Light-Dependency of Growth and Secondary Metabolite Production in the Captive Zooxanthellate Soft Coral Sinularia flexibilis

The branching zooxanthellate soft coral Sinularia flexibillis releases antimicrobial and toxic compounds with potential pharmaceutical importance. As photosynthesis by the symbiotic algae is vital to the host, the light-dependency of the coral, including its specific growth rate (μ day⁻¹) and the physiological response to a range of light intensities (10–1,000 μmol quanta m⁻² s⁻¹) was studied for 12 weeks. Although a range of irradiances from 100 to 400 μmol quanta m⁻² s⁻¹ was favorable for S. flexibilis, based on chlorophyll content, a light intensity around 100 μmol quanta m⁻² s⁻¹ was found to be optimal. The contents of both zooxanthellae and chlorophyll a were highest at 100 μmol quanta m⁻² s⁻¹. The specific budding rate showed almost the same pattern as the specific growth rate. The concentration of the terpene flexibilide, produced by this species, increased at high light intensities (200–600 μmol quanta m⁻² s⁻¹).     

Early development of germlings of <Emphasis Type="Italic">Sargassum thunbergii</Emphasis> (Fucales,Phaeophyta) under laboratory conditions

Morphology and culture studies on germlings of Sargassum thunbergii (Mertens et Roth) Kuntze were carried out under controlled laboratory conditions. Growth characteristics of these germlings grown under different temperatures (from 10 to 25°C), irradiances (from 9 to 88 μmol photons m⁻² s⁻¹), and under blue and white light conditions are described. The development of embryonic germlings follows the classic “8 nuclei 1 egg” type described for Sargassaceae. Fertilized eggs spent 5–6 h developing into multicellular germlings with abundant rhizoids after fertilization. Under conditions of 20°C, 44 μmol photons m⁻² s⁻¹ and photoperiod of 12 h, young germlings with one or two leaflets reached 2–3 mm in length after 8 weeks. Temperature variations (10, 15, 20, 25°C) under 88 μmol photons m⁻² s⁻¹ significantly influenced the growth rate within the first week, although this effect became less obvious after 8 weeks, especially at 15 and 20°C. Variation in germling growth was highly significant under different irradiances (9, 18, 44, 88 μmol photons m⁻² s⁻¹) at 25°C. Low temperature (10°C) reduced germling growth. Growth of germlings cultured under blue light was lower than in white light. Optimal growth of these germlings occurred at 25°C and 44 μmol photons m⁻² s⁻¹.     

Characterization of PSI recovery after chilling-induced photoinhibition in cucumber (<Emphasis Type="Italic">Cucumis sativus</Emphasis> L.) leaves

By simultaneously analyzing the chlorophyll a fluorescence transient and light absorbance at 820 nm as well as chlorophyll fluorescence quenching, we investigated the effects of different photon flux densities (0, 15, 200 μmol m⁻² s⁻¹) with or without 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) on the repair process of cucumber (Cucumis sativus L.) leaves after treatment with low temperature (6°C) combined with moderate photon flux density (200 μmol m⁻² s⁻¹) for 6 h. Both the maximal photochemical efficiency of Photosystem II (PSII) (F _v/F _m) and the content of active P700 (ΔI/I _o) significantly decreased after chilling treatment under 200 μmol m⁻² s⁻¹ light. After the leaves were transferred to 25°C, F _v/F _m recovered quickly under both 200 and 15 μmol m⁻² s⁻¹ light. ΔI/I _o recovered quickly under 15 μmol m⁻² s⁻¹ light, but the recovery rate of ΔI/I _o was slower than that of F _v/F _m. The cyclic electron transport was inhibited by chilling-light treatment obviously. The recovery of ΔI/I _o was severely suppressed by 200 μmol m⁻² s⁻¹ light, whereas a pretreatment with DCMU effectively relieved this suppression. The cyclic electron transport around PSI recovered in a similar way as the active P700 content did, and the recovery of them was both accelerated by pretreatment with DCMU. The results indicate that limiting electron transport from PSII to PSI protected PSI from further photoinhibition, accelerating the recovery of PSI. Under a given photon flux density, faster recovery of PSII compared to PSI was detrimental to the recovery of PSI or even to the whole photosystem.     

