The density-dependency of dark- and low-background radiation effects on water and water solution properties

The effects of dark -(Ev=0 lux) and low-background radiation (BGR), where R<1μRongen/h, on physicochemical properties (specific electrical conductivity, heat fusion, hydrogen peroxide (H2O2), and oxygen contents) of distilled water (DW) and physiological solution (PS) at 4°C and 18°C were studied. The incubation of DW and PS samples in dark and in low BGR (under dark) medium at 4°C and 18°C brings to changes of their physicochemical properties compared with DW and PS samples incubated in light and normal BGR condition (Ev=500-550 lux and R=17 μRoentgen/h). The observed changes of DW and PS properties depended on their initial temperature, density and ionic composition. It is suggested that water molecules dissociation and ions hydration are sensitive to illumination and BGR. Therefore, the cell-bathing medium can be considered as a messenger through which direct and non direct (by modulating of others factors-induced effects) influences of illumination and BGR on cell metabolism are realized.     

Photoabatement by Anthocyanin Shields Photosynthetic Systems from Light Stress

Leaves and other chlorophyllous tissues of plants often show transient or permanent anthocyanin coloration. The question of whether anthocyanins can function as effective light screens to modulate photosynthesis in plants was addressed by comparing photosynthetic responses in reddish-purple pods with those in green pods of the ornamental leguminous tree Bauhinia variegata. For these comparisons the actinic radiation employed was either red radiation (RR) which was poorly absorbed by anthocyanin or blue-green radiation (BGR) which was strongly absorbed by anthocyanin. Photon yields of photosystem 2 (PS2) photochemistry and photochemical chlorophyll fluorescence quenching coefficients (q_p), measured over a range of photon flux densities (PFD) up to 1200 μmol m^-2 s^-1 at 23 °C and at five temperatures from 8 to 28 °C at a PFD of 260 μmol m^-2 s^-1, were almost identical in green pods irradiated with either RR or BGR and in purple pods irradiated with RR. However, q_p values remained much higher in purple pods irradiated with BGR, e.g., 0.80 in BGR versus 0.29 in RR at a PFD of 1200 μmol m-2 s^-1 at 23 °C, and 0.67 in BGR versus 0.28 in RR at a PFD of 260 μmol m^-2 s-1 at 8 °C. The higher values of q_p in BGR compared to RR indicated that photoabatement by anthocyanin allowed the first stable acceptor of PS2, Q_A, to be kept in a more oxidized state, thus decreasing the likelihood of photoinhibition. This was confirmed by demonstrating a lower susceptibility to photoinhibition in purple pods than in green pods in the sunlight, either naturally in pods on trees or in detached pods exposed to photoinhibitory conditions. We conclude that photoabatement by anthocyanin is a mechanism for allowing maintenance of higher oxidative levels of PS2 acceptor during episodes of high radiation stress, thereby minimizing photodamage to photosynthetic tissues. This revised version was published online in June 2006 with corrections to the Cover Date.     

The Non Thermal Effect of Weak Intensity Millimeter Waves on Physicochemical Properties of Water and Water Solutions

The comparative study of the effects of 5.8mW/cm² Millimeter Waves (MMW) and near Infrared (IR) irradiation on thermal properties, specific adsorption rate (SAR), specific electrical conductivity (SEC) and hydrogen peroxide (H₂O₂) content of distilled water (DW), and physiological solutions (PS) was performed. The thermal effect of MMW irradiation appeared only after the first minute of irradiation, while the IR heating started from the first minute of irradiation. The heat fusion of frozen MMW-treated DW and PS was significantly less than sham and IR-treated DW and PS. MMW irradiation had time-dependent elevation effect on water SEC and SAR, which was accompanied by the increase of H₂O₂ formation in it. We suggest that the MMW-induced vibration of water dipole molecules caused the non thermal changes of physicochemical properties of DW and PS, which promote the formation of H₂O₂ in water.     

