Aspects of plant growth such as height, branch number, leaf number, leaf area, pod area, 100-seed mass, etc., were correlated with biochemical changes such as contents of chlorophyll (Chl), proteins, DNA, and RNA, and protease activity during development and senescent phases in leaves, flowers, and pods of Cajanus cajan L. cv. UPAS-120 after treatments with kinetin (Kn). A significant increase was noticed in branch number, leaf number, leaf area, and seed mass while other growth processes registered a small increase after Kn application. Effectiveness of 5 µM Kn was also noticed in minimizing the loss of Chls, proteins, and nucleic acids as well as reducing the protease activity during maturity and senescence. Chl a/b ratio maintained a high value up to 30-d followed by a decline in leaves while flowers registered much lower ratio at 20-d-age. Pods were unique in having relatively lower ratio of Chl a/b in comparison to leaves.

The relationship between photosynthetic rate and yield formation processes of the newer and older maize hybrids were investigated. Leaf area at flowering (source) and kernel number (sink) of the newer hybrids were greater than the older ones although their light-saturated photosynthetic rate (P_sat) were not greater than the older ones before flowering. After flowering, P_sat and chlorophyll content of the newer hybrids declined more slowly than the older ones. They not only distributed almost all photosynthates produced after flowering to grain but also reallocated some reserved photosynthates produced before flowering to grain. The newer hybrids exhibited greater grain mass than the older ones mostly because they could optimally regulate the photosynthetic rate and yield formation processes to maximize grain mass.

Cytophysiological and cytogenetic characteristics of cell suspension culture of Panax japonicus var. repens were studied in relation to the accumulation of ginsenosides (GSs). The minimal time of cell number doubling was 1.3 ± 0.1 d and cell number increased 7 to 8-fold during growth cycle. The cell culture can be considered as aneuploid with about tetraploid (46-60 chromosomes) modal class. Upon long-term cultivation, the total content of GSs considerably increased and maximal concentration of GSs was 2.2 %(d.m.). The ratio of seven major GSs only slightly altered both over each and different subcultures. The overall amount of GSs of Rg-group significantly exceeded that of Rb-group. Cell volume and the number of large cellular aggregates with the higher proportion (by 20 %) of parenchymal cells increased late in the subculture. In this time the population contained about 20 % of the cells with doubled amount of nuclear DNA and accompanied with elevation in the GS content. These data prompted us to suggest that biosynthesis of GSs has a link with cell differentiation.

Differential mRNA display was used to identify pathogen-responsive, stress-related genes in potato cell suspensions treated with salicylic acid and a cell wall-derived elicitor from Phytophthora infestans. Among the positive clones identified, one was found to be expressed at a significantly higher level in elicited cells than in control cells. DNA sequencing of this amplicon revealed high homology and identified it as a potato cyclophilin cDNA. The maximum amount of the cyclophilin mRNA was found 9 to 12 h after elicitation. Cyclophilin (CyP) mRNA synthesis was also up-regulated from 12 to 24 h in potato leaves locally infected with zoospores from Phytophthora infestans. However, untreated leaves responding systemically to the pathogen showed only a weak, delayed response at 24 h post infection. The observed accumulation of potato CyP mRNA in response to salicylic acid, P. infestans elicitor and P. infestans infection, suggest that CyPs play an important role in plant stress responses.

Cytokinins N⁶-benzyladenine (BA) and 1-(2-chloropyridin-4-yl)-3-phenylurea (4PU-30) delayed the senescence of detached leaves (3^rd to 7^th leaf node) of wild and ethylene insensitive eti5 mutant of Arabidopsis thaliana. The novel anticytokinins, structural analogues of purine and phenylurea cytokinins also affected the senescence of detached rosette leaves of A. thaliana. They diminished to a significant extent the cytokinin-induced delay of chlorophyll destruction, but without a considerable difference in their action against both types of cytokinins. These results correlated with changes observed in ribonuclease (RNase) activity.

