Multiple shoot regeneration from the cut plumular ends of embryo axes of chickpea (Cicer arietinum L.) was evaluated on Murashige and Skoog medium having different concentrations of thidiazuron (TDZ) (0.1 to 10.0 mg dm^-3) 6-benzylaminopurine (BAP) (0.5 and 1.0 mg dm^-3), kinetin (0.5 and 1.0 mg dm^-3) or zeatin (2.0 and 4.0 mg dm^-3). TDZ (0.2 mg dm^-3) was found to be the most effective cytokinin as it produced multiple shoots in 100 % of the explants from genotypes C235, ICC5166, ICC12269, ICC4951, ICC11531, BG256 and a local cultivar. Shoots were elongated on growth regulator-free medium, and rooted on growth regulator-free medium containing 1/4 MS salts + full vitamins + 3 % sucrose. Plantlets formed were acclimatized for 12 - 15 d in MS medium with a gradual reduction in sucrose concentration and transferred into pots filled with soil and kept in the field; this resulted in more than 70 % survival. The plants developed normally and produced fertile flowers and set seeds. Low temperatures, maximum 19.0 °C, and minimum 8.2 °C, during the first 15 d of transfer favoured survival on transfer to pots.

This study assessed the effects of salinity and drought upon leaf construction (CC) and maintenance (MC) costs in Avicennia germinans growing under contrasting soil salinities in the field. Additionally, an analysis of leaf chemical composition was carried out to determine the underlying reasons for possible variation in CC. With the increase of external salinity, leaf osmotic potential (ψ_s) declined and a significant negative correlation was found between both variables. Changes in average CC were about 15 % when ψ_s changed by 85 %. Despite this, leaf ash-free heat of combustion per unit of leaf dry mass remained relatively constant at 21.3 ± 0.9 kJ g^-1 in all conditions. The estimated maximum and minimum CC for the synthesis of protein present in leaves increased significantly with total leaf CC. Conversely, the estimated CC for lipid synthesis shows a tendency to decrease with total leaf CC. Under conditions of high salinity, ash content increased by 78 % compared to that of leaves collected at low salinity. It is concluded that undergoing changes in proteins and lipids (expensive) and ash (cheap) explain the changes in CC under contrasting salinities. Additionally, the average MC remained almost constant at moderate salinity and declined by 6 % under hyper-saline conditions. Thus, differences in CC and MC with salinity are explained in part by changes in the chemical composition of different compound fractions.

Influence of supra-optimal concentrations of N on growth and accumulation of N, K, P and Ca in the shoots and roots in Pennisetum glaucum (L.) R.Br. under water stress was assessed in a pot experiment under glasshouse conditions. Thirty four-day-old plants of two lines, ICMV94133 and WCA-78, were subjected to 224, 336, or 448 mg(N) kg^-1(soil) and soil moisture 100 or 30 % of field capacity for 30 d. Increasing soil N supply decreased growth of both lines under water deficit. Nitrogen content in the shoots of both lines was not affected by supra-optimal levels of N or different watering regimes, but in contrast, the root N content was increased consistently in WCA-78 with increase in soil N content. Shoot P content increased considerably in WCA-78 at the two higher N contents, but it was significantly lower at drought stress than at well-watered treatment. In contrast, shoot or root P content in ICMV94133 did not differ under both watering regimes. Potassium content in the shoots of WCA-78 was considerably increased at the two higher N contents under drought conditions. Root K content was increased in WCA-78 at the highest N content under well-watered conditions, whereas the reverse was true in ICMV94133. Calcium content in the shoots of ICMV94133 was higher under drought stress compared with that at well-watered conditions, but such pattern was not observed in WCA-78. However, root Ca content increased in both lines with increase in N supply.

