Effects of Progesterone, β-Estradiol,and Androsterone on the Changes of Inorganic Element Content in Barley Leaves

The present study was performed to determine the changes in inorganic element content in barley leaves of mammalian sex hormones (MSH). Barley leaves were sprayed with 10⁻⁴, 10⁻⁶, 10⁻⁹, 10⁻¹², 10⁻¹⁵ M concentrations of progesterone, β-estradiol, and androsterone at 7th day after sowing. The plants were harvested at the end of 18 days after treatment with MSH solutions. The inorganic element concentrations were determined using wavelength dispersive X-ray fluorescence spectroscopy technique. Although the all MSH concentrations significantly (p < 0.05) increased the concentrations of calcium, magnesium, phosphorus, sulfur, copper, manganese, aluminum, zinc, iron, potassium, and chlorine, it decreased those of sodium concentration in barley leaves. The maximum changes in the element concentrations were obtained at 10⁻⁹ M for plant leaves treated with progesterone, 10⁻⁶ M for plant leaves treated with β-estradiol and androsterone. The present study elucidated that MSH significantly (p < 0.05) affected the inorganic element concentrations in barley leaves.     

Mammalian sex hormones stimulate antioxidant system and enhance growth of chickpea plants

In the present study, it was aimed to investigate the influence of exogenous mammalian sex hormones (MSH) (progesterone, β-estradiol and androsterone) on the morphological (root and shoot growth) and biochemical parameters (protein and sugar content, antioxidant enzyme activities, and lipid peroxidation and H₂O₂ levels) of chickpea (Cicer arietinum L.) plants growing under control conditions. The solutions of hormones prepared at different concentrations (10⁻⁴, 10⁻⁶, 10⁻⁹, 10⁻¹² and 10⁻¹⁵ M) were sprayed once on the leaves of 7-day plants. The plants were harvested on 18 days after the hormone treatment. Although all of the hormones at the tested concentrations significantly increased plant growth, soluble protein and sugar contents, and antioxidant enzyme activities [superoxide dismutase (SOD), peroxidase (POX) and catalase (CAT)], they decreased H₂O₂ content and lipid peroxidation level when compared with control plants. The activities of SOD, POX and CAT reached to the highest levels at 10⁻⁶ M for progesterone, and 10⁻⁹ M for β-estradiol and androsterone, which maximum growth, protein and sugar contents were determined. The same concentrations also resulted in the lowest levels for H₂O₂ content and lipid peroxidation. It can be interpreted that the MSH improve plant growth and development by affecting some biochemical parameters including antioxidative system.     

Stimulation of generative development in partly vernalized winter wheat by animal sex hormones

The influence of selected animal steroid sex hormones, on generative development of winter wheat var. Grana was investigated. Wheat plants of this variety necessitate 63-day long vernalization. Mature, isolated embryos of wheat were cultured in vitro on media containing androsterone, androstenedione, estriol, estrone, 17β-estradiol and progesterone in concentrations 10⁻⁵ and 10⁻⁶ M. They were not vernalized or vernalized for 7, 14, 21 and 28 days (5 °C, 8 h photoperiod). Investigated steroids stimulated the generative development of winter wheat by an increasing in the percentage of heading plants and accelerating the heading. The strongest effect was observed when plants were treated with steroids during the suboptimal, 21 and 28 day, vernalization. After 28 days of vernalization, 100 % heading were observed in plants obtained on the media containing androsterone and androstenedione in concentrations 10⁻⁵ and 10⁻⁶ M or estrone, 17β-estradiol and progesterone in concentration 10⁻⁵ M. Control plants showed only 8 % heading. An erratum to this article is available at .     

