We aimed to assess the potential effects of fumigation by methyl salicylate (MeSA) on plant monoterpene production and emissions. We evaluated monoterpene production and emissions both by chromatographic and proton transfer reaction mass spectrometry at the whole plant-and leaf-scales, in MeSa-fumigated (ca. 60 mm³ m^-3 in air) and control (without MeSa fumigation) holm oak (Quercus ilex L.) plants exposed to temperatures ranging from 25 to 50 °C. The MeSa-fumigated plants showed ca. 3-4-fold greater leaf monoterpene concentrations and emission rates than the control plants between the temperatures of 25 to 45 °C.

High efficiency shoot regeneration was achieved through leaflet and cotyledon derived calli in Cassia angustifolia - an important medicinal plant. Dark brown compact callus was induced at the cut ends of the explants on Murashige and Skoog's (MS) medium augmented with 1 µM N⁶-benzyladenine (BA) + 1 µM 2,4-dichlorophenoxyacetic acid (2,4-D). Such callus pieces on transfer to cytokinins (BA or kinetin) supplemented medium differentiated shoots within 10 - 15 d. Of the two cytokinins, 5 µM BA was optimum for eliciting morphogenic response in 83.33 and 70.83 % cultures with an average of 4.16 ± 0.47 and 3.70 ± 0.56 shoots in cotyledon and leaflet derived calli, respectively. The addition of 0.5 µM α-naphthaleneacetic acid (NAA) to MS + 5 µM BA further elevated the maximum average number of shoots to 12.08 ± 1.04 and 5.37 ± 0.52 for cotyledon and leaflet calli, respectively. The excised shoots were transferred to a rooting medium containing either IAA (indole-3-acetic acid), IBA (indole-3-butyric acid) or NAA. Nearly 95 % shoots developed an average of 5.4 ± 0.41 roots on half strength MS medium supplemented with 10 µM IBA.

A short time effects of 25 and 150 µM Cu²⁺ or 50 µM methyl jasmonate (MJ) on growth of roots and leaves of Phaseolus coccineus, Allium cepa and Zea mays were investigated. Both Cu²⁺ and MJ inhibited root growth. Jasmonate synthesis inhibitors (ibuprofen, IB, salicylhydroxamic acid, SHAM, and propylgallate, PG) partially reversed the inhibitory effect of Cu²⁺ in P. coccineus, but in A. cepa this effect was not clear. Pretreatment with NADPH oxidase inhibitor (20 mM imidazole, IM), and especially ethylene inhibitor (silver thiosulphate, STS) mostly weakened Cu²⁺ effect on root growth in P. coccineus and A. cepa. The growth of P. coccineus leaves also slowed down by Cu²⁺ and this effect was partially ameliorated by IB, PG and IM, and completely by SHAM and STS. In Z. mays the effect of STS was considerably lower than that of PG and SHAM which reversed the effect of Cu²⁺. These results indicate that jasmonate, ethylene and NADPH oxidase activity may be involved in Cu²⁺ inhibitory action on the roots of dicotyledon plants, but in A. cepa only ethylene and NADPH oxidase are involved. However, leaf growth inhibition induced by excess Cu²⁺ is connected in Z. mays especially with jasmonate, and in P. coccineus with ethylene, NADPH oxidase and, to a minor degree, with jasmonate.

Embryonal axis explants from 2-d-old in vitro germinated seeds were used to induce multiple shoot production. The combination of 4.44 µM BA and 1.59 µM NAA in MS medium triggered the initiation of adventitious shoot buds. The explants with shoot buds produced maximum number of shoots (10.6 per explant) in MS medium supplemented with 4.44 µM BA and 0.065 mM L-glutamine in three successive transfers. The elongated shoots were rooted on MS medium with 4.92 µM IBA. Rooted plants were transferred to soil with a survival rate of 65 %.