Hairy roots of Rhamnus fallax Boiss. were induced using Agrobacterium rhizogenes strain A4M70GUS. The culture established on Woody plant media (WPM) showed a typical hairy root phenotype: rapid growth, reduced apical dominance and root plagiotropism. Seven clones of R. fallax were selected on the basis of their differences in colour and the root branching. The growth of hairy root culture, measured through gain in fresh mass, was done under 16-h photoperiod or in the dark. An increase in anthraquinone (AQ) content was obtained in clones with yellow and less branched roots, like clone 1 [16.43 mg g^-1(d.m.)] and clone 7 [14.21 mg g^-1(d.m.)], compared with other analysed transformed and non-transformed tissue. This study presents the first report of successful transformation of any species from family Rhamnaceae by A. rhizogenes and analysis of AQ production in transformed tissue.

Purpose of this study was to investigate different responses of two wheat genotypes (Triticum aestivum L.) from the wet and dry climate regions to exogenous abscisic acid (ABA) application under well-watered and water-stressed conditions. Exogenous ABA was applied to the leaves by spraying and changes in dry matter accumulation and allocation, endogenous ABA content and carbon isotope ratio (δ¹³C) were monitored. The ABA application significantly decreased stem height, total biomass, total leaf area, total grain mass and leaf area/mass ratio, and significantly increased root/aboveground biomass ratio, endogenous ABA content and δ¹³C under well-watered and water-stressed conditions. Compared with the wet climate genotype, the dry climate genotype was more responsive to exogenous ABA application, resulting in lower stem height, total biomass, total leaf area, total grain mass and leaf area/mass ratio, and higher root/aboveground biomass ratio, endogenous ABA content and δ¹³C under all experimental treatments.

For screening tolerance of six cultivars of rice (Oryza sativa) to different concentrations of nickel (0, 6, 12, 18, 24 and 30 μg dm3), growth parameters (root and shoot length, root and shoot dry matter production) and root and shoot tolerance indexes were tested after 7, 10 and 13 d of treatment. In presence of nickel in nutrient solution, root growth of cv. Nilgiri was enhanced, while in cvs. Subhadra, Khandagiri, Rudra, Sankar and Annapurna it decreased. The root tolerance index (RTI) and shoot tolerance index (STI) in cv. Nilgiri were high; cvs. Sankar and Khandagiri, however, showed a low RTI. Based on the standard growth parameters, six cultivars of rice were ranked in respect of their tolerance to nickel: Nilgiri > Annapurna > Subhadra > Khandagiri > Rudra > Sankar.

Organogenetic buds were induced from hypocotyl and cotyledon explants of oil crop Perilla frutescens in Murashige and Skoog (MS) medium supplemented with 5.7 μM indole-3-acetic acid (IAA) and 8.9 - 13.3 μM 6-benzylaminopurine (BA). Shoots were rooted on MS medium with 2.9 μM IAA and 1.4 μM gibberellic acid (GA3) and the regenerated plants flowered and set seeds normally.

The effect of N-(2-chloro-4-pyridyl)-N'-phenylurea (4PU-30) on the growth and content of endogenous cytokinins of adenine type in tobacco (Nicotiana tabacum L.) callus was investigated. Biomass accumulation in calli grown on Murashige and Skoog (MS) medium with 4PU-30 was higher than that on MS medium with kinetin. The obvious presence of isopentenyladenine type cytokinins and traces of zeatin type cytokinins supposes modification in the endogenous cytokinin metabolism of the tobacco callus grown on 4PU-30.

In vitro proliferated shoot culture of six grape genotypes (Vitis vinifera L.) were screened for tolerance to NaCl (0 to 200 mM). The cv. Perlette was found to tolerate 175 mM NaCl followed by cvs. Pusa Seedless and Beauty Seedless 150 mM NaCl. Na, K, Cl, Ca and Mg content increased upto 100 mM NaCl in most of the genotypes. Total sugar and proline content of stem tissue gradually increased under NaCl stress while leaf chlorophyll a+b content declined. Studies suggest that the in vitro screening procedure can be used for ranking the grape genotypes for salinity tolerance.

