Effect of fluoride (10 and 50 mM) on the activities of sucrose metabolizing enzymes, alkaline inorganic pyrophosphatase, and transaminases in relation to the accumulation of free sugars, starch, and soluble protein was studied in detached ears of wheat cultured in a liquid medium. Culturing for 5 d in the presence of fluoride reduced the amount of grain starch whereas contents of total free sugars, particularly sucrose, and soluble protein increased. Fluoride inhibited the activities of soluble acid and neutral invertases, as well as sucrose synthase acting in the cleavage direction. Uptake of uniformly labelled ¹⁴C-sucrose or fructose was also drastically reduced by fluoride. Glutamate-oxaloacetate transaminase (GOT) and glutamate-pyruvate transaminase (GPT) activities also increased with fluoride addition in correspondence with an increase in soluble protein. Apparently, the wheat grain responds to fluoride-mediated disruption of carbon metabolism by a compensatory effect on nitrogen metabolism.

The influence of the applied photoperiod during floral stem development has been studied. A clear relationship between the length of the photoperiod, the frequency of light/dark switches, and the dynamic of growth and the subsequent morphology and architecture has been found. An increase in the day length (L/D = 16/8) or an increase in frequency of light/dark transitions (L/D = 7/7) stimulated the inflorescence growth. The two treatments induced the same enlargement in the early inflorescence, but the L/D = 7/7 condition causes a greater increase in the late one. The treatments induced also a diminution in the number of lateral inflorescences, principally with the high frequency light-on/light-off condition. Moreover than differences in the growth along the early inflorescence, at the level of the internode, a gradual growth was observed from the basal part to the upper part of the internode. Finally, the floral stem growth dynamic showed that it could be adapted to the environmental light/dark signalling by a good extension rate synchronisation.

Changes in ribonucleases (RNases) and glucose-6-phosphate dehydrogenase (G6P DH) activities, their content and subcellular localisation were studied in relation to virus multiplication in susceptible (cv. Samsun) or resistant (transgenic breeding line NCTG 83) tobacco plants infected with the potato virus Y^N (necrotic strain of PVY). Activities of RNases and G6P DH from diseased susceptible tobacco plants were markedly increased during the experimental period and significantly correlated with the multiplication curve of the PVY^N. In contrast, the activities of RNases and G6P DH were not changed after PVY inoculation of resistant breeding line NCTG 83 producing the CP mRNA of PVY. Changes in the content and in the subcellular localisation of RNases and G6P DH isozymes were also determined in mesophyll protoplasts isolated from healthy as well as PVY^N infected plants of both cultivars by differential centrifugation of broken protoplasts on day eight post inoculation (the culmination of multiplication curve of PVY and enhanced activity of both enzymes). The chloroplasts fraction from infected protoplasts showed an enhanced content of RNases (192.4% when compared with that from healthy control ones), and of G6P DH (174.4 %). The cytosol fraction from infected protoplasts contained slightly enhanced levels of G6P DH (117.4 %) and considerably enhanced levels of RNases (141.7 %). No significant differences in the activities, contents and subcellular localisation of RNases and/or G6P DH isozymes were observed in the resistant line NCTG 83. This is in accordance with no detectable contents of PVY.

Gemma morphology, histology and ultrastructure before and after germination in vitro were studied in Drosera pygmaea. The histology of the gemma is similar to that of a seed, being characterized by an embryo-like structure and storage tissue, although no seed coat is formed. One embryo-like structure within the gemma, which gives origin to a new plant, expresses polar organisation with distinct meristematic regions. Storage tissue surrounding the embryo-like structure resembles endosperm and it is built of parenchyma cells possessing plastids with starch grains and dense material within vacuoles. The regeneration from the gemma may provide useful system to study plant morphogenesis under stress conditions including in vitro culture.

Expression of esterase isoenzymes in 1272 seeds was used to estimate hybridity of Lycopersicon esculentum Mill. Individual seeds (440) of the parental cultivars taken from different experimental stations in Bulgaria were also analysed. The banding patterns were obtained by means of vertical block electrophoresis in polyacrylamide gels. It was established that quantitative variation of locus Est-1 can be applied to prove hybridity of F₁ tomato seeds. This marker is related to the genetic nature of tomatoes and is not the result of the environmental influence. The reason for this conclusion is the fact that the isoenzymes of the Est-1 locus are indicative for hybridity determination of all examined seeds taken from different regions in Bulgaria. Use of this locus is to be recommended for both its universality and the fact that the reagents for esterase isoenzymes staining are not expensive.