Effects of temperature and light on the growth and geosmin production of Lyngbya kuetzingii (Cyanophyta)

The effects of temperature and light on the growth and geosmin production of Lyngbya kuetzingii were determined. Of the three temperatures tested, 10, 25 and 35°C, the maximal geosmin concentration and geosmin productivity were yielded at 10°C, while the highest chl a production was observed at 25°C. In the studies on light intensity, the maximal geosmin concentration and geosmin productivity were observed at 10 μmol m⁻² s⁻¹, while the highest chl a production was at 20 μmol m⁻² s⁻¹. It was suggested that more geosmin was synthesized with lower chl a demand. Meanwhile, the relative amounts of extra- and intracellular geosmin were investigated. Under optimum growth conditions (20 μmol m⁻² s⁻¹, 25°C; BG-11 medium), the amounts of extracellular geosmin increased as the growth progressed and reached the maximum in the stationary phase, while the intracellular geosmin reached its maximum value in the late exponential phase, and then began to decline. However, under the low temperature (10°C) or light (10 μmol m⁻² s⁻¹) conditions, more intracellular geosmin was synthesized and mainly accumulated in the cells. The proportions of extracellular geosmin were high, to 33.33 and 32.27%, respectively, during the stationary phase at 35°C and 20 μmol m⁻² s⁻¹. It was indicated that low temperature or light could stimulate geosmin production and favor the accumulation of geosmin in cells, while more intracellular geosmin may be released into the medium at higher temperatures or optimum light intensity.     

Combined effect of temperature and light intensity on growth and extracellular polymeric substance production by the cyanobacterium Arthrospira platensis

The effects of light intensity and temperature on Arthrospira platensis growth and production of extracellular polymeric substances (EPS) in batch culture were evaluated using a three-level, full-factorial design and response surface methodology. Three levels were tested for each parameter (temperature: 30, 35, 40°C; light intensity: 50, 115, 180 μmol photons m⁻² s⁻¹). Both growth and EPS production are influenced mainly by the temperature factor but the interaction term temperature*light intensity also had a significant effect. In addition, conditions optimising EPS production are different from those optimising growth. The highest growth rate (0.414 ± 0.003 day⁻¹) was found at the lowest temperature (30°C) and highest light intensity (180 μmol photons m⁻² s⁻¹) tested, no optima were detectable within the given test range. Obviously, optima for growth must be at a temperature lower than 30°C and a light intensity higher than 180 μmol photons m⁻² s⁻¹. For EPS production, light intensity had a positive linear effect (optimum obviously higher than 180 μmol photons m⁻² s⁻¹), but for the temperature parameter a maximum effect was detectable at 35°C.     

Light acclimatisation of Elodea nuttallii grown under ambient DIC conditions

Light acclimatisation capabilities of Elodea nuttallii at nearly ambient DIC conditions were investigated by determining growth characteristics, main photosynthetic parameters and pigmentation of plants incubated at 5 different irradiances (10–146 μmol photons m⁻² s⁻¹). Positive net growth was observed under all light treatments tested. Maximum ratio root versus shoot (r:s) of 1.86 was achieved at medium irradiances (72–94 μmol photons m⁻² s⁻¹), whereas at low (10 μmol photons m⁻² s⁻¹) and high irradiances (146 μmol photons m⁻² s⁻¹) r:s was significantly lower (0.39 and 1.05, respectively). With respect to main photosynthetic parameters, an increase of light compensation points (E _c), attended by decreasing ratios of light saturation points of photosynthesis (E _k)/irradiance were observed. E _c values were comparable to other low-light adapted macrophytes, which indicate that E. nuttallii can be regarded as a low-light adapted plant, under photorespiratory conditions. This was also confirmed by maximum E _k values of just 73 μmol photons m⁻² s⁻¹. Further support was achieved from pigmentation and non-photochemical quenching (NPQ) data, both indicating rather limited acclimatisation ability at light treatments above 90 μmol photons m⁻² s⁻¹. These results are discussed with respect to the competitive abilities of E. nuttallii under nearly ambient (photorespiratory) DIC conditions, especially in dense stands and turbid phytoplankton-dominated waters.     

