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At least 19 records

Advantages of Microgreen Carotenoid Composition for Space Travel

Microgreens are attractive space crop candidates for their ease of maintenance and nutrient density. These tender greens are particularly rich phytochemicals like carotenoids. To determine the effect of space conditions on carotenoid synthesis, this NASA funded research used Gilroy’s Life Sciences TOAST database to compare transcriptomic data of Arabidopsis thaliana grown on previous spaceflight missions. Of the eight genes studied that involve carotenoid biosynthesis, it was determined that most were upregulated.

microgreens↗

Carotenoid diagenesis in a marine sediment

The major carotenoids at three levels (3, 40, and 175 m below the sediment-water interface) in a core from a marine sediment (Cariaco Trench, off Venezuela) have been examined. Mass and electronic spectral data have provided evidence for the onset of a progressive reduction of carotenoids in the geological column. The time scale of the process appears to depend on the particular carotenoid. Reduction of up to two double bonds is observed for the diol, zeaxanthin, in the oldest sediment (about 340,000 years old) but no reduction is observed in the younger samples (about 5000 and 56,000 years old). The diketone, canthaxanthin, shows evidence of reduction of up to two double bonds in the 56,000-yr sample and up to five double bonds in the oldest sample. No reduction of beta-carotene was observed in any of the samples.

Watts, C. D.↗

Chlorophyll and carotenoid pigments of prochloron (prochlorophyta)

High-performance liquid chromatography (HPLC) with a gradient-elution technique was utilized to separate and quantify chlorophylls a and b as well as major carotenoid pigments present in freeze-dried preprations of prochloron-didemnid associations and in Prochloron cells separated from host colonies. Results confirm earlier spectrophotometric evidence for both chlorophylls a and b in this prokaryote. Chlorophyll a:b ratios range from 4.14 to 19.71; generally good agreement was found between ratios determined in isolated cell preprations and in symbiotic colonies (in hospite). These values are 1.5 to 5-fold higher than ratios determined in a variety of eukaryotic green plants. The carotenoids in Prochloron are quantitatively and qualitatively similar to those found in various freshwater and marine blue-green algae (cyanopbytes) from high-light environments. However, Prochloron differs from cyanophytes by the absence of myxoxanthophyll and related glycosidic carotenoids. It pigment characteristics are considered sufficiently different from those of cyanophytes to justify its assignment to a separate algal division.

Paerl, H. W.↗

Graviresponsiveness and abscisic-acid content of roots of carotenoid-deficient mutants of Zea mays L

The abscisic-acid (ABA) content of roots of the carotenoid-deficient w-3, vp-5, and vp-7 mutants of Z. mays was analyzed using gas chromatography-mass spectrometry with an analysis sensitivity of 6 ng ABA g-1 fresh weight (FW). Roots of normal seedlings of the same lines were characterized by the following amounts of ABA (as ng ABA g-1 FW, +/- standard deviation): w-3, 279 +/- 43; vp-5, 237 +/- 26; vp-7, 338 +/- 61. We did not detect any ABA in roots of any of the mutants. Thus, the lack of carotenoids in these mutants correlated positively with the apparent absence of ABA. Primary roots of normal and mutant seedlings were positively gravitropic, with no significant differences in the curvatures of roots of normal as compared with mutant seedlings. These results indicate that ABA 1) is synthesized in maize roots via the carotenoid pathway, and 2) is not necessary for positive gravitropism by primary roots of Z. mays.

NASA Discipline Plant Biology↗

Root graviresponsiveness and columella cell structure in carotenoid-deficient seedlings of Zea mays

Root graviresponsiveness in normal and carotenoid-deficient mutant seedlings of Zea mays was not significantly different. Columella cells in roots of mutant seedlings were characterized by fewer, smaller, and a reduced relative volume of plastids as compared to columella cells of normal seedlings. Plastids in columella cells of mutant seedlings possessed reduced amounts of starch. Although approximately 10 per cent of the columella cells in mutant seedlings lacked starch, their plastids were located at the bottom of the cell. These results suggest that (i) carotenoids are not necessary for root gravitropism, (ii) graviresponsiveness is not necessarily proportional to the size, number, or relative volume of plastids in columella cells, and (iii) sedimentation of plastids in columella cells may not result directly from their increased density due to starch content. Plastids in columella cells of normal and mutant seedlings were associated with bands of microtubule-like structures, suggesting that these structures may be involved in 'positioning' plastids in the cell.

