A non-destructive method of total vertical projections for estimating the length of rigid root systems, not introduced yet in plant sciences, is described. It is demonstrated on measuring less and more dense root systems of seedlings of Zea mays grown at hypoxic or control conditions. Photographs of six vertical projections (30° apart) of each root system were taken and evaluated. The method being based on proved mathematical formula offers unbiased estimation of the length of a rigid root system, curved in three-dimensional space, by non-destructive means. Thus, it is applicable during ongoing experimentation on plants grown in a solution culture. It was shown that less than 120 intersections between the root projection and test lines in one photograph ensured sufficient precision of the method and that the observer subjectivity could be overcome by presented instructions.

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.

Molecular markers were used to detect the influence of high concentrations of 2,4 dichlorophenoxyacetic acid (2,4-D) in the callusing media on DNA variations in regenerated rice plants. Restriction fragment length polymorphism (RFLP), random amplified polymorphic DNA (RAPD) and polymerase chain reaction (PCR) based RFLP analysis were carried out on 12 somaclones of Oryza sativa L. cv. B-370. In vitro culture induced DNA variations were detected in the regenerated plants but the effect of high auxin concentration in the medium could not be revealed. In a second study, fingerprinting of 15 semi-dwarf, high yielding somaclones of B-370 was carried out using RAPD technique. Amplification using 20 random primers produced a total of 167 DNA bands out of which 97 bands were polymorphic. A total of 32 unique DNA bands were detected across all the somaclones and they could be grouped based on their similarity to B-370. RAPD analysis helped to reveal similarity or differences among the somaclones while fingerprinting using additional RAPD markers was not successful.

Somatic hybridization of Brassica oleracea (cauliflower, cabbage, kohlrabi) and Armoracia rusticana (horseradish) protoplasts was performed to test the possibility of the formation of intergeneric hybrids. Out of three B. oleracea crops tested only cauliflower (B. oleracea var. botrytis) yielded viable hybrids when fused with horseradish protoplasts. Regenerated plants were asymmetric hybrids as revealed by morphological, karyological, and DNA analysis. In these plants, extensive elimination of A. rusticana genome occurred during subsequent cultivation. Using a chloroplast DNA probe no A. rusticana-specific hybridization signals were detected thus indicating a rapid elimination of horseradish cytoplasmic (chloroplast) components.

Mutagenesis provoked by exposure to increased concentration of antibiotics of five indigenous Rhizobium galegae strains resulted in the generation of several antibiotic-resistant mutants. The mutants differed from the wild type and one from another in respect to the nodulation capacity, the nitrogenase activity, the nodule ultrastructure, and the plant growth response. Galega plants inoculated with mutants resistant to streptomycin and rifampicin formed nodules with higher nitrogenase activity and accumulated more shoot dry biomass than plants inoculated with the parent strains. Resistance to kanamycin and nalidixic acid was associated with significant decrease of nitrogenase activity. A correlation between nitrogen-fixing efficiency and nodule infected cell ultrastructure was found. When the bacteroids occupied about 10 times higher area in infected cells of nodule than peribacteroid spaces and host cytosol had electron dense and homogenous structure, the nitrogenase activity was the highest.

In Kalanchoë blossfeldiana cv. Tom Thumb the initial rate of ATP-dependent H+-transport into tonoplast vesicles was stimulated up to three times if the H+-ATPase (EC 3.6.1.3) was energized a few minutes after pre-energization of the H+-PPase (EC 3.6.1.1). H+-PPase-activated ATP-dependent H+-transport was observed in plants of K. blossfeldiana cultivated in short day (SD) or long day (LD) conditions expressing different degrees of crassulacean acid metabolism (CAM). However, based on the higher activity and protein amount of H+-PPase and H+-ATPase present in the vacuolar membrane of SD plants the maximum H+-transport activity in the stimulated mode of the H+-ATPase was significantly higher in tonoplast vesicles of SD plants than of LD plants. Hence, a co-ordinated action of the H+-PPase and H+-ATPase at the tonoplast of Kalanchoë could allow a higher transport capacity at the vacuolar membrane when plants perform high CAM. Immunoprecipitation experiments with an antiserum raised against the A-subunit of the vacuolar H+-ATPase of Mesembryanthemum crystallinum L. showed that in SD and LD plants of K. blossfeldiana the H+-PPase was co-precipitated with the vacuolar H+-ATPase holoenzyme. The co-percipitation of the two transport proteins indicates a close structural localization of the H+-PPase and the A-subunit of the vacuolar H+-ATPase.