To study the defense mechanism of the resistance to the disease of Chinese wild Vitis species and offer powerful bases for the molecular breeding of highly disease-resistant grape cultivars, using mRNA differential display reverse transcript-PCR (DDRT-PCR) and RACE, a full-length cDNA was isolated from Chinese wild Vitis pseudoreticulata clone Baihe-35-1 inoculated with Uncinula necator by pressing infected leaves under natural field conditions. The cDNA designated as GLOXrg is 1708 bp in length with an open reading frame of 1572 bp encoding 523 amino acids, containing the conserved domain, glyoxaloxid N domain (pfam07250). No homologous nucleotide sequence was found, but the deduced amino acid sequence of GLOXrg shows 74 % identity with a putative protein from Arabidopsis thaliana (accession No. CAB88357). GLOXrg was cloned into pGEX-4T-1 vector. The recombinant vector expressed an about 83-kDa GST-GLOXrg fusion protein as insoluble inclusion bodies in Escherichia coli BL21. Fusion protein GST-GLOXrg was isolated and used to raise the polyclonal anti-(GST-GLOXrg) in rabbits. Western blot analysis showed a high titer (10 000) and the high specificity of the polyclonal anti-(GST-GLOXrg). This is the first report on the cloning of the full-length GLOXrg cDNA from the plant except Arabidopsis thaliana. Through the differential expression, GLOXrg probably plays a crucial role in the resistance to Uncinula necator of Chinese wild Vitis species. An optimized system on prokaryotic expression, protein purification and preparation of the polyclonal antibodies of GLOXrg was also established.

An in vitro method for propagation of Holarrhena antidysenterica Wall. has been developed using nodal explants from mature trees growing in the field. Irrespective of concentrations and combinations of growth regulators used, the axillary and terminal buds sprouted and elongated when inoculated on Murashige and Skoog (MS) medium. The highest numbers of shoots were formed when sprouted shoots were subcultured from MS basal medium onto MS medium containing 2 mg dm^-3 N⁶-benzyladenine (BA) and 0.5 mg dm^-3 α-naphthalene acetic acid (NAA). The shoot number further increased upon subculture on MS medium containing 0.5 mg dm^-3 BA. By repeated sub-culturing of shoots derived from nodal axillary buds, a high frequency multiplication rate was established. The elongated shoots were excised and rooted in auxin free MS basal medium. Ex vitro rooting of in vitro formed shoots was achieved upon dipping the microshoots for 2 min in 2 mg dm^-3 of indole-3-butyric acid solution. Successful field establishment and high (80-90 %) survival of plants was observed.

Two wheat cultivars, HF9703 (drought tolerant) and SN215953 (drought sensitive) were used to examine the effects of glycinebetaine (GB, 100 mM) on lipid composition and function of thylakoid membranes under drought stress. GB application mitigated negative effect of drought on Ca²⁺-ATPase and Hill reaction activities, chlorophyll content, gas exchange and photosynthesis. These positive effects of GB application maybe, in part, correlated with improving the lipid composition of the thylakoid membranes.

Maize (Zea mays L.) seedlings of two cultivars (cv. Bastion adapted to W. Europe, and cv. Batan 8686 adapted to the highlands of Mexico), raised in a glasshouse (19-25 °C), were transferred to 4.5 or 9 °C at photon flux density (PPFD) of 950 µmol m^-2 s^-1 with 10-h photoperiod for 58 h and then allowed to recover at 22 °C for 16 h (14 h dark and 2 h at PPFD of 180 µmol m^-2 s^-1). The ultrastructural responses after 4 h or 26 h at 4.5 °C were the disappearance of starch grains in the bundle sheath chloroplasts and the contraction of intrathylakoid spaces in stromal thylakoids of the mesophyll chloroplasts. At this time, bundle sheath chloroplasts of cv. Batan 8686 formed peripheral reticulum. Prolonged stress at 4.5 °C (50 h) caused plastid swelling and the dilation of intrathylakoid spaces, mainly in mesophyll chloroplasts. Bundle sheath chloroplasts of cv. Batan 8686 seedlings appeared well preserved in shape and structure. Batan 8686 had also higher net photosynthetic rates during chilling and recovery than Bastion. Extended leaf photobleaching developed during the recovery period after chilling at 4.5 °C. This was associated with collapsed chloroplast envelopes, disintegrated chloroplasts and very poor staining.