Studies on synchronously dividing cultures of Euglena gracilis Klebs (strain Z). 3. Circadian components of cell division

Euglena gracilis Klebs (Z) was grown axenically and autotrophically in four-liter serum bottles at 25°C on an aerated, continuously stirred, inorganic salt medium. Four fluorescent illumination regimes were employed: (1) continuous bright light of 3500 lux (LL_b); (2) continuous dim light of 800 lux (LL_d); (3) a L_bD: 14, 10 (3500 lux) light-dark cycle; and (4) a L_dD: 14, 10 (800 lux) light-dark cycle. Cell number was automatically monitored throughout all experiments. In LL_b the generation time (G.T.) of the population was about 12 hours, whereas in LL_d following LL_b it was approximately five days; exponential growth occurred in either case. In L_bD: 14, 10 synchronous growth occurred with a doubling of cell number every cycle of 24 hours. In L_dD: 14, 10, however, although rhythmic cell division took place every 24 hours, the average increase in cell number during the division burst which occurred in each dark period was only 13.4%, so that the G.T. of the culture was about five days, as was the case for LL_d. In the constant conditions of temperature and continuous dim light (LL_d), following synchronous growth in L_bD: 14, 10, small (17.0%) rhythmic division bursts lasting 14.5 hours continued to occur for at least ten days, with a period of 24.2 hours. The overall G.T. of the culture was about five days. These data demonstrating the circadian, endogenous nature of rhythmic cell division under certain conditions of continuous dim illumination were discussed in relation to the synchronous division observed in temperature and light-dark cycles.     

Photosystem 2 is more tolerant to high temperature in apple (<Emphasis Type="Italic">Malus domestica</Emphasis> Borkh.) leaves than in fruit peel

Tolerance of photosystem 2 (PS2) to high temperature in apple (Malus domestica Borkh. cv. Cortland) leaves and peel was investigated by chlorophyll a fluorescence (OJIP) transient after exposure to 25 (control), 40, 42, 44, and 46 °C in the dark for 30 min. The positive L-step was more pronounced in a peel than in leaves when exposed to 44 °C. Heat-induced K-step became less pronounced in leaves than in peel when exposed to 42 °C or higher temperature. Leaves had negative L-and K-steps relative to the peel. The decrease of oxygen-evolving complex (OEC) by heat stress was higher in the peel than in the leaves. OJIP transient from the 46 °C treated peel could not reach the maximum fluorescence (F_m). The striking thermoeffect was the big decrease in the relative variable fluorescence at 30 ms (V_I), especially in the leaves. Compared with the peel, the leaves had less decreased maximum PS2 quantum efficiency (F_v/F_m), photochemical rate constant (K_P), F_m and performance index (PI) on absorption basis (PI_abs) and less increased minimum fluorescence (F₀) and non-photochemical rate constant (K_N), but more increased reduction of end acceptors at PS1 electron acceptor side per cross section (RE₀/CS₀) and per reaction center (RE₀/RC₀), quantum yield of electron transport from Q_A ⁻ to the end acceptors (ϕ _R0) and total PI (PI_abs,total) when exposed to 44 °C. In conclusion, PS2 is more thermally labile than PS1. The reduction of PS2 activity by heat stress primarily results from an inactivation of OEC. PS2 was more tolerant to high temperature in the leaves than in the peel.     

Fruiting ofLyophyllum tylicolor in plate culture on Soytoneglucose agar and urea-treated soil extract agar

Lyophyllum tylicolor, which forms mycelial basidia (and basidiospores), produced fruit-bodies when cultivated at 20°C under continuous illumination of 400–700 lux on agar plates containing Bacto-Soytone and glucose or an extract from urea-treated soil. Under these conditions, mycelial basidia were also observed on the Soytone-glucose agar, but not on the soil extract agar. In darkness, fruit-bodies and mycelial basidia were not observed on either medium. In culture on the soil extract agar, fruit-body primordia were produced at the position of the margin of the colony when it was transferred from darkness to continuous light; stipes did not elongate under illumination of ca. 2000 lux; and mycelial basidia and basidiospores, but not fruit-bodies, developed when glucose concentration in the medium was as high as 1% (w/v).     

