Gas exchange in paclobutrazol-treated triticale plants during water stress and rehydration was studied. Seed treatments with the retardant (1 and 25 mg dm^-3) alleviate negative effect of PEG-induced water stress. Net photosynthetic rate, transpiration rate, stomatal conductance, relative water content, and leaf water potential were higher while peroxidase activity and free proline concentration were lower in the paclobutrazol-treated plants than in control plants. This confirmed our assumption that paclobutrazol possessed a protective effect against water stress.

Primer screening and optimization for random amplified polymorphic DNA (RAPD) analysis of cashew (Anacardium occidentale L.) was investigated. Among four series (A, B, D and N) of 10-mer primers, A-series performed better amplification of fragments than other series. The maximum amplification fragments was obtained using OPA-02, OPA-03, OPA-09, OPB-06, OPB-10, OPD-03, OPD-05 and OPN-03 primers. The primers OPA-02 and OPN-03 produced maximum number of DNA fragments in Anacardium occidentale cv. H-320. Primers (OPB-08 and OPN-05 performed a least number of amplification fragments. RAPD profile also indicate that some primer did not produce good amplification. The primer OPA-02 amplified 12 number of polymorphic bands in 20 cultivars of cashew. Only one DNA fragment was produced in A. occidentale cv. Vridhachalam - 2 (M-44/3) by using the primer OPA-02.

Tortula ruralis is an important experimental system for the study of plant desiccation tolerance. EST gene discovery efforts utilizing desiccated gametophytes have identified a cDNA TrRpt2 encoding a predicted polypeptide with significant similarity to the 26S proteasome regulatory subunit IV. TrRPT2, the 446 amino acid deduced polypeptide, has a predicted molecular mass of 49.6 kDa, and a predicted pI of 8.15. Phylogenetic analysis demonstrated that previously characterized RPT2 polypeptide sequences could be reproducibly grouped into 3 major clades and that TrRPT2 forms a discrete evolutionary group. RNA blot hybridizations were used to analyze TrRpt2 expression in response to: 1) desiccation and rehydration, 2) abscisic acid-treatment, 3) increased NaCl concentration, and 4) NaCl-shock. TrRpt2 steady-state mRNA transcript levels are unchanged in response to all treatments and the gene is constitutively expressed.

Aluminium effect on the mobility of apoplast protein in root tips was studied. Two-day seedlings of soybean (Glycine max. (L.) Merr. cv. Tsurunoko) were treated with 50 μM AlCl₃ for 2 h. Using infiltration method, the apoplast protein in root tips was extracted with 20 or 100 mM MgCl₂. When 20 mM MgCl₂ was used to collect weakly bound protein to apoplast, the amount of protein extracted was reduced to be about 20 % compared with that of control and the band of 97 kDa disappeared in SDS-PAGE gel. However, the 97 kDa protein could be extracted by 100 mM MgCl₂, which were used for extraction of more tightly bound protein to apoplast, and the amount was estimated to be the same as that of control. When the protein was further developed in two-dimensional electrophoresis, three spots were found between pI 6.4 and 6.5. This is the first report of an Al effect on the mobility of apoplast protein.

A rapid micropropagation protocol through induced multiple shoots from the cotyledonary explant of mulberry (Morus alba L) is described. The highest number of shoots (20.3) was obtained when explants from 14-d-old embryos were cultured on Murashige and Skoog (MS) medium supplemented with 7 μM thidiazuron for 45 d. Of the three cultivars used, cv. S-36 was the best followed by cv. K-2 and S-1. The shoots were transferred to MS medium supplemented with 5 μM 6-benzylaminopurine for elongation. The elongated shoots were rooted on half strength MS medium containing 1 - 7 μM indole 3-butyric acid or 1-naphthalene acetic acid. The rooted plants were transplanted to soil with 90 % success. The emerged shoot primordia probably initiated from the pre-existing meristems since the shoot bud show definite vascular connection to the major vascular tissue.

Supplementary UV-B (12.2 kJ m^-2 d^-1 UV-B_BE) provided to Vigna radiata for 2 h d^-1 suppressed the length of root, shoot and whole plants, number of leaves, total leaf area, leaf area index, specific leaf mass, fresh and dry mass of leaves and shoot, relative growth rate and net productivity. In unstressed green gram plants (10 kJ m^-2 d^-1 UV-B_BE), triadimefon (TRIAD) (20 mg dm^-3) enhanced growth in all parameters over control. The growth promoting effect of TRIAD enabled the UV-B impacted plants to overcome the growth inhibitions to varying degrees indicating its protective potential against UV-B stress.