The Exogenous Application of Brassinosteroids to Zea mays (L.) Stressed by Long-Term Chilling Does Not Affect the Activities of Photosystem 1 or 2

The effect of various concentrations of exogenously applied 24-epibrassinolide (E) and 2α,3α,17β-trihydroxy-5α-androstan-6-one (A) on the activities of Photosystem 1 and the Hill reaction, the contents of photosynthetic pigments, and the growth of plants was examined in young maize (Zea mays L.) plants subjected to long-term chilling stress or grown in normal-temperature conditions. Neither the activity of Photosystem 1 nor the Hill reaction activity of plants was in any way affected by the treatment with brassinosteroids (BRs), which suggests that the photosynthetic complexes of thylakoid membranes are not the primary site of the influence of BRs on photosynthesis. An extremely low (10⁻¹⁴ M) concentration of A applied to the nonstressed plants significantly increased the length of their 4th to the 7th leaves and their height, as well as the contents of chlorophylls a and b and total carotenoids. However, under chilling conditions, this positive effect was significant for the chlorophyll content only and higher concentrations of BRs (10⁻¹², 10⁻¹⁰, 10⁻⁸ M) usually had no effect at all.     

Regulation of Sorus Formation by Auxin in Laminariales Sporophyte

Young sporophytes of Laminaria japonica Areshoug were cultured in six indole-acetic acid (IAA) concentrations (0, 10⁻⁸, 10⁻⁷, 10⁻⁶, 10⁻⁵, 10⁻⁴ M) to examine the effect of auxin on growth. The effects of auxin on sorus formation were also examined by using discs taken from the adult sporophyte. The auxin contents and IAA oxidase activities in the thallus and sorus parts of the sporophyte were determined with the blade and sporophyll of other Laminariales plants, Undaria pinnatifida (Harvey) Suringar and Alaria crassifolia Kjellman. The young sporophytes of L. japonica showed highest elongation rate in 10⁻⁵ M IAA. In contrast, the sorus formation on the discs cultured in 10⁻⁵ M IAA was markedly delayed in comparison with other concentrations, indicating that sorus formation was suppressed by IAA. Free and conjugated auxin contents were lower in the reproductive parts than in the vegetative parts. In three Laminariales sporophytes, IAA oxidase activity was about 3–9 times higher in the reproductive parts than in the vegetative parts. Taken together these results suggest that the growth and reproduction of Laminariales sporophytes are regulated by internal auxin levels. Elucidating the regulation mechanism is likely to provide information that is important for the management of plant production and the assessment of the physiological status of plants in the field.     

Growth-Regulating Activity of Three 4-Hydroxycoumarin Derivatives on Inoculated Soybean Plants

Three 4-hydroxycoumarin derivatives (ethyl 2-[(4-hydroxy-2-oxo-2H-chromen-3-yl)(4-hydroxyphenyl)methyl]-3-oxobutanoate (SS-14), ethyl 2-[(4-hydroxy-2-oxo-2H-chromen-3-yl)(3-nitrophenyl)methyl]-3-oxobutanoate (SS-21), and 2-[(3,4,5-trimethoxyphenyl)(4-hydroxy-2-oxo-2H-chromen-3-yl)methyl]-3-oxobutanoate (T-2)] were tested for growth-regulating activity on nitrogen-fixing soybean plants in different concentrations: 10⁻⁵, 10⁻⁴, and 10⁻³ M. They revealed growth-regulating activity in a concentration-dependent manner. The most powerful suppression effect of T-2 on shoot and root fresh and dry biomass accumulation, length of roots, and height of plants was found. Shoot fresh biomass was suppressed in an equal extent at 10⁻³ M of the three compounds but the order of inhibition regarding the three applied concentrations was T2 > SS-14 ≈ SS-21. The compound SS-14 inhibited nodule number and nodule biomass mainly at the highest applied concentration, 10⁻³ M. The highest inhibition of nitrogenase activity was established at the three applied concentrations of the compound SS-14.     