Total genomic DNA isolated from maize hygromycin B resistant cell line(hygr-G204) was used to transform the maize hygromycin B sensitive cell line(hygs-G204) to the hygr-phenotype using polyethyleneglycol treatment and the transformed calli were selected using hygromycin B. The primary transformant maize plants were regenerated and analysed at the molecular level using DNA hybridization, transgenome rescue and histochemical β-glucuronidase assay. The results indicated that genomic DNA-mediated transformation can lead to transfer, expression and stable integration of a DNA fragment into the host genome.

A screening of leaves of 25 taxa of angiosperms was made for their volatile toxicity against damping-off fungi. The volatile substances fromHyptis suaveolens andOcimum canum were toxic againstPythium aphanidermatum, P. debaryanum andRhizoctonia solani. The fungitoxicity of the leaves persisted for 15 d of storage. The volatile substances from the leaves ofO. canum were thermostable, while those fromH. suaveolens were thermolabile. The essential oils exhibited strong potency against the pathogens tested, non-phytotoxic nature to the host plants and superiority over commonly used synthetic fungicidesAgrosan G.N. andCaptan. The findings indicate the possibility to use these essential oils as potential natural fungicides in management of damping-off pathogens.

Accumulation of the extracellular proteins localized in intercellular spaces of barley primary leaves was examined after inoculation with powdery mildew (Erysiphe graminis f. sp. hordei) as biotic stress factor and after abiotic stresses such as heat shock, low temperature and heavy metal (Mg,Zn, Cu, Al, Cd and Co) treatment using native polyacrylamide gel electrophoresis. Six to eight major pathogen-induced proteins (bands on native gel) have been identified. Their accumulation at host-parasite incompatibility was more expressive than at compatibility interaction. Elevated temperature did not induce pathogenesis-related (PR) proteins while low temperature induced three of them. Cu, Al, Cd and Co induced accumulation pattern of extracellular proteins was very similar to that in powdery mildew inoculated leaves. Mg and Zn had no effect on the induction of protein accumulation in the intercellular spaces of leaves. Induction of PR proteins by different stresses indicated their general function in the resistance of plants to changing environment.

Ten-days-old bean plants (Phaseolus vulgaris L., cv. Cheren Starozagorski) were treated with simulated acid rain (pH 2.4, 2.2, 2.0 and 1.8). Anatomical changes in the primary leaves were studied 3, 48 and 168 h after a single treatment. This treatment induced: 1) change in the shape of palisade cells, contraction of their contact surfaces and expansion of spongy cells (pH 1.8, 3 h after treatment); 2) reduction of symplast connections among palisade cells and of apoplast in the spongy mesophyll (pH 1.8, 48 h after treatment); 3) destruction of adaxial epidermis and portions of palisade mesophyll, plasmolysis of spongy cells (pH 1.8, 168 h after treatment); 4) full destruction of mesophyll (pH 2.4, 2.2, 2.0 and 1.8, 168 h after treatment). The structure of abaxial epidermis was more stable than that of the adaxial one. With respect to anatomical parameters the studied species could be considered as comparatively resistant to acid rain.

An efficient protocol was developed for in vitro plant regeneration via somatic embryogenesis from cell suspension cultures of metal tolerant grass Echinochloa colona (L.) Link. Callus was obtained by culturing leaf base on MS medium supplemented with 0.5 mg dm^-3 of 6-benzylaminopurine (BAP) and 2.0 mg dm^-3 of 1-naphthaleneacetic acid (NAA). Cell suspensions were initiated and established in MS liquid medium containing 0.5 mg dm^-3 BAP, 1.0 mg dm^-3 NAA and 2.0 mg dm^-3 2,4-dichlorophenoxyacetic acid (2,4-D). A reduction in the concentration of 2,4-D to 0.5 mg dm^-3 induced formation of somatic embryos. The embryos developed and grew into normal plants in the presence of half strength MS medium without growth regulators. The regenerated plants were hardened in the greenhouse and subsequently grown in the open. This system may be also used for isolation and culture of protoplasts as a first step in somatic hybridization.

Changes in hexokinase particulate and soluble isozyme composition and activities in leaves of 65- and 115-d-old tobacco plants were determined by ion exchange chromatography on DEAE cellulose. During plant ageing, the activities of glucose and of fructose phosphorylating isozymes of particulate hexokinase decreased to 9.9 and 9.2 % of initial value, respectively. The activity of soluble hexokinase decreased to a lesser extent: that of glucose phosphorylating isozyme to 49.8 % and of fructose phosphorylating isozyme to 37.8 %. The activity of soluble fructokinase isozyme dropped to 34.8 %. Thus also the ratio of particulate and soluble isozymes was dependent on the age of leaf tissue.

