Retention of shoot regeneration capacity of tobacco callus by Na2SO4

Callus of tobacco (Nicotiana tabacum L. cv. Wisconsin 38) was grown in the light on shoot-forming medium in the presence of Na₂SO₄ for over a year. An increase in Na₂SO₄ concentration resulted in decreasing callus growth, decreasing percentage of calli producing shoots and number of shoots per callus, and increasing callus percent dry weight. Regeneration of shoots from callus grown in the absence of Na₂SO₄ began to decline after 14 months in culture, and shoot regeneration capacity was completely lost after 18 months. In contrast, 18-month old callus continuously grown in the presence of Na₂SO₄ retained the ability to form shoots. The highest percent of callus pieces that formed shoots and the maximum number of shoots per callus occurred at 70 mM (1%) Na₂SO₄. All plants arising from the 18-month old callus were polyploid. Both 9- and 18-month old callus exhibited more negative water and osmotic potentials in the presence of increasing Na₂SO₄ concentration.Abbreviations water potential - _S osmotic potential - CGI callus growth index     

NaCl and Na2SO4 alter responses of jack pine (Pinus banksiana) seedlings to boron

Using sand culture, we examined the responses of 6-month-old jack pine (Pinus banksiana Lamb.) seedlings to boron and salinity (sodium chloride and sodium sulfate) treatments. During 4 weeks of treatments, 60 mM NaCl and 60 mM Na₂SO₄ significantly decreased survival, new shoot length, number of new roots, shoot to root dry weight ratio and transpiration rates. When applied in absence of the salts, B had little effect on the measured variables. However, when applied together with salts, B decreased seedling survival, increased needle injury and altered tissue elemental concentrations in jack pine seedlings. In 2 mM B treatment, B concentration was higher in the shoots than in the roots. However, when 2 mM B was present in NaCl and Na₂SO₄ treatments, shoot boron concentration declined and greater proportion of B accumulated in the roots. This shift corresponded to a decline in transpiration rates. In plants treated with NaCl, Na accumulated primarily in the shoots, while in Na₂SO₄-treated plants Na accumulated mostly in the roots. Based on the electrolyte leakage and needle necrosis data, Cl^– appears to be the major factor contributing to seedling injury and B aggravates the injurious effects of NaCl. We suggest that Cl^– may contribute to Na and B toxicity in jack pine by altering cell membrane permeability leading to increased Na concentration in the shoots.     

Some morphogenic effects of sodium sulfate on tobacco callus

Callus cultures of Nicotiana tabacum L cv. Wisconsin 38 were initiated and grown on shoot-forming (SF) and callus proliferation (CP) medium with or without Na₂SO₄. Two cultures were maintained on SF medium with 0, 0.75, 1 or 1.5% Na₂SO₄ for 2.5 and 3.5 years. In the older culture only callus grown on salt formed shoots throughout the maintenance period, while in the younger culture the control responded best and Na₂SO₄ was inhibitory. Callus from the older culture which had been grown on salt continued to form shoots in the absence of salt. Na₂SO₄ caused adventitious shoot formation in three cultures on CP medium. These shoots were present for 7 subcultures after removal of Na₂SO₄; but established, control callus, did not form shoots when transferred to Na₂SO₄. Callus initiated and maintained on NaCl or mannitol showed a slight increase in shoot initiation. On NaCl, Na₂SO₄ or mannitol, the tissue osmotic potential became more negative and proline concentration increased.     

Physiological effects of a long term exposure to low concentrations of NH3, NO2 and SO2 on Douglas fir (Pseudotsuga menziesii)

