DNA variations of forty-eight Eucalyptus globulus plants, regenerated by successive culture from seven different explants were assessed by AFLP analysis using 18 primer combinations. At least one variation showed 66.7 % of the analyzed plants, and the numbers of polymorphic bands per plant ranged from 1 to 22. The proportion of polymorphic fragments did not correlate with the numbers of the regenerated plants. However, the more times of successive culture were done the more of polymorphic bands were found within the groups. On average, between 97.39 and 99.88 % of all fragments were shared within the same group. AMOVA analysis showed 39.33 % of the variation was found among the accessions that originated from different calli while 60.67 % was from same calli.

An efficient in vitro plant regeneration system from leaves of Ophiorrhiza japonica Blume was established for the first time. Callus formation rate was more than 90.4 % from leaf segments on Murashige and Skoog (MS) supplemented with either α-naphthaleneacetic acid (NAA) alone or in combination with 6-benzyladenine (BA). The highest shoot regeneration (78.9 %) was achieved on MS medium containing 2.0 mg dm^-3 BA and 0.2 mg dm^-3 NAA, with an average of 9.4 shoots developed per leaf segment. Shoot regeneration was also improved when the leaf explants were cultured in MS basal medium supplemented with 0.5 % (m/v) polyvinylpyrrolidone (PVP). The leaf explants from seedlings with age of about 18-27 d showed the highest shoot regeneration. The regenerated shoots were rooted on half-strength basal MS medium supplemented with 0.5 mg dm^-3 indole-3-butyric acid (IBA), which averagely produced 24.8 roots per shoot. The plantlets were transferred to soil, where 100 % survived after 1 month of acclimatization.

Yellow lupin (Lupinus luteus L.) plants were grown in hydroponic solution for 15 d under different copper concentrations (0.1, 0.5, 1.0, 10, 25 and 50 µM). With increasing Cu concentration total biomass was not affected, leaf area slightly decreased, while chlorophyll content decreased considerably. Cu content increased significantly both in roots and in leaves, but the contents of other ions were only slightly affected at the highest Cu concentration (Mn content decreased both in roots and in leaves, P content decreased only in leaves and Zn content increased in roots). Superoxide dismutase (SOD) activity increased up to day 7 after copper application. Peroxidase (GPOD) and polyphenol oxidase (PPO) activities also increased, while catalase (CAT) activity remained constant.

The effects of salicylic acid (SA) and salinity on the activity of apoplastic antioxidant enzymes were studied in the leaves of two wheat (Triticum aestivam L.) cultivars: salt-tolerant (Gerek-79) and salt-sensitive (Bezostaya). The leaves of 10-d-old seedlings grown at nutrient solution with 0 (control), 250 or 500 mM NaCl were sprayed with 0.01 or 0.1 mM SA. Then, the activities of catalase (CAT), peroxidase (POX) and superoxide dismutase (SOD) were determined in the fresh leaves obtained from 15-d-old seedlings. The NaCl applications increased CAT and SOD activities in both cultivars, compared to those of untreated control plants. In addition, the NaCl increased POX activity in the salt-tolerant while decreased in the salt-sensitive cultivar. In control plants of the both cultivars, 0.1 mM SA increased CAT activity, while 0.01 mM SA slightly decreased it. SA treatments also stimulated SOD and POX activity in the salt-tolerant cultivar but significantly decreased POX activity and had no effect on SOD activity in the saltsensitive cultivar. Under salinity, the SA treatments significantly inhibited CAT activity, whereas increased POX activity. The increases in POX activity caused by SA were more pronounced in the salt-tolerant than in the salt-sensitive cultivar. SOD activity was increased by 0.01 mM SA in the salt-tolerant while increased by 0.1 mM SA treatment in the salt-sensitive cultivar.

