Nuclear DNA characterization of two species ofVicia:Vicia bithynica L. andVicia narbonensis L.

The speciesVicia bithynica andVicia narbonensis, from the same subgeneric section ofVicia faba, show variations in nuclear DNA content Nuclear DNAs, extracted from root tips of the twoVicia species, were characterized by thermal denaturation, analytical ultracentrifugation and reassociation kinetics. The thermal denaturations of DNA, the number of DNA components reassociating with second order kinetics, the proportion of repeated DNA sequences, the frequency of the repeated DNA classes are reported and compared to previous data onVicia faba DNA. Feulgen absorptions at different thresholds of optical density⁺ of interphase nuclei in cytological preparations of the root meristems ofV. bithynica andV. narbonensis are determined and compared withV. faba analogous determinations. The results, confirming that plant genome is highly flexible, are discussed in relation to other data on the interspecific variations of the nuclear DNA content.     

Genetic effects of root extracts of Glycyrrhiza glabra L. in different test systems

The antimutagenic and geroprotective activities of licorice (Glycyrrhiza glabra) root extracts were established in cells of plant test systems of Allium fistulosum L., Allium cepa L., and Vicia faba L. and of laboratory animals (Vistar rats). The prospects for the practical application of licorice Glycyrrhiza glabra root extracts as antimutagenic substances are discussed.     

Abscisic Acid Accumulation by in Situ and Isolated Guard Cells of Pisum sativum L. and Vicia faba L. in Relation to Water Stress

Isolated guard cells, prepared by sonication of epidermal peels, were used to investigate the endogenous level of abscisic acid (ABA) in the guard cells of turgid and stressed leaves of Vicia faba L. and the argenteum (arg) mutant of Pisum sativum L. The guard cells of V. faba and arg were found to contain 18 and 8 times more ABA, respectively, when isolated from stressed leaves than from turgid leaves. Isolated guard cells of V. faba were also directly stressed with the osmoticum Aquacide III. These guard cells were capable of producing stress-induced ABA to at least 3 times their ABA level when non-stressed.     

Position and extent of the elongation zone in the root tip of the broad beanVicia faba L.

It was found by measuring the length of the cortex cells of the root tips of the broad beanVicia faba L. that the beginning of the elongation zone lies at about 1–2 mm from the initials and its end at about 7–8 mm from the initials. Shrinkage of the object during microtechnical treatment was negligible. The autonomy of the individual tissues of the root tip was taken into account.     

Mitochondrial plasmid-like molecules in fertile and male sterileVicia faba L

Biochemical analysis and electron microscopy showed that mitochondria of both the fertile and the male sterile 350 and 447 cytoplasms ofVicia faba. L. contain two small supercoiled DNA molecules of mean length of 1 700 and 1 420 base pairs in addition to the main mitochondrial DNA of high molecular weight. By agarose gel electrophoresis, the male sterile cytoplasm 350 is distinguished from the fertile cytoplasm and from the male sterile cytoplasm 447 by the presence of an additional supercoiled DNA molecule of approximately 1 540 bp.     

Electrophoretic investigation of ribonuclease in roots and leaves ofVicia faba L.

Isozyme patterns and specific activity of ribonuclease (ribonucleate pyridinenucleotido-2′-transferase, E. C. 2.7.7.16) were followed in the extracts of segments from three growth zones of the root and in extracts of young and senescent leaves ofVicia faba L. Electrophoreograms of extracts from all three investigated root zones were identical, in the electrophoreograms of extracts from senescent leaves however one new ribonuclease occurred which could not be detected in the electrophoreograms of extracts from young leaves. Extracts from senescent leaves had higher specific activity of ribonuclease than extracts from young leaves. Extracts from the enlargement zone of the root and those from the maturation zone had a three times higher specific activity of RNase than extracts from the division zone.     