The above-ground parts of two years old seedlings of Douglas fir (Pseudotsuga menziesii) were exposed to filtered air, NH₃, NO₂₊, SO₂ (66, 96 and 95 μg m^?3, respectively), to a mixture of NO₂+NH₃ (55 + 82 μg m^?3) or SO₂+NO₂ (128 + 129 μg m^?3), for 8 months in fumigation chambers. Both chlorophyll fluorescence and gas exchange measurements were carried out on shoots which had sprouted at the beginning of the exposure period. The chlorophyll fluorescence measurements were performed after 3 and 5 months of exposure (average shoot age 70 and 140 days, respectively). Light response curves of electron transport rate (J) were determined, in which J was deduced from chlorophyll fluorescence. In addition, light response curves of net CO₂ assimilation were determined after 5 months of exposure. After 3 months of exposure (average shoot age 70 days) all exposure treatments showed a lower maximum electron transport rate (J_max) as compared to the control shoots (filtered air). A large reduction (45%) was observed for shoots exposed to SO₂+NO₂. During the exposure period between 3 and 5 months (average shoot age 70 and 140 days, respectively) a decrease of J_max was observed for all treatments. J_max had further declined some time after termination of the exposure, when average shoot age was 310 days. Shoots exposed to SO₂ and SO₂+NO₂ also showed a reduction in maximum net CO₂ assimilation (P_max) as compared to the control shoots. However, shoots exposed to NO₂ showed no reduction and even a higher P_max was observed for shoots exposed to NH₃ or NO₂+NH₃. Needles of these treatments also showed a higher chlorophyll content which might explain the contradictory results obtained for these treatments: the increased amount of photosynthetic units counteracts the reduction in J_max and consequently no reduction in P_max is measured. Shoots exposed to SO₂ and SO₂+NO₂ also showed a reduction in maximum stomatal conductance (g_s). However, the stomatal opening was larger than could be expected on basis of their (maximum) CO₂ assimilation rate. Consequently, water use efficiency of these shoots was lower than that of the control shoots. Also shoots exposed to NO₂ had a lower water use efficiency due to a significantly higher maximum g_s. Shoots exposed to NH₃ showed a high transpiration rate in the dark, indicating imperfect stomatal closure.     

Overcoming salinity inhibition of sorghum seed germination by hydration-dehydration treatment

Summary Sorghum (Sorghum bicolar (L) Moench cv EA-955 Serena) seeds, subjected or not to hydration-dehydration treatment were planted in distilled water, NaCl, and Na₂SO₄. When seeds were not hydrated-dehydrated the salts inhibited both seed germination and seedling vigour, and caused an increase in the per cent of abnormal seedlings. The salts neither inhibited seed germination nor increased the per cent of abnormal seedlings if the seeds were treated by hydration-dehydration. However, this treatment was not effective in overcoming the inhibitory effect of either NaCl or Na₂SO₄ on seedling vigour. Application of these principles in growing plants under saline environment is discussed.     

In vitro clonal propagation of annatto (Bixa oreliana L.)

Summary A protocol for in vitro propagation of Bixa orellana is described. Plants were regenerated from shoot apex and nodal explants on B5 medium supplemented with 4.9 μM 2-isopentenyl adenine. The multiplication factor of shoot apex explants was higher (nine shoots per explant) than that of the nodal explants (five shoots per explant). Regardless of the position of the nodes, all the nodal explants gave similar responses. However, the size of the nodal explant was an important factor in producing multiple shoots: 0.5 cm nodal explants produced the maximum multiple shoots. Regenerated shoots from shoot apex explants rooted best on MS medium supplemented with 0.05 μM α-naphthalene acetic acid (NAA). whereas shoots regenerated from nodal explants needed 2.7 μM NAA for rooting. Eighty per cent survival of in vivo transferred plants occurred on the best potting substrate, coco peat. Since the multiplication factor was nine per explant, this protocol can be use for commercial microprogation. However, the regeneration capacity declined after 10 subcultures. Approximately, 3350 rooted plants could be generated in 10 mo. after eight subcultures, from one shoot with a shoot apex and four nodes.     

Salt resistance of chickpea genotypes in solutions salinized with NaCl or Na2SO4

Summary To assess the potential for developing a salt resistant cultivar of chickpea (Cicer arietinum L.) 160 genotypes were screened for percent survival after 9 weeks in greenhouse solution cultures, with 50 mM NaCl or 25 mM Na₂SO₄. All plants grew well in the sulfate treatment but only cv. L-550 survived the chloride treatment. Salt damage appeared and developed slowly. To check these apparent effects of cultivar and kind of anion, three genotypes including cv. L-550 were then grown in solutions with isoosmotic NaCl or Na₂SO₄ at three levels (−0.044, −0.088, and −0.132 MPa), and in a separate experiment cv. L-550 was grown with NaCl and Na₂SO₄ at four levels: 10, 20, 30 and 50 mM Na. Salt composition affected shoot weight less than salt level or cultivar did. Shoot dry weight was only slightly less in chloride treatments than in isoosmotic sulfate, and for the least sensitive cultivar (L-550) this held only at the highest salt level, corresponding to that in the screening trial. Further, sensitivity to sulfate and to chloride was equal when sodium concentrations in shoots were equal, regardless of anion compositions of media. Shoot Na concentration was a useful negative indicator of growth under salt stress regardles of cultivar, and may be a useful tolerance indicator also for other species that neither accumulate nor efficiently exclude Na.     