An efficient protocol has been developed for rapid micropropagation of Ocimum basilicum. Multiple shoots were induced by culturing shoot tip explants excised from mature plants on a liquid Murashige and Skoog (MS) medium supplemented with 5-100 µM of thidiazuron (TDZ) for different treatment duration (4, 8, 12 and 16 d). The optimal level of TDZ supplementation to the culture medium was 50 µM for 8 d induction period followed by subculturing in MS medium devoid of TDZ as it produced maximum regeneration frequency (78 %), mean number of shoots (11.6 ± 1.16) and shoot length (4.8 ± 0.43 cm) per explant. A culture period longer than 8 d with TDZ resulted in the formation of fasciated or distorted shoots. The regenerated shoots rooted best on MS medium containing 1.0 µM indole-3-butyric acid (IBA). The micropropagated shoots with well developed roots were successfully established in pots containing garden soil and grown in greenhouse with 95 % survival rate. The regenerated plants were morphologically uniform and exhibited similar growth characteristics and vegetative morphology to the donor plants.

We measured PAR-saturated CO₂ exchange rate (CER), and leaf N, P and chlorophyll (Chl) concentrations in 21 plant species, selected to encompass as broad a range in specific leaf area (SLA) as possible, and encompassing non-succulent C3 as well as succulent CAM plants. We worked with plants growing under uniform conditions in the facilities of a biological research station to ensure that any correlations found were due to inherent, genetically controlled, relationships between the measured parameters and not due to variations in resource availability in different habitats. We found CER to be strongly correlated to SLA, leaf N concentration and Chl concentration. CER increased much faster with increasing leaf N concentration (CER ≈ N^3.1) than with increasing SLA (CER ≈ SLA^1.2). CER also increased much faster with leaf N concentration than with increasing Chl concentration (CER ≈ Chl^1.3), indicating the photosynthetic N-use efficiency (NUE) to be higher for plants with high N concentration than for plants with low N concentration (NUE ≈ N^2.1). Analysis of covariance showed that these relationships exist even when comparing plants of widely different growth forms - succulent or non-succulent, and of different photosynthetic pathways, as the C3 and CAM plants compared here. Testing against scaling coefficients calculated using dimensional analysis, showed that the scaling of N, Chl and CER against SLA was not merely a result of diluting N and Chl with carbon in thicker leaves but that SLA, probably through influencing light absorptio and/or CO2 diffusion pathway, played an independent role in controlling CER.

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

The in vitro studies with Cardiospermum halicacabum indicated that the different explants, i.e cotyledon, hypocotyl, cotyledonary node, leaf, internode and node had the potential to produce calli on Murashige and Skoog (MS) medium supplemented with benzylaminopurine (BAP) and napthalene acetic acid (NAA). Calli of different explant origin showed variable growth responses on different BAP concentrations. The shoots were favourably formed from the calli of leaf and cotyledon explants. The maximum number of shoots were produced from calli subcultured on MS + BAP (17.8 µM). The roots were initiated on growth regulator free MS medium.

Methods for induction of callus and cell suspension cultures have been developed for the medicinally important herb Centella asiatica (L.) Urban. Thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) analysis showed the presence of asiaticoside in the in vitro grown leaves, callus and cell suspension cultured cells.

Ferredoxin-dependent glutamate synthase (Fd-Gogat; EC 1.4.7.1) in leaf and root plastids is the last enzyme involved in the pathway of nitrate assimilation in higher plants. Arabidopsis thaliana expresses two different genes: the first, light regulated, specific of green tissues and the second expressed in other tissues. In this work, we investigated whether in our clone, OsGog2 AC Y12595, this gene is up-regulated by light or it is expressed under darkness. Fd-Gogat specific activity, protein and mRNA increased after light treatment in rice shoots. In roots, the activity and the protein content remained constant, whereas the mRNA is repressed by light treatment. The results obtained using a specific probe, situated in the 3' untranslated region of the OsGog2 cDNA, indicated that OsGog2 gene is up-regulated by light and that its expression is tissue specific and suggested that a dark expressed Fd-Gogat gene could be present in rice similarly as in Arabidopsis.