Nodulation was inhibited in plants of green gram (Vigna radiata, cvs. ADT-1 and CO-5) exposed to different levels of simulated acid rain using a mixture of H2SO4, HNO3 and HCl (6:3:1) of pH 2.5, 4.0 and 5.5 in comparison with control (pH 7.0). Protein content of leaves increased in cv. CO-5 but decreased in cv. ADT-1 whereas the nitrate content of leaves increased in cv. ADT-1 but lowered in cv. CO-5. Nitrate reductase activity was increased in the nodular roots of cv. ADT-1 but was decreased in leaves. In cv. CO-5 it was increased in leaves but was insignificantly reduced in the nodules at pH 2.5. The nodule nitrogenase activity increased at pH 4.0 and 2.5 in cv. ADT-1.

The specificity of a monoclonal antibody IgG1, raised against trans-zeatin riboside-keyhole limpet hemocyanin conjugate, was investigated by means of inhibition experiments with soluble competing antigens. A competitive enzyme immunoassay was set up, with immobilized antigen. The analysis of the cross reaction profile enabled a study of the specificity of the antigen-antibody interaction. The antibody was able to distinguish the trans form of zeatin riboside from the cis form (cross reaction index = 1 %); cross reactions with ribose, adenine, adenosine and other related heterologous antigens were not detectable over the range of concetration tested. The recognition centres for the antibody seem to be the purine ring and the R substituent, especially in its hydroxymethyl group. Employment of this monoclonal antibody to localize cytokinins in control and shedding affected fruits of Malus domestica Borkh. evidenced high content of trans-zeatin riboside in developing seeds, differences in its content in embryo and endosperm, and a strong reduction of its content in the tissues of drop fruits. This decrease may be an important component responsible for early fruit abscission.

Gerbera jamesonii plants were subjected to a drying and rewatering for 10 d under greenhouse conditions. Transpiration rate and leaf water potential decreased with the application of stress and recovered to a level similar to that observed in the control plants. Leaf abscisic acid concentration increased while ethylene production decreased under stress. After rewatering, each of the parameters recovered, to similar levels, as in the control.

An attempt to reduce the production cost on tissue cultured plants, photoautotrophic culture of a high value orchid Dendrobium was established under CO2-enriched conditions. The shoot length and the number of leaves were almost equal in plantlets grown on medium with 2 % sucrose or without sucrose and under normal or enhanced (40 g m-3) CO2 concentration, whereas the fresh and dry masses were higher in cultures grown in sucrose containing media or under CO2 enrichment. Development of roots was observed only on media without sucrose, but CO2 enrichment did not have significant effects on in vitro rootings.

Growth and Na⁺, K⁺, Cl^-, proteins, sugars and proline concentrations were measured in three triticale genotypes M2A, DF99 and Asseret grown on nutrient solution with or without 75 mM NaCl. In saline conditions, leaf area of the three triticales was reduced by 50 % and dry to fresh mass ratio increased. Total protein concentration was diminished by 10 %. K⁺ concentration decreased whereas Na⁺ and Cl^- accumulated in roots and shoots of salt-stressed plants. This ion accumulation was greater in roots of Asseret than in roots of the other triticales. Soluble sugar concentration increased in M2A and Asseret and decreased in DF99. Proline concentration increased in M2A and DF99 and decreased in Asseret. Osmotic adjustment was essentially realized by Na⁺ and Cl^- uptake. Non-reducing sugars and proline contributed too, but to a lesser extent.

The effect of 2 mM AlCl3 on NAD+ kinase (E.C. 2.7.1.23) activity was studied using Euglena gracilis strain Z grown heterotrophically in darkness at pH 3.5 in the presence of lactate as sole carbon source. The Al-treatment slowed down the culture growth and suppressed the peak of NAD+ kinase activity, which characterizes the beginning of the exponential phase of growth of the control cell cultures. There are two possible explanations of the Al effect: it 1) either prevents the enzyme activation by the Ca-calmodulin (CaM) complex; or 2) suppresses the CaM-dependent NAD+ kinase form. In Euglena cells, a part of the NAD+ kinase activity is enhanced by EGTA and lowered by Ca2+: this peculiar NAD+ kinase activity is unaffected by the Al treatment.