NASA Discipline Number 40-10↗

Method of producing purified carotenoid compounds

A method of producing a carotenoid in solid form includes culturing a strain of Chlorophyta algae cells in a minimal inorganic medium and separating the algae comprising a solid form of carotenoid. In one embodiment f the invention, the strain of Chlorophyta algae cells includes a strain f Chlamydomonas algae cells.

Eggink, Laura↗

Non-invasive in vivo measurement of macular carotenoids

A non-invasive in vivo method for assessing macular carotenoids includes performing Optical Coherence Tomography (OCT) on a retina of a subject. A spatial representation of carotenoid levels in the macula based on data from the OCT of the retina can be generated.

Lambert, James L.↗

Root growth, secondary root formation and root gravitropism in carotenoid-deficient seedlings of Zea mays L

The effect of ABA on root growth, secondary-root formation and root gravitropism in seedlings of Zea mays was investigated by using Fluridone-treated seedlings and a viviparous mutant, both of which lack carotenoids and ABA. Primary roots of seedlings grown in the presence of Fluridone grew significantly slower than those of control (i.e. untreated) roots. Elongation of Fluridone-treated roots was inhibited significantly by the exogenous application of 1 mM ABA. Exogenous application of 1 micromole and 1 nmole ABA had either no effect or only a slight stimulatory effect on root elongation, depending on the method of application. The absence of ABA in Fluridone-treated plants was not an important factor in secondary-root formation in seedlings less than 9-10 d old. However, ABA may suppress secondary-root formation in older seedlings, since 11-d-old control seedlings had significantly fewer secondary roots than Fluridone-treated seedlings. Roots of Fluridone-treated and control seedlings were graviresponsive. Similar data were obtained for vp-9 mutants of Z. mays, which are phenotypically identical to Fluridone-treated seedlings. These results indicate that ABA is necessary for neither secondary-root formation nor for positive gravitropism by primary roots.

NASA Discipline Plant Biology↗

Protein and carotenoid synthesis and turnover in gravistimulated root caps

In certain cultivars of corn gravitropic bending occurs only after the root cap, the site of gravity perception, is exposed to light. Light appears to trigger or to remove some block in the gravity translation process. Using light sensitive cultivars of corn, it was shown that light affects various processes in the cap. The roles of these light-induced processes in gravitropic bending in roots were studied.