The aim of this research was to determine whether exogenous abscisic acid (ABA) applied immediately after ex vitro transfer of in vitro grown plants can improve their acclimatization. Tobacco (Nicotiana tabacum L.) plantlets were transferred into pots with Perlite initially moistened either by water or 50 µM ABA solution and they were grown under low (LI) or high (HI) irradiance of 150 and 700 µmol m^-2 s^-1, respectively. Endogenous content of ABA in tobacco leaves increased considerably after ABA application and even more in plants grown under HI. Stomatal conductance, transpiration rate and net photosynthetic rate decreased considerably 1 d after ex vitro transfer and increased thereafter. The gas exchange parameters were further decreased by ABA application and so wilting of these plants was limited. Chlorophyll (a+b) and β-carotene contents were higher in ABA-treated plants, but the content of xanthophyll cycle pigments was not increased. However, the degree of xanthophyll cycle pigments deepoxidation was decreased what also suggested less stress in ABA-treated plants. No dramatic changes in most chlorophyll a fluorescence parameters after ex vitro transfer suggested that the plants did not suffer from restriction of electron transport or photosystem damage.

Maize (Zea mays L.) is generally considered to be a plant with aerenchyma formation inducible by environmental conditions. In our study, young maize plants, cultivated in various ways in order to minimise the stressing effect of hypoxia, flooding, mechanical impedance or nutrient starvation, were examined for the presence of aerenchyma in their primary roots. The area of aerenchyma in the root cortex was correlated with the root length. Although 12 different maize accessions were used, no plants without aerenchyma were acquired until an ethylene synthesis inhibitor was employed. Using an ACC-synthase inhibitor, it was confirmed that the aerenchyma formation is ethylene-regulated and dependent on irradiance. The presence of TUNEL-positive nuclei and ultrastructural changes in cortical cells suggest a connection between ethylene-dependent aerenchyma formation and programmed cell death. Position of cells with TUNEL-positive nuclei in relation to aerenchyma-channels was described.

An efficient, highly reproducible system for plant regeneration via somatic embryogenesis was developed for Cenchrus ciliaris genotypes IG-3108 and IG-74. Explants such as seeds, shoot tip segments and immature inflorescences were cultured on Murashige and Skoog (MS) medium supplemented with 2.0-5.0 mg dm^-3 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5 mg dm^-3 N⁶-benzyladenine (BA) for induction of callus. Callus could be successfully induced from all the three explants of both the genotypes. But the high frequency of embryogenic callus could be induced only from immature inflorescence explants. Somatic embryos were formed from nodular, hard and compact embryogenic calli when 2,4-D concentration was gradually reduced and BA concentration increased. Histological studies of somatic embryos indicated the presence of shoot apical meristem with leaf primordia. Ultrastructural details of globular and scutellar somatic embryos further validated successful induction and progression of somatic embryogenesis. Shoots were differentiated upon germination of somatic embryos on MS medium containing 2,4-D (0.25 mg dm^-3) and BA or kinetin (1-5 mg dm^-3). Roots were induced on 1/2 MS medium containing charcoal (0.8 %), and the regenerated plants transferred to pots and established in the soil showed normal growth and fertility.

Exposure of barley plants (Hordeum vulgare L.) to soil flooding for 72 - 120 h led to decrease in the content of the both subunits of ribulose-1,5-bisphosphate carboxylase/oxygenase. The effect was more pronounced on the small subunit. Further, the changes in protein pattern were observed, mainly proteins with molecular masses 30 - 85 kD were down-regulated.

Effects of zinc [0 and 5.0 mg Zn kg^-1 (soil)] on photosynthetic rate (P_N), and chlorophyll fluorescence in leaves of maize (Zea mays L.) cv. Zhongdan 9409 seedlings grown under different soil moisture regimes (40-45 % and 70-75 % of soil saturated water content) were studied. Zn application did not enhance maize plant adaptation to drought stress. The relative water content and the water potential of leaves were not affected by Zn treatment. Moreover, The P_N of drought-stressed plants was not improved by Zn supply. The increases of plant biomass, stomatal conductance and quantum yield of photosystem 2 due to Zn addition were notable in well-watered plants.