Development of regulation mechanisms for SO42- influx in spring wheat roots

The development of SO₄^2- influx in roots and sulfur transport to shoots was followed in ³⁵S-tracer experiments for sulfur-deficient spring wheat (Triticum aestivum L. cv. Svenno) seedlings pretreated for various time periods (0–24 h) in nutrient solutions with SO₄^2-. Effects of the metabolic inhibitor 2,4-dinitrophenol (DNP) and the protein synthesis inhibitor cycloheximide (CH) on SO₄^2- influx were also evaluated. The SO₄^2- influx appears feedback-regulated by the internal sulfur level of the roots. Regulation may be achieved solely by a rapidly changed SO₄^2- carrier activity through an allosteric effect by the intracellular SO₄^2- concentration of the roots, followed first by induction of carrier synthesis and then by repression of carrier synthesis after transfer of the roots from SO₄^2--deficient nutrient solutions to solutions with SO₄^2-. A Hill plot of the partly sigmoidal relationship between SO₄^2- influx and intracellular sulfur concentration in the roots gave a Hill coefficient of -4.2, indicating negative cooperativity between a minimum number of four interacting allosteric binding sites for sulfur on each carrier entity. DNP-experiments showed that SO₄^2- influx was mainly metabolic, especially after short pretreatment in SO₄^2- at an external SO₄^2- concentration of 0.1 mM. Pretreatment with CH rapidly prevented new SO₄^2- carriers from being formed. Long CH pretreatment (24 h) and different SO₄^2- pretreatments reduced SO₄^2- influx below the non-metabolic level obtained by uptake experiments with DNP, indicating the existence of SO₄^2- carriers mediating passive SO₄^2- transport across the plasmalemma of the root cells. SO₄^2- influx was further decreased for the CH pretreated (24 h) plants by the presence of both CH and DNP in the experimental nutrient solution. This probably indicates the diffusive part of the non-metabolic SO₄^2- influx in the present experiments. Finally, it is suggested that there is a feedback signal between root and shoot, regulating sulfur transport upwards.     

Isolated Thellungiella shoots do not require roots to survive NaCl and Na2SO4 salt stresses

Shoots of Thellungiella derived by micropropagation were used to estimate the plants'' salt tolerance and ability to regulate Na⁺ uptake. Two species with differing salt tolerances were studied: Thellungiella salsuginea (halophilla), which is less tolerant, and Thellungiella botschantzevii, which is more tolerant. Although the shoots of neither ecotype survived at 700 mM NaCl or 200 mM Na₂SO₄, micropropagated shoots of T. botschantzevii were more tolerant to Na₂SO₄ (10–100 mM) and NaCl (100–300 mM). In the absence of roots, Na₂SO₄ salinity reduced shoot growth more dramatically than NaCl salinity. Plantlets of both species were able to adapt to salt stress even when they did not form roots. First, there was no significant correlation between Na⁺ accumulation in shoots and Na⁺ concentration in the growth media. Second, K⁺ concentrations in the shoots exposed to different salt concentrations were maintained at equivalent levels to control plants grown in medium without NaCl or Na₂SO₄. These results suggest that isolated shoots of Thellungiella possess their own mechanisms for enabling salt tolerance, which contribute to salt tolerance in intact plants.Key words: Thellungiella salsuginea, Thellungiella botschantzevii, salt tolerance, isolated shoots, growth, rhizogenesis, ion accumulation     

Micropropagation of Hemidesmus indicus for cultivation and production of 2-hydroxy 4-methoxy benzaldehyde