Polyamines increase carpospore output and growth during in vitro cultivation of Hydropuntia cornea

Carpospore output and development in the marine red alga Hydropuntia cornea J. Agardh. were increased by adding polyamines (PAs) (putrescine, spermidine and spermine) singly or in combinations at 10⁻⁹, 10⁻⁶ and 10⁻³ M. Cell divisions after spore release and development of apical axis between 17 and 21 days characterized carpospore development. PAs increased carpospore development by promoting cell divisions to form cell masses between day 2 and 3. Morphogenesis to develop apical axes occurred at day 7. Spermine at 10⁻⁶ M and a combination of putrescine 10⁻⁹ M + spermidine 10⁻⁹ M + spermine 10⁻⁹ M gave a higher number of carpospores and enhanced their further development to sporelings.     

Effect of nitrite on growth and microcystins production of <Emphasis Type="Italic">Microcystis aeruginosa</Emphasis> PCC7806

As a common pollutant, nitrite concentrations can approach 15 mg NO₂⁻-N L⁻¹ in some aquatic systems. Microcystis aeruginosa blooms are common and widespread in eutrophic freshwater bodies. In this study, M. aeruginosa was exposed to nitrite concentrations ranging from 0 to 15 mg NO₂⁻-N L⁻¹, and the responses of M. aeruginosa were investigated. The specific growth rates, maximum cell densities, light-saturated photosynthetic rates (P_m ^chla ), dark respiration rates (R_d ^chla ), and apparent photosynthetic efficiencies (α^chla ) showed a significant decline with nitrite concentrations increasing. Electrical conductivity and malondialdehyde contents investigation revealed cell membrane damage and apparent leakage of intracellular contents under high nitrite level conditions due to oxidative stress enhancement. Intracellular microcystin (MC)-LR content reached the highest value at 10 mg NO₂⁻-N L⁻¹; however, extracellular MC-LR contents showed a continuous increase until 15 mg NO₂⁻-N L⁻¹ owing to the increasing leakage of intracellular contents. These results elucidated that the high-level nitrite inhibited M. aeruginosa growth by rising oxidative stress, damaging cell membrane, and reducing photosynthesis. However, the moderate increase in nitrite concentrations promoted toxin production and release of toxin.     

A comparative effect of IAA and 4-Cl-IAA on growth,nodulation and nitrogen fixation in <Emphasis Type="Italic">Vigna radiata</Emphasis> (L.) Wilczek

The plants of mung bean (Vigna radiata L. Wilczek) were raised from the seeds soaked in water (control), IAA or 4-C-IAA (10⁻⁶, 10⁻⁸ or 10⁻¹⁰ M) for 8 or 12 h. The plants were allowed to grow in a net house and were sampled at 30 and 45 days after sowing (DAS). Both IAA and 4-Cl-IAA significantly affected the growth (length, fresh and dry mass of roots and shoots), the number of nodules, their fresh and dry mass and the activity of nitrogenase. However, the contents of nitrogen and carbohydrate exhibited a decrease in response to both the auxins. 4-Cl-IAA, at a concentration of 10⁻⁸ M, generated the best response. Moreover, 4-Cl-IAA at other two concentrations (10⁻⁶ and 10⁻¹⁰ M) was much more active than any of the IAA concentration used.     

Exposure to airborne microorganisms,hyphal fragments,and pollen in a field of organically grown strawberries

Many working environments are predisposed for larger than average amounts of fungi and other microorganisms often due to organic material being handled. From 2003 to 2007, the area used for strawberry production in Denmark increased by 62%. The purpose of this study was to determine the levels of exposure to microorganisms, endotoxin, (1→3)-β-d-glucan (β-glucan), and pollen in a field of strawberries. The study was carried out in eastern Denmark from the middle of June to the beginning of August 2008. The strawberries were grown organically, and microbiological pest control agents (MPCAs) were applied during this and former growth seasons. In order to measure exposure to inhalable bioaerosol components, we used stationary filter samplers. Bioaerosol sampling was performed during 4 working days, and a total of 57 samplings were performed. The filters were analysed for contents of fungi, MPCAs, endotoxin, β-glucan, and pollen. The mean exposure was 6,154 CFU Cladosporium sp. m⁻³, 1.0 × 10⁵ fungal spores m⁻³, 4.1 × 10⁴ hyphal fragments m⁻³, 5.8 × 10³ pollen m⁻³, 57.3 ng β-glucan m⁻³, and 8.9 endotoxin units (EU) m⁻³. A significant and positive correlation was found between β-glucan and fungal spores and between CFU of Cladosporium sp. and CFU of fungi. We selected specifically for Metarhizium anisopliae, Beauveria bassiana, and the applied MPCAs Trichoderma harzianum, T. polysporum, and Bacillus thuringiensis but found none of these species. In conclusion, our study shows that berry pickers in this organic strawberry field were potentially subjected to higher levels of fungal spores, Cladosporium sp., hyphal fragments, pollen, and thus also β-glucan than is usually seen in outdoor air. Exposure to MPCAs was not seen. The exposure to endotoxin was only slightly higher than e.g. in a town.     

