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Rothschild, L. J.

Publications and source records attributed to Rothschild, L. J..

Biology as a Key Technological Foundation for Settlement Beyond Earth

Moving materials beyond Earth, whether spacecraft, living organisms, or both, is limited by mass constraints. Yet human survival requires an extensive infrastructure, from environmental regulation to life support. In practice this means habitats, food, oxygen, waste recycling, medicine and so on. Thus, there is a mismatch between what will be required in transit and at destination to fulfill dreams of human settlements and what can realistically moved there. Further, settlement off planet with current transportation systems requires the ability to operate independently of the Earth for prolonged periods of time, requiring long-term storage of supplies and the flexibilityto satisfy new needs.

resource utilization↗

Life in the Clouds of Venus? An Experimental Synthetic Biology Approach

The surface of Venus constitutes the most hellish and biologically inhospitable planetary surface in our solar system, boasting a pH of 0, blistering winds that can melt lead, and pressures of 60 atm. However, during the earlier years of the solar system, without the runaway greenhouse effect that has plagued the planet, Venus potentially housed oceans and perhaps even life. There is a possibility that microbes could have retreated into hospitable niches in the atmosphere, as suggested by Carl Sagan as early as 1967 [1]. For example, 50 km above the raging hell of the Venusian surface, exists a relatively temperate environment that might serve as reservoir for life. This astrobiology project seeks to explore life at the extremes and to theorize whether microbial communities could not only survive but also reproduce in the Venusian atmosphere. Specifically, we ask: are aerosols viable microbial environments? But before we can test for life in the clouds, we have to develop a proper reporter to visualize cell growth in situ. For this purpose, we aimed to develop cell-growth dependent reporters to serve as remote biosensors for cell growth. We developed two using the polA promoter, a DNA-replication dependent promoter, and nrd operon promoter, a cell-cycle dependent promoter. Using these cell-growth reporters, the next step is to aerosolize microbes expressing these reporters in a suspension chamber adapted from a Millikan Drop Apparatus to assay reproduction in an aerosolized environment. Better yet is to test the reproduction of microbes in a microgravity regime such as on ISS.Approach: We engineered two cell-cycle dependent genetic reporters. One was the polA promoter which codes for DNA Polymerase I, a gene active in DNA replication [2]. The other was the nrdP. The activation of ribonucleotide reductase reduces ribonucleotides into deoxyribonucleotides and is involved in the bacterial cell cycle [3]. This promoter began activation during the initiation of DNA replication and is cell-cycle dependent [4]. These promoters were fused to a GFP reporter, transformed into E. coli. The constructs were deposited in the iGEM registry as K847210: Escherichia coli DNA-replication dependent polA promoter K847211: Escherichia coli cell-division dependent nrd promoter Results: Our constructs displayed fluorescence when transformed into NEB-5alpha competent cells. While nrdP-E0840 displayed sufficient fluorescence as verified by fluorescent microscopy, the original polAP-E0840 construct (which uses mut3b GFP) exhibited low expression; while fluorescence was visible under the microscope, the signal was too weak for the camera to recognize. The polA promoter was therefore digest-ed with EcoRI and SpeI then ligated into plasmid pNCS containing a RBS, Clover, and a terminator. Clover is a highly engineered green fluorescent protein that exhibits extreme brightness [5] Fluoresence time course data demonsrated that the genes were induced in a cell cycle dependant manner [6]. Our assays via microscopy and the bulk assay shows that our promoters are functional as cell cycle reporters.Conclusions: The application of such tools are widespread and not limited to astrobiology; nrdP could be used to determine doubling times empirically and could possibly extrapolate DNA content from intensity of signals expressed by polAP. However, we are pri-marily interested in its use in astrobiology.

Escherichia↗

Radiation: microbial evolution, ecology, and relevance to mars missions

Ultraviolet (UV) radiation has been an important environmental parameter during the evolution of life on Earth, both in its role as a mutagen and as a selective agent. This was probably especially true during the time from 3.8 to 2.5 billion years ago, when atmospheric ozone levels were less than 1% of present levels. Early Mars may not have had an "ozone shield" either, and it never developed a significant one. Even though Mars is farther away from the Sun than the Earth, a substantial surficial UV flux is present on Mars today. But organisms respond to dose rate, and on Mars, like on Earth, organisms would be exposed to diurnal variations in UV flux. Here we present data on the effect of diurnal patterns of UV flux on microbial ecosystems in nature, with an emphasis on photosynthesis and DNA synthesis effects. These results indicate that diurnal patterns of metabolism occur in nature with a dip in photosynthesis and DNA synthesis in the afternoon, in part regulated by UV flux. Thus, diurnal patterns must be studied in order to understand the effect of UV radiation in nature. The results of this work are significant to the success of human missions to Mars for several reasons. For example, human missions must include photosynthetic organisms for food production and likely oxygen production. An evolutionary approach suggests which organisms might be best suited for high UV fluxes. The diurnal aspect of these studies is critical. Terraforming is a potential goal of Mars exploration, and it will require studies of the effect of Martian UV fluxes, including their diurnal changes, on terrestrial organisms. Such studies may suggest that diurnal changes in UV only require mitigation at some times of day or year.

Review, Tutorial↗

The influence of UV radiation on protistan evolution

Ultraviolet radiation has provided an evolutionary challenge to life on Earth. Recent increases in surficial ultraviolet B fluxes have focused attention on the role of UV radiation in protistan ecology, cancer, and DNA damage. Exploiting this new wealth of data, I examine the possibility that ultraviolet radiation may have played a significant role in the evolution of the first eukaryotes, that is, protists. Protists probably arose well before the formation of a significant ozone shield, and thus were probably subjected to substantial ultraviolet A, ultraviolet B, and ultraviolet C fluxes early in their evolution. Evolution consists of the generation of heritable variations and the subsequent selection of these variants. Ultraviolet radiation has played a role both as a mutagen and as a selective agent. In its role as a mutagen, it may have been crucial in the origin of sex and as a driver of molecular evolution. As a selective agent, its influence has been broad. Discussed in this