Caulogenic responses of various explant types from 12-month-old plants of Hemidesmus indicus were tested. Second and third visible nodes (0.5 cm) from the apex and root segments (0.5 cm) were the most and least regenerative respectively, with the formation of 9.37 and 2.6 shoots in 4 weeks on half strength MS medium supplemented with 2.22 μM BA and 1.07 μM NAA and 4.44 μM BA and 2.69 μM NAA respectively. Caulogenic ability of the nodes decreased with increasing maturity. Shoot buds initiated upon the young nodes on day 10 developed into 7.2 cm long shoots within 4 weeks thereby making a shoot elongation phase unnecessary. Nodal explants of the in vitro raised shoots subcultured in the same medium produced 9.32 shoots of 7.1 cm length in 3–4 weeks, similar to those of the mature plant-derived nodes. Multiplication through subculture of the nodes up to 25 passages of 4 weeks each was achieved without decline. Shoot cultures were rooted in quarter salt strength MS medium containing 9.8 μM IBA and the rooted plants were hardened for establishment in pots at 96% rate. Four months after establishment, the micropropagated plants were stable and showed uniform morphological and growth characteristic. After 12 months of cultivation in the field, on an average micropropagated plant consisted of 4–5 shoots, 5–8 branches per shoot and increased root biomass (13.5 g) compared to the poor growth (single shoot and 2–3 branches) and root production (4.6 g) values obtained with plants raised from conventional rooted stem cuttings. The concentration of the root specific compound, 2-hydroxy 4-methoxy benzaldehyde per plant was 2–3 fold higher in micropropagated plants though on unit dry root biomass (0.12% per g dry wt) basis it remained the same between two sources of plants. This revised version was published online in June 2006 with corrections to the Cover Date.     

Different apparent CO2 compensation points in nitrate- and ammonium-grown Phaseolus vulgaris and the relationship to non-photorespiratory CO2 evolution

The classical theory of the relationship between gas fluxes and photosynthetic electron fluxes was extended by two additional terms: J_L describing flux to electron sinks other than the Calvin cycle, and R_L accounting for light-induced changes in non-photorespiratory CO₂ evolution. R_L comprises two main components, R_r resulting from light-induced decrease in tricarboxylic acid activity, and R_S related to extra CO₂ evolution resulting from citrate-to-2-oxoglutarate conversion for N-assimilation in NO₃^– grown leaves. This extended theory was applied to two experiments. First, A–C_i curves (dependence of CO₂ flux on stomatal CO₂ concentration) revealed a higher apparent CO₂ compensation point (Γ*_app) in NO₃^–-grown plants than in NH₄⁺-grown plants. Secondly, photosynthetic electron fluxes at different light intensities were determined by means of the Genty parameter of chlorophyll fluorescence and compared with those calculated from measured CO₂ uptake. Curve-fitting based on the extended theory provided a coincidence of these two measurements and resulted in higher R_S in NO₃^–-grown than in NH₄⁺-grown plants. This difference in R_S (about 15% of the CO₂ flux bound by carboxylation) is the same as that obtained from the analysis of Γ*_app. Further, the analysis suggests that J_L related to the extra electron flux used for N-assimilation in NO₃^–-grown plants is diverted to other sinks in NH₄⁺-grown plants. SHAM decreased photosynthetic electron flow and O₂ evolution in NH₄⁺-grown plants, antimycin A in NO₃^–-grown plants. The effect of oligomycin was small. The results are discussed in terms of different mechanisms of chloroplast/mitochondrion interaction in NO₃^–- and NH₄⁺-grown plants, their effects on non-photorespiratory CO₂ evolution and on Γ*_app.     

Effects of NaCl and Na2SO4 on red-osier dogwood (Cornus stolonifera Michx) seedlings

Sodium chloride and sodium sulfate are commonly present in extraction tailings waters produced as a result of surface mining and affect plants on reclaimed areas. Red-osier dogwood (Cornus stolonifera Michx) seedlings were demonstrated to be relatively resistant to these high salinity oil sands tailings waters. The objectives of this study were to compare the effects of Na₂SO₄ and NaCl, on growth, tissue ion content, water relations and gas exchange in red-osier dogwood (Cornus stolonifera Michx) seedlings. In the present study, red-osier dogwood seedlings were grown in aerated half-strength modified Hoagland's mineral solution containing 0, 25, 50 or 100 mM of NaCl or Na₂SO₄. After four weeks of treatment, plant dry weights decreased and the amount of Na⁺ in plant tissues increased with increasing salt concentration. Na⁺ tissue content was higher in plants treated with NaCl than Na₂SO₄ and it was greater in roots than shoots. However, Cl^– concentration in the NaCl treated plants was higher in shoots than in roots. The decrease in stomatal conductance and photosynthetic rates observed in presence of salts is likely to contribute to the growth reduction. Our results suggest that red-osier dogwood is able to control the transport of Na⁺ from roots to shoots when external concentrations are 50 mM or less.     