Improved in vitro shoot multiplication and rooting of <Emphasis Type="Italic">Dendrocalamus hamiltonii</Emphasis> Nees et Arn. Ex Munro: production of genetically uniform plants and field evaluation

An efficient in vitro regeneration protocol and field performance of a multipurpose bamboo species Dendrocalamus hamiltonii Nees et Arn. Ex Munro has been demonstrated using single node cuttings taken from the lateral branches of a 20-year-old bush. Axillary buds on the nodal explant sprouted within 10 days of culture on Murashige and Skoog (MS) medium without any plant growth substance. High-frequency proliferation was induced on the propagules (small clusters with 3–5 multiple shoots and rhizomatous portions). Subsequent removal of the shoots (about 1.5 cm) from the rhizomatous portion of propagules (shoot cut) influenced the plantlet formation capacity. A multiplication of about 20-folds was achieved on MS medium supplemented with 8 μM BAP and 1 μM NAA. Rooting efficiency was also markedly enhanced (>90%) when the propagules, following shoot cut, were placed on to MS medium supplemented with 100 μM IBA for 10 days and then transferred to IBA-free medium. This is the first report from this species where 20-fold increment in multiplication was observed at the end of second subculture followed by >90% rooting. The hardened plants, established in the field, exhibited normal growth; their physiological performance has been monitored at 6-month intervals. The rate of photosynthesis increased from 3.55 μmol CO₂ m⁻² s⁻¹ (hardened, ready for field transfer) to 5.44 μmol m⁻² s⁻¹ (6 months of field transfer); following a year of plantation net photosynthesis recorded was 14.0 μmol CO₂ m⁻² s⁻¹ while after 1.5 years it was 12.76 μmol CO₂ m⁻² s⁻¹. These values were compared with those observed for the mother bush. Genetic fidelity of these regenerants was established by RAPD analysis advocating clonal propagation of this species through nodal segment culture and its commercial cultivation.     

Effects of light and temperature on the attachment and development of <Emphasis Type="BoldItalic">Gloiopeltis tenax</Emphasis> and <Emphasis Type="BoldItalic">Gloiopeltis furcata</Emphasis> tetraspores

Two 60-day experiments were conducted to study the influence of photon flux density (PFD) and temperature on the attachment and development of Gloiopeltis tenax and Gloiopeltis furcata tetraspores. In the first experiment, tetraspores of the two Gloiopeltis species were incubated at five temperature ranges (8°C, 12°C, 16°C, 20°C, 24°C) under a constant PFD of 80 μmol photons m⁻² s⁻¹ with a photoperiod of 12:12. In a second experiment, tetraspores were incubated under five PFD gradients (30, 55, 80, 105, 130 μmol photons m⁻² s⁻¹) at a constant temperature of 16°C with a photoperiod of 12:12. Maximum density of attached tetraspores was observed at 16°C for both species. Maximum per cent of spore germinating into disc was recorded at 12–16°C for G. tenax and 8–12°C for G. furcata. Maximum per cent of discs producing erect axes for G. tenax and G. furcata were recorded at 24°C and 20°C, respectively. Light had no significant effect on tetraspore attachment and developing into disc, but it affected the growth, sprouting and survival of its discs. Under 30–55 μmol photons m⁻² s⁻¹, the discs of the two species of Gloiopeltis did not form thallus until the end of the experiment. Optimum PFD range for G. tenax discs was 80–105 μmol photons m⁻² s⁻¹, whilst it was 80–130 μmol photons m⁻² s⁻¹ for G. furcata. Results presented in this study are expected to assist the progress of artificial seeding of Gloiopeltis.     