An experiment was conducted with two contrasting pigeon pea (Cajanus cajan L.) genotypes, ICPL 84023 (tolerant) and ICP 7035 (susceptible), to study the physiological and molecular basis of waterlogging tolerance in relation to oxidative stress and antioxidant enzyme activities. Waterlogging resulted in visible yellowing and premature senescence of leaves, and greater decline in relative water content, chlorophyll content, and membrane stability index in ICP 7035 than in ICPL 84023. Superoxide radical and hydrogen peroxide contents increased at day 4 and 6 of waterlogging probably due to activation of NADPH-oxidase. O₂ ^.- production was inhibited, by diphenylene iodonium chloride, a specific inhibitor of NADPH oxidase and expression of NADPH oxidase-mRNA was increased under waterlogging condition in ICPL 84023. ICP 7035 showed higher contents of ROS in control condition and after recovery, however, during waterlogging the O₂ ^.- production was higher in ICPL 84023. Activities of antioxidant enzymes superoxide dismutase (SOD), ascorbate peroxidase (APX), glutathione reductase and catalase increased under waterlogging more in ICPL 84023 than in ICP 7035. Cu/Zn-SOD and APX-mRNA expression in 24-h waterlogged plants showed enhanced expression in ICPL 84023 compared to ICP 7035. The cloning and sequencing of APX gene of tolerant and susceptible genotypes yielded cDNAs of 622 and 623 bp, having 95 % homology with each other and 92 % with the corresponding sequences of Vigna unguiculate APX-gene.

Na⁺ accumulation was investigated in the roots of 11-d-old cowpea [Vigna unguiculata (L.) Walp.] plants. The relative contribution of different membrane transporters on Na⁺ uptake was estimated by applying Ca²⁺, K⁺, NH₄ ⁺, and pharmacological inhibitors. Na⁺ accumulation into the root symplast was decreased by half in the presence of 1 mM Ca²⁺ and it was almost abolished by 100 mM K⁺. The inhibitory effect of external NH⁴⁺ on Na⁺ accumulation was more pronounced in the roots of NH₄ ⁺-free growing plants. Na⁺ accumulation was reduced about 73 % by 0.1 mM flufenamate and it was almost blocked by 2 mM quinine. In addition, 20 mM tetraethylammonium and 1.0 mM Cs⁺ decreased Na⁺ accumulation by 28 and 30 %, respectively. These results evidenced that low-affinity Na⁺ uptake by cowpea roots depends on Ca²⁺-sensitive and Ca²⁺-insensitive pathways. The Ca²⁺-sensitive pathway is probably mediated by nonselective cation channels and the Ca²⁺-insensitive one may involve K⁺ channels and to a lesser extent NH₄ ⁺-sensitive K⁺ transporters.

Oat (Avena sativa L.) plants cultured in soil and hydroponic culture were treated with cadmium [0.154 mg g^-1 (dry soil) and 100 μM CdSO₄, respectively] for 21 d and growth rate and various biochemical processes were studied. Applied cadmium reduced plant growth and chlorophyll content. Changes in activity of enzymes involved in C, N and S metabolism and in guaiacol peroxidase activity were observed. In particular, O-acetylserine sulphydrylase (OASS; EC 4.2.99.8) activity was increased by Cd exposure in both growth conditions, probably as a resistance mechanism to cadmium based on the production of phytochelatins. Results show that both field and hydroponic conditions represent suitable systems for investigating Cd effects on plant growth and metabolism.

We used a random amplified polymorphic DNA (RAPD) amplification method to identify molecular markers associated with high quercetin accumulation in the leaves of Psidium guajava L. trees, selected from four different Mexican agronomic regions. We identified six polymorphic RAPD fragments of 620, 590, 370, 690, 480 and 460 bp among individuals of P. guajava. Genetic linkage disequilibrium analysis revealed that three RAPD profiles considered as DNA markers (620/590 bp, 370 bp and 480/460 bp) had a positive, direct association with quercetin content. These informative molecular markers can be used for selective identification of plants with the highest accumulation of flavonoids.

This study examined the impacts of elevated CO₂ or O₃ on indole-3-acetic acid (IAA) content, activities of IAA oxidase (IAAO) and peroxidase (POD) in Ginkgo biloba leaves. Plants grown in open-top chambers were exposed to ambient atmosphere (control; C), elevated CO₂ and elevated O₃ from 1 June to 30 September. An increase in IAA content and decrease in IAAO and POD activities were observed in plants exposed to elevated CO₂ compared with C. Elevated O₃ had no significant effect on IAA content and IAAO activity, but increased POD activity during the early days. When trees pre-exposed to elevated CO₂ were transferred to elevated O₃ or C, the increase in IAAO activity resulted in the decrease in IAA content. When trees pre-exposed to elevated O₃ were transferred to elevated CO₂ or C, IAA content, IAAO and POD activities showed no significant changes. The influence of POD activity on the IAA activity was low.