Estrogen mitogenic action. I. Demonstration of estrogen-dependent MTW9/PL2 carcinogen-induced rat mammary tumor cell growth in serum-supplemented culture and technical implications

Summary The MTW9/PL cell line was established by our laboratory in culture from the carcinogen-induced hormone-responsive MT-W9A rat mammary tumor of a Wistar-Furth (W/Fu) rat. This tumor formed estrogen, androgen, and progesterone responsive tumors in W/Fu rats (Sirbasku, D. A., Cancer Res. 38:1154–1165; 1978). It was later used to derive the MTW9/PL2 cell population which was also estrogen-responsive in vivo (Danielpour, D., et al., In Vitro Cell. Dev. Biol. 24∶42–52; 1988). In the study presented here, we describe serum-supplemented culture conditions in which the MTW9/PL2 cells demonstrate≥80-fold steroid hormone growth responses. All sera used were steroid hormone-depleted by charcoal-dextran treatment at 34°C. The studies were done with horse serum as well as serum from other mammalian species. The growth of the MTW9/PL2 cells was biphasic in response to hormone-depleted serum. Concentrations of ≤5% (v/v) promoted optimum growth. Above this concentration, serum was inhibitory. Concentrations ≥40% (v/v) inhibited growth altogether. Addition of 1.0×10⁻¹³−1.0×10⁻⁸ M 17β-estradiol (E₂) reversed the inhibition completely. At 1.0×10⁻⁸ M, estrone, estriol and diethylstilbestrol promoted growth as well as E₂. Testosterone and dihydrotestosterone promoted growth only at ≥10⁻⁷ M. Progesterone was effective only at≥10⁻⁶ M. Cortisol was ineffective. Labeled-hormone-binding analysis and Western immunoblotting documented that MTW9/PL2 cells had estrogen and progesterone receptors but not androgen or cortisol receptors. Estrogen treatment of MTW9/PL2 cells induced a concentration and time dependent increase in progesterone receptors. We conclude (1) the MTW9/PL2 population is the first highly steroid hormone-responsive rat mammary tumor cell line to be established in culture from a carcinogen-induced tumor, and (2) sera from a number of species including horse, rat and human contain an inhibitor which mediates estrogen sensitive MTW9/PL2 cell growth in culture.     

Triacontanol-mediated regulation of growth and other physiological attributes,active constituents and yield of <Emphasis Type="Italic">Mentha arvensis</Emphasis> L.

Triacontanol (TRIA) has been realized as a potent plant growth promoting substance for a number of agricultural and horticultural crops. Out of a large number of essential oil bearing plants, mint (Mentha arvensis L.) constitutes the most important source of therapeutic agents used in the alternative systems of medicine. The mint plant has marvelous medicinal properties. In view of enhancing growth, yield and quality of this medicinally important plant, a pot experiment was conducted according to simple randomized block design. The experiment was aimed at studying the effect of four concentrations of TRIA (10⁻⁰, 10⁻⁷, 10⁻⁶ and 10⁻⁵ M) on the performance of mint with regard to growth and other physiological attributes, crop yield and quality attributes and the yield and contents of active constituents of the plant. The growth and other physiological parameters as well as yield and quality attributes were studied at 100 and 120 DAP. The foliar application of TRIA at 10⁻⁶ M concentration significantly enhanced most of the growth and other physiological attributes, crop herbage yield and the yield and content of active constituents (menthol, l-methone, isomenthone and menthyl acetate) of mint at both the stages. However, the next higher concentration of TRIA (10⁻⁵ M) exhibited slightly negative effect and did not further increase the values of the attributes studied, but it proved significantly better than the control. Application of TRIA significantly enhanced the yield and content of all the active constituents determined by GLC technique.     