Effects of high osmotic potential of a medium on mitotic cycle in roots ofVicia faba L

The present paper deals with the effects of osmotic potential of a medium on cell reproduction and elongation of the roots ofVicia faba L. andVicia sativa L. As the osmoticum polyethylene glycol (PEG 4000) in various concentrations ranging from 5 % to 25 % (i.e. fromca.-0.11 up to -1.27 MPa) has been used. The results show that at higher concentrations than 7.5 %, the growth of roots is slowed down and at the concentration of 25 % PEG this decrease in growth rate is as much as 6 fold compared with the control. The mitotic cycle is prolonged, however, only 1.86 times. Thus, the inhibition of root growth is caused mainly by the inhibition of cell elongation. Concerning the effect of high osmotic potential on mitotic cycle it was found that the roots after immersion into 25% PEG are able to overcome this osmotic stress and after 6 h to renew the mitotic activity. The S phase of the cycle is the most sensitive to this factor and even after mitotic activity was renewed it showed a slower rate in comparison with the control.     

Genetic diversity of Vicia faba L. and Pisum sativum L. nodulating rhizobia in the central Black Sea region of Turkey

In this study, we obtained a total of 60 rhizobial isolates from root nodules of Vicia faba L. (n = 30) and Pisum sativum L. (n = 30) grown in the Central Black Sea region of Turkey. The 16S rDNA PCR-RFLP analysis with enzymes CfoI, HinfI, NdeII and MspI revealed a single pattern. Moreover, nucleotide sequence phylogenies based on both the 16S rDNA and recA suggested that these isolates belonged to Rhizobium leguminosarum. Phylogenetic analysis showed that some of our V. faba L.-originated isolates were closely related, indicating molecular evidence for the selection of some special R. leguminosarum bv. viciae isolates by V. faba L., as suggested in previous studies. Network analysis based on recA sequences revealed a common evolutionary history for Turkish, European, North and South American, and Jordanian R. leguminosarum bv. viciae isolates. We isolated four haplotypes using nodA and nifH nucleotide sequence data, i.e. four types of sym plasmids. Two of these types were common to rhizobial isolates from both V. faba L. and P. sativum L., indicating that nodulation factors may not be the mechanism for selection of the special R. leguminosarum bv. viciae populations by V. faba L.     

Bemerkungen zur Taxonomie vonVicia sepium L.

In der ArtVicia sepium L. kann man in Mitteleuropa und in südlichen Teilen von Nordeuropa drei Varietäten unterscheiden: 1. var.sepium mit breiten Blättchen und einem kahlen bis angedrückt behaarten Kelch, 2. var.montana Koch mit schmalen Blättchen und einem kahlen oder zerstreut angedrückt behaarten Kelch, und 3. var.eriocalyx ?elak. mit breiten bis mittelbreiten Blättchen und einem abstehend langbehaarten Kelch, noch mit einer niedrigen und einer hohen Wuchsform. Bei allen Proben wurde die Chromosomezahl 2n=14 festgestellt.     

Cellular pathway of photosynthate transport in coats of developing seed of Vicia faba L. and Phaseolus vulgaris L. I. Extent of transport through the coat symplast

The extent of post-phloem solute transport through the coatsymplasts of developing seeds of Vicia faba L. and Phaseolusvulgaris L. was evaluated. For Vicia seed coats, the membrane-impermeantfluorochrome, CF, moved radially from the chalazal vein to reachthe chlorenchyma and thin-walled parenchyma transfer cell layers.Thereafter, the fluorochrome moved laterally in these two celllayers around the entire circumference of the seed coat. Transferof CF from the chalazal vein was inhibited by plasmolysis ofattached ‘empty’ seed coats. In contrast, the spreadof phloem imported CF was restricted to the ground parenchymaof Phaseolus seed coats. Fluorochrome loaded into the outermostground parenchyma cell layer was rendered immobile followingplasmolysis of excised seed-coat halves. Phloem-imported [¹⁴C]sucroseand the slowly membrane permeable sugar, L-[¹⁴C]glucose, werepartitioned identically between the vascular and non-vascularregions of intact Vicia seed coats. For ¹⁴C-photosynthates,these partitioning patterns in attached ‘empty’Vicia seed coats were unaffected by PCMBS, but inhibited byplasmolysis. Tissue autoradiographs of intact Phaseolus seedcoats demonstrated that a pulse of ¹⁴C-photosynthate moved fromthe veins to the grounds tissues. In excised Vicia seed coats,preloaded with ¹⁴C-photosynthates, the cellular distributionof residual ¹⁴C-label was unaffected by PCMBS. In contrast,PCMBS caused the ¹⁴C-photosynthate levels to be elevated inthe veins and ground parenchyma relative to the branch parenchymaof Phaseolusseed coat halves. Based on the above findings, itis concluded that the phloem of Vicia seed coats is interconnectedto two major symplastic domains; one comprises the chlorenchyma,the other the thin-walled parenchyma plus thin-walled parenchymatransfer cells. For Phaseolusseed coats, the phloem forms amajor symplastic domain with the ground parenchyma. Key words: Phaseolus vulgaris L, phloem unloading, photosynthate transport, seed coat, symplast, Vicia faba L     