Two Coffea arabica - Hemileia vastatrix incompatible interactions (I₁: coffee cv. Caturra - rust race VI and I₂: coffee cv S4 Agaro - rust race II) and a compatible interaction (coffee cv. Caturra - rust race II) were compared in relation to the infection process and chitinase activity. In the two incompatible interactions the fungus ceased growth in the early infection stages, while in the compatible interaction no fungus growth inhibition was observed. A high constitutive level of chitinase activity was detected in the intercellular fluid of healthy leaves. Upon infection, chitinase isoforms were more abundant in incompatible interactions than in the compatible interaction. Immunodetection showed that class I chitinases are particularly relevant in the incompatible interactions and might participate in the defence response of the coffee plants.

Strawberry plants (Fragaria×ananassa Duch.) cvs. Nyoho and Toyonoka were exposed to temperatures of 20, 33, and 42 °C for 4 h, and protein patterns in leaves and flowers was analyzed by 2-dimensional polyacrylamide gel electrophoresis and immunoblotting. In leaves and flowers of both cultivars, the content of most proteins decreased, but a few new proteins appeared in response to heat stress. These heat shock proteins (Hsps) were detected in the range of 19 - 29 kDa in leaves, and 16 - 26 kDa in flowers. The intensity of a 43 kDa protein spot increased in response to heat stress in Nyoho flowers, but not in Toyonoka flowers. The peaHsp17.7 antibody recognized one band at approximately 26 kDa in leaves, and two bands at approximately 16 and 17 kDa in flowers of both cultivars. These results show that the effects of heat stress on Hsp synthesis in strawberry plants differ between plant organs and between cultivars.

The objective of this work was to test whether Ca²⁺, a second messenger in stress response, is involved in ABA-induced antioxidant enzyme activities in Stylosanthes guianensis. Plants were sprayed with abscisic acid (ABA), calcium channel blocker, LaCl₃, calcium chelator, ethylene glycol-bis(β-amino ethyl ether)-N,N,N', N'-tetraacetid acid (EGTA), and ABA in combination with LaCl₃ or EGTA. Their effects on superoxide dismutase (SOD) and ascorbate peroxidase (APX) activities and chilling resistance were compared. The results showed that ABA decreased electrolyte leakage and lipid peroxidation but increased maximum photochemical efficiency measured as variable to maximum fluorescence ratio (F_v/F_m) under chilling stress. Treatment with LaCl₃ or EGTA alone and in combination with ABA increased electrolyte leakage and lipid peroxidation, decreased Fv/Fm, suggesting that the block in Ca²⁺ signalling decreased chilling resistance of S. guianensis and the ABA-enhanced chilling resistance. ABA-induced SOD and APX activities were suppressed by LaCl₃ or EGTA. The results suggested that Ca²⁺ is involved in the ABA-enhanced chilling resistance and the ABA-induced SOD and APX activities in S. guianensis.

In pot experiments performed on maize seedlings chilled at 5 °C, leaf injury was diminished by the application of elevated temperature (1 or 5 h at 15 or 20°C, "warm breaks" treatment) in a dose-dependent manner. The lower the injury count, the higher the catalase (CAT) activity. In a separate experiment, the application of 100 % relative humidity also protected the plants from chilling injury and water loss, increased their gas exchange and variable to maximum chlorophyll fluorescence ratio (F_v/F_m), but did not influence CAT activity. Another protective environmental factor, elevated atmospheric CO₂ concentration [700 µmol(CO₂) mol^-1(air)] diminished CAT activity inhibition, but only in plants of chilling-resistant cultivar. The positive impact of specific environmental factors accompanying chilling is not obviously related to the suppression of the inhibition of CAT activity, although the enzyme is considered as chilling-sensitive.