The free amino acid, protein, water and dry matter contents were determined during the seed development of Araucaria angustifolia. Soluble and insoluble proteins in the mature seed represent 4.2 % of the fresh matter. The embryonic axis stored the greatest amount of soluble proteins, while cotyledons both with the embryonic axis showed the largest quantities of insoluble proteins in the mature seed. The greatest concentration of free amino acids was detected during the stage when cotyledons start to develop. Glutamic acid, aspartic acid, alanine and serine were predominant in the whole seed while arginine, lysine and γ-aminobutyric acid were present in great amounts only in cotyledons and embryonic axis. Although megagametophyte was important as a source of free amino acids, it was not the major protein storage organ in the mature seed. In the embryogenetic process, the rise of cotyledons is closely related to physiological and biochemical changes.

Using apical and axillary nodes as explants, a rapid and efficient method for propagation of Amorpha fruticosa L. has been developed. When grown on Murashige and Skoog (MS) medium supplemented with 8 mg dm^-3 benzyladenine, 100 % explants responded with 4.94 shoots per explant after 6-weeks culture, and explants taken from the in vitro proliferated shoots subsequently produced multiple shoots when cultured on the same medium. The addition of casein hydrolysate (200 mg dm^-3) enhanced the number of shoots up to 8.77 per subculture, and coconut milk was found to promote the shoot elongation and make them grow more vigorously, 82.53 % excised shoots were rooted on half-strength MS medium containing 2.0 mg dm^-3 indoleacetic acid after 3 weeks of incubation. After acclimatization, all of the rooted plantlets established in soil, exhibiting uniform morphological and growth characteristics.

Electrophoretic patterns of glutamate oxaloacetic transaminase (Got) and malate dehydrogenase (Mdh) of Zea mays L. × Tripsacum dactyloides L. hybrids and their parents have been compared. The results suggested that Got and Mdh isozymes may be used as markers for genic regions on 5 S and 6 L maize chromosomes and for linkage groups D and L on T. dactyloides chromosomes, syntenic to genic regions on 5 S and 6 L maize chromosomes. The latter have a regulatory effect on fertility and on the apomictic mode of reproduction.

Transgenic rice plants harbouring Bacillus subtilis protoporphyrinogen oxidase (Protox) gene, which is targeted into plastid, were generated by Agrobacterium-mediated transformation using a rice (Oryza sativa L. cv. Dongjin) and their gene integration at T₁ generation by Southern and mRNA expression in T₂ generation by Northern blotting were analyzed. Their herbicide-resistant trait was further confirmed by in vitro leaf segment assay and in planta bioassays such as seed germination assay and measurement of growth inhibition. The herbicide oxyfluorfen resistance in transgenic rice plants was not very high. The results showed that the Protox from B. subtilis can not be applicable as a gene source to generate a high level oxyfluorfen tolerance in plants.

The alterations of organization of leaf tissues and cell ultrastructure as a consequence of salt stress (75 and 150 mM NaCl) were studied in two tomato (Lycopersicum esculentum Mill.) cultivars showing different salinity tolerance. The salinity brought changes in cell shape, volume of intercellular spaces and chloroplast number, shape and size. These characteristics were specific in each cultivar. The ultrastructural changes were also different in the two tomato cultivars studied and the most important ones were in the number and size of starch granules in chloroplasts, the number of electron-dense corpuscules in the cytoplasm, the structure of mitochondria, and number of plastoglobuli.

Exposure of groundnut seeds and seedlings to 25, 100, and 250 μM concentrations of jasmonic acid resulted in a reduction of germination percentage, growth, fresh mass, dry mass, chlorophyll content, chlorophyll stability index, proteins and an increase in free proline content and cell membrane injury. The results suggest the inhibitory effect of jasmonic acid on growth of the groundnut seedlings.

The effect of exogenous brassinosteroids (BR) on the flowering induction of Pharbitis nil was examined. Generally plants treated with brassinolide and castasterone form less number of flowers than control plants, but degree of flowering inhibition was depended on the concentration and the method of BR application as well as the length of the inductive dark period. In plants regenerated from sub-induced apices treated with brassinolide at concentration of 1 and 10 μM the flower formation was inhibited completely.