Protein,fatty acid,and pigment content of <Emphasis Type="Italic">Chlorella vulgaris</Emphasis> under different light regimes

The effects of irradiance and photoperiod on growth rates, chlorophyll a, β-carotene, total protein, and fatty acid content of Chlorella vulgaris were determined. The maximum growth rate (1.13 day⁻¹) was at 100 μmol photons m⁻² s⁻¹ and 16:8-h light/dark photoperiod. Chlorophyll a and β-carotene contents significantly differed under different light regimes with chlorophyll a content lower at high irradiance and longer light duration, while β-carotene showed the inverse trend. The total protein and fatty acid content also significantly differed in different light regimes; the maximum percentage of protein (46%) was at 100 μmol photons m⁻² s⁻¹ and 16:8 h photoperiod, and minimum (33%) was at 37.5 μmol photons m⁻² s⁻¹ and 8:16 h photoperiod; the total saturated fatty acids increased, while monounsaturated and polyunsaturated fatty acids decreased with increasing irradiance and light duration.     

Physiological differences in the growth of <Emphasis Type="Italic">Sargassum horneri</Emphasis> between the germling and adult stages

The effects of temperature, irradiance, and daylength on Sargassum horneri growth were examined at the germling and adult stages to discern their physiological differences. Temperature–irradiance (10, 15, 20, 25, 30°C × 20, 40, 80 μmol photons m⁻²s⁻¹) and daylength (8, 12, 16, 24 h) experiments were carried out. The germlings and blades of S. horneri grew over a wide range of temperatures (10–25°C), irradiances (20–80 μmol photons m⁻²s⁻¹), and daylengths (8–24 h). At the optimal growth conditions, the relative growth rates (RGR) of the germlings were 21% day⁻¹ (25°C, 20 μmol photons m⁻²s⁻¹) and 13% day⁻¹ (8 h daylength). In contrast, the RGRs of the blade weights were 4% day⁻¹ (15°C, 20 μmol photons m⁻²s⁻¹) and 5% day⁻¹ (12 h daylength). Negative growth rates were found at 20 μmol photons m⁻²s⁻¹ of 20°C and 25°C treatments after 12 days. This phenomenon coincides with the necrosis of S. horneri blades in field populations. In conclusion, we found physiological differences between S. horneri germlings and adults with respect to daylength and temperature optima. The growth of S. horneri germlings could be enhanced at 25°C, 20 μmol photons m⁻²s⁻¹, and 8 h daylength for construction of Sargassum beds and restoration of barren areas.     

Effect of sucrose,light, and carbon dioxide on plantain micropropagation in temporary immersion bioreactors

In vitro physiology and carbon metabolism can be affected by the sink–source relationship. The effect of different sucrose concentrations (10, 30, and 50 g L⁻¹), light intensities (80 and 150 μmol m⁻² s⁻¹), and CO₂ levels (375 and 1,200 μmol mol⁻¹) were tested during plantain micropropagation in temporary immersion bioreactors. Activities of pyruvate kinase, phosphoenol pyruvate carboxylase, and the photosynthesis rate were recorded. From the morphological and practical point of view, the best results were obtained when plants were cultured with 30 g L⁻¹ sucrose, 80 μmol m⁻² s⁻¹ light intensity, and 1,200 μmol mol⁻¹ CO₂ concentration. This treatment improved leaf and root development, reduced respiration during in vitro culture, and increased starch level at the end of the hardening phase. In addition to that, the number of competent plants was increased from 80.0% to 91.0% at the end of the in vitro phase and the survival percentage from 95.71% to 99.80% during ex vitro hardening.     