Waterlogging of soybean plants (Glycine max L.) led to impaired symbiotic N₂ fixation and a marked decline in glutamine (Gln) concentration in xylem bleeding sap. Xylem Gln concentration increased during the growth cycle of the plant and was correlated with nodule formation. Treatments known to impair N₂ fixation, such as exposing the root system to pure N₂ gas or a mixture of Ar and O₂ (80:20; v/v), led to specific declines in xylem sap Gln. The decrease in Gln observed during waterlogging was also seen on transfer of nodulated plants to aerated hydroponics, where the decline was highly correlated with ureide content in the xylem sap. Upon flooding the nodulated root system, the specific decline in xylem sap Gln could be detected within 10 min and reached a minimum within 60 min, indicating that waterlogging has an immediate effect on N₂ fixation. It is concluded that xylem Gln arises directly from N₂-fixation and is a useful indicator of N₂ fixation activity of symbiotic soybean plants.

The effects of sodium nitroprusside (SNP; nitric oxide donor) treatment on drought stress induced by PEG for different periods of time in wheat seedlings were investigated. Our results suggested that treatment for 2, 4 and 6 d with 15 % PEG could be termed as mild, moderate and severe stress, respectively. Drought stress induced accumulation of hydrogen peroxide and resulted in lipid peroxidation. On the other hand, activities of SOD, CAT and PAL increased under mild stress to counteract the oxidative injury and then decreased when the stress became severe (6 d). As the effect of SNP treatment, 0.2 mM enhanced wheat seedlings growth and kept high relative water content and alleviated the oxidative damage. However, 2 mM SNP aggravated the stress as a result of uncontrolled generation of reactive oxygen species and ineffectiveness of antioxidant systems.

The effects of NaCl and ABA on the respiration of N₂-fixing nodules were analysed in common bean (Phaseolus vulgaris) inoculated with Rhizobium tropici the reference strain CIAT899. Shoot and nodule growth was more inhibited by NaCl than root growth. The O₂ uptake by nodulated roots at 21 kPa O₂ was significantly inhibited by salinity. Raising pO₂ stimulated nodule respiration more under NaCl treatment than for the control, although it did not compensate totally for the inhibitory effect of NaCl. Short NaCl application was less destructive than long term application. Also, the external application of ABA inhibited nodule respiration, and this inhibition was partly compensated by raising pO₂.

Salix paraplesia was used as an experimental model to investigate the effect of short day photoperiod (SD) and low temperature (LT) on development of freezing tolerance and on endogenous abscisic acid (ABA) contents. We characterized differences in SD and LT-induced cold acclimation in three ecotypes from different altitudes. The results demonstrated that cold acclimation could be triggered by exposing the plants to SD or LT alone, and that a combination of the different treatments had an additive effect on freezing tolerance in all ecotypes studied. However, the high altitudinal ecotype was more responsive to SD and LT than the low altitudinal ecotype. Development of freezing tolerance induced by SD and LT was accompanied by changes in ABA contents which were ecotype-dependent. Although the stem had higher initial freezing tolerance, the leaves developed freezing tolerance more quickly than the stem and thus leaves may provide an interesting experimental system for physiological and molecular studies of cold acclimation in woody plants.

Cytokinin-like activity of orotic acid and its 1-cyclohexyl and 1-phenyl derivatives was tested estimating the anthocyanin accumulation and inhibition of root formation in isolated buckwheat cotyledons (Fagopyrum esculentum Moench.). The anthocyanin content was stimulated most by 1-phenylorotic acid. Strong synergetic effect of the phenylurea cytokinin 4PU-30 was found.

Contents of total free [PA(S)] and conjugated polyamines [PA(SH), PA(PH)] were higher in turnip (Brassica rapa L. cv. Rapa) seeds during imbibition (0-36 h) and radicle protrusion (36-48 h) than during the further growth (10 d). Ethylene production was activated with the protrusion, reaching a maximum at the second day of germination and dropping afterwards. The application of ethrel accelerated radicle emergence but the direct intervention of ethylene in the breaking of the seed coat was not clear from the use of ethylene-biosynthesis inhibitors (CoCl₂ and AVG). Finally, in this work the gene BrACO2 was characterized. Although its expression was not detected in seeds through zygotic embryogenesis, it increased concomitantly with the germination process.