Fluorescent in situ hybridisation (FISH) was used to determine the number and distribution of the 18S-25S and 5S rDNA sites on mitotic chromosomes of 6 wild and 2 edible diploid (2n=22) accessions belonging to the two banana species, Musa acuminata and M. balbisiana. FISH with the 18S-25S probe resulted in signals on one pair of chromosomes, the position of signals corresponded to the secondary constriction at the end of a short arm. The intensity of labelling was different between the homologues and the larger site corresponded to a larger secondary constriction. This labelling pattern was observed consistently in all genotypes. On the other hand, differences in the number of 5S sites were observed between the accessions. While in some of the wild seeded species, the 5S rDNA was localised on two pairs of chromosomes, hybridisation signals appeared on three pairs of chromosomes in other wild accessions. Quite unexpectedly, only five sites of 5S rDNA were reproducibly observed in the two vegetatively propagated diploid edible cultivars, Pisang Mas and Niyarma Yik, evidence for structural heterozygosity. A dual colour FISH showed that in all accessions, the satellite chromosomes carrying the 18S-25S loci did not carry the 5S loci. The results demonstrate that molecular cytogenetics can be applied to Musa and that physical cytogenetic maps can be generated.

Adventitious root formation in vitro in 1-mm stem slices cut from microshoots of apple cv. Jork 9 was studied using light and electron microscopy. When indole-3-butyric acid (IBA) had been added to the medium, starch grains accumulated during the first 24 h of culture in cells of the cambial region and in cells in the vicinity of vascular tissue and in the primary rays. This accumulation occurred only in the basal part of explants. After that, the nuclei in these cells were activated, and the density of the cytoplasm and the number of cell organelles increased, whereas starch was broken down. Cambium cells started to divide transversely and at 96 h, after several divisions, a continuous ring of isodiametric cytoplasmic cells had appeared around the xylem near the basal cutting surface. The cells in this ring were rich in cell structures, and did not contain large starch grains and a central vacuole. Root meristemoids regenerated from the portions of the ring that were localized in the primary rays. From the other cells in the ring, callus developed. The meristemoids did not grow into the direction of the epidermis as in shoots, but along the vascular bundles. After emergence from the cutting surface, the meristemoids were transformed into small, dome-like primordia. They developed a typical root apex with root cap, root ground meristem and tracheid connection with shoot vascular tissue.

We studied the influence of ethephon (2-chloroethylphosphonic acid) on the first stages of development of the main apex of Liatris spicata cv. Callilepis and its relation to the mobilization of soluble saccharides in the corm. Ethephon enhanced cell expansion during the first weeks of cultivation but did not affect the timing of flower initiation. Soluble saccharide concentration in the corm reached a maximum after sprouting, probably due to rapid growth of the shoot. There was no change in the concentrations of soluble saccharides immediately prior to flower initiation, which suggested that quantitative changes in these saccharides were not responsible for activating flower initiation. When corms were treated with ethephon there was a greater degree of soluble saccharide mobilization during the first weeks of cultivation than in untreated corms. However, this did not seem to be the direct result of the addition of this phytohormone and was more probably the consequence of cell growth being encouraged.

The content of cytokinins especially zeatin nucleotide decreased in shoots as a result of temperature increase. Simultaneously the cytokinins accumulated in roots. The changes in cytokinins distribution were followed by a decline of evapotranspiration after its initial temperature-induced uprising.

Developing fruits on lower nodes were major reproductive sink for photosynthates in bell pepper (Capsicum annuum L.). The removal of these fruits resulted in faster growth of other fruits on upper nodes. Sink manipulated plants have the higher rate of net photosynthetic rate at the later stages of plant development.

Four clones of Malus domestica Borkh. cv. Golden Delicious - namely Smoothee, Crielaard, Reinders and Golden B - were cultured in vitro from single-node microcuttings placed on solid medium under irradiance (PPFD) of 50 µmol m-2 s-1. After 9 months an average shoot proliferation of 5.3 was achieved; Crielaard showed the highest rate (7.1), followed by Golden B(5.4), Smoothee and Reinders (4.4). Proliferating shoots were then exposed to higher PPFD (80 µmol m-2 s-1) and different spectral composition of radiation using PMMA-B and PMMA-R/FR filters. High PPFD decreased the average proliferation rate to 4.5, in particular in Crielaard and Golden B, while it increased proliferation in Reinders. When a PMMA-R/FR filter was interposed, the mean proliferation rate slightly increased. PMMA-B filters decreased the overall proliferation rate to 3.0; only in Crielaard it was increased, but shoots were very small. Thus PPFD and spectral composition influenced in vitro shoot proliferation and growth and the responses were different among the clones.