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 physiological responses to NaCl salinity were investigated in two floating aquatic macrophytes, Pistia stratiotes L. and Salvinia molesta L. With the increasing NaCl concentration a decrease in chlorophyll and carotenoid contents was recorded in Salvinia as compared to Pistia. Also a greater increase in H₂O₂ accumulation and lipid peroxidation was observed in the shoot and root tissues of Salvinia as compared to Pistia. The superoxide dismutase, glutathione reductase, catalase and guaiacol peroxidase activities, and ascorbate and glutathione contents increased in Salvinia and Pistia shoot and root tissues in response to NaCl.

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.

Gender based differences in response to low temperature stress in leaf chlorophyll (Chl), and carotenoids (Car) contents and chlorophyllase (Chlase) activity were monitored in male (Kapoori Vellaikodi and Madras Pan Kapoori) and female (Bangla Mahoba, Desi Bangla and Kaker) betel vine landraces. Although female plants contained nearly two fold more Chl than male counterparts, the low temperature induced Chl loss was comparable, however, male plants showed higher Chl a/b ratio than females. Chlase activity increased due to cold stress in all the landraces. Male plants always showed higher activities of Chlase, which may be one of the reasons for the rather low Chl contents in male plants.

The plants of pigeonpea (Cajanus cajan L.) cv. H77-216 were subjected to moderate [soil moisture content (SMC) = 7.3 ± 0.5 %] and severe (SMC = 4.3 ± 0.5 %) drought by withholding the irrigation at vegetative stage (45 d after sowing). The control plants were maintained at SMC of 11.0 ± 0.5 %. Half of the stressed plants were re-irrigated and their recovery was studied after 2 d. Leaf water potential, osmotic potential, and relative water content of leaf and root decreased significantly while a sharp rise in proline and total soluble sugars contents were noticed. Drought induced a significant increase in 1-aminocyclopropane 1-carboxylic acid (ACC) content and ACC oxidase activity which caused a considerable increase in ethylene evolution. Malondialdehyde content and relative stress injury were increased under drought whereas reverse was true for ascorbic acid content. The membrane integrity of roots decreased during stress and recovered on rehydration. The specific activity of total superoxide dismutase, ascorbate peroxidase, glutathione reductase, and glutathione transferase decreased to 37 - 78 %, 17 - 62 %, 29 - 36 % and 57 - 79 % at moderate and severe drought, respectively. The increase in activity of catalase and peroxidase could not overcome the accumulation of H₂O₂ content in the roots.

Azolla caroliniana was exposed to 5 °C in darkness for 1, 2, 3, 5 or 7 d and then recovered for 7 d. Plants previously chilled for 2 or 3 d exhibited higher growth rates when transferred to normal temperature than either the control plants or those previously chilled for 5 or 7 d. Increased plant growth may be related to increased contents of chlorophyll, sucrose, and reducing sugars, due to increased photosynthetic capacity. In another experiment Azolla plants were chilled at 5 °C for 7 d and then transferred for 0, 4, 8, 12, or 16 d recovery to the N-free Hoagland solution or Hoagland solution containing 5 mM KNO₃. In previously chilled plants, the growth rate was decreased. In the medium supplemented with nitrogen, the growth rate was greater than in the N-free medium in both chilled and nonchilled plants. In chilled plants the decrease in growth rate may be related to the disturbance of Anabaena azollae cells where the protecting envelope of the heterocysts was deorganized. During the recovery the rate of N₂-fixation increased in both chilled and nonchilled plants up to 12 d after which both rates were similar. However, during the first 4 d the rate of the nonchilled plants was approximately 4-fold that of the previously chilled plants. Nitrate reductase and nitrite reductase activities in control plants were higher than in those previously chilled for 7 d. Both activities increased in nonchilled and previously chilled plants up to 12 d then decreased. The total protein content increased up to 12 d in chilled and nonchilled plants after which it decreased. Under all treatments, the values were higher in nonchilled plants than in those previously chilled ones and were also higher in presence of N than in its absence. Thus the presence of N-source in the medium counteracts the effect of chilling injury particularly during prolonged recovery.