Explants obtained from in vitro-propagated plantlets of two potato cultivars, Shepody and Atlantic, were treated with five doses of γ-radiation (0, 2, 4, 6 and 8 Gy) to investigate the stimulating effects of low irradiation on the production and quality of microtubers in vitro. Microtubers of both cultivars treated with γ-radiation initiated 5 d earlier than in the non-irradiated control. The whole period of microtuberization was prolonged by 10 - 15 d with 4, 6 and 8 Gy irradiation treatment for cv. Atlantic. Irradiation of the plantlets (4 Gy) led to a significant increase not only in the microtuber number (116.7 and 34.5 % over the control) but also in the fresh mass (77.6 and 23.2 % in Shepody and Atlantic, respectively). Low dose irradiation (2 - 4 Gy) increased the starch content of microtubers. High doses (6 - 8 Gy) enhanced ascorbic acid and reducing sugar contents. 4 - 6 Gy doses also effectively increased the protein contents of microtubers.

Two sets of wheat (Triticum aestivum L.) substitution lines for the homoeologous group 5 chromosomes, 5A, 5B and 5D, carrying vernalization genes (Vrn-A1, Vrn-B1, Vrn-D1) were used to study the relationship between vernalization requirement and winter survival, with respect to the induction and maintenance of frost tolerance. Substitution lines carrying dominant Vrn loci substituted from the spring cultivars Zlatka (5A), Chinese Spring (5D) and the alternative cultivar Česká Přesívka (5B) into three different winter wheat backgrounds, Vala, Košútka and Zdar, showed lower winter survival by 20, 36, and 41 % for substitutions of 5B, 5A and 5D, respectively, compared to the original winter cultivars. Reciprocal substitution lines between two winter cultivars Mironovskaya 808 and Bezostaya 1 carrying different recessive alleles, vrn-A1, vrn-B1, vrn-D1, did not exhibit a modified induction of frost tolerance, but the duration of good frost tolerance, as well as the ability to survive the whole winter, was changed. In accordance with the model suggesting that genes for vernalization act as a master switch regulating the duration of frost tolerance, substitutions of homoeologous group 5 chromosomes induced, at first, frost tolerance at a level equal to the parental cultivar, and then, relative to the different extent of saturation of vernalization requirement, they gradually lost both frost tolerance and their ability to re-induce significant frost tolerance with a drop in temperature following warm periods in the winter.

The effects of boron and NaCl induced salinity on growth and mineral composition of the pear (Pyrus communis L.) rootstock OH × F 333 shoots cultured in vitro were investigated. Shoots were grown in vitro for seven weeks on a Murashige and Skoog medium containing two B concentrations (0.1 and 2 mM) combined with five NaCl concentrations (0, 10, 20, 40, and 80 mM). The longest shoots were produced at 0.1 mM B and 80 mM NaCl, but highest number of shoots were produced at 0.1 mM B and 0-20 mM NaCl. Inclusion of 20 and 40 mM NaCl in the culture medium significantly increased fresh mass of cultures compared to 0 mM NaCl for all B concentrations tested. The concentrations of P, K, Ca, Mg, Na, Fe, Mn and Zn of plants were affected by B and NaCl concentration of the medium.

The phytopathogenic fungi Colletotrichum circinans, Fusarium solani, and Rhizoctonia solani were incubated in aerated (0, 0.5, 1 dm³ min^-1) potato dextrose broth (PDB) or Czapek-Dox broth (CDB), under 0-, 12- or 24-h photoperiods. Greater dry mass was produced in PDB. Higher air flows improved dry mass of F. solani and R. solani. The 24-h photoperiod improved F. solani dry mass. Except for F. solani, which was not affected, incubation in PDB increased protein content. The no air treatment increased protein content in F. solani, 0.5 dm³ min^-1 produced the highest protein content in R. solani, but air flow-rate did not affect C. circinans. Incubation in the dark produced the lowest protein content in C. circinans, the highest under the 24-h photoperiod for R. solani, and photoperiod did not affect protein content in F. solani. Protein content in R. solani, incubated in CDB, was lowest at the 0 dm³ min^-1 air flow at all photoperiods. Molecular masses of most proteins were under 30 kDa, and numbers of bands in SDS-PAGE gels varied with air flow. In CDB, especially under 12- or 24-h photoperiods, proteins in F. solani were between 1.6 and 310 kDa. For R. solani in PDB, at 0.5 dm³ min^-1 air flow and 12-h light, proteins were between 6 and 81 kDa.