The effect of root temperature on the uptake of nitrogen and the relative size of the root system in Lolium perenne. I. Solutions containing both NH+4 and NO3−

Abstract Lolium perenne L. cv. S23 was grown in flowing culture solution, pH 5, in which the concentrations of NH₄⁺, NO₃^? and K⁺ were frequently monitored and adjusted to set values. In a pre-experimental period, plants were acclimatized to a regime in which roots were treated at 5°C with shoots at 25°C. The root temperature was then changed to one of the following, 3, 7, 9, 11, 13, 17 or 25°C, while air temperature remained at 25°C. When root temperature was increased from 5X, the relative growth rate of roots increased immediately while that of shoots changed much less for a period of approximately 9 d (phase 1). Thus, the root: shoot ratio increased, but eventually approached a new, temperature-dependent, steady value (phase 2). The fresh: freeze-dried weight ratio (i.e. water content) in shoots (and roots) increased during the first phase of morphological adjustment (phase 1). In both growth phases and at all temperatures, plants absorbed more NH₄⁺ than NO₄⁺, the tendency being extreme at temperatures below 9° where more than 85% of the N absorbed was NH₄⁺. Plants at different root temperatures, growing at markedly different rates, had very similar concentrations of total N in their tissues (cells) on a fresh weight basis, despite the fact that they derived their N with differing preference for NH₄⁺. Specific absorption rates for NH₄⁺, NO_x^?, K⁺ and H₂PO₄^? showed very marked dependence on root temperature in phase 1, but ceased to show this dependence once a steady state root: shoot ratio had been established in phase 2. The results indicate the importance of relative root size in determining ion fluxes at the root surface. At higher temperatures where the root system was relatively large, ‘demand’ per unit root was low, whereas at low temperatures roots were small relative to shoots and ‘demand’ was high enough to offset the inhibitory effects of low temperature on transport processes.     

HCO−3 uptake through the roots and its effect on the productivity of willow cuttings

Abstract Young willow plants (Salix‘aquatica gigantea’) were grown in hydroponic culture media, and ¹⁴C–labelled sodium bicarbonate was fed to the roots. Uptake of ¹⁴C-label in the leaves and shoots was assayed after two different feeding periods (6 h, 48 h). Even during the shortest feeding period, ¹⁴C-label had been transferred to the leaves and shoots. Compared with the longer feeding period, after the 6 h feeding period more label was in the form of acid-labile products, whereas after the 48 h feeding period most of the label was in acid-stable products. A second experiment was designed to test whether carbon uptake by roots affects the growth of young willow plants. Uniform rooted cuttings were grown in hydroponic cultures at five different levels of bicarbonate: 0, 0.015, 0.147 0.737, and 1.473 mol m^?3 NaHCO₃. After a 4-week growing period we determined the biomass of leaves, shoots, roots and cuttings. Production of total dry matter (shoots, leaves and roots) increased with increasing bicarbonate concentration. Saturation of dry matter production was reached at 0.737 mol m^?3 NaHCO₃, but a higher concentration of NaHCO₃ (1.470 mol m^?3) caused a slight decrease in the dry matter production. At 0.737 mol m^?3 NaHCO₃ the total dry weight increased by 31.1%, which suggests that uptake of dissolved carbon dioxide through the roots might affect carbon budgeting in young willow plants.     

