Dania Esposito

Ionizing radiation impacts photochemical quantum yield and oxygen evolution activity of Photosystem II in photosynthetic microorganisms.

Purpose: Long-term space exploration requires biological life support systems capable of coping with the deleterious space environment. The use of oxygenic photosynthetic microorganisms represents an intriguing topic in this context, mainly from the point of view of food and O2 production. The aim of the present study was to assess the effects of space ionizing radiation exposure on the photosynthetic activity of various microorganisms. Materials and methods: Ground-based irradiation experiments were performed using fast neutrons and gamma rays on microorganisms maintained at various light conditions. A stratospheric balloon and a European Space Agency (ESA) flight facility were used to deliver organisms to space at the altitude of 38 and 300 km, respectively. During the balloon flight, the fluorescence activity of the organisms was real-time monitored by means of a special biosensor. Results: The quantum yield of Photosystem II (PSII), measured directly in flight, varied among the microorganisms depending on the light conditions. Darkness and irradiation of cells at 120 and 180 μmol m−2 s−1 enhanced the radiation-induced inhibition of photosynthetic activity, while exposure to weaker light irradiance of 20 and 70 μmol m−2 s−1 protected the cells against damage. Cell permanence in space reduced the photosynthetic growth while the oxygen evolution capacity of the cells after the flight was enhanced. Conclusions: A potential role of PSII in capturing and utilizing ionizing radiation energy is postulated.

A new miniaturized multiarray biosensor system for fluorescence detection

A miniaturized biosensor-based optical instrument has been designed and fabricated for multiarray fluorescence measurements of several biomediators in series, with applications in environmental monitoring and agrofood analysis. It is a multicell system featuring two arrays of five static cells (1 × 1 × 2 cm 3 ) which are sealed to avoid contamination. Every cell is made up by two modular sections: the bottom compartment with optical LED light excitations and a photodiode detector for fluorescence emission capture, and the top biocompatible compartment where the biosample is deposited. The system (0.250 kg without batteries and case, 100 x 100 x 150 mm 3 internal case dimensions) is equipped with electronic control boards, a flash memory card for automatic data storage, and internal batteries, thus being portable and versatile. The instrument allows one to perform simultaneous and multiparametric analyses and offers a large applicability in biosensor technology. The first prototype has been implemented with genetically modified oxygenic photosynthetic algae that were employed in the instrument experimental testing by monitoring pesticide pollution in water. Pesticides modify the photosystem II (PSII) activity in terms of fluorescence quenching. The PSII complex features a natural nanostructure and can be considered a sophisticated molecular device. Results from measurements employing several PSII mutants and six different pesticides at increasing concentrations and incubation times are presented and discussed.

The effect of ionising radiation on photosynthetic oxygenic microorganisms for survival in space flight revealed by automatic photosystem II-based biosensors

Photosynthetic microorganisms are expected to be useful to maintain an oxygenic atmosphere and to provide biomass for astronauts in the International Space Station as well as in future long-term space flights. However, fluxes of complex ionizing radiation of various intensities and energies make space an extreme environment for the microorganisms, affecting their photosynthetic efficiency. To automatically monitor the photosynthetic Photosystem II (PSII) activity of microorganisms under space conditions an optical biosensor, which utilizes chlorophyll fluorescence as biological transduction system, was built; the PSII activity was monitored by the biosensor during balloon flights at stratospheric altitudes of about 40 km. The effect of space stress on quantum yield of PSII varied among the tested species depending on the growth light conditions at which they were exposed during the flights.

Photosystem II stress tolerance in the unicellular green algaChlamydomonas Reinhardtii under space conditions

Photosynthesis was established on the earth 3.5 billion years ago. Due to the absence of the ozone layer in the early atmosphere it was most likely adapted to the presence of ionizing radiation continuously emitted by solar and stellar flares. That complex radiation spectrum comprises protons, alpha particles, heavy charged particle-HZE, electrons, X-ray and neutrons. Such spectrum has a significant impact on biological systems which capture light energy for e.g. photosynthesis. Oxygenic photosynthesis of plants, algae and cyanobacteria initiates at the level of photosystem II (PSII), a multisubunit protein complex embedded in the thylakoid membrane inside chloroplasts. PSII uses sunlight to power the unique photo-induced oxidation of water to atmospheric oxygen which is indispensable for most life forms. It is an especially sensitive component if exposed to space radiation and thus an important target for research aimed at improving bioregenerative life-support systems. The unicellular green algae Chlamydomonas reinhardtii is a long standing model organism for photosynthesis research. It was exposed to ionizing radiation in the ESA facility Biopan located in the Foton capsule brought to space by the Russian Soyuzfor 15 days. The algae were tested in space under shielded conditions in the past, but they were never exposed to direct ionizing radiation such as in Biopan. Conditions for survival were identified. It was observed that the effect of space stress on the survival of the algae varied depending on the light conditions to which they were exposed during the flight. In some cases the flight experience caused a stimulation of the photosystem II oxygen evolution of the cells.

Biodevices for Space Research

This review focuses on the realisation of optical sensors able to monitor the effect of complex space radiation on biological components, based on the biosensor concept. A biosensor is a device that can reveal a biochemical variable using a biological component interfaced with a transducer. It issues an electric signal which is easy to process, depending on the analysed variable. Biosensors are useful to study the effect of stress conditions on living organisms. One of the goals of this research was to develop two types of biosensors able to monitor directly the response of oxygenic photosynthetic organisms to radiation present in space in view of their importance for future space colonization. In ground experiments and in balloon stratosphere flights, the photosynthetic process has been analysed at the level of photosystem II (PSII), the supramolecular pigment-protein complex in the chloroplast which catalyses the light-induced transfer of electrons from water to plastoquinone; PSII splits water into molecular oxygen, protons and electrons, thereby sustaining an aerobic atmosphere on Earth and providing the reducing equivalents necessary to fix carbon dioxide to organic molecules, creating biomass, food and fuel. The results indicated that presence of space radiation in the dark has a synergistic effect on photosystem II activity, suggesting that PSII D1 protein turnover may be involved in resistance to space stress. The resistance of the tested microorganisms to space stress seems to be related to their position on the evolutive scale of photosynthesis. The present studies allow to establish a regular and reliable correlation between measured physical characteristics of space radiation and biological radiation effect.

Comparison of Photosynthetic Organisms at Various Evolutionary Stages for Protein Biochips

Many chromophore molecules, such as bacteriochlorophylls, bacteriopheophytins and quinones, are arranged in Reaction Centers with a relevant distance and energy status such as to ensure unidirectional electron transfer. Therefore even a single Reaction Center is a sophisticated molecular device suitable for technological approaches.