Feldman, L. J.↗

Microbial Pigments and Their Degradation Products as Biosignatures

Carotenoids are a class of vibrant biological pigments that have a characteristic chemical structure centered around a polyene core (Lu et al. 2018). Carotenoids and their derivatives are candidate biosignatures because they can persist in the terrestrial geologic record for up to 1.73 billion years (Vinnichenko et al. 2020), have specific structures that are likely the result of complex pathways, mediate the survival of many microorganisms in Mars and Ocean Worlds analog environments, and are detectable with multiple techniques, including Raman spectroscopy. In this project, we aim to investigate the detectability of carotenoid pigments with different spectroscopic methods to inform future instrument selection. We compare the spectra of five unaltered carotenoids, two model compounds, and carotenoid-forming archaeon with visible and deep UV Raman spectroscopy and UVVis absorption spectrophotometry. We then use one model pigment, beta-carotene, to evaluate the likelihood that unique spectral properties of carotenoids, or their refractory byproducts, would be preserved and detectable on a remote planetary surface by exposing it to simulated conditions for Mars. Sample Acquisition. Pigments betacarotene, lutein, zeaxanthin, astaxanthin, and lycopene were purchased from Sigma Aldrich. Halobacterium salinarum NRC-1 was acquired from Carlina Biological and grown in Halobacterium media. Mineral salts including sodium sulfate, sodium carbonate, and halite were used to form matrices in which the beta-carotene was embedded before exposure. Pigment-mineral mixes were at a 1:10 ratio in water. Analytical Techniques. Deep UV Raman data were collected on a custom laboratory mapping spectrometer called MOBIUS (Mineral and Organic Based Investigations using Ultraviolet Spectroscopy), which is an analog to the SHERLOC instrument on the Mars 2020 Perseverance rover (Bhartia et al. 2021). It features a 248.56 nm NeCu pulsed laser, liquid nitrogen-cooled detector, and tunable optical setup. Visible Raman data were collected using a Horiba Jobin Yvon LabRam HR spectrometer with a frequencydoubled Nd:YAG laser (532 nm) and a HeNe laser (633 nm). A VWR 6300 PC UV/Visible Spectrophotometer was used to collect absorption data for carotenoid solutions, model compounds, and solvents in UVpermissible capped cuvettes. Data were collected from 190-1100 nm at 1 nm increments. All spectral data were analyzed using Igor Pro 9 (Wavemetrics). Irradiation. We used a vacuum chamber equipped with a cryostat and a flood electron gun to simulate Martian surface temperatures, low pressures, and ionizing radiation (10keV, 10μA for 6h at 200K for our initial tests). The samples were prepared by drying the pigment-mineral mix onto polished metal tabs, then mounted on the cryostat for processing. Samples were then analyzed directly on the tabs after exposure. Results: In comparing the visible and deep UV Raman spectra of unaltered pigments, we found that they differed drastically. Carotenoids are often studied with visible Raman and typically have peaks at 1525 cm-1 and 1157 cm-1, due to the stretching of the C=C and C-C bonds in the polyene structure. However, in deep UV, the strongest feature is at ~1630 cm-1 and is broad, possibly indicating that multiple peaks are forming this feature. This stark difference is likely due to different preresonant enhancement effects. The UV-Vis results show that there is an absorption band in the deep UV <300 nm, which supports the hypothesis that the 248.6 nm excitation is interrogating another aspect of carotenoids than visible Raman. Our preliminary exposure tests indicated that pigments – even without minerals present - were largely unaltered in the applied conditions, with only a slight broadening in the primary polyene peaks apparent in the visible Raman data. Figure 1. A) Visible vs. deep UV Raman spectra of unaltered beta carotene. B) Schematic of exposure. Conclusions: Our results to date indicate that deep UV and visible Raman spectroscopy, both techniques with planetary mission heritage from Mars 2020 (Wiens et al. 2021, Bhartia et al. 2021), may be used in a complementary manner to observe carotenoids. In addition, we find that beta-carotene is largely resistant to our current exposure conditions, though there may be some amount of amorphization of the material which could cause the broadening of the peaks at 1525 and 1157 cm-1. As a next step, we aim to increase the dosage and duration of exposure to observe degradation of the parent pigment, possibly add UV as a factor via an Ar mini-arc UV lamp and use GC-MS to characterize possible degradation products.

pigments↗

Engineered Yeast to Test Risks for Human Exploration of the Lunar Surface

Jessica W. Chau, Natalie N. Ball, Aditya Hindupur, Sandra T. Vu, Jennifer Gil Acevedo, Lauren C. Liddell, Chinmayee Govinda Raj, Gentry, Sergio R. Santa Maria, A. Mark Settles Crewed exploration of the Moon carries risks of long duration exposure to reduced gravity and to deep space radiation. The Lunar Explorer Instrument for space biology Applications (LEIA) investigates the effects of increased radiation and reduced gravity on yeast viability and growth in a Commercial Lunar Payload Services (CLPS) surface mission to the south polar region. LEIA conducts yeast genetics experiments to quantify growth, metabolism, and synthetic biology-enabled production of human nutrients, while taking real time measurements of biologically relevant radiation exposure on the lunar surface. We have engineered beta-carotene producing yeast strains to test the importance of selected DNA damage repair and reactive oxygen species (ROS) defense pathways in mitigating cellular damage from lunar surface radiation. Carotenoids are important dietary antioxidants, and beta-carotene is pro-vitamin A, which is needed for vision and immune function. Carotenoids are sensitive to ROS produced by ionizing radiation and NASA is testing on-demand production of carotenoids from yeast in the BioNutrients space flight experiments. In LEIA, we test the effects of deep space on carotenoid yield in engineered yeast strains. The LEIA team uses CRISPR-Cas9 to engineer yeast to express carotenoids as well as to generate loss-of-function mutations. We are generating mutations in the RAD51 DNA damage repair locus and three genes that function to reduce oxidative damage to the cell: SOD1, SOD2, and TSA1. These strains are tested for carotenoid production using microfluidics and LED spectroscopy to allow remote sensing of cellular growth and carotenoid levels. Keywords: synthetic biology, oxidative stress tolerance, biosensors, space radiation, beyond low Earth orbit, lunar surface, CRISPR/Cas9, gene editing, desiccation, carotenoids.