Tobacco (Nicotiana tabacum L.) plantlets were grown on Murashige and Skoog medium in ventilated Magenta boxes and for the last subculture 10 µM ABA was added to the medium. After three weeks plantlets were transferred into pots with Perlite moistened with water and grown in controlled conditions (16-h photoperiod, day/night temperature 25/20 °C, air humidity about 45 %) either under low or high irradiance of 150 (LI) and 700 (HI) µmol m^-2 s^-1, respectively. Content of endogenous ABA was 271.7 pmol g^-1(f.m.) in ABA treated plantlets, while in control plantlets it was only 53.3 pmol g^-1(f.m.). After ex vitro transfer, stomatal conductance and transpiration rate decreased considerably in comparison with in vitro grown plantlets and remained lower also 7 d after ex vitro transfer, especially in ABA-treated plants and so wilting of plants was practically eliminated. Net photosynthetic rate also decreased 1 d after ex vitro transfer but after 7 d it was mostly higher than that of in vitro grown plantlets. Water use efficiency significantly increased in ABA-treated plants. Chlorophyll a+b content did not change immediately after ex vitro transfer, nevertheless, after 7 d chlorophyll content was higher in ABA-treated plants. Pool of xanthophyll cycle pigments (XCP) and the degree of their deepoxidation (DEPS), which are connected with harmless dissipation of light energy, increased under high irradiance. Contents of XCP and ABA precursors (neoxanthin and violaxanthin) were lower in ABA-treated plants than in control plants indicating less stress in these plants. Most chlorophyll a fluorescence parameters did not change considerably after ex vitro transfer and so the photoinhibition was not observed even under HI. Slight increase in non-photochemical quenching under HI in ABA-treated plants suggested their better photoprotection. Thus application of ABA to the last subculture can improve acclimatization of in vitro grown plants to ex vitro conditions

The involvement of plant hormones in the very early response of plants to virus infection was studied in potato plants (Solanum tuberosum L.) infected with potato virus Y^NTN (PVY^NTN). Endogenous plant hormones and compounds mediating a stress response (JA-jasmonic acid, OPDA-12-oxo phytodienoic acid, SA-salicylic acid, IAA-indole-3-acetic acid, ABA-abscisic acid) were simultaneously quantified in susceptible cv. Désirée and resistant cv. Santé, one and three hours after inoculation. Of the hormones analysed, only the contents of endogenous JA and its precursor OPDA changed in a way that could be clearly connected with the early resistant response. In comparison to susceptible cultivar, a much more pronounced increase of JA was detected in virus-inoculated leaves of resistant cultivar at both time points. The same trend of changes was also observed with OPDA. However, there were no significant changes of JA and its precursor in upper intact systemic leaves and roots, at either time point. These findings implicate the jasmonate signalling pathway in a very early local but not systemic resistant defence of potato to PVY^NTN.

Cuttings of Populus kangdingensis and Populus cathayana, originating from high and low altitudes in the eastern Himalaya, respectively, were examined during one growing season in a greenhouse to determine their responses to drought stress (soil moisture decreased from 100 to 55 or 25 % field capacity). Compared to control plants grown under 100 % field capacity, those poplars grown under 55 and 25 % field capacity possessed lower increases in height and stem diameter, and higher contents of soluble sugars, free proline, malondialdehyde (MDA) and hydrogen peroxide, and higher activities of catalase (CAT), superoxide dismutase (SOD), peroxidase (POD), ascorbate peroxidase (APX) and glutathione reductase (GR). Compared with P. cathayana with greater leaf area, P. kangdingensis with greater root/shoot ratio exhibited lower MDA and H₂O₂ contents, higher soluble sugar and free proline contents, and higher activities of CAT, SOD, POD, APX and GR. These results suggested that P. kangdingensis was more drought tolerant than P. cathayana.

Plantago algarbiensis and P. almogravensis are endemic Al tolerant species from the Western-centre of the Algarve region (South of Portugal) and Portuguese Southwest coast, respectively, which are in risk of global extinction. The aim of this work was to establish an efficient protocol to in vitro propagate these species using shoots obtained from in vitro germinated seeds. The best results in terms of multiplication response were afforded in Murashige and Skoog's (MS) medium supplemented with 6-benzyladenine (8.5 and 9.2 shoots per explant in P. algarbiensis and P. almogravensis, respectively). Shoots of both species showed a great rooting capacity (100 and 80 % for P. algarbiensis and P. almogravensis, respectively) that was not significantly influenced by the concentration of MS macronutrients or auxins. Plants were acclimatized to ex vitro conditions, exhibited normal development (survival rate of 95 and 80 % in P. algarbiensis and P. almogravensis, respectively), and were successfully reintroduced in their natural habitat.