Effects of light intensity and nitrogen starvation on growth, total fatty acids and arachidonic acid in the green microalga Parietochloris incisa

The effects of light and nitrogen deficiency on biomass, fatty acid content and composition were studied in Parietochloris incisa, the unicellular freshwater chlorophyte accumulating very high amounts of arachidonic-acid-rich triacylglycerols. P. incisa cultures grown on complete nutrient medium and under high light (400 μmol photons m^{− 2} s⁻¹) showed the highest rate of growth in comparison to medium (200 μmol photons m⁻² s⁻¹) and low (35 μmol photons m⁻² s⁻¹) light intensity. Cultures grown under high light (on complete BG-11 medium) attained higher volumetric contents of total fatty acids and arachidonic acid due to greater increase in biomass. Nitrogen starvation brought about a strong increase in the arachidonic acid proportion of total fatty acids. Thus, adjustments to cultivation conditions could serve as an efficient tool for manipulation of yield and relative content of arachidonic acid in P. incisa. The significance of the changes in lipid metabolism for adaptation of P. incisa to high-light stress and nitrogen deficiency is also discussed.     

Eco-physiological responses of nitrogen-fixing cyanobacteria to light

The eco-physiological responses of three nitrogen-fixing cyanobacteria (N-fixing cyanobacteria), Aphanizomenon gracile, Anabaena minderi, and Ana. torques-reginae, to light were assessed under nutrient saturation. The N-fixing cyanobacteria were isolated into monocultures from a natural bloom in a shallow colored lake and their growth irradiance parameters and pigment composition were assessed. The different ecological traits related to light use (μ_max, α, I _k) suggest that these N-fixing cyanobacteria are well adapted to low light conditions at sufficient nutrients, yet interspecific differences were observed. Aphanizomenon gracile and Anabaena minderi had high relative growth rates at low irradiances (ca. 70% of those in high light), low half saturation constant for light-limited growth (I _k < 9.09 μmol photon m⁻² s⁻¹) and high efficiency (α < 0.11 day⁻¹ μmol photon⁻¹ m² s). Conversely, Ana. torques-reginae showed poorer light competitiveness: low relative growth rates at low irradiances (ca. 40% of those in high light), low α (0.009 day⁻¹ μmol photon⁻¹ m² s) and higher I _k (35.5 μmol photon m⁻² s⁻¹). Final densities in Aphanizomenon gracile and Anabaena minderi reached bloom densities at irradiances above 30 μmol photon m⁻² s⁻¹ with different hierarchy depending on irradiance, whereas Ana. torques-reginae never achieved bloom densities. All species had very low densities at irradiances ≤17 μmol photon m⁻² s⁻¹, thus no N-fixing blooms would be expected at these irradiances. Also, under prolonged darkness and at lowest irradiance (0 and 3 μmol photon m⁻² s⁻¹) akinetes were degraded, suggesting that in ecosystems with permanently dark sediments, the prevalence of N-fixing cyanobacteria should not be favored. All species displayed peaks of phycocyanin, but no phycoeritrin, probably due to the prevailing red light in the ecosystem from which they were isolated.     

