioning circuit and measuring circuit with an infra-red CO₂ analyser. Capabilities of the whole equipment were tested by growing barley plants under 330 and 1000 cm³ (CO₂)m^-3 and in combination with high or low nitrate level.

Haploid tobacco plants (cv. Samsun) form inflorescences with a larger number of flowers than diploid plants. Leaves of haploid plants were shown to have lower free IAA level (by 40 %), higher peroxidase (by 160 %) and IAA-oxidase (by 70 %) activities and produce less ethylene (by 25 %) than leaves of corresponding diploid plants. The increase of peroxidase activity in haploids was due to the increase in the activity of the cathodic isozyme which is known to have high IAA-oxidase activity. It is proposed that higher peroxidase/IAA-oxidase activity in haploid plants may take part in IAA catabolism, at least duringin vitro culture of haploid explants. Lowered IAA level and ethylene production may then be directly correlated with a larger number of flower buds; as a higher IAA level is generally considered to act as a background inhibitor of flowering.

Chlorophyll deficient plantlets were obtained after the treatment of sugar cane callus cells with 3-azido-l,2-propanediol (AG) and sodium azide with approximately the same frequency. Both mutagenic agents postponed the regeneration of plantlets from calli, the total number of regenerated plantlets following the AG treatment being, however, higher than after the sodium azide treatment and control.

Germinated tubers of selected cultivars Kera, Resy, Nicola and Oreb were made healthier by heat treatment. They were derived from germ explants on MS media with the addition of BAP 1 mg 1^-1 and IAA 0.2 mg 1^-1. After sufficient multiplication of stems, optimum conditions of the photoperiod were followed for the induction of axillary microtubers on stem segments in media with BAP 10 mg 1^-1 and 8% sucrose. The ability of tuberization is different: the early cvs. Kera and Resy induce earlier tubers at a long photoperiod and late cvs. Nicola and Oreb tuber rather at a short photoperiod. In suitable photoperiods the inhibitory substances accelerate the induction of axillary tubers and limit the formation of adventitious roots. Synthetic inhibitors applied in induction media increase the number of the tubers.
The quality of tubers was affected by the addition of a mixture of amino acids: aspartic, glutamic, lysine and proline in concentrations of 12.5, 25 and 50 mg.1^-1 into the induction media. In the course of cultivation the types which were growing well, formed tubers and increased the volume were selected. The representation of amino acids in the tubers was not significantly affected, there was only an increase in proline. The higher content of amino acids was reflected in the increase of proteins in the tubers. The selected clones are further multiplied.

Plants regenerated from callus cultures derived from leaf discs and mesophyll protoplasts ofPetunia hybrida cv. Rose of Heaven exhibit a high frequency of genetic and chromosomal variation. Of twelve leaf disc-derived plants examined, only three had the normal diploid chromosome number (2n=14) while seven were tetraploid and two were aneuploid (16 and 27 chromosomes). Of seventeen plants derived from two protoplasts, none had the diploid chromosome number. Most had 28 chromosomes, one 29, two 27, one 26 and one had variable numbers (14-28) in different root tip cells. In all cases aneuploidy was associated with developmental abnormality. In addition, heritable differences in growth, morphology and flower pigmentation were observed in callus-derived tetraploids and diploids, including one diploid which differed from parent plants in at least four characters. These results are discussed in terms of the importance ofPetunia in genetics research and for studies of somaclonal variation.

Lemon plants (Citrus limonum L. cv. Verna) were grown in the field under two different flood irrigation treatments. The dry treatment received four irrigations per year (March, July, September and November) and the wet treatment one monthly. The amounts of water applied per year for dry and wet treatments were 340.0 mm and 1020.0 mm, respectively. The effects of the two treatments on certain aspects of the plant water relations during the period between flowering and the end of rapid fruit growth (critical period) were studied. Soil matric potential (ψm) and leaf water potential (ψi) values in the dry treatment revealed development of water stress during the experimental period. The water supply in the wet treatment seems sufficient to achieve the crop water requirements. The g₁ values in July were higher in the wet than dry treatments. Pronounced oscillations in g₁ from sunrise to afternoon were found especially in the dry treatment.