The relative water content (RWC), cell membrane integrity, protein pattern and the expression of late embryogenesis abundant proteins (LEA; group 1, 2, 3 and 4) under different levels of salt stress (0, 1.0, 1.5 and 2.0 % NaCl) were investigated in mulberry (Morus alba L.) cultivars (S1 and ATP) with contrasting salt tolerance. RWC and membrane integrity decreased with increase in NaCl concentration more in cv. ATP than in cv. S1. SDS-PAGE protein profile of mulberry leaves after the NaCl treatments showed a significant increase in 35, 41, 45 and 70 kDa proteins and significant decrease in 14.3, 18, 23, 28, 30, 42, 47 and 65 kDa proteins. Exposure of plants to NaCl resulted in higher accumulation of LEA proteins in S1 than ATP. The maximum content of LEA (group 3 and 4) was detected in S1 at 2.0 % NaCl, which correlates with its salt tolerance.

A novel method of organogenesis in neem (Azadirachta indica A. Juss.) from unfertilized ovaries is described. The Murashige and Skoog's (MS) medium with 9 % sucrose, 1 µM 2,4-dichlorophenoxyacetic acid (2,4-D) and 5 µM 6-benzylaminopurine (BAP) was the best for callus induction from unfertilized ovaries. However, further proliferation of callus occurred better on MS medium supplemented with 0.5 µM 2,4-D either alone or in combination with 4.5 µM kinetin. Maximum shoot regeneration (78 %) was observed when calli, induced from ovaries of 4 mm size flower buds and proliferating on MS + 0.5 µM 2,4-D, were subcultured to MS medium containing 5 µM BAP. Histological analysis revealed that 4 mm sized flower bud corresponds to a 2-nucleate stage of embryo sac. The shoots were then multiplied by forced axillary branching on MS medium supplemented with 1.0 µM BAP and 250 mg dm^-3 casein hydrolysate. The shoots could be rooted on 1/4 strength MS medium supplemented with 0.5 µM indole-3-butyric acid (IBA) at a frequency of 79 %. Cytological analysis by root tip squash preparations revealed that all the plantlets were diploids. These plants were subsequently hardened and established in soil with transplantation rate of 81.8 %.

Axillary buds sampled from a mature 27-year-old Cornus mas cv. Macrocarpa were grown in vitro on modified woody plant medium (WPM). Adventitious rooting performance of microshoots was assayed on half-strength WPM supplemented with 1-naphthaleneacetic acid (NAA) or indole-3-butyric acid (IBA) under various pH. NAA induced significantly higher rooting frequencies than IBA. The pH of 6.8 inhibited rooting, and differentiated roots were extremely thick and fragile. The highest rooting frequency was recorded on half-strength WPM supplemented with 5.37 µM NAA at the pH value adjusted to 6.2 (73 % of rooted shoots). In the presence of IBA, the formation of adventitious roots was observed only in the basal part of the microshoot dipped into rooting medium. In the case of NAA, however, adventitious roots arose also from the parts of microshoots that were not in contact with medium. The growth of aerial roots was always positively gravitropic. The nuclear microsatellite Cf-G17 gave a monomorphic fingerprinting pattern across the mother shrub and micropropagated plantlets. Acclimatized plants did not show any visually detectable morphological variation and the aerial adventitious root formation was no longer observed.