The effect of CO₂ concentration elevated to 575 - 620 µmol mol^-1 on growth, tillering, grain yield, net photosynthetic rate, dark respiration rate, stomatal conductance, sugar content and protein profile of two rice (Oryza sativa L.) cultivars Pusa Basmati-1 and Pusa-677 at flowering stage was studied using open top chambers. The cultivar Pusa Basmati-1 responded more markedly for most of the growth and physiological parameters compared to Pusa-677. The increase in grain yield in Pusa Basmati-1 attributed largely to increased grain number. The increased net photosynthetic rate and greater accumulation of sugar contributed significantly to the accelerated development of leaves and tillers in both the cultivars. The reduction in the low molecular mass proteins including Rubisco and increase in high molecular mass photosystem 2 proteins was observed in both the cultivars. Additional sugars may possibly help in balancing the profile of photosynthetic proteins and sustain greater growth and productivity in rice cultivars.

The changes induced by 80 and 120 mM NaCl during emergence and growth of sprouts in salt-tolerant (CPF-213) and sensitive (L-116) genotypes of sugarcane were determined. The rate and percentage of emergence of sprouts, length and dry mass of shoot and root, and number of nodal roots decreased under salinity. Concentrations of Na and Cl increased and those of K, Ca, N and P decreased with a rise in substrate salinity. A greater salinity tolerance ability of CPF-213 than L-116 was attributable to greater root mass and higher nutrient concentrations in the sprouts of the former genotype.

A simple protocol for mass multiplication of Crataeva nurvala, a medicinal tree, from seedling-derived explants is described. Six different types of explants (cotyledonary nodes, epicotyl nodes, hypocotyl segments, first pair of leaves, cotyledons, and root segments) developed shoots on Murashige and Skoog's (MS) basal medium or the same supplemented with different concentrations of 6-benzylaminopurine (BAP). Among the explants tested for caulogenic potential, only the epicotyl and cotyledonary nodal explants developed shoots on MS basal medium, while on BAP (0 - 2.0 mg dm^-3) adjuvated media all the explants exhibited caulogenesis. The optimum concentration of BAP varied for these explants. The shoots could be rooted on half strength MS with 0.02 mg dm^-3 α-naphthalene acetic acid to get plants, which have been transferred to soil. The explants from in vitro regenerated shoots also possessed a similar caulogenic potential.

Distribution of cadmium between roots and shoots of barley was manipulated by the cadmium concentration (0.01 and 0.005 mM Cd²⁺), pH (4.6 and 5.9) as well as treatment duration. The prolongation of treatment increased dry mass and content of cadmium in plants. The cadmium is accumulated mainly in roots. Presence of both, 0.005 mM Cd²⁺ and 100 mM NaCl in medium at pH 5.9 (Cd-NaCl plants) resulted in the most severe growth inhibition of plants, but about one half accumulation of cadmium in roots then in a case of only Cd-treated plants. In the Cd-NaCl plants, the net photosynthetic and transpiration rates were less reduced then in a case of only NaCl-treated plants. The treatments also influenced uptake of Ca, Cd, Cu, K, Mg, Na and Zn predominantly in roots.

A spontaneously embryogenic cell line of the coumarin producing angelica [Angelica archangelica (L.) subsp. archangelica] was established via callus formation from seedlings grown from sterilized seeds on semi-solid, hormone-free modified B5 medium. The cell line has retained its embryogenic capacity for 5 years. The highest coumarin production for the cell line after 3 weeks of cultivation was achieved in the medium containing 3.0 % sucrose. Jasmonic acid had no statistically significant effect on the biomass or coumarin production. The established embryogenic cell line could be stored using cryopreservation. Plantlets grown in an air-sparged bioreactor were transferred directly to soil and vermiculite, and 63 % of them grew to maturity through two growth seasons. The coumarin content in the regenerated plants was comparable to that in wild plants. Thus this cell line could be used for in vitro propagation.