The organogenic competence of leaf explants of eleven Carthamus species including C. tinctorius on Murashige and Skoog (MS) medium supplemented with different concentrations of thidiazuron (TDZ) + α-naphthaleneacetic acid (NAA) and 6-benzyladenine (BA) + NAA was investigated. Highly prolific adventitious shoot regeneration was observed in C. tinctorius and C. arborescens on both growth regulator combinations and the shoot regeneration frequency was higher on medium supplemented with TDZ + NAA. Nodal culture of nine Carthamus species on media supplemented with BA and kinetin (KIN) individually revealed the superiority of media supplemented with BA over that of KIN in facilitating a higher shoot proliferation index. Proliferating shoots from axillary buds and leaf explants were transferred to medium supplemented with 1.0 mg dm^-3 KIN or 0.5 mg dm^-3 BA for shoot elongation. Elongated shoots were rooted on half-strength MS medium supplemented with 1.0 mg dm^-3 each of indole-butyric acid (IBA) and phloroglucinol. The plantlets thus obtained were hardened and transferred to soil.

This work focuses on the comparison of field characteristics and amounts of reducing sugars in cold-stored tubers of transgenic plants derived from two potato cultivars. The bacterial gene coding for phosphofructokinase under the tuber-specific promoter was used to support the glycolysis in stored tubers. While the tubers from untransformed control plants steadily accumulated reducing sugars during cold storage, the tubers from transformed plants regardless the genotype were characterized by subsequent decrease in the sugar content. After long period of cold storage the greatest reduction in the reducing sugar content was by more than 60 % compared to control. Before the storage, however, the content of reducing sugars was in 80 % of transgenic lines higher than in control ones. The plants evaluated in field trials for their appearance showed any changes in growth characteristics in about 25 % of the transgenic lines. Despite the introduced modification of sugar metabolism the yield of transgenic plants with normal appearance did not differ significantly from the yield of control plants.

Four rice indica genotypes of local importance were transformed with RC7, rice chitinase cDNA clone through Agrobacterium-mediated gene transfer method using mature seed derived calli as explants. The putative hygromycin resistant calli showed varied level of regeneration efficiency ranging from 2.0 to 7.6 %. The stable integration and expression of RC7 was confirmed through polymerase chain reaction (PCR) and Western analysis. Transformation efficiency ranged from 0.9 to 5.2 %. The expression of RC7 (35 kDa chitinase) in different tissues of transgenic plant (root, sheath and leaf) was proved through Western analysis and in terms of increased chitinase activity. The inheritance of transgene was studied through PCR and Western analysis in transgenic plants of Pusa Basmati 1. Bioassays with transgenic plants of local cultivars exhibited enhanced resistance up to 33.3 % to rice sheath blight pathogen Rhizoctonia solani under glasshouse conditions. Enhanced expression or 3-to 4-fold increased activity of chitinase in transgenic plants was correlated with sheath blight resistance.

Potted 2-year-old lemon trees [Citrus limon (L.) Burm. fil, cv. Verna] grafted on sour orange (C. aurantium L.) rootstock were subjected to flooding for 3 d. Control plants were irrigated daily to field capacity. Continuously (sap flow, trunk diameter fluctuations) and discretely (predawn and midday leaf water potential, leaf conductance) measured plant-based water status indicators were compared. The sensitivity of the maximum daily trunk shrinkage signal intensity to flooding and its behaviour during the recovery period demonstrated that this indicator is more feasible than the others for use in automatic irrigation. The responses to flooding of continuously and discretely measured plant-based water status indicators were very similar to those observed in response to drought stress indicating that it necessary to use soil water measurement automatic sensors to detect the cause of the stress. The results underlined the robustness of the compensation heat-pulse technique for estimating instantaneous and daily transpiration rates on flooding stress and recovery.