Effects of temperature and light on germination and early seedling development of the pine pink orchid (Bletia purpurea)

Orchid seed physiology is a poorly understood phenomenon owing to an emphasis on production and the challenges associated with propagating orchids from minute seed. We investigated the role of simulated south Florida temperatures and illumination (dark and 12 h photoperiod) in regulating germination and seedling development using asymbiotic seed germination assays of Bletia purpurea. Our objectives were to determine whether in situ germination is limited by seasonal temperatures and to determine whether temperature alters responses to illumination. Bletia purpurea seeds were able to germinate to > 90% under all treatments. The greatest germination after 3 weeks was observed at 29/19°C under continual darkness and at 25°C under dark and illuminated conditions. The slowest germination was observed at simulated winter temperatures (22/11°C). Illumination initially inhibited germination and development, but resulted in equal or greater development by week six. Germination under 22/11°C was strongly inhibited by illumination, indicating an interaction between temperature and light sensing systems.     

Some effects of light and heat on the feeding and resting behaviour of tsetse flies,Glossina morsitans Westwood

A laboratory investigation was carried out into the effects of light and heat on the feeding and resting behaviour of Glossina morsitans morsitans Westwood. Flies placed in small cages on the ears of rabbits were tested for feeding responsiveness at light intensities of 0, 10, 100, 1000 and 25 000 lux. Feeding occurred at the same rate in darkness as it did in light at up to 1000 lux, but at 25 000 lux was largely inhibited. At 26° C, flies flew to and settled in the light end of a 1.2 m-long choice chamber. As the temperature was raised (by 10° C/hour) they eventually took off and flew directly to the dark end. The temperature at which this skototaxis occurred was strongly negatively correlated with the light intensity: each log unit increase in light intensity (in lux) caused a 2.2° C fall in take-off temperature. Skototaxis occurred at higher temperatures in recently fed female flies than in starved ones.     

Functional characterisation of a purified homogeneous Photosystem II core complex with high oxygen evolution capacity from spinach

The functional properties of a purified homogeneous spinach PS II-core complex with high oxygen evolution capacity (Haag et al. 1990a) were investigated in detail by measuring thermoluminescence and oscillation patterns of flash induced oxygen evolution and fluorescence quantum yield changes. The following results were obtained:

1. Depending on the illumination conditions the PS II-core complexes exhibit several thermoluminescence bands corresponding to the A band, Q band and Z_v band in PS II membrane fragments. The lifetime of the Q band (T_max=10°C) was determined to be 8s at T=10°C. No B band corresponding to S₂Q_B ^? or S₃Q_B ^? recombination could be detected.
2. The flash induced transient fluorescence quantum yield changes exhibit a multiphasi relaxation kinetics shich reflect the reoxidation of Q _A ^? . In control samples without exogenous acceptors this process is markedly slower than in PS II membrane fragments. The reaction becomes significantly retarded by addition of 10 μM DCMU. After dark incubation in the presence of K₃[Fe(CN)₆
3. Excitation of dark-adapted samples with a train of short saturating flashes gives rise to a typical pattern dominated by a high O₂ yield due to the third flash and a highly damped period four oscillation. The decay of redox states S₂ and S₃ are dominated by short life times of 4.3 s and 1.5 s, respectively, at 20°C.

The results of the present study reveal that in purified homogeneous PS II-core complexes with high oxygen evolution isolated from higher plants by β-dodecylmaltoside solubilization the thermodynamic properties and the kinetic parameters of the redox groups leading to electron transfer from water to Q_A are well preserved. The most obvious phenomenon is a severe modification of the Q_B binding site. The implications of this finding are discussed.     