Seasonal changes in foliage nitrogen (N) and carbon (C) concentrations and δ¹⁵N and δ¹³C ratios were monitored during a year in Erica arborea, Myrtus communis and Juniperus communis co-occurring at a natural CO₂ spring (elevated [CO₂], about 700 μmol mol^-1) and at a nearby control site (ambient [CO₂], 360 μmol mol^-1) in a Mediterranean environment. Leaf N concentration was lower in elevated [CO₂] than in ambient [CO₂] for M. communis, higher for J. communis, and dependent on the season for E. arborea. Leaf C concentration was negatively affected by atmospheric CO₂ enrichment, regardless of the species. C/N ratio varied concomitantly to N. Leaves in elevated [CO₂] showed lower δ¹³C, and therefore likely lower water use efficiencies than leaves at the control site, regardless of the species, suggesting substantial photosynthetic acclimation under long-term CO₂-enriched atmosphere. Leaves of E. arborea showed lower values of δ¹⁵N under elevated [CO₂], but this was not the case of M. communis and J. communis foliage. The use of the resources and leaf chemical composition are affected by elevated [CO₂], but such an effect varies during the year, and is species-dependent. The seasonal dependency and species specificity suggest that plants are able to exploit different available water and N resources within Mediterranean sites.

During germination and post-germinative growth of Pinus pinaster Ait. seeds, triglycerides are hydrolysed and concurrently the embryo accumulates starch. In this study, the spatio-temporal variation of starch accumulation was described in zygotic embryos associated (ZE⁺) or not (ZE^-) to their megagametophyte and in somatic embryos (SE). In germinating ZE⁺, starch was accumulated in the growing tissues, following closely the spatio-temporal pattern of triglycerides depletion. In contrast, in ZE^- and SE, starch was only found in cortical cells close to the culture medium. In germinating ZE⁺, the spatio-temporal variations of starch accumulation can be thus interpreted as the result of the changing contact between the megagametophyte and the growing tissues and also of the existing interactions between triglyceride hydrolysis and the allocation of sucrose exported from the megagametophyte.

Image analysis has become a powerful tool for the quantification of histochemical detections mainly in animal and medical sciences. There is not, however, much information about the accuracy of image analysis quantification of histochemical products and their comparison to biochemical analyses. Our study focused on the quantification of histochemically detected amounts of phenolic compounds, lignin and activity of peroxidases in the two youngest age classes of Norway spruce (Picea abies (L.) Karst) needles with the use of image analysis and its comparison with biochemical assays. The image analysis-determined amount of lignin and peroxidase activity was well correlated with its biochemical assay. On the other hand, no optical parameters of the reaction product of the histochemical reaction of phenolics correlated with the biochemical quantification. This proves that results of quantification of histochemical detection with image analysis greatly depend on the character and procedure of the histochemical proof. If it is done in full respect of the conditions of a histochemical reaction it can gain reliable and interesting results. Great advantage of the image analysis approach is quantification of the studied substance in different tissues within single sample as well as among different samples, which in many cases cannot be detected with biochemical tools.

The influence, was investigated, of abiotic parameters on the isolation of protoplasts from in vitro seedling cotyledons of white lupin. The protoplasts were found to be competent in withstanding a wide range of osmotic potentials of the enzyme medium, however, -2.25 MPa (0.5 M mannitol), resulted in the highest yield of protoplasts. The pH of the isolation medium also had a profound effect on protoplast production. Vacuum infiltration of the enzyme solution into the cotyledon tissue resulted in a progressive drop in the yield of protoplasts. The speed and duration of orbital agitation of the cotyledon tissue played a significant role in the release of protoplasts and a two step (stationary-gyratory) regime was found to be better than the gyratory-only system.

The cyanobacterial crust occurring in desiccated state exposed to high temperature and solar radiation on the rock surface contained several low molecular mass (LMM) proteins (10.5, 13, and 25 kDa), water stress protein (wsp, 39 kDa), additional proteins (43 and 49 kDa), a chaperonin (58 kDa), and a stress-induced protein (84 kDa). When the crust was exposed to UV-C radiation, it counteracted the UV-B damage by overproduction of certain proteins and synthesis of two high molecular mass (HMM) proteins. Exposure of the crust to heat had an adverse effect on the survival of the organism there-in which was due to repression of few proteins. Unlike the modification pattern of protein synthesis in the cyanobacterium inhabiting the crust, the same organism grown in culture could tolerate heat by synthesizing two HMM proteins. Several proteins of diverse groups were repressed in the cyanobacterium in culture upon exposure to UV-C, which was counteracted by induction of three new polypeptides (chaperonin and HMMs), and overproduction of one 41 kDa protein.