Effect of alginate matrix composition on regrowth performance of encapsulated microcuttings of hybrid aspen (Populus tremula L. × P. tremuloides Mincx.) was studied. Both high regrowth frequency and viability of explants were registered in all encapsulation mixtures tested. Some ingredients of the matrix (nutrient medium salts, sugars, growth regulators) significantly affected the initial development of the microcuttings. Sucrose appeared to play an important role in the starting stage of the regrowth event.

A rapid plantlet regeneration system for Perilla frutescens was established from cotyledon and hypocotyl explants. A maximum of 91.06 % cotyledon and 76.4 % hypocotyl explants could directly produce shoots (3.09 ± 0.18 shoots per explants) on Murashige and Skoog (MS) medium. The optimum hormone combinations were 4.44 μM 6-benzylaminopurine (BA) for cotyledon and 2.22 μM BA + 2.85 μM indole-3-acetic acid (IAA) for hypocotyls. Rooting was induced on half-strength hormone-free MS medium. After transplantation to soil, approximate 80 % of the regenerated plantlets could survive, flower and fruit. Moreover, some morphological abnormalities were found among the regenerated plants.

A protocol of high frequency shoot organogenesis and plant establishment from stem derived callus has been developed for Tylophora indica (Burm. f.) Merrill. - an endangered medicinal plant. Callus was developed on Murashige and Skoog (MS) medium supplemented with 10 μM 2,4,5-trichlorophenoxy acetic acid (2,4,5-T). Multiple shoot induction was achieved from the surface of the callus after transferring onto shoot induction medium. The highest rate (80 %) of shoot multiplication was achieved on MS medium containing 5.0 μM kinetin. The developed shoots rooted best on half-strength MS medium supplemented with 0.5 μM indole-3-butyric acid (IBA). The in vitro raised plantlets with well developed shoot and roots were acclimatized successfully and grown in greenhouse.

The effect of elevated CO₂ concentration (CE) on leaf chlorophyll (Chl) and nitrogen (N) contents and photosynthetic rate (P_N) was evaluated during the post-flowering stages of rice grown at CE (570 ± 50 µmol mol^-1) in open top chamber (OTC), at ambient CO₂ concentration (∼ 365 µmol mol^-1) in OTC and at open field. Thirty-five day old seedlings were transplanted in OTCs or in field and allowed to grow till maturity. Chl and N contents were highest at the time of flowering and thereafter it started to decline. The rate of decline in Chl and N contents was faster in plants grown under CE mostly in later part of growth. Irrespective of treatment difference, flag leaf contained the highest amount of Chl and N than penultimate and third leaf. The higher P_N was observed in leaves under CE than in the leaves in other two growing conditions. Considering growth stage, P_N was the highest at flowering which reduced at the later part of growth due to degradation of Chl and N content of the leaf. Under CE it was 40.02 µmol m^-2 s^-1 at flowering and it reduced to only 14.77 µmol m^-2 s^-1 at maturity stage. The beneficial effect of CE in increasing leaf P_N may be maintained by applying extra dose of nitrogen at the later stages of plant growth.

The in vitro response of two Prunus rootstocks: GF 677 (Prunus persica × Prunus amygdalus), and Nemared (Prunus persica) to increasing concentrations of KCl of the culture medium was studied. Shoots were grown in vitro for 8 weeks on an Murashige and Skoog medium supplemented with 0, 5, 10, 15, 20, 40 or 80 mM KCl. By increasing KCl concentration from 0 to 40 mM, the number of shoots per explant was not significantly affected for both rootstocks. However, Nemared rootstock formed more shoots per explant than GF 677 under respective KCl concentrations of the medium. Inclusion of 80 mM KCl in the medium resulted in a reduction of growth of both rootstocks. Sodium, Fe, Mn, and Zn concentration in tissues of Nemared rootstock were significantly higher than the respective values of GF 677.