Light and temperature regulation of greening in dark‐grown ginkgo (Ginkgo biloba)

The last steps of chlorophyll (Chl) biosynthesis were studied at different light intensities and temperatures in dark‐germinated ginkgo (Ginkgo biloba L.) seedlings. Pigment contents and 77 K fluorescence emission spectra were measured and the plastid ultrastructure was analysed. All dark‐grown organs contained protochlorophyllide (Pchlide) forms with similar spectral properties to those of dark‐grown angiosperm seedlings, but the ratios of these forms to each other were different. The short‐wavelength, monomeric Pchlide forms were always dominating. Etioplasts with small prolamellar bodies (PLBs) and few prothylakoids (PTs) differentiated in the dark‐grown stems. Upon illumination with high light intensities (800 μmol m^?2 s^?1 photon flux density, PFD), photo‐oxidation and bleaching occurred in the stems and the presence of ¹O₂ was detected. When Chl accumulated in plants illuminated with 15 μmol m^?2 s^?1 PFD it was significantly slower at 10°C than at 20°C. At room temperature, the transformation of etioplasts into young chloroplasts was observed at low light, while it was delayed at 10°C. Grana did not appear in the plastids even after 48 h of greening at 20°C. Reaccumulation of Pchlide forms and re‐formation of PLBs occurred when etiolated samples were illuminated with 200 μmol m^?2 s^?1 PFD at room temperature for 24 h and were then re‐etiolated for 5 days. The Pchlide forms appeared during re‐etiolation had similar spectral properties to those of etiolated seedlings. These results show that ginkgo seedlings are very sensitive to temperature and light conditions during their greening, a fact that should be considered for ginkgo cultivation.     

Multicellular genesis of leaf primordium was demonstrated via chimaeric transgenic plant of maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.) regenerated from Type II calli

Type-II embryonic calli were induced from immature embryos of maize (Zea mays L.) genotype YD and bombarded with beta-glucuronidase gene. Bombarded calli were proliferated on normal N6 medium for 2 weeks at 26°C in the dark and selected on N6 medium containing 1 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) and 5 mg/l phosphinothricin (PPT) but without casamino acids and proline under the same conditions for 14 days. Regeneration was carried out on hormone-free MS medium containing 5 mg/l phosphinothricin at 26°C under 3000 lux illumination. Plants over 8 cm were transplanted into soil and sprayed with 250 mg/l phosphinothricin when two new leaves appeared. Except normal transgenic plants, chimaeric transgenics also were regenerated in the present work. The expression pattern of beta-glucuronidase gene in leaves of chimaeric transgenic plant revealed that more than one cell formed leaf primordium at the initial stage, and filial cells stemed from each cell in leaf primordium arranged in a row longitudinally from leaf base to leaf apex. There was a clear boundary as a straight line between the area formed by transformed cells and the area formed by normal cells. A hypothesis was put forward that the primitive cells in leaf primordium divided in a longitudinal style, resulted in leaf elongation, then the filial cells divided transversally and synchronously toward the outside to broaden the leaf.     

Germination responses of four native terrestrial orchids from south-west Western Australia to temperature and light treatments