The growth retardant paclobutrazol (PBZ) inhibited stem internode growth of in vitro cloned potato plants. The extent of growth inhibition caused by10-9 M PBZ in Murashige and Skoog medium was genotype-specific, varying between 10 - 60% of the stem growth of untreated controls in ten cultivars examined. An increase in percentage of de novo bud regenerating stem internode segments (SIS) as well as in the total number of buds per explant was observed in SIS taken from PBZ pretreated plants. PBZ applied directly into the regenerative media had no stimulatory effect on there generation process. We assume that the enhancing effect of PBZ on regeneration may be attributed to its interaction with cytokinin metabolism.

The physiological responses of alfalfa (Medicago sativa L. cv. Gilboa) to salinity (100 mM NaCl) and some inorganic nutrients (K+, Ca2+ and N as NO3-) were investigated. Salinity caused a substantial reduction in biomass, carbon assimilation rate, stomatal conductance, water use efficiency, leaf area, relative growth rate, NO3- content and nitrate reductase activity, whereas, transpiration rate was slightly affected. Inclusion of K+, Ca2+ and N as NO3- in plant nutrient medium in combination or alone brought about a marked stimulation in control plants and moderated the salinity caused reductions in NaCl treated plants. In addition, plants also exhibited differences in these parameters at two growth stages.

The ultrastructure of chloroplasts in the primary leaf of 10-d-old bean plants (Phaseolus vulgaris L., cv. Cheren Starozagorski) was studied 3, 5, 24, 48, 72 and 168 h after a single treatment with simulated acid rain (pH 2.4, 2.2, 2.0 and 1.8). Different changes in chloroplast structure till full destruction of organelles were traced. A determining factor for these changes was the histological localization of chloroplasts. In the chloroplasts of palisade parenchyma different degrees of expansion of thylakoids (3, 5, and 24 h after the single treatment), and conformational changes of the inner membrane system (48, 72 and 168 h) were observed. The chloroplasts of spongy parenchyma showed a significantly higher degree of structure resistance. The expansion of thylakoids was weak and did not depend on the duration of treatment. The ultrastructural changes of chloroplasts confirmed relative resistance of this species till pH 2.0.

Dry matter production of shoots and roots and the diurnal fluctuation of titratable acidity of single leaves were investigated in the CAM plant Agave attenuata during the first 70 d after germination. The plants were grown either in vermiculite sub-irrigated with a nutrient solution or in in vitro cultures on an inorganic nutrient agar. Two types of culture tube covers were used: either airtight closures or polypropylene caps with membranes permeable to air.
In the earliest ontogenetic phases of development (cotyledon and primary leaf stage), the plants were already able to carry out considerable nocturnal organic acid accumulation. In vitro cultivated plants, from the beginning of their development, were also capable of diurnal acid fluctuation, though of distinctly weaker activity than the pot plants. The mean relative growth rates (RGR) of pot culture plantlets approached a third of perennial herbaceous plants. Plantlets grown in in vitro culture reached only half to the one quarter of the RGR of pot plants. The reduced yield could be attributed to the low CO2 supply in the culture tubes and the less than optimal water and nutrient supply in the agar medium.

Changes in growth and anatomical structure of vascular tissues in stem, root and leaf of safflower plants grown in NaCl and CaCl2 solutions having different osmotic potentials (ΨS from 0 to -0.9 MPa) with addition of 0, 10 and 20 mg dm-3 zinc were studied. Shoot and root lengths, fresh and dry masses and fresh/dry mass ratio were lower in salt-stressed plants compared to unstressed plants. Salinity induced structural changes in stem, root and leaf tissues; few xylem vessels with smaller size were noticed in stressed plants. The higher concentration of Zn improved growth especially in roots and enhanced xylem formation in comparison to stressed plants grown at the same osmotic potential without Zn. Zn also protected xylem distructure by salinity in leaves.

Transient expression of the β-glucuronidase (GUS) gene introduced into Arabidopsis thaliana intact plants by T-DNA after vacuum infiltration of Agrobacterium tumefaciens was followed. The first incidence of GUS activity was found 2 - 3 d after treatment and a peak of activity one week after treatment in both A. thaliana races, Columbia and C24. GUS activity was sharply increased by cultivation of Arabidopsis plants at elevated temperature (29 °C) compared to cultivation at 25 °C. The density of inocula also influenced the GUS activity.