In vitro salt tolerant rice plants established by step up treatment with 0.5, 1.0, 1.5 and 2.0 % NaCl at 3-week intervals were examined to determine whether they could grow in potted paddy soil containing 0, 0.55 or 0.75 % NaCl till harvesting. All the control plants were necrotic by the 4^th week in the culture. At the 10^th week of culture, 100 % of the salt-tolerant plants subjected to 0 or 0.55 % NaCl survived, and 78 % of the plants at 0.75 % NaCl. The Na⁺ and Cl^- contents in the leaves of salt-tolerant plants grown at 0.55 and 0.75 % NaCl were about 4 times of those without NaCl. The ion contents in non-tolerant plants and seedling plants were 10 to 12 times of those in 0 % NaCl treatment. One of the hypotheses to explain the present data is that the in vitro step up salt selection induces the capability to maintain no lethal concentration of NaCl in the leaves.

A high frequency adventitious shoot regeneration protocol was developed for henbane (Hyoscyamus niger L.) using thidiazuron (TDZ). Hypocotyl, cotyledon and stem explants were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations of N⁶-benzylaminopurine and TDZ. MS medium supplemented with 16 μM TDZ was the most effective for providing 100 % regeneration frequency associated with a 19.53 shoots per hypocotyl explant. Plantlets were rooted on MS medium supplemented with different concentrations of indole-3-butyric acid (IBA) and α-naphthaleneacetic acid. High rooting and survival was achieved using half strength MS medium supplemented with 8 μM IBA.

Wild-type and the handshake (has) mutants in Arabidopsis thaliana were analyzed. Compared to the wild-type, has mutants display a number of morphological alterations, which can largely be traced back to altered meristem function. Analyses of apical meristem of mutant plants showed that mutation affected meristem structure and patterns of STM expression.

An efficient protocol of direct somatic embryogenesis (without involving intermediate callus) has been developed from stem segments and shoot tips of Capsicum annuum L. Explants were cultured on Murashige and Skoog (MS) medium supplemented with thidiazuron (TDZ). Among the various concentration of TDZ tested, 0.5 μM was proved to be best for induction of somatic embryos. Induction, maturation and germination were achieved on the same medium. The shoots developed from somatic embryos were transferred for rooting to MS medium supplemented with indole-3-butyric acid (IBA). All the regenerated plants with 85 % survival rate were normal with respect to morphology and growth characteristics.

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.

Rice (Oryza sativa L.) was grown in pots with pyridine N-oxide (PNO), 4-morpholino pyridine N-oxide (MNO), and sodium meta silicate as the sources for silicon. Aliquots of these were added in fortnightly intervals to seedlings through anthesis stage. The plants were monitored for plant growth characteristics, chlorophyll content (SPAD values), photosystem 2 activity (variable to maximum fluorescence ratio of dark adapted leaves), and for blast and yellow stem borer resistance. Deposition of silica in the leaves was monitored by scanning electron microscopy and silicon mapping. PNO or MNO application resulted in significant silicon accumulation in leaf bundle sheath cells. Application of PNO and MNO imparted disease and pest resistance by increasing silicon uptake of rice plants.

Common polypody (Polypodium vulgare L.) belongs to desiccation-tolerant ferns. The structure of storage parenchyma of their rhizome was examined by transmission electron microscopy after dehydration and subsequent rewetting. Analysis revealed that treatment with supplemental abscisic acid resulted in protection of cells against ultrastructural damage compared to untreated ones. Dehydration rate appears to modify the ability of rhizome parenchyma to stand water stress.

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.