Leaf explants of Phalaenopsis amabilis var. formosa formed clusters of somatic embryos directly from epidermal cells without an intervening callus within 20 - 30 d when cultured on 1/2-strength modified Murashige and Skoog medium supplemented with 0.1, 1 and 3 mg dm^-3 TDZ. Repetitive production of embryos involved secondary embryogenesis could be obtained by culturing segments of embryogenic masses on TDZ-containing media. Plantlet conversion from embryos was successfully achieved on regulator-free growth medium.

Mycorrhizal and nonmycorrhizal Pinus halepensis plants were subjected to water stress by withholding irrigation for four months and then rehydrated for 30 d. Water stress affected plants growth and mycorrhizal association was unable to avoid the effects of drought on plant growth. However, when irrigation was re-established the increase in height, number of shoots, total dry mass, and chlorophyll content in the mycorrhizal plants were greater than in non-mycorrhizal plants. The decrease in soil water content decreased the leaf water potential, leaf pressure potential and stomatal conductance. These decreases were higher for nonmycorrhizal than for mycorrhizal plants, indicating that the mycorrhizal fungi permit a higher water uptake from the dry soils. The total content of inorganic solutes was not changed by presence of mycorrhizae.

Effect of two Ni concentrations (10 and 200 μM) on growth, Ni accumulation, chlorophyll and proline contents, relative water content (RWC) as well as the activities of superoxide dismutase (SOD), catalase (CAT), peroxidase (POD) and glutathione S-transferase (GST) were studied in shoots of wheat plants. Treatments caused a considerable accumulation of Ni in the shoots. However, exposure of plants to 10 μM Ni did not lead to significant alterations in shoot growth except for a slight increase in fresh mass. The other parameters studied were not affected by treatment of plants with 10 μM Ni. In contrast, 200 μM Ni caused inhibition of shoot growth, a decline in RWC and chlorophyll content, accumulation of proline and occurrence of visible symptoms of Ni toxicity. The activities of SOD and CAT decreased in response to 200 μM Ni. Conversely, several-fold enhancements of POD and GST activities were observed following the 3^rd day of 200 μM Ni treatment.

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.

Gas exchange in Gossypium hirsutum L. cv. H-777 as affected by water deficit and growth regulators (IAA, GA₃, BAP, ABA, ethrel) was examined. Sixty days after sowing, growth regulators in concentration 50 µM were applied as foliar spray and irrigation was withheld to get desired (moderate and severe) water deficit. All the parameters were measured on the third leaf from the top between 10:00 and 11:00. Net photosynthetic rate (P_N), transpiration rate (E), stomatal conductance (g_s), carboxylation efficiency (CE), and water potential (ψ_w) decreased significantly with the increasing water stress, however, water use efficiency (WUE) was unaffected. Foliar spray with IAA, GA₃ and BAP partially counteracted the effect of water deficit on the above parameters except ψ_w, which became more negative. ABA and up to some extent ethrel increased WUE and maintained higher ψ_w, however, caused further decrease in P_N, E, and g_s.

The results demonstrate the benefits of using filter-paper-based system for cultivation the germinating barley seeds for Al toxicity or Al tolerance analyses. Due to the high affinity of filter paper to Al monomeric forms, milimolar Al concentrations were required to cause similar Al toxicity symptoms of roots as micromolar Al concentrations in hydroponics: 1 mM Al had no effects on the emerging barley roots, 2 mM Al was moderately toxic but roots showed good recovery, 4 mM Al was highly toxic and 8 mM Al even lethal. Screening of eight barley cultivars revealed different rank of their tolerance to Al. The root growth inhibition positively correlated with the Al concentration in root tips.