We report an investigation into the impact of temperature and illumination on in vitro symbiotic and asymbiotic germination of the threatened taxon Caladenia huegelii, and three other orchid spp. namely—Caladenia latifolia, Microtis media and Pterostylis sanguinea, all species from south-west Western Australia, a recognized biodiversity hotspot. High symbiotic germination on oatmeal agar (OMA + fungal symbionts specific to each species) was recorded in three species in continuous dark incubation i.e. C. huegelii seeds (98 % germination at 25 °C), and M. media and P. sanguinea (93 and 98 % respectively at 20 °C). Highest symbiotic germination for C. latifolia (100 %) was observed at 15 and 20 °C under light treatment (12/12 h light/dark). Low temperature incubation (10 °C) significantly suppressed symbiotic germination/development of seedlings across all species. Asymbiotic media treatments assessed (OMA minus fungal symbionts, Pa₅ and ½ MS), failed to stimulate any germination with C. latifolia seeds at 20 °C in either light or dark treatments after an 8 week incubation period. Seeds of M. media sown onto ½ MS medium resulted in higher germination in all developmental stages (3–5) in dark treatment than OMA and Pa₅. Seeds of P. sanguinea sown onto ½ MS medium resulted in higher overall germination in all developmental stages (3–5) in light and dark incubation compared to OMA and Pa₅. OMA supported the highest asymbiotic germination (100 %) in both light and dark incubation with M. media (only to stage 3) but did not support germination and development with other spp. tested. Caladenia huegelii seeds reached developmental Stage 3 (i.e. germinated), but only on Pa₅ medium and only at a relatively low rate in either light (2.6 %) or dark (2.1 %). Germination was higher and development of seedlings faster overall in all test species in symbiotic compared with asymbiotic media treatments. P. sanguinea seeds demonstrated the best response (among species tested) to asymbiotic germination on ½ MS with 40–53 % of germinated seeds spread over developmental stages 3–5 in light or dark incubation (at 20 °C) respectively. Illumination had no effect on fungal symbiont growth across all species, however incubation temperature treatments (10, 15, 20 and 25 °C) affected fungal growth rate. Growth of the fungal symbionts of C. huegelii, M. media and C. latifolia demonstrated significantly lower activity at 10 °C, but the cumulative radial growth rate of the P. sanguinea fungal symbiont reached 64 cm² after only 2 weeks at all temperatures tested, including 10 °C. The study highlights differences in symbiotic and aysmbiotic germination and early protocorm development in vitro between co-occurring herbaceous terrestrial Australian orchid taxa in response to variations in basal media, temperature and light.     

Effects of root temperature on leaf gas exchange and xylem sap abscisic acid concentrations in six Cucurbitaceae species

Roots of six Cucurbitaceae species were exposed to low (14 °C), middle (24 °C), and high (34 °C) temperatures while aerial parts of plants were maintained at ambient temperatures between 23 and 33 °C. The highest dry mass (DM), photon-saturated rate of net photosynthesis (P _Nsat), and stomatal conductance (g _s) were found at 14 °C in figleaf gourd and turban squash plants, at 24 °C in cucumber and melon plants, while bitter melon and wax gourd plants had lower DM, P _Nsat, and g _s at 14 °C than at 24 or 34 °C. Sub-or supra-optimum root temperatures did not induce photoinhibition but induced slight changes in the quantum efficiency of photosystem 2, PS2 (Φ_PS2) and photochemical quenching (q_p). Meanwhile, xylem sap abscisic acid (ABA) concentration followed a contrasting change pattern to that of g _s. Thus the change in P _Nsat was mainly due to the change in g _s and roots played an important role in the regulation of stomatal behaviour by delivering increased amount of ABA to shoots at sub-or supra-optimum root temperatures.     

Effects of Temperature,Photoperiod, and Light Intensity on the Eclosion Rhythm of the High-Altitude Himalayan Strain of Drosophila ananassae

Eclosion rhythm of the high-altitude Himalayan strain of Drosophila ananassae from Badrinath (altitude 5123 m) was temperature-dependent and at 21°C, it was entrained by cycles of 12 h light: 12 h darkness (LD 12:12) and free-ran in constant darkness, however, it was arrhythmic at 13°C or 17°C under identical experimental conditions (Khare, P. V., Barnabas, R. J., Kanojiya, M., Kulkarni, A. D., Joshi, D. S. (). Temperature dependent eclosion rhythmicity in the high altitude Himalayan strains of Drosophila ananassae. Chronobiol. Int. 19:1041–1052). The present studies were designed to see whether or not these strains could be entrained at 13°C, 17°C, and 21°C by two types of LD cycles in which the photoperiod at 100 lux intensity varied from 6 h to 18 h, and the light intensity of LD 14:10 cycles varied from 0.001 lux to 1000 lux. All LD cycles entrained this strain at 21°C but not at 13°C or 17°C. These results demonstrate that the entrainment of eclosion rhythm depends on the ambient temperature and not on the photoperiod or light intensity of LD cycles. Thus the temperature has taken precedence over the light in the entrainment process of eclosion rhythm of the high altitude Himalayan strain of D. ananassae. This may be the result of natural selection in response to the environmental temperature at Badrinath that resembles that of the sub-Arctic region but the photoperiod or light intensity are of the subtropical region.     

