ABSTRACT
A field population of Ulva pseudocurvata Koeman et C. Hoek (hereafter termed Ulva) at Sylt Island (North Sea, Germany) exhibited biweekly peaks of gametophytic reproduction during the colder seasons and approximately weekly peaks during summer. The reproductive events lasted 1-5 d and were separated from each other by purely vegetative phases. Under constant conditions in the laboratory, a free-running rhythm was observed with reproductive peaks occurring approximately every 7 d. When artificial moonlight was provided every 4 weeks, fewer reproductive events occurred, and the reproductive rhythm became synchronized to the environmental artificial moonlight rhythm. In the laboratory, apical disks were entirely converted into reproductive tissue after 8 d cultivation, while almost all basal disks stayed vegetative, which prevented the entire loss of the vegetative thallus during reproductive events. Seasonal size reduction of the thallus occurred from late autumn onward and was determined to be controlled by a genuine photoperiodic response, since size reduction could be induced from May onward by experimental short-day (SD) treatment but was prevented in a long-day (LD) or night-break regime (NB). A daily fine-tuning occurred with gamete release early in the morning at the first sign of daylight, following an obligatory dark ("night") period of at least 1 h duration. No release took place if the overnight dark phase was replaced by continuous light. Blue, green, or red light all triggered gamete release after a dark phase at an irradiance of 0.1 µmol photons · m(-2) ·s(-1) , while 0.001 µmol photons · m(-2) · s(-1) was equivalent to a dark control.
ABSTRACT
Melatonin, the chief secretory product of the vertebrate pineal gland is also known to occur in numerous photoautotrophic organisms. The indoleamine is suspected to act as a transducer of photoperiodic information and/or to participate in antioxidative protection. In higher plants and other photoautotrophic organisms, contradictory results for melatonin content for samples from the same species show that further improvement of methods for reliable quantification is required. In the present study, melatonin was quantified in tomatoes, ginger and the marine green macroalga, Ulva lactuca, after extraction with three different extraction methods based on ether, acetone or perchloric acid. Melatonin was determined by enzyme-linked immunosorbent assay (ELISA) in high-performance liquid chromatography (HPLC)-purified extracts. The same HPLC system used for purification of extracts was used for parallel quantifications after derivatization of melatonin under alkaline conditions in the presence of hydrogen peroxide (HPLC-PD). Both quantification methods gave similar results with a high correlation [f(x) = 0.99x + 3.01; R(2) = 0.99]. In ginger, the melatonin concentration was below 5 pg/g (fresh weight, f.w.), whereas in tomatoes about 1200 pg/g (f.w.) were found, and in the green alga, U. lactuca, approximately 12 pg/g (f.w.). Taking into account the recovery rates for synthetic melatonin added prior to extraction, no substantial differences were observed in melatonin quantification between different extraction methods. The demonstrated methods based on HPLC purification and subsequent quantification by ELISA and HPLC-PD allow highly sensitive melatonin determinations in diverse photoautotrophic organisms with a low risk of overestimations by false-positive results.