Electrophoretic study on peroxidase,indoleacetic acid oxidase,and o-diphenol oxidase fractions in extracts from different growth zones ofvicia faba L. roots

Using electrophoresis in acrylamide gel, fractions of peroxidase, indoleacetic acid oxidase, and o-diphenol oxidase were investigated in extracts from three growth zones ofVicia faba L. roots. Three peroxidase fractions (zones) moving towards the anode were revealed as well as four peroxidase fractions (zones) migrating towards the cathode. Three peroxidase fractions showed detectable indoleacetic acid oxidase activity. The o-diphenol oxidase activity was revealed in all peroxidase fractions moving towards the anode, in those moving towards the cathode the o-diphenol oxidase activity differred according to the substrate used. One fraction with both peroxidase and o-diphenol oxidase activity occurred only in electrophoreograms of extracts from the maturation zone; in this fraction no indoleacetic acid oxidase activity was demonstrable.     

High Levels of Malic Enzyme Activities in Vicia faba L. Epidermal Tissue

Specific activities of NADP-malic enzyme, NAD-malic enzyme, phosphoenolpyruvate carboxykinase and pyruvate, orthophosphate dikinase in various cells of Vicia faba L. leaflets were determined. Expressed on dry weight, chlorophyll or protein basis, the averages for NADP- and NAD-malic enzyme specific activities were higher in guard cells than in photosynthetic parenchyma cells. Malic enzyme-specific activities were also high in epidermal cells. Phosphoenolypyruvate carboxykinase activity was not detected in Vicia leaf extracts or guard cells; the assay techniques were validated by mixed Vicia-Brachiaria leaf extraction and assays on nanogram samples of Brachiaria bundlesheath cells. It was inferred from these data that guard cell malate depletion is by decarboxylation to pyruvate in the epidermal layer, but how the various epidermal cells interact remains obscure.     

Metabolism of Abscisic Acid in Guard Cells of Vicia faba L. and Commelina communis L

Metabolism of abscisic acid (ABA) was investigated in isolated guard cells and in mesophyll tissue of Vicia faba L. and Commelina communis L. After incubation in buffer containing [G-³H]±ABA, the tissue was extracted by grinding and the metabolites separated by thin layer chromatography. Guard cells of Commelina metabolized ABA to phaseic acid (PA), dihydrophaseic acid (DPA), and alkali labile conjugates. Guard cells of Vicia formed only the conjugates. Mesophyll cells of Commelina accumulated DPA while mesophyll cells of Vicia accumulated PA. Controls showed that the observed metabolism was not due to extracellular enzyme contaminants nor to bacterial action.

Metabolism of ABA in guard cells suggests a mechanism for removal of ABA, which causes stomatal closure of both species, from the stomatal complex. Conversion to metabolites which are inactive in stomatal regulation, within the cells controlling stomatal opening, might precede detectable changes in levels of ABA in bulk leaf tissue. The differences observed between Commelina and Vicia in metabolism of ABA in guard cells, and in the accumulation product in the mesophyll, may be related to differences in stomatal sensitivity to PA which have been reported for these species.