Environmentally relevant cadmium concentrations affect development and induce apoptosis of <Emphasis Type="Italic">Paracentrotus lividus</Emphasis> larvae cultured in vitro

Sea urchin embryos and larvae represent suitable model systems on where to investigate the effects of heavy metals on development and cell viability. Here, we tested the toxic effects of low (10⁻¹² M), medium (10⁻⁹ M), and high (10⁻⁶ M) cadmium chloride concentrations, mimicking unpolluted, moderately and highly polluted seawaters, respectively, on Paracentrotus lividus sea urchins offspring. Larvae were continuously treated from fertilization and inspected at time intervals comprised between 10 and 30 days of development. Delays and/or morphological abnormalities were firstly evident in larvae treated for 15 days with high cadmium (10⁻⁶ M) and for 25 days with medium cadmium (10⁻⁹ M). Major defects consisted in the reduction and lack of arms and skeleton elongation. No obvious differences with respect to controls were observed in embryos/larvae exposed to low cadmium (10⁻¹² M), even after 30 days of exposure. Using in situ terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay (TUNEL) assay on larvae whole mounts, we detected apoptosis after 10 days of treatment with 10⁻⁶ and 10⁻⁹ M CdCl_2, when no morphological abnormalities were recognizable yet. Supernumerary apoptotic cells were found in arm buds, ciliary bands, and apex. In conclusion, echinoderm embryos and larvae represent candidates of choice for the study of stress and defense mechanisms activated by cadmium exposure.     

Effects of gibberellin and auxin on the synthesis of abscisic acid and ethylene in buds of dormant and sprouting potato tubers

Gibberellic and β-indolylacetic acids at concentrations of 10⁻⁷-10⁻⁵ M were shown to change the hormonal status and duration of true dormancy in potato tubers. Gibberellic acid shortened the true dormancy and decreased the contents of abscisic acid and ethylene in the apical meristem. β-Indolylacetic acid elongated the true dormancy and decreased abscisic acid production, but caused a more than tenfold increase in the production of ethylene by apical tissues. The data suggest that β-indolylacetic acid and ethylene, as well as gibberellic and abscisic acids, are involved in the regulation of true dormancy in potato tubers.     

Bioelectrical activity in the heart of the lugworm Arenicola marina

Standard microelectrode technique was used to study electrical activity of the isolated heart of the polychaete annelid, Arenicola marina. Typical pacemaker activity with slow diastolic depolarization was observed in all recordings. The average maximum diastolic potential (−58.4 ± 3.2 mV), the average amplitude of the action potential (28.7 ± 4.7 mV) and the average total duration of the action potential (2,434 ± 430 ms) were determined. There has been no gradient of automaticity observed in our studies, which suggests that all regions of the Arenicola heart could possess pacemaker functions. Acetylcholine (ACh) produced a concentration dependent (5 × 10⁻⁸–5 × 10⁻⁵ M) increase of the beating rate via increase in the rate of the diastolic depolarization. ACh (5 × 10⁻⁵ M) increased beating rate by 2.5-fold compared to the control rate. A stronger action of ACh resulted in depolarization, block of action potential generation and contracture of the heart. The non-hydrolysable ACh analog carbacholine (10⁻⁸–10⁻⁶ M) produced similar effects. All effects of ACh and carbacholine were abolished by 5 × 10⁻⁶ M atropine. d-Tubocurarine (5 × 10⁻⁵ M) did not significantly alter effects of ACh or carbacholine. Epinephrine (10⁻⁸–10⁻⁶ M) caused the slowing of pacemaker activity and marked decrease of action potential duration. 10⁻⁶ M epinephrine produced complete cardiac arrest. The effects of epinephrine were not significantly altered by the β-blocker propranolol (5 × 10⁻⁶ M). The β-agonist isoproterenol (10⁻⁷–10⁻⁵ M) and the α-agonist xylometazoline (10⁻⁶–10⁻⁵ M) did not produce significant effects. Thus, cholinergic effects in the Arenicola heart are likely to be mediated via muscarinic receptors, while the nature of adrenergic effects needs further investigation.     