Flow cytometric analysis with 4,6-diamidino-2-phenylindole (DAPI) staining was used to screen for chromosomal changes in Quercus robur during in vitro culture. The initiated cell lines (1992 until 1999) were maintained via secondary embryogenesis on P24 medium with 0.9 μM 6-benzylaminopurine (BAP) in regular subculture intervals of 6 weeks. Regenerated plants established in the greenhouse and in vitro plantlets derived from encapsulated somatic embryos were screened. The embryogenic cell lines were characterized as individual clones by isoenzyme analysis. Flow cytometric relative DNA content analysis of the first screening period revealed that somaclonal variation in form of tetraploidy occurred in two out of 26 tested somatic embryo clones (Alt and Jung). These two clones lost their ability to convert into plantlets. Intraspecific relative DNA content variation including technical variation was below 3 %. In the second screening period, however, 3 out of 37 clones (Alt, E4.31H9 and P3.27H) contained tetraploid cells leading to the assumption that the frequency of tetraploidy seems to be correlated with the duration of in vitro culture. No chromosomal differences were detected in regenerated plants. However, tetraploidy occurred in 8 % of the tested clones over a culture period of 7 years.

We describe the multi-step regeneration system of medicinal plant Schisandra chinensis (Turcz.) Baill. The seeds were pre-treated with 0.005 μM thidiazuron. Subsequently the zygotic embryos of the early heart stage were cultured on medium with 50 μM of 2,4-dichlorophenoxyacetic acid (2,4-D) and after three weeks the embryogenic calli were transferred to a medium with 10 μM of 2,4-D and 4 μM of 6-benzyladenine and were sub-cultured at the 4-week intervals. Abscisic acid (30 μM) and polyethyleneglycol (3 %) significantly influenced the synchronization of development of the somatic embryos (SEs) to the globular stage. The following culture on a medium without growth regulators resulted in full developed cotyledonary stage SEs. Indole-3-butyric acid (0.05 μ) contributed to their rapid conversion to plantlets.

Competition is a major density-dependent factor structuring plant populations and communities in both natural and agricultural systems. Seedlings of the model plant species Arabidopsis thaliana cv. Columbia, and the Columbia-derived stomatal mutants sdd1 and tmm1, were grown under controlled conditions at increasing densities of 1, 10, 20, and 50 plants per pot. We demonstrate significant effects of time (days after planting), density, genotype, density and genotype, and the three-way interaction with time upon several fitness components (plant height, silique number, leaf biomass and flowering stalk biomass) in Columbia and these mutants.

In vitro morphogenesis of sweet potato (Ipomoea batatas) shoot explants after cultures in callus initiation medium (CIM) with two sucrose contents and plant regeneration medium (PRM) with three growth regulator combinations for different durations was studied. After 4 weeks, explants on 5 % sucrose CIM had significantly more shoots but similar or lower root fresh mass and callus fresh mass than those on 3 % sucrose CIM subsequent to transfer for 6 weeks on all three PRM. Cultures transferred to growth regulator-free PRM after 4 and 12 weeks on 5 % sucrose CIM formed plants through organogenesis and embryogenesis, respectively. Embryogenic cultures from 4 weeks on CIM + 10 weeks on callus proliferation medium when transferred to PRM without growth regulator for 4 and 8 weeks produced multiple embryos in the prior and both embryos and shoot buds in the later.

The induction of haploid plants by in vitro gynogenesis is a promising practice in onion breeding. In order to increase the frequency of embryo regeneration and haploid plant production in Valcatorce INTA, Cobriza INTA and Navidena INTA cultivars, putrescine and CCC were used, either as a component of the culture media or by spraying or injecting them to the umbels. Additionally, two concentration of gellam gum were tested. A higher number of gynogenic embryos was achieved by using 7 g dm^-3 gellam gum, and this number was not affected by the addition of putrescine to the media. CCC sprayed at the umbels significantly increased the gynogenic embryo rate, which was more than three times higher than the control. Cobriza INTA showed the highest induced embryo rate (4.76 %).