Dedifferentiation was monitored in Rubia tinctorum L. leaves over a 14-d period after callus induction using transmission electron microscope (TEM), high performance liquid chromatography (HPLC), spectroscopy and thin layer chromatography (TLC). Photosynthetic pigment loss of leaves took 3-5 d coinciding with the first period of anthraquinone accumulation. Callus cells were discernible in the region of the vascular bundles and wounded edges of leaves after 10-14 d. Characteristic ultrastructural alterations were manifested in vacuolization, appearance of mitochondria, amount of smooth endoplasmatic reticulum and cytoplasm, caryolympha density of nuclei and cytoplasm content of cells. There were special events in the transfer cells: unequal divisions of dedifferentiated plastids and lytic activity in the cell wall. Our results show that mesophyll cells seem to be stopped at a particular level of dedifferentiation, while transfer cells embodied in veins of leaves pass through further alterations and lead to callus formation. Findings suggest that a sort of dedifferentiation drift manifests in the various cells of R. tinctorum leaves during callus induction and depending on their specialized status they achieve different levels of dedifferentiation. Approximately 4 weeks after callus induction, root growth has started from the young calli.

Accumulation of extracellular chitinases in Brassica napus plants infected with Turnip yellow mosaic virus (TYMV) and fungal pathogen Leptosphaeria maculans was studied in both compatible and incompatible interaction. Analysis of apoplast fluid by means of non-denaturing anodic and cathodic PAGE followed by in-gel detection of chitinase activity revealed a number of chitinase isozymes. TYMV induced 8 acidic and 4 basic isozymes in a systemic way. Except for one acidic and one basic isozyme, all other chitinases were also constitutively present in low amounts in mock inoculated control. In TYMV systemically infected plants, chitinases were detected in leaves expressing symptoms as well as in symptomless ones. Both virulent and avirulent L. maculans isolates induced production of chitinase isozymes in cotyledons in a time dependent manner. Some of them were present in plants constitutively and their content increased after inoculation. Three of five acidic and two of three basic isozymes responded to L. maculans infection. Chitinases started to accumulate before symptom appearance. First two acidic isozymes were detected 24 h after inoculation. The difference between compatible and incompatibe interaction reflected two basic isozymes.

Foliar sprays of water or 1, 10 and 100 µM aqueous solutions of gibberellic acid (GA₃) or kinetin (KIN) were applied to 40-d-old plants of Nigella sativa (L.) to study their effects on net photosynthetic rate, nitrogen metabolism, and the seed yield. 10 µM solutions of both the hormones, especially GA₃, appreciably increased the activities of nitrate reductase and carbonic anhydrase, chlorophyll and total protein contents and net photosynthetic rate in the leaves, along with capsule number and seed yield plant^-1, at harvest.

To evaluate the genetic diversity of some Vicia species, seed proteins of 160 accessions (30 of Vicia faba, 15 of V. narbonensis, 82 of V. sativa and 25 of V. ervilia and 8 accessions of other Vicia species) were analysed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The dendrogram showed that the two outcrossing species V. faba and V. villosa were the most distant among all species (average percent disagreement value PDV 0.47 and 0.45, respectively). The tree was divided into small clusters of two species each. V. narbonensis fell in one cluster with V. michausai (at PDV = 0.35) and V. lutea (var. hirta) fell in one cluster with V. serococorpes (at PDV = 0.32) whereas, V. ervilia fell in one cluster with V. sativa (at PDV = 0.27). The V. sativa subspecies, however, were closely related (PDV < 0.1). In general, this study did not prove any relationship between the studied storage proteins and the geographical distribution or ecological needs of the studied accessions.

Embryogenic cultures of Dactylis glomerata L. were subjected to NaCl stress by culturing for 4 passages in Schenk and Hildebrandt (SH) medium containing 30 µM dicamba and 200 mM NaCl. Ultrastructural studies indicated invaginations and disruptions of the plasma membrane. Membrane-bound vesicles were observed in the cytoplasm of NaCl treated cells and their occurrence were increased with the culture age.