synthetic biology↗

Yeast strain development to test in-space bioproduction in the Lunar Explorer Instrument for space biology Applications (LEIA) mission

The Lunar Explorer Instrument for space biology Applications (LEIA) is investigating the effects of lunar radiation and gravity on yeast viability, growth, and metabolism. LEIA is part of the CP-22 Commercial Lunar Payload Services (CLPS) surface mission to the south polar region of the Moon. The biological payload will test genetic factors that are likely to influence the tolerance of yeast for deep space and lunar surface radiation. LEIA is also investigating in-space production of β-carotene in bioengineered yeast. This carotenoid is both an antioxidant and pro-vitamin A- an essential human micronutrient. We report progress on engineering carotenoid-expressing strains to test the impacts of strain background and specific genetic variants on growth and production of β-carotene. To test for enhanced sensitivity to the LEIA mission environment, we generated gene knockouts for the RAD51 DNA damage repair locus as well as the SOD1, SOD2, and TSA1 reactive oxygen species (ROS) defense enzymes. We are also generating strains expected to increase tolerance to abiotic stressors and ROS. To be included in the biology payload, each strain needs to satisfy a series of requirements to be compatible with the mission concept of operations. The LEIA mission will conduct experiments using an autonomous light emitting diode optical detection system and microfluidics incubator to quantify growth, metabolism, and carotenoid production. Strains must produce sufficient carotenoids for bioproduction to be detectable with this optical system. Cells will be loaded into microfluidics cards, desiccated, and stored for 8-12 months prior to the initiation of lunar surface operations. The CLPS lander will operate for one lunar day, and strains will need to grow to stationary phase within 96 hours to ensure that telemetry of LEIA data to Earth can be completed. Genetic variants also need to display expected phenotypes within these optical detection, storage, and growth cycle constraints.

Yeast Engineering↗

Lipophilic pigments from cyanobacterial (blue-green algal) and diatom mats in Hamelin Pool, Shark Bay, Western Australia

Lipophilic pigments were examined in microbial mat communities dominated by cyanobacteria in the intertidal zone and by diatoms in the subtidal and sublittoral zones of Hamelin Pool, Shark Bay, Western Australia. These microbial mats have evolutionary significance because of their similarity to lithfied stromatolites from the Proterozoic and Early Paleozoic eras. Fucoxanthin, diatoxanthin, diadinoxanthin, beta-carotene, and chlorophylls a and c characterized the diatom mats, whereas cyanobacterial mats contained myxoxanthophyll, zeaxanthin, echinenone, beta-carotene, chlorophyll a and, in some cases, sheath pigment. The presence of bacteriochlorophyll a within the mats suggest a close association of photosynthetic bacteria with diatoms and cyanobacteria. The high carotenoids : chlorophyll a ratios (0.84-2.44 wt/wt) in the diatom mats suggest that carotenoids served a photoprotective function in this high light environment. By contrast, cyanobacterial sheath pigment may have largely supplanted the photoprotective role of carotenoids in the intertidal mats.

NASA Discipline Exobiology↗

Lipophilic pigments from the benthos of a perennially ice-covered Antarctic lake

The benthos of a perennially ice-covered Antarctic lake, Lake Hoare, contained three distinct 'signatures' of lipophilic pigments. Cyanobacterial mats found in the moat at the periphery of the lake were dominated by the carotenoid myxoxanthophyll; carotenoids: chlorophyll a ratios in this high light environment ranged from 3 to 6.8. Chlorophyll c and fucoxanthin, pigments typical of golden-brown algae, were found at 10 to 20 m depths where the benthos is aerobic. Anaerobic benthic sediments at 20 to 30 m depths were characterized by a third pigment signature dominated by a carotenoid, tentatively identified as alloxanthin from planktonic cryptomonads, and by phaeophytin b from senescent green algae. Pigments were not found associated with alternating organic and sediment layers. As microzooplankton grazers are absent from this closed system and transformation rates are reduced at low temperatures, the benthos beneath the lake ice appears to contain a record of past phytoplankton blooms undergoing decay.

NASA Discipline Number 52-30↗