Rapid multiplication of Dalbergia sissoo Roxb.: a timber yielding tree legume through axillary shoot proliferation and ex vitro rooting

An efficient and improved method for in vitro propagation of mature tree of Dalbergia sissoo, an ecologically and commercially important timber yielding species, has been developed through axillary shoot proliferation. Bud breaking occurred from nodal shoot segments derived from rejuvenated shoots produced during early spring from a 20–25-year-old lopped tree, on MS medium containing 8.88 μM benzylaminopurine (BAP). Multiple shoots differentiated (20–21shoots/node) on re-culture of explants on half-strength agar gelled amended MS medium with a combination of 2.22 μM of BAP and 0.002 μM of thidiazuron (TDZ) with 1.0 mM each of Ca(NO₃)₂, K₂SO₄, KCl, and NH₄(SO₄)₂. The maximum shoot multiplication (29–30 shoots/node) was achieved on subculturing in the above mentioned but liquid medium. Furthermore, the problem of shoot tip necrosis and defoliation observed on solid medium were overcome by the use of liquid medium. Ex vitro rooting was achieved on soilrite after basal treatment of microshoots with 984 μM of indole-3-butyric acid (IBA) for 2 min. About 90 % microshoots were rooted on soilrite within 2–3 weeks under the greenhouse conditions. From 20 nodal shoot segments, about 435 hardened plants were acclimatized and transplanted. This is the first report for rapid in vitro propagation of mature trees of D. sissoo on liquid medium followed by ex vitro rooting.     

Seed germination and in vitro propagation of Maytenus canariensis through regeneration of adventitious shoots from axillary and apical buds

Seed germination and micropropagation protocols of the medicinal species Maytenus canariensis (Loes.) G. Kunkel & Sunding were optimized. In vitro seed germination occurred (86 to 94.7 %) only after treatment of the seeds with H₂SO₄, followed by surface sterilization and culture on solid nutrient medium without any growth regulators. Micropropagation failed when explants were taken from mature trees, and browning of the nutrient medium frequently occurred despite testing many growth media. Nonetheless, adventitious shoot regeneration was achieved employing axillary or apical buds taken from 2–2.5 months old plantlets obtained after in vitro germination of seeds, following culture on nutrient media supplemented with benzylaminopurine, kinetin and naphthalenacetic acid (NAA), attaining up to 3.9 shoots per explant, after 4–6 months. Root induction was best on a medium containing 4.0 mg dm⁻³ NAA, achieving a 100 % induction. After hardening of rooted plants, survival after transfer to soil was 71.43 %.     

Osmotic and ionic effects of NaCl and Na2SO4 salinity on Phragmites australis

Osmotic and ion-specific effects of NaCl and Na₂SO₄ on Phragmites australis (Cav.) Trin ex. Steud. were investigated in a laboratory experiment by examining effects of iso-osmotic solutions of NaCl and Na₂SO₄ on growth, osmolality of cell sap, proline content, elemental composition and gas exchange. Plants were supplied with a control standard nutrient solution (Ψ = −0.09 MPa) or solutions of NaCl or Na₂SO₄ at water potentials of −0.50, −1.09 or −1.74 MPa. Salt treatments increased root concentrations of Na and S or Cl, whereas P. australis had efficient mechanisms for exclusion of Na and S and partly Cl ions from the leaves. Incomplete exclusion of Cl from the leaves may affect aboveground biomass production, which was significantly more reduced by NaCl than Na₂SO₄. Stomatal conductance was negatively influenced by decreasing water potentials caused by NaCl or Na₂SO₄, implying that a non-significant photosynthetic depression observed in plants grown at −1.74 MPa was mainly due to osmotically induced stomatal closure. This was supported by decreasing internal CO₂ concentrations. Saline conditions increased the intrinsic water use efficiency and did not alter photosynthetic parameters derived from light response curves, supporting the assumption of a well-functioning CO₂ utilization in salt stressed plants. The leaf proline concentration increased equally in NaCl and Na₂SO₄-treated plants, and may play an important role as a compatible organic solute. P. australis possesses a range of mechanisms conferring tolerance to both NaCl and Na₂SO₄ stress and except in terms of growth the phytotoxicity of NaCl and Na₂SO₄ are comparable.     