Metabolism of N-alkylated spermine analogues by polyamine and spermine oxidases

N-alkylated polyamine analogues have potential as anticancer and antiparasitic drugs. However, their metabolism in the host has remained incompletely defined thus potentially limiting their utility. Here, we have studied the degradation of three different spermine analogues N,N′-bis-(3-ethylaminopropyl)butane-1,4-diamine (DESPM), N-(3-benzyl-aminopropyl)-N′-(3-ethylaminopropyl)butane-1,4-diamine (BnEtSPM) and N,N′-bis-(3-benzylaminopropyl)butane-1,4-diamine (DBSPM) and related mono-alkylated derivatives as substrates of recombinant human polyamine oxidase (APAO) and spermine oxidase (SMO). APAO and SMO metabolized DESPM to EtSPD [K _m(APAO) = 10 μM, k _cat(APAO) = 1.1 s⁻¹ and K _m(SMO) = 28 μM, k _cat(SMO) = 0.8 s⁻¹, respectively], metabolized BnEtSPM to EtSPD [K _m(APAO) = 0.9 μM, k _cat(APAO) = 1.1 s⁻¹ and K _m(SMO) = 51 μM, k _cat(SMO) = 0.4 s⁻¹, respectively], and metabolized DBSPM to BnSPD [K _m(APAO) = 5.4 μM, k _cat(APAO) = 2.0 s⁻¹ and K _m(SMO) = 33 μM, k _cat(SMO) = 0.3 s⁻¹, respectively]. Interestingly, mono-alkylated spermine derivatives were metabolized by APAO and SMO to SPD [EtSPM K _m(APAO) = 16 μM, k _cat(APAO) = 1.5 s⁻¹; K _m(SMO) = 25 μM, k _cat(SMO) = 8.2 s⁻¹; BnSPM K _m(APAO) = 6.0 μM, k _cat(APAO) = 2.8 s⁻¹; K _m(SMO) = 19 μM, k _cat(SMO) = 0.8 s⁻¹, respectively]. Surprisingly, EtSPD [K _m(APAO) = 37 μM, k _cat(APAO) = 0.1 s⁻¹; K _m(SMO) = 48 μM, k _cat(SMO) = 0.05 s⁻¹] and BnSPD [K _m(APAO) = 2.5 μM, k _cat(APAO) = 3.5 s⁻¹; K _m(SMO) = 60 μM, k _cat(SMO) = 0.54 s⁻¹] were metabolized to SPD by both the oxidases. Furthermore, we studied the degradation of DESPM, BnEtSPM or DBSPM in the DU145 prostate carcinoma cell line. The same major metabolites EtSPD and/or BnSPD were detected both in the culture medium and intracellularly after 48 h of culture. Moreover, EtSPM and BnSPM were detected from cell samples. Present data shows that inducible SMO parallel with APAO could play an important role in polyamine based drug action, i.e. degradation of parent drug and its metabolites, having significant impact on efficiency of these drugs, and hence for the development of novel N-alkylated polyamine analogues.     

Fast cyclic electron transport around photosystem I in leaves under far-red light: a proton-uncoupled pathway?

Fast cyclic electron transport (CET) around photosystem I (PS I) was observed in sunflower (Helianthus annuus L.) leaves under intense far-red light (FRL) of up to 200 μmol quanta m⁻² s⁻¹. The electron transport rate (ETR) through PS I was found from the FRL-dark transmittance change at 810 and 950 nm, which was deconvoluted into redox states and pool sizes of P700, plastocyanin (PC) and cytochrome f (Cyt f). PC and P700 were in redox equilibrium with K _e = 35 (ΔE _m = 90 mV). PS II ETR was based on O₂ evolution. CET [(PS I ETR) − (PS II ETR)] increased to 50–70 μmol e⁻ m⁻² s⁻¹ when linear electron transport (LET) under FRL was limited to 5 μmol e⁻ m⁻² s⁻¹ in a gas phase containing 20–40 μmol CO₂ mol⁻¹ and 20 μmol O₂ mol⁻¹. Under these conditions, pulse-saturated fluorescence yield F _m was non-photochemically quenched; however, F _m was similarly quenched when LET was driven by low green or white light, which energetically precluded the possibility for active CET. We suggest that under FRL, CET is rather not coupled to transmembrane proton translocation than the CET-coupled protons are short-circuited via proton channels regulated to open at high ΔpH. A kinetic analysis of CET electron donors and acceptors suggests the CET pathway is that of the reversed Q-cycle: Fd → (FNR) → Cyt c_n → Cyt b_h → Cyt b_l → Rieske FeS → Cyt f → PC → P700 →→ Fd. CET is activated when PQH₂ oxidation is opposed by high ΔpH, and ferredoxin (Fd) is reduced due to low availability of e⁻ acceptors. The physiological significance of CET may be photoprotective, as CET may be regarded as a mechanism of energy dissipation under stress conditions.     