Growth inhibitory activity of ABA-β-<Emphasis Type="SmallCaps">d</Emphasis>-glucopyranosyl ester and ABA-β-<Emphasis Type="SmallCaps">d</Emphasis>-glucosidase

Exogenously applied ABA-β-d-glucopyranosyl ester (ABA-GE) inhibited shoot growth of alfalfa (Medicago sativa L.), cress (Lepidium sativum L.), lettuce (Lactuca sativa L.), Digitaria sanguinalis L., timothy (Pheleum pratense L.) and ryegrass (Lolium multiflorum Lam.) seedlings at concentrations greater than 0.1 μM. The growth inhibitory activity of ABA-GE on these shoots was 26–40% of that of (+)-ABA. ABA-β-d-glucosidase activities in these seedlings were 11–31 nmol mg⁻¹ protein min⁻¹. These results suggests that exogenously applied ABA-GE may be absorbed by plant roots and hydrolyzed by ABA-β-d-glucosidase, and liberated free ABA may induce the growth inhibition in these plants. Thus, although ABA-GE had been thought to be physiologically inactive ABA conjugate, ABA-GE may have important physiological functions rather than an inactive conjugated ABA form.     

Pb2+ reduces PKCs and NF-κB in vitro

The mechanism of lead (Pb²⁺)-induced neurotoxicity has not yet been fully elucidated. The purpose of this study was to examine the effects of Pb²⁺ on several protein kinase C (PKC) isoforms and the nuclear factor-κB (NF-κB)–I-κB kinase-alpha (IKK-α) axis in cultured neuronal cells. Neurons were isolated from rat fetal brain at the 18th day of gestation of pregnant Sprague Dawley rats and cultured for 10 days before use. Neurons were exposed to Pb²⁺ at concentrations of 10⁻¹⁰, 10⁻⁹, 10⁻⁸, and 10⁻⁷ mol/L for 14 h and antigens of typical PKC-α,β,γ; novel PKC (ε, δ), atypical PKC (λ), NF-κB (p50), and IKK-α were enriched by immunoprecipitation and determined by western blotting. Total, calcium-dependent and independent PKC activities were also determined by counting the transferred γ-³² P in the substrate-histone. The results indicated that inorganic Pb²⁺ significantly reduced all PKC isoforms (α,β,γ, ε, λ) except δ, inhibiting the total, calcium-dependent and calcium-independent PKC activities in a dose-dependent manner. Additionally, Pb²⁺ gradually reduced NF-κB (p50) and IKK-α protein levels. This suggests that Pb²⁺ exhibits varying preference for individual PKC isoforms but reduces the NF-κB–IKK-α axis to a similar extent.     

Gene cloning,expression and characterization of a neoagarotetraose-producing β-agarase from the marine bacterium <Emphasis Type="Italic">Agarivorans</Emphasis> sp. HZ105