An extent of helper component protein (HC-Pro) sequence variability within virus population of single Czech isolate of potato virus Y^NTN (PVY^NTN) Nicola was identified by temperature-gradient gel electrophoresis (TGGE) and heteroduplex analysis. HC-Pro region was approximated with 6 pairs of primers derived from Hungarian PVY^NTN isolate (sequence AC M95491). Immunocapture reverse transcription - polymerase chain reaction (RT-PCR) was used to obtain six mixtures of individual overlapping polymerase chain reaction (PCR) products. cDNA libraries were prepared by cloning of purified PCR products in pCR-Script vector and screened by heteroduplex analyses. 15 different subfragments within the HC-Pro region were isolated and sequenced. In comparison to AC M95491, 19 nucleotide changes were identified, 13 led to amino acid (aa) changes. In comparison to available post-transcriptional gene silencing (PTGS) specific suppressor HC-Pro sequences of potyviruses we found out 4 aa changes in conserved regions.

A simple and rapid method for multiple shoot formation in vitro from immature embryo axis explants of Carica papaya L. cvs. Honey Dew, Washington and Co2 is described. Multiple shoot regeneration was achieved by culture of the explants on modified Murashige and Skoog (MS) medium supplemented either with thidiazuron (TDZ; 0.45-22.7 μM) or a combination of benzylaminopurine (BAP; 0.2 - 8.84 μM) and naphthalene acetic acid (NAA; 0.5 - 2.64 μM). Highest frequency of shoot regeneration occurred on medium supplemented either with 2.25 μM TDZ or a combination of BAP (4.4 μM) and NAA (0.5 μM). Composition of the basal media influenced the frequency of multiple shoot initiation. Stunted shoots regenerated at 4.5 μM and higher concentrations of TDZ. Such shoots could, however, be elongated by transfer to medium containing 5.7 μM GA₃. Rooting of the regenerated shoots was achieved in presence of indolebutyric acid (IBA; 4.92 - 19.68 μM), however, least response was in presence of 14.7 μM IBA. Rooted plants were hardened and transferred to pots.

The report describes in vitro plant regeneration from embryo axis explants of six cultivars of cotton. Induction of a maximum number of multiple shoots in all six cultivars could be achieved on Murashige and Skoog's (MS) salts and Gamborg's (B5) vitamins supplemented with 0.4 μM benzyladenine (BA) and 0.1 μM napthaleneacetic acid (NAA). Elongated shoots could be rooted on half strength medium supplemented with 0.5 μM NAA. Rooted shoots survived (92 %) after hardening in the greenhouse and grew to maturity (100 %) after transfer to field.

An Agrobacterium rhizogenes-mediated transformation system for Aesculus hippocastanum L. has been developed. Wounded androgenic embryos of A. hippocastanum were inoculated with bacteria containing the pRiA4 plasmid, with the uid A sequence as a reporter gene. The hairy roots emerging from the wounded sites of androgenic embryos were isolated and maintained in Murashige and Skoog's (MS) liquid hormone-free medium. Five hairy root lines have been maintained in vitro for 4 years with unchanged growth rate and might be a suitable source for secondary metabolite production. The transformation events have been confirmed by a polymerase chain reaction specific to the rol A, B, C and D genes. The absence of residual contaminating bacteria has been shown by a polymerase chain reaction specific to the vir D1 sequence.

The effects of different paraquat, alachlor and metolachlor concentrations on superoxide dismutase (SOD), catalase (CAT), guaiacol peroxidase (GPX) activities and pigment contents in lettuce, bean and pea seeds and leaves were studied. Under paraquat (1.0 and 2.0 μM) treatment declined SOD and CAT activities were observed in seeds and undetectable ativity of GPX. Germination of all investigated seeds was completely inhibited. All used concentrations of alachlor and metolachlor inhibited antioxidant enzyme activities in seeds but did not prevent germination and growth. In leaves, lower concentrations of these herbicides increased activities of antioxidant enzymes but at the highest herbicide concentrations (200 μM) activities of investigated enzymes declined. The pigment contents the leaves decreased due to alachlor and metolachlor treatment in a concentration dependent manner.