Photosynthetic Response of Carrots to Varying Irradiances

Response to irradiance of leaf net photosynthetic rates (P _N) of four carrot cultivars: Cascade, Caro Choice (CC), Oranza, and Red Core Chantenay (RCC) were examined in a controlled environment. Gas exchange measurements were conducted at photosynthetic active radiation (PAR) from 100 to 1 000 μmol m⁻² s⁻¹ at 20 °C and 350 μmol (CO₂) mol⁻¹(air). The values of P _N were fitted to a rectangular hyperbolic nonlinear regression model. P _N for all cultivars increased similarly with increasing PAR but Cascade and Oranza generally had higher P _N than CC. None of the cultivars reached saturation at 1 000 μmol m⁻² s⁻¹. The predicted P _N at saturation (P _Nmax) for Cascade, CC, Oranza, and RCC were 19.78, 16.40, 19.79, and 18.11 μmol (CO₂) m⁻² s⁻¹, respectively. The compensation irradiance (I _c) occurred at 54 μmol m⁻² s⁻¹ for Cascade, 36 μmol m⁻² s⁻¹ for CC, 45 μmol m⁻² s⁻¹ for Oranza, and 25 μmol m⁻² s⁻¹ for RCC. The quantum yield among the cultivars ranged between 0.057–0.033 mol(CO₂) mol⁻¹(PAR) and did not differ. Dark respiration varied from 2.66 μmol m⁻² s⁻¹ for Cascade to 0.85 μmol m⁻² s⁻¹ for RCC. As P _N increased with PAR, intercellular CO₂ decreased in a non-linear manner. Increasing PAR increased stomatal conductance and transpiration rate to a peak between 600 and 800 μmol m⁻² s⁻¹ followed by a steep decline resulting in sharp increases in water use efficiency. This revised version was published online in August 2006 with corrections to the Cover Date.     

Thidiazuron-induced high-frequency direct shoot organogenesis of <Emphasis Type="Italic">Cannabis sativa</Emphasis> L.

Induction of high-frequency shoot regeneration using nodal segments containing axillary buds from a 1-yr-old mother plants of Cannabis sativa was achieved on Murashige and Skoog (MS) medium containing 0.05–5.0 μM thidiazuron. The quality and quantity of regenerants were better with thidiazuron (0.5 μM thidiazuron) than with benzyladenine or kinetin. Adding 7.0 μM of gibberellic acid into a medium containing 0.5 μM thidiazuron slightly increased shoot growth. Elongated shoots when transferred to half-strength MS medium supplemented with 500 mg l⁻¹ activated charcoal and 2.5 μM indole-3-butyric acid resulted in 95% rooting. The rooted plants were successfully acclimatized in soil. Following acclimatization, growth performance of 4-mo-old in vitro propagated plants was compared with ex vitro vegetatively grown plants of the same age. The photosynthesis and transpiration characteristics were studied under different light levels (0, 500, 1,000, 1,500, or 2,000 μmol m⁻² s⁻¹). An increase in photosynthesis was observed with increase in the light intensity up to 1,500 μmol m⁻² s⁻¹ and then decreased subsequently at higher light levels in both types of plants. However, the increase was more pronounced at lower light intensities below 500 μmol m⁻² s⁻¹. Stomatal conductance and transpiration increased with light intensity up to highest level (2000 μmol m⁻² s⁻¹) tested. Intercellular CO₂ concentration (C _i) and the ratio of intercellular CO₂ concentration to ambient CO₂ (C _i/C _a) decreased with the increase in light intensity in both in vitro as well as ex vitro raised plants. The results show that in vitro propagated and hardened plants were functionally comparable to ex vitro plants of same age in terms of gas and water vapor exchange characteristics, within the limits of this study.     