The objective of this study was to examine the role of antioxidant enzymes in waterlogging tolerance of pigeonpea (Cajanus cajan L. Halls) genotypes ICP 301 (tolerant) and Pusa 207 (susceptible). Waterlogging resulted in visible yellowing and senescence of leaves, decrease in leaf area, dry matter, relative water content and chlorophyll content in leaves, and membrane stability index in roots and leaves. The decline in all parameters was greater in Pusa 207 than ICP 301. Oxidative stress in the form of superoxide radical, hydrogen peroxide and thiobarbituric acid reactive substances (TBARS) contents initially decreased, however at 4 and 6 d of waterlogging it increased over control plants, probably due to activation of DPI-sensitive NADPH-oxidase. Antioxidant enzymes such as superoxide dismutase, ascorbate peroxidase, glutathione reductase and catalase also increased under waterlogging. The comparatively greater antioxidant enzyme activities resulting in less oxidative stress in ICP 301 could be one of the factor determining its higher tolerance to flooding as compared to Pusa 207. This study is the first to conclusively prove that waterlogging induced increase in ROS is via NADPH oxidase.

There are significant changes in the structure of the upper tobacco (Nicotiana tabacum L.) leaves systemically infected with tobacco mosaic virus (TMV) especially in the light green tissue (LGT). Dark green areas (DGI) had intermediate status between healthy tissue and LGT. DGI contained significantly less infectious TMV and viral antigen than the LGT. The DGI, LGT and healthy tissues did not differ in the permeability of cell membranes and in the set of acidic pathogenesis-related (PR) proteins but the total content of PR-proteins in the healthy plants was higher than in the infected ones with the DGI being intermediate between healthy tissue and LGT. The crude leaf extracts from DGI and LGT showed less total ribonuclease activity and ribonuclease isozymes in comparison with control.

The transcription factors CBF/DREB play an important role during low temperature, drought and high-salt stress in higher plants. A new CBF (CRT/DRE binding factor) gene was cloned from trifoliate orange [Poncirus trifoliata (L.) Raf.] by RT-PCR with degenerate primers and rapid amplification of cDNA ends (RACE) techniques. The full-length cDNA of CBF gene from trifoliate orange (designated as Ptcbfb) was 847 bp containing a 732 bp open reading frame (ORF), encoding a 243 amino acid protein. The predicted protein (designated as PtCBFb) had over 60 % identity to CBFs from some other plant species. Bioinformatical analysis showed that PtCBFb contained N-terminal bipartite nuclear targeting sequence, potential C-terminal acid domain and high conserved AP2 domain. Some other loci such as phosphorylation sites of several protein kinases, N-myristoylation site, tyrosine sulfation site and amidation site were also conserved in PtCBFb. Predicted three-dimentional structure of PtCBFb was similar to CBF from Arabidopsis thaliana. Expression pattern analysis revealed Ptcbfb expression in every tested organ, and Ptcbfb was cold induced.

Fatty acid content and composition of chloroplast membranes, ethylene production associated with thylakoid lipids degradation as well as photosynthetic electron transport involving photosystems 1 and 2 were used to determine the effects of increasing Cd concentrations in the growth medium [0, 14, 28, and 42 mg (Cd) kg^-1(sand)] on the photosynthetic performance of barley plants (H. vulgare L., cv. CE9704). High concentrations of Cd triggered serious disturbances of the chloroplast membranes. Ethylene production increased whereas a drop of 18:3 fatty acid content occurred, indicating that Cd mediates lipid peroxidation in the thylakoids. The enhanced ethylene production could be used as an early indicator of Cd-induced membrane degradation, yet at very high concentration (42 mg kg^-1) Cd decreased ethylene production.

The leaf water potential, gas exchange and chlorophyll fluorescence were evaluated in five common bean (Phaseolus vulgaris) genotypes A222, A320, BAT477, Carioca and Ouro Negro subjected to moderate water deficit. At the maximum water deficit (10 d of water withholding), the leaf water potential of genotypes A320 and A222 was higher (-0.35 and -0.50 MPa) when compared to the other genotypes (-0.67 to -0.77 MPa). The stomatal conductance and net photosynthetic rate were significantly reduced in all genotypes due to the water deficit. The greater reduction in stomatal conductance of A320 under drought resulted in high intrinsic water use efficiency. Mild water deficit affected the photochemical apparatus in bean genotypes probably by down-regulation since plants did not show photoinhibition. The photochemical apparatus of A222 and A320 genotypes was more sensitive to drought stress, showing reduced apparent electron transport even after the recovery of plant water status. On the other hand, even after 10 d of water withholding, the maximum efficiency of photosystem 2 was not affected, what suggest efficiency of the photoprotection mechanisms.