Eichhornia crassipes plants brought from the River Nile were cultured in jars containing river water supplemented with various concentrations of Cd, Pb, and Sr (0 to 100 μg cm-3), added simultaneously. Treatment continued for 20 d during which each cultivation solution was being replaced with fresh one every 3 d. The growth of Eichhornia was drastically retarded at heavy metal concentrations higher than 15 μg cm-3. At concentrations 15 or 25 μg cm-3, the accumulation of Cd and Pb to levels several times higher than those in control plants was found. More than 50 % of the uptaken metals were retained by roots alone. Leaves and leaf petiols received around 30 and 20 % of the accumulated metals, respectively. X-ray microanalysis indicated the presence of the three heavy metals in Ca oxalate crystals. Content of metals in the crystals increased progressively over time of exposure in a way similar to those in whole plant tissues. These results suggest a possible role for Ca oxalate crystalization in toxic heavy metal deposition and thus tolerance by Eichhornia.

Standard methods of isolation of chromatin histones from Arabidopsis thaliana yield strongly degraded proteins when applied to plants grown from seeds in axenic liquid media. For isolation of undegraded histones from Arabidopsis grown in liquid media we used extraction with guanidine hydrochloride followed by selective binding of histones on BioRex 70 resin in the batch system. The quality of obtained proteins is confirmed by SDS polyacrylamide gel electrophoresis.

The effects of Cd, Ni and Pb on the growth, chlorophyll (Chl) and protein contents, and content of proteases of potted weed plants Cyperus difformis, Chenopodium ambrosioides and Digitaria sanguinolis were determined. The three heavy metals inhibited the shoot growth but were less suppresive to root growth. They also lowered leaf Chl content. The changes in root and shoot protein and proteases contents of weeds were interrelated. The heavy metal additions to soil increased their contents in both roots and shoots, several times more in roots than in shoots.

The effect of low irradiance on three rice cultivars (shade tolerant cvs. Swarnaprabha and CO 43 and shade susceptible cv. IR 20) was studied. The large subunit (LSU) of ribulose-1,5-bisphosphate carboxylase/oxygenase with molecular mass of 55 kDa was reduced in cv. IR 20 grown under low irradiance (LI). Native protein profile studied showed, under LI, reduction in the contents of proteins with RF values 0.03, 0.11 and 0.37. Analysis of chloroplast polypeptides revealed an induction of light-harvesting chlorphyll-protein 2 (LHCP2) under shade. The induction was more expressed in cv. CO 43 than in cv. IR 20. Under LI, in vivo labelled protein bands in the molecular range of 26 - 27 kDa were induced. These proteins were highly turned over in the LI-grown plants of cv. CO 43 than in cv. IR 20. A signal for rbcL gene sequences in EcoRI digested lanes was also found. Isozyme analysis of peroxidase showed an induction of a new band with RF 0.43 in cv. IR 20 subjected to LI.

Transfer of chloroplasts from Solanum tuberosum into S. nigrum cell resulted in an atrazine sensitive cybrid plant. The shoots of this cybrid were bleached under atrazine stress. The cybrid displayed identical isoenzyme patterns that have been found in S. nigrum, and thus nuclei of the cybrid plant cells did not integrate any chromosomes or chromosome fragments from S. tuberosum nuclei. Under atrazine stress, differences in the isoenzyme expression were found in both the cybrid and original plants. In the peroxidase patterns, POX-4 was detected while POX-1 disappeared. Esterase patterns were less influenced, EST-4 was expressed in both plants under the influence of atrazine. Atrazine treatment of both cybrid and original plant shoots had specific effects on carbonic anhydrase, malate dehydrogenase, and glutamate oxaloacetate transaminase. The number and/or the staining intensity of bands corresponding to these isoenzymes decreased under atrazine stress more in cybrid plants (atrazine sensitive) than in S. nigrum (atrazine resistant).

Shoot and root fresh and dry matters and their Cu content were determined in 7-d-old seedlings of Triticum aestivum L. cv. Alkora treated with Cu (10,20, 40 µg cm-3) and Si (500 µg cm-3). Si significantly reduced the toxic effect of Cu on fresh and dry matter production of wheat seedlings. Moreover, plants treated with Cu and Si absorbed less Cu from the solution and had higher water content in shoots and roots than that treated with Cu only.