A mathematical computer-aided model CELLOP was constructed in which the desirability functions in a three-dimensional experimental design can be used to find the optimal growing conditions for plant cells. CELLOP is intended for the optimisation of 3 to 6 physical, chemical, or biological variables in the cultivation conditions of plant cell cultures. The model was used to optimise the culturing conditions (calcium, inorganic nitrogen, and sucrose concentrations) for coumarin-producing, spontaneously embryogenic cell lines of angelica Angelica archangelica L. subsp. archangelica and hogs fennel Peucedanum palustre (L.) Moench. For A. archangelica the overall optimum concentrations were 0.47 mM Ca²⁺, 5.06 mM NO₃ ^-, 0.40 mM NH₄ ⁺, and 96.25 mM sucrose. The dry mass was 24.7 % higher and the coumarin content 40.5 % higher than those achieved in the standard 75 % Gamborg B5 medium. For A. archangelica the highest embryogenic activity was reached in the media containing 1.25 mM Ca²⁺. In the case of P. palustre the overall optimum concentrations were 1.60 mM Ca²⁺, 2.84 mM NO³ _-, 0.23 mM NH₄ ⁺, and 85.10 mM sucrose. For P. palustre the dry mass production increased by 61.8 % and the coumarin content by 58.1 % compared to the values achieved in the Gamborg B5 medium. For P. palustre the highest embryogenic activity was reached in the presence of 50.0 mM NO₃ ^- and 4.01 mM NH₄ ⁺.

The present work investigated the effect of NaCl and seawater salinity (SWS) on accumulation of proline and glycinebetaine (betaine) and expression of two key enzymes of their biosynthetic pathway, betainealdehyde dehydrogenase (BADH) and Δ¹-pyrroline-5-carboxylate synthetase (P5CS) in two aquatic macrophytes, salt-tolerant Najas gramenia and moderately-salt-tolerant Najas indica. N. gramenia did not show any accumulation of betaine or expression of BADH in response to NaCl or SWS treatments. The content of proline increased significantly in response to the salt treatments. High expression of P5CS was also observed, however, the transcript abundance did not correlate with the amount of proline. N. indica, exhibited significantly greater accumulation of proline in response to SWS, which contained Ca²⁺ in addition to NaCl, than in response to NaCl alone.

Ultraviolet-B radiation (biologically effective dose 2.6 kJ m^-2 d^-1) had negative influence on morphology and physiology of the young triticale plants. Plants exposed to UV-B were of lower height than control plants, their leaves were narrow, and the rate of net photosynthesis was decreased. The line RAH 336, which wax cover is lesser than that of traditional cultivar Magnat, was more susceptible to UV-B radiation, considering primary photosynthesis reactions, recorded by chlorophyll a fluorescence. An activation of protective mechanisms was observed: plants responded to UV-B by an increase of the content of UV-B absorbing compounds, and changes of antioxidant enzyme activities.

The human papillomavirus type 16 (HPV 16) oncogene E7 fused with the gene for β-glucuronidase (gus) was used in plant transformation experiments. The E7 gene modified for lower cancerogenicity and fused with the 5' end of the gus in cassettes with cauliflower mosaic virus 35S promoter and transcription terminator produced high contents of fusion proteins in potato protoplasts. Expression vectors harbouring E7 fusion cassettes were used for Agrobacterium tumefaciens LBA4404 mediated transformation of either potato (Solanum tuberosum L. cv. Bintje) or tomato (Lycopersicon esculentum Mill. cv. Moneymaker). A fusion gene was found in all rooted regenerants using polymerase chain reaction with primers providing amplified fragments from E7 and gus genes. GUS activity was revealed in all regenerants obtained. Nevertheless, the level of GUS expression in different constructs varied much more than in transient expression experiments with potato protoplasts. Especially, expression level in plants carrying vectors with the whole E7 gene fused with gus was lowered by 2-3 orders of magnitude comparing with fusion of the first 41 codons of E7 and gus. Southern hybridisation of 18 tomato and 23 potato regenerants revealed mostly multiple tandem integration of T-DNA into the plant genome and Western blot proved the presence of the fusion protein in 9 tomato and 11 potato plants out of 41 tested individuals.