Differences in hypericin synthesis between experimentally induced seedling shoot cultures of Hypericum hookerianum Wight & Arn.

During seed germination trials of Hypericum hookerianum, seedlings of Lake View accession from Palni hills of Southern India segregated into green- (97.44 %) and red-pigmented (2.56 %) types. Seedlings cultured in Murashige and Skoog (1962) basal medium developed into fast growing green and slow growing red plant types in 6 weeks, the latter showing increased concentrations of total phenols, anthocyanins and flavonoids and 19-fold higher concentration of hypericin. Hypocotyls/cotyledons of red seedlings cultured using 2.325 μM kinetin (KIN) produced hypericin-rich (4.38 ± 0.18 mg/g DW), stunted (0.5–1.2 cm) shoots which ceased to grow in 8 weeks. Segments (4–6 mm) of these shoots sub-cultured in the dark for 4 weeks followed by 2-week light exposure and repeated subculture enabled mass multiplication of productive (3.93 ± 0.06 mg hypericin/g DW) shoots. Green hypocotyls and cotyledons subjected to 4 + 2 weak dark–light treatment also produced 9.18 ± 2.44 and 4.25 ± 0.96 comparable hypericin-rich (3.73 ± 0.21 mg/g DW) shoots. Red and green seedling explants cultured in basal medium in the dark produced 6.82 ± 0.75 cm etiolated shoots with reduced leaves which synthesized 2.27 ± 0.15 mg hypericin/g DW on illumination. Green cotyledons cultured in the dark using 2.45 μM indole-3 butyric acid (IBA) formed calluses which on illumination formed 12.64 ± 3.8 productive (3.86 ± 0.31 mg hypericin/g DW) 0.5- to 1.5-cm-long shoot clusters. Phenotypic segregation of seedlings, the ability of both red and green seedling explants to multiply in the dark and produce hypericin on illumination, and IBA-induced indirect shoots producing significant amounts of metabolite compared to wild plants (0.35 ± 0.09 mg/g DW) and green shoot cultures (0.91 ± 0.03 mg/g DW) are new to Hypericum.     

Changes in photosynthetic apparatus during dark incubation of detached leaves from control and ultraviolet-B treatedVigna seedlings

Changes in various components of photosynthetic apparatus during the 4 d dark incubation at 25°C of detached control and ultraviolet-B (UV-B) treatedVigna unguiculata L. leaves were examined. The photosynthetic apparatus was more degraded in younger control seedlings and for a longer time UV-B treated seedlings than in the older or for a shorter time UV-B treated seedlings. This was shown by determining the losses in chlorophyll (Chl) and protein contents, variable fluorescence yield, photosystem (PS) 2, PS1 and ribulose-1,5-bisphosphate carboxylase (RuBPC) activities, and photosynthetic¹⁴CO₂ fixation. In contrast, the Car/Chl ratio increased during the dark incubation due to less expressed degradation of Car.     