Indoor experiments demonstrated that allelopathic potential of rosette and flowering plants of qort is an important factor explaining the growth reduction of its associated species. Aqueous tissue extracts of flowering plants exhibited strong inhibitory effects on the germination percentage and radicle growth rate of the tested species as compared with those of vegetative plants. Under laboratory conditions, this inhibition was in agreement with toxicity assessments of soil samples collected from the rhizosphere of T. resupinatum L., where shoot and root dry mass of the tested species were significantly reduced. Detoxification of allelochemicals by presence of activated carbon can eliminate the inhibitory effects of the different extracts. This technique clarifies the occurrence of allelopathic interference by qort on seed germination and seedling growth, and hence suspects the allelopathic potential of qort in the growth reduction of associate species under field conditions along with competition.

Pot experiments were conducted to evaluate the possible interaction of salinity (osmotic potential -0.3, -0.9 and -1.2 MPa) and occurrence of Azospirillum lipoferum or exogenous gibberellic acid (GA₃) (100 µg g^-1) on growth and some physiological parameters of maize. ¹⁵N-uptake as well as the percentage of nitrogen derived from ¹⁵N-fertilizer were decreased by increasing the NaCl concentrations and completely inhibited at concentrations corresponding to osmotic potentials -0.9 and -1.2 MPa. The percentage of nitrogen originating from N₂ fixation was significantly correlated to the total counts of Azospirillum cells that colonized the histosphere. At high NaCl concentrations although no significant changes in N % in shoot dry mass either in inoculated or uninoculated plants were observed, the total N-yield [mg(N) pot^-1] was decreased. Fresh and dry shoot mass significantly increased by Azospirillum inoculation. Azospirillum and GA₃ treatments were positively correlated with most of the parameters analysed. Azospirillum inoculation or GA3 application at NaCl concentrations up to -1.2 MPa significantly increased the chlorophyll, K, Ca, soluble saccharides and protein contents as compared with control plants.

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.

A reliable protocol for the regeneration of onion through repetitive somatic embryogenesis was established. Embryogenic callus was derived from mature seeds on Murashige and Skoog (MS) medium supplemented with 2 mg dm-3 2,4-dichlorophenoxyacetic acid (2,4-D). Somatic embryos aroused on the surface of calli cultures and formed plantlets after the removal of 2,4-D or its substitution with 1 mg dm-3 kinetin (Kin). Reculturing the somatic embryos on 2,4-D containing medium led to secondary embryos formation. The embryogenic cultures which were preserved for five months on maintenance medium containing 2 mg dm-3 2,4-D + 0.5 mg dm-3 Kin have retained their ability for regeneration, while those kept on 2,4-D only, failed to form plantlets. Electrophoretic analysis of total soluble proteins revealed that the competence for successful conversion of somatic embryos into plantlets is associated with the expression of new set of proteins (112, 58 and 30 kD). The regenerated plants were successfully transferred to the soil.

During the germination, wheat and sorghum plants tolerated salinity up to 10 mM NaCl. Seedling growth and dry matter production remained more or less unchanged up to 10 mM NaCl in case of wheat and up to 5 mM NaCl in case of sorghum. The water content of test plant species exhibited nearly constant values irrespective of the salinity level applied. The proline and carbohydrate content increased with elevating NaCl, while free amino acids content decreased. Irrigating of seedlings with tryptophan had non significant effect at all salinity levels used.

Differences in anatomy and morphology of the kiwifruit leaves and leaf petioles might play a considerable role in the sex-determination. Three months after bud break (June), the kiwifruit leaves of both male and female plants, grown on the vegetative and generative shoots showed different leaf area (128.6 ± 13.45 cm2 in male and 104.5 ± 4.02 cm2 in female plants) and shape. The most frequently leaf shape was determined as "folium cordatum" and "folium rotundato-cordatum". Higher values of total leaf thickness of the female leaves (190 ± 3.84 µm) in comparison to male leaves (174 ± 3.52 µm) were estimated, resulting in the thicker adaxial leaf epidermis and especially in thicker palisade parenchyma in female leaves (136 ± 2.76 µm in comparison to 104 ± 1.61 µm in male leaves). Typically bifacial leaves were observed in both male and female leaves. Anomocytic stomata in hypostomatic leaves were found. The reticulate venation appears to be the main type of leaf venation. Stalked stellate multicellular trichomes on the abaxial leaf side were frequently observed in the leaves of both sexes. No important differences between male and female plants were found in the structures of vascular system in leaves and leaf petioles. Thus leaf thickness and surface morphology of adaxial leaf epidermis can be considered as important structural parameters in the sex determination.