The uptake of K⁺ by the leafy shoots of four submersed higher aquatic plants (Elodea canadensis, Ranunculus aquatilis, R. trichophyllus, and Callitriche hamulata) with different HCO₃ ^- affinity was measured in successive 2-h periods under the conditions of high or low photosynthetic rates (i.e. at pH 7.5 or 10). At pH 7.5 the uptake of K⁺ by species with the higher HCO₃ ^- affinity (E. canadensis, R. trichophyllus) was significantly faster than that by species with a lower HCO₃ ^- affinity (R. aquatilis, C. hamulata). In the former group of species, the K⁺ uptake rate at pH 7.5 was 1.7 - 3.5 times higher than at pH 10. At pH 10, the soft-water species, R. aquatilis, had the lowest net photosynthetic rate (PN) of the three HCO₃ ^- users but, in contrast to the relative hard-water species, R. trichophyllus, showed a small K⁺ efflux (47 nmol kg^-1 s^-1). Thus, K⁺ uptake by shoots was not strictly correlated with PN. A significant K⁺ efflux (73 - 86 nmol kg^-1 s^-1) occurred from all HCO₃ ^- users in darkness. The relatively low K⁺ uptake by the strict CO₂ user, C. hamulata, was quite independent of PN and light or darkness. It may be suggested that uptake of K⁺ by shoots of submersed plants depends on their HCO₃ ^- affinity.

Forty-one accessions belonging to ten genera of the tribe Triticeae representing both wild and cultivated species were analyzed by polymerase chain reaction (PCR). Of two consensus tRNA primers tested, one primer revealed characteristic amplification products for all of the species. A total of 35 tRNA markers were scored across all accessions. Five genus-specific and three species-specific markers were obtained. Genomic fingerprints were largely conserved within a genus. The phylogram obtained using parsimony has separated most of the accessions into their prevailing taxonomic species and genus groups. The phylogram showed close association among the three genera Secale, Triticum and Hordeum as expected. The Triticum-Secale relationship was closer than the Triticum-Hordeum or the Secale-Hordeum relationships. The tree also reflected the close associations among the forage grass species belonging to Leymus and Elymus. Thus tDNA-PCR helped to identify species and genera.

Plant regeneration was achieved from coleoptile tissue of wheat (Triticum aestivum L. cv. Kharachia-65). Coleoptiles (1.0 - 3.5 cm long) were excised from 2- to 5-d-old seedlings and cultured on Murashige and Skoog's (MS) medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D - 0.5, 2.5, and 5.0 mg dm^-3). Cream, friable callus was obtained after 6 weeks of inoculation. This callus was sub-cultured on MS medium supplemented with 2,4-D (2.5 mg dm^-3) and 5 % coconut water. After 6 weeks of sub-culturing white, cream or pale, friable, nodular callus was obtained. Plant regeneration occurred when this callus was sub-cultured on MS medium supplemented with 0.2 mg dm^-3 1-naphthalene acetic acid + 1.0 mg dm^-3 6-benzylaminopurine. For rooting, regenerated shoots or plantlets were transferred on MS medium supplemented with 0.5 mg dm^-3 indole-3-acetic acid. Rooted plantlets were directly transferred into pots and grown under field conditions. Seed setting invariably occurred in all plants.

Plants of teak (Tectona grandis L.f.) growing after transplantation to fertile soil for 5 - 6 months were subjected to water stress by withholding watering continuously for 3 weeks. The growth rates of both plants in height and developing leaves in length were unaffected during the first week after withholding watering, but they were decreased by about 50 % during the second week and became negligible during the third week of water stress treatment. The rate of leaf production and internodal elongation were also decreased in plants experienced 2 weeks of water stress. However, after rewatering, these plants regained growth potential and exhibited high rates of leaf expansion and plant growth comparable to those of well-watered plants. Diurnal course of net photosynthetic rate (PN) of plants subjected to water stress for 2 weeks was similar as that of well-watered plants. However, PN of plants subjected to water stress for 3 weeks was reduced in the afternoon. Similarly, stomatal conductance (gs) and transpiration rate (E) of plants experiencing 3-week water stress were decreased in the afternoon. Soon after rewatering, PN, gs and E reached similar values to those of well-watered plants.