THERMOREGULATORY EFFECT IN HUMANS OF SUPPRESSED ENDOGENOUS MELATONIN BY PRE-SLEEP BRIGHT-LIGHT EXPOSURE IN A COLD ENVIRONMENT

This study investigated the physiological function of suppressed melatonin through thermoregulation in a cold environment. Interactions between thermoregulation directly affected by exposure to a cold environment and indirectly affected by endogenous melatonin suppression by bright-light exposure were examined. Ten male subjects were exposed to two different illumination intensities (30 and 5000 lux) for 4.5?h, and two different ambient temperatures (15 and 27°C) for 2?h before sleep under dark and thermoneutral conditions. Salivary melatonin level was suppressed by bright light (p?<?0.001), although the ambient temperature condition had no significant effect on melatonin. During sleep, significant effects of pre-sleep exposure to a cold ambient temperature (p?<?0.001) and bright light (p?<?0.01) on rectal temperature (T_re) were observed. Pre-sleep, bright-light exposure led to an attenuated fall in T_re during sleep. Moreover, T_re dropped more precipitously after cold exposure than thermoneutral conditions (cold: ?0.54?±?0.07°C/h; thermoneutral: ?0.16?±?0.03°C/h; p?<?0.001). Pre-sleep, bright-light exposure delayed the nadir time of T_re under thermoneutral conditions (p?<?0.05), while cold exposure masked the circadian rhythm with a precipitous decrease in T_re. A significant correlation between the T_re nadir and melatonin level (r?=??0.774, p?<?0.05) indicated that inter-individual differences with higher melatonin levels lead to a reduction in T_re after cold exposure. These results suggest that suppressed endogenous melatonin inhibits the downregulation of the body temperature set-point during sleep. (Author correspondence: ishibasi@design.kyushu-u.ac.jp)     

Further studies on the flowering ofLemna paucicostata in Japan

Flowering behavior of 22 strains ofLemna paucicostata collected in Japan by Yukawa and Takimoto (1976) was re-examined. The critical dark periods of the short-day strains (N-1 and N-2 types) were shorter than those determined by Yukawa and Takimoto except for that of one strain. Particularly in strains 391, 381 and 321, the differences were as large as 2.25, 1.75 and 1.5 hr, respectively. Such differences were found to be due at least partly to the difference in night temperature; 25 C for the light and 23 C for the dark periods in the present experiment, and 25 C throughout the light and dark periods in the previous experiment. The S type strains did not flower under our experimental conditions (fluorescent light of 6,000 lux at 25 C) at any photoperiod tested, but flowered as a quantitative long-day plant under natural daylight or high-intensity light (12,000 lux). Addition of sucrose or ammonium ion to the medium suppressed the flowering of these strains under high-intensity light. Addition of benzoic acid (1–5 μM) to 0.5 strength NH₄ ⁺-free Hutner's medium caused daylength-independent flowering in some N-1 type strains and in all N-2 type strains tested. S type strains cultured under fluorescent light of 6,000 lux also flowered rapidly in response to benzoic acid.     

Interactive effect of light,temperature and TDZ on the regeneration potential of leaf discs of <Emphasis Type="Italic">Fragaria x ananassa</Emphasis> Duch

This is the first report where shoot regeneration in strawberry cultivar Chandler has been achieved simultaneously through both somatic embryogenesis and shoot bud formation. Direct somatic embryogenesis was observed in leaf discs which were cultured on medium containing MS salts + B₅ vitamins + 2% glucose + 18.16 μM thidiazuron (TDZ) and given both chilling and dark treatment for 2 wk at 4 ± 2°C followed by incubation at 25 ± 2°C under 16-h photoperiod for third wk. After 3 wk, these explants were then subcultured on medium containing MS salts + B₅ vitamins + 2% glucose and incubated under 16-h photoperiod at 25 ± 2°C for further growth and development. Direct regeneration via de novo shoot bud formation was observed in leaf disks which were given dark treatment and were cultured on medium containing MS salts + B₅ vitamins + 2% glucose supplemented with 9.08 μM TDZ. There was a synergistic effect of photoperiod, dark, and chilling treatments on somatic embryogenesis, whereas chilling treatment had an inhibitory effect on shoot organogenesis.