A β-agarase gene hz2 with 2,868 bp was cloned from the marine agarolytic bacterium Agarivorans sp. HZ105. It encoded a mature agarase HZ2 of 102,393 Da (920 amino acids). Based on the amino acid sequence similarity, agarase HZ2 was assigned to the glycoside hydrolase family 50. The β-agarase shared a gene sequence identity of 98.6% with the reported but much less characterized β-agarase agaB from Vibrio sp. JT0107. Its recombinant agarase rHZ2 was produced in E. coli cells and purified to homogeneity. The agarase rHZ2 degraded agarose and neoagarooligosaccharides with degrees of polymerization above four, to yield neoagarotetraose as the dominant product, which was different from β-agarase agaB of Vibrio sp. JT0107. The agarose hydrolysis pattern suggested that rHZ2 was an endo-type β-agarase. Beta-mercaptoethanol (90 mM) and dithiothreitol (9 mM) increased the agarase activity of rHZ2 by 72.9% and 17.3% respectively, while SDS (9 mM) inhibited the activity completely. The agarase activity was independent of Na⁺, K⁺, Mg²⁺ and Ca²⁺. The maximal enzyme activity was observed at 40°C and pH 7. The kinetic parameters K _m, V _max, K _cat, and K _cat/K _m values toward agarose of agarase rHZ2 were 5.9 mg ml⁻¹, 235 U mg⁻¹, 401 s⁻¹ and 6.8 × 10⁵ M⁻¹ s⁻¹, respectively. Agarase rHZ2 could have a potential application in the production of bioactive neoagarotetraose.     

Effects of Brassinolide with Naphthalene Acetic Acid on the Formation of Adventitious Roots,Trichome-Like Roots and Calli from Cultured Tobacco Leaf Segments,and the Expression Patterns of <Emphasis Type="Italic">CNT103</Emphasis>

We treated cultured tobacco leaf segments with brassinolide (BL) and naphthalene acetic acid (NAA) and determined that optimum concentrations of NAA for adventitious root, trichome-like root, and calli formation were, respectively, 10⁻⁶, 10⁻⁵, and 10⁻⁴ M. In the adventitious root formation group, the number and length of adventitious roots were increased at lower concentrations of BL; however, they became trichome-like roots at higher levels of BL. The trichome-like root formation group showed better development when a low concentration of BL was added. However, at higher concentrations of BL, trichome-like root production was reduced, forming calli instead. In the calli formation group, more calli were formed at low BL concentrations and after persistent exposure to BL regardless of BL concentration, and the size of the leaf segments increased. The CNT103 gene, which is expressed at the root tips showed increased levels of expression at BL concentrations up to 10⁻⁹ M and decreased levels of expression at BL concentrations over 10⁻⁹ M in the adventitious roots, trichome-like roots, and calli formation groups.     

Changes in fatty acids and sterols during batch growth of <Emphasis Type="Italic">Pavlova viridis</Emphasis> in photobioreactor

Changes in the composition of fatty acids and sterols of Pavlova viridis cultured in an air-lift photobioreactor were studied using gas chromatography-mass spectrometry (GC-MS). The results show that radical changes in fatty acid and sterol contents and compositions occurred during growth phase transitions: the total lipid increased along with the culture age, from 166.4 mg g⁻¹ (late exponential phase) to 232.7 mg g⁻¹ (linear phase), and increased further to be 235.1 mg g⁻¹ in the stationary phase. Polyunsaturated fatty acids (PUFAs), especially eicosapentaenoic acid (EPA), decreased along with the culture time, PUFAs, and EPA contents maximized in the late exponential phase to become 46.2 mg g⁻¹ and 22.1 mg g⁻¹ respectively; there was no significant change in docosahexaenoic acid (DHA) content during the whole growth phase, although it reached the peak in the linear phase with 3.5 mg g⁻¹. As for the sterols, two unique sterols with two hydroxyl groups, termed pavlovols, were observed. 4α,24-Dimethylcholestan-3β,4β-diol, one of the pavlovols, increased almost 2-fold from the late exponential phase (2.5 mg g⁻¹) to the stationary phase (4.3 mg g⁻¹). On the contrary, the contents of stigmasterol and sitosterol decreased with culture age, with the maximum content of 2.4 mg g⁻¹ and 3.1 mg g⁻¹, both obtained in the late exponential phase, respectively. The results indicate that growth phase control could be used as a methodology to optimize the total lipid, EPA, PUFA, and sterol contents with the potential for both aquaculture feeds and nutraceutical applications, especially for further research into unique pavlovols.