Responses of Photosynthetic and Defence Systems to High Temperature Stress in Quercus suber L Seedlings Grown under Elevated CO2

Abstract: Growth in elevated CO₂ led to an increase in biomass production per plant as a result of enhanced carbon uptake and lower rates of respiration, compared to ambient CO₂-grown plants. No down-regulation of photosynthesis was found after six months of growth under elevated CO₂. Photosynthetic rates at 15°C or 35 °C were also higher in elevated than in ambient CO₂-grown plants, when measured at their respective CO₂ growth condition. Stomata of elevated CO₂-grown plants were less responsive to temperature as compared to ambient CO₂ plants. The after effect of a heat-shock treatment (4 h at 45 °C in a chamber with 80% of relative humidity and 800–1000 tmol m^-2 s^-1 photon flux density) on A_max was less in elevated than in ambient CO₂-grown plants. At the photochemical level, the negative effect of the heat-shock treatment was slightly more pronounced in ambient than in elevated CO₂-grown plants. A greater tolerance to oxidative stress caused by high temperatures in elevated CO₂-grown plants, in comparison to ambient CO₂ plants, is suggested by the increase in superoxide dismutase activity, after 1 h at 45 °C, as well as its relatively high activity after 2 and 4 h of the heat shock in the elevated CO₂-grown plants in contrast with the decrease to residual levels of superoxide dismutase activity in ambient CO₂-grown plants immediately after 1 h at 45 °C. The observed increase in catalase after 1 h at 45 °C in both ambient and elevated CO₂-grown plants, can be ascribed to the higher rates of photorespiration and respiration under this high temperature.     

Extracellular Matrix in Early Stages of Direct Somatic Embryogenesis in Leaves of Drosera spathulata

The growth and water relations of Paulownia fortunei in photoautotrophic cultures (nutrient medium lacking sucrose and growth regulator) with CO₂ enrichment (PWAH) or without CO₂ enrichment (PWAL) were compared with those in photomixotrophic shoot (PWC; 30 g dm⁻³ sucrose and 0.3 mg dm⁻³ N⁶-benzyladenine) and root cultures (PWR; 0.3 mg dm⁻³ indole-3-butyric acid). The photoautotrophic and photomixotrophic cultures were incubated under photosynthetic photon flux 125 and 60 μmol m⁻² s⁻¹, respectively. 100 % sprouting and significantly higher number of shoots (1.6) were obtained with PWAH as compared to PWAL and PWC. PWAH and PWAL stimulated spontaneous rooting from the cut end of axillary shoots. In PWAH, 84 % of shoots rooted with an average of 5.9 roots per shoot and 4.0 cm of root length in 21 d. Rooting of photomixotrophic shoot cultures were stimulated by an auxin treatment. In this case, 98.3 % of shoots were rooted with an average of 4.6 roots per shoot and 1.9 cm length. A microscopic observation on leaf abaxial surface prints from photomixotrophic shoot and root cultures showed widely open (6 – 8 μm) spherical stomata (12 – 14 μm) and from photoautotrophic cultures elliptical stomata (10 – 12 μm) with narrow openings (3 – 4 μm). Leaves from photomixo-trophic cultures had higher stomatal index as compared to photoautotrophic cultures. The rate of moisture loss from detached leaves was not varying significantly in different cultures. This revised version was published online in July 2006 with corrections to the Cover Date.     

A micropropagation protocol forCinnamomum camphora

Summary A micropropagation protocol was developed forCinnamomum camphora (L.) Sieb., using as initial explants 3–5-mm shoot tips from newly emerged laterals of 2-yr-old trees. Performance of small shoot tips was compared with that of 2.0-cm nodal segments during subculture. Murashige and Skoog medium (MS) supplemented with different concentrations of N₆-benzyladenine (BA) or thidiazuron (TDZ) was used to examine shoot proliferation. In separate experiments, MS was supplemented with 1-naphthaleneacetic acid (NAA) for rooting of shoots, and the commercial preparation EM₂ for prevention of hyperhydricity. BA stimulated shoot formation and callus development, whereas TDZ promoted only callus development. Both cytokinins induced hyperhydricity when small shoot tips were used, with severity being directly related to concentrations. Hyperhydricity was avoided in subcultures by using larger nodal segments. EM₂ did not alter degree of hyperhydricity but suppressed callus development and strongly promoted shoot multiplication. The number of new shoots after a 6-wk subculture was 9 per nodal segment when supplemented solely with 4.4 μM BA and 18 per segment when further supplemented with 1000 mg EM₂ per I. Rooting of shoots occurred best when supplemented solely with 0.54 μM NAA, averaging 7 roots per shoot in 4 wk. Ninety percent of rooted shoots survived transfer to the greenhouse.