     

Anchoring of genetic linkage maps to the chromosome complement of Vicia faba L.

Using amplification of marker sequences with DNA from a set of distinct microdissected Vicia faba L. chromosomes covering the entire genome, we could unambiguously show that the linkage group I.B, which includes the pseudogene of legumin B4 (ψ1) and was previously ascribed to the metacentric chromosome I, actually belongs to chromosome IV. By considering the breakpoints of the translocated BKH chromosomes III and IV, even the subchromosomal position of loci LG085 and CNGC4 could be inferred. Anchoring all linkage groups to distinct faba bean chromosomes will facilitate quantitative trait locus fine mapping and gene identification using synteny, and will boost the development of efficient markers for selection in breeding programs.     

Comparative electrophoretical analysis of plasmid-like mitochondrial DNAs in Vicia faba and in some other legumes

Minicircular DNAs found in mitochondria of 6-d-old etiolated seedlings of Vicia faba L. were also present in mitochondria isolated from cell suspension cultures and from green leaves of this plant. These results support the suggestion that the plasmid-like molecules found in mitochondria of V. faba are an essential component of the mitochondrial genome. The minicircular DNAs were, apparently, peculiar for the species V. faba since they were found in all three cultivars of this species which were studied. The distribution pattern of plasmid-like DNAs in Vicia villosa L. was completely different and mitochondria from Medicago sativa L. also contained specific minicircular DNAs. Thus, minicircular DNAs are typical for the mitochondrial genomes of several legumes and different plant species have their specific mitochondrial plasmid-like DNAs     

Fractions of non-specific esterase in root tip ofVicia faba L. revealed by disc electrophoresis in acrylamide gel

Fractions of non-specific esterase were studied in homogenates ofVicia faba L. root tips using disc electrophoresis in acrylamide gel. With α-naphthyl acetate, 7 bands were revealed in electrophoreograms, whereas only 5 bands appeared with naphthol AS acetate; the position of bands detected with naphthol AS acetate corresponds to 5 of 7 bands which appear when using α-naphthyl acetate. If incubated with α-naphthyl acetate, 1 band is totally blocked by the inhibitor E 600, whereas the other bands are weakened slightly and—more or less— proportionally, similarly, as if incubated with naphthol AS acetate. No bands appear after detection with α-naphthyl caprylate and with α-naphthyl myristate. Electrophoreograms developed with α-naphthyl propionate differ both from those treated with α-naphthyl acetate and from those treated with α-naphthyl butyrate. Remarkable, but only quantitative differences were revealed when comparing electrophoreograms from division, enlargement and maturation zones, detected with either α-naphthyl acetate, or α-naphthyl propionate. The non-specific esterase fractions revealed by disc electrophoresis are correlated with protein fractions distinguished in the same material by the same technique using xylene brilliant cyanine G staining and with fractions of the same enzyme found in sections of the same object treated histochemically.     

Histological Compartmentation of Phosphate in Vicia faba L. Leaflet : Possible Significance to Stomatal Functioning

Guard cells, epidermal cells, palisade parenchyma cells, and spongy parenchyma cells of Vicia faba L. leaflet were analyzed for inorganic phosphate and phosphorus. On a molar basis, cells in the epidermal layer contained about 15-fold more inorganic phosphate than mesophyll cells did. Although a metabolic role for this asymmetric distribution cannot be defined unequivocally, we note that high epidermal inorganic phosphate would buffer against pH changes in the epidermis during stomatal movements.     

The cytotaxonomic study of theVicia cracca complex 2. The taxa of the southern part of northern Europe

In the southern parts of Scandinavia there occur two species of theVicia cracca L. complex:Vicia cracca L., represented by the chromosome race 2n=28, andV. oreophila ?ertová. WhereasV. oreophila predominantly inhabits habitats of primary character, especially near the coast, the tetraploid race ofV. cracca, which is the commonest in other parts of area, occurs predominantly on sites of secondary character, such as the borders of paths, roads, meadows and bushy places. The two species differ in several characters.     

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.