Development of laboratory-scale photobioreactor for water purification by use of a biofilter composed of the aerial microalga Trentepohlia aurea (Chlorophyta)

Biofilms formed by the green alga Trentepohlia aurea could be a useful tool in the removal of nitrate and phosphate from water. When a prepared biofilter was dampened with medium and incubated under low light intensity (10 μmol photons m⁻² s⁻¹) between 5 and 50 μmol photons m⁻² s⁻¹, the efficiency of removal of inorganic compounds from water was higher without the decomposition of chlorophylls in the cells. Algal cells immobilized on a glass fiber filter could be kept for 12 weeks under dark conditions at 4°C in the refrigerator. We tried to construct a laboratory-scale photobioreactor for the removal of inorganic nitrogen and phosphate from water by the biofilm. In this study, the synthetic wastewater was prepared by diluting 18-fold Bold’s basal medium with deionized water. The photobioreactor could efficiently remove nitrate and phosphate from the synthetic wastewater under continuous illumination. The removal ability of nitrate and phosphate per sheet of the biofilter in the photobioreactor exhibited about an 8- and 16-fold increase, respectively, in 3 days, compared with the bath experimental results. This study showed that the cycling of wastewater in the reactor by the pump led to a significant improvement in the efficiency of the inorganic ion uptake from water.     

Changes in PAL,CHS, DAHP synthase (DS-Co and Ds-Mn) activity during anthocyanin synthesis in suspension culture of Fragaria ananassa

The activity of 3-deoxy-D-arabino-heptulosonate 7-phosphate (DAHP) synthase (DS-Mn, DS-Co), phenylalanine ammonia-lyase (PAL), and chalcone synthase (CHS) was monitored at various light intensities (dark, 8.88 μmol m⁻² s⁻¹, 88.8 μmol m⁻² s⁻¹) using a strawberry cell suspension culture. DS-Mn, PAL, and CHS were found to increase significantly (p>0.05) under light intensitie of 88.8 μmol m⁻² s⁻¹ compared to those of 8.88 μmol m⁻² s⁻¹ and dark. The activity of DS-Mn, PAL, and CHS were maximum at 88.8 μmol m⁻² s⁻¹. Anthocyanin content reached a maximum after 48–60 h of culturing at 88.8 μmol m⁻² s⁻¹. DS-Co showed greater activity than DS-Mn during cell culturing, but showed no correlation with anthocyanin production and light intensity. The CHS gene expression was continuous at a light intensity of 88.8 μmol m⁻² s⁻¹. This revised version was published online in June 2006 with corrections to the Cover Date.     

Palm oil mill effluent (POME) cultured marine microalgae as supplementary diet for rotifer culture

Malaysia is the world’s leading producer of palm oil products that contribute US$ 7.5 billion in export revenues. Like any other agro-based industries, it generates waste that could be utilized as a source of organic nutrients for microalgae culture. Present investigation delves upon Isochrysis sp. culture in POME modified medium and its utilization as a supplement to Nanochloropsis sp. in rotifer cultures. The culture conditions were optimized using a 1 L photobioreactor (Temp: 23°C, illumination: 180 ∼ 200 μmol photons m⁻²s⁻¹, n = 6) and scaled up to 10 L outdoor system (Temp: 26–29°C, illumination: 50 ∼ 180 μmol photons m⁻²s⁻¹, n = 3). Algal growth rate in photobioreactor (μ = 0.0363 h⁻¹) was 55% higher compared to outdoor culture (μ = 0.0163 h⁻¹), but biomass production was 1.3 times higher in outdoor culture (Outdoor = 91.7 mg m⁻²d⁻¹; Photobioreactor = 69 mg m⁻²d⁻¹). Outdoor culture produced 18% higher lipid; while total fatty acids (FA) was not significantly affected by the change in culture systems as both cultures yield almost similar concentrations of fatty acids per gram of sample (photobioreactor = 119.17 mg g⁻¹; outdoor culture = 104.50 mg g⁻¹); however, outdoor cultured Isochrysis sp. had 26% more polyunsaturated fatty acids (PUFAs). Rotifers cultured in Isochrysis sp./ Nanochloropsis sp. (1:1, v/v) mixture gave similar growth rate as 100% Nanochoropsis sp. culture (μ = 0.40 d⁻¹), but had 45% higher counts of rotifers with eggs (t = 7, maximum). The Isochrysis sp. culture successfully lowered the nitrate (46%) and orthophosphate (83%) during outdoor culture.