A protocol for induction of direct somatic embryogenesis, secondary embryogenesis and plant regeneration of Dendrobium cv. Chiengmai Pink was developed. Thidiazuron (TDZ) at 0.3, 1 and 3 mg dm^-3 induced 5-25 % of leaf tip segments of in vitro grown plants to directly form embryos after 60 d of culture, and 1 mg dm^-3 TDZ was the best treatment. Somatic embryos mostly formed from leaf surfaces near cut ends, and occasionally found on leaf tips. Higher frequency of embryogenesis was obtained in light than in darkness. During subculture, secondary embryos developed from outer cell layers of primary embryos. All combinations of NAA (0, 0.1, 1 mg dm^-3) and TDZ (0, 0.3, 1, 3 mg dm^-3) increased the multiplication rate of embryos. It takes about 8 months from embryo induction, plantlet formation to eventually acclimatization in greenhouse.

Water potential (ψ_w) and water saturation deficit (WSD), and several reflectance (R) indexes were assessed in an aerophytic lichen Umbilicaria hirsuta (Sw. ex Westr.) Hoffm. The water index (WI, R₉₀₀/_R970) and normalized difference vegetation index, NDVI [(R₉₀₀-R₆₈₀)/(R₉₀₀+R₆₈₀)] were strongly correlated both with the ψ_w and the WSD of lichen thalli. No significant changes during desiccation were found in structural independent pigment index, SIPI [(R₈₀₀-R₄₄₅)/(R₈₀₀-R₆₈₀)]. Sensitivity of the spectral detection of water status was rather small at high hydration level (WSD < 25 %, or ψ_w > -1 MPa), but this is not much limiting its value and potential use, because physiological processes in lichens are usually inhibited at much lower values of ψ_w than in leaves of vascular plants.

Impact of ultraviolet-B (UV-B) and Cd, applied individually and in combination, measured in terms of oxygen-evolution, chlorophyll (Chl) and protein contents, lipid peroxidation, and enzymatic and non-enzymatic antioxidants of Anabaena doliolum, revealed a greater oxidative damage induced by UV-B than by Cd. While superoxide dismutase (SOD) showed a greater stimulation by UV-B than Cd, the activities of catalase (CAT) and glutathione reductase (GR) declined after UV-B treatment. Cd treatment, however, enhanced the activities of ascorbate peroxidase (APX) and GR. CAT activity increased at low but decreased at high dose of Cd. Increase in carotenoid (Car) content in UV-B treated cells suggested a shielding effect of Car against UV-B stress. A 15-and 10-fold rise in α-tocopherol (α-TOC) content at high dose of Cd and/or UV-B offered testimony to the antioxidant role of α-TOC.

The phytoremediation is an environment friendly, green technology that is cost effective and energetically inexpensive. Metal hyperaccumulator plants are used to remove metal from terrestrial as well as aquatic ecosystems. The technique makes use of the intrinsic capacity of plants to accumulate metal and transport them to shoots, ability to form phytochelatins in roots and sequester the metal ions. Harbouring the genes that are considered as signatures for the tolerance and hyperaccumulation from identified hyperaccumulator plant species into the transgenic plants provide a platform to develop the technology with the help of genetic engineering. This would result in transgenics that may have large biomass and fast growth a quality essential for removal of metal from soil quickly and in large quantities. Despite so much of a potential, the progress in the field of developing transgenic phytoremediator plant species is rather slow. This can be attributed to the lack of our understanding of complex interactions in the soil and indigenous mechanisms in the plants that allow metal translocation, accumulation and removal from a site. The review focuses on the work carried out in the field of metal phytoremediation from contaminated soil. The paper concludes with an assessment of the current status of technology development and its future prospects with emphasis on a combinatorial approach.

Mature apricot (Prunus armeniaca L. cv. Búlida) trees, growing under field conditions, were submitted to two drip irrigation treatments: a control (T1), irrigated to 100 % of seasonal crop evapotranspiration (ETc), and a continuous deficit (T2), irrigated to 50 % of the control throughout the year. The behaviour of leaf water potential and its components, leaf conductance and net photosynthesis were studied at three different times during the growing season, when they revealed a diurnal and seasonal pattern in response to water stress, evaporative demand of the atmosphere and leaf age. The deficit-irrigated trees showed, among other effects, a pronounced decrease in leaf water potential (ψ_w), decreased in leaf conductance (g_s) and no osmotic adjustment. For this reason, g_l and ψ_w can be considered good indicators of mature apricot tree water status and can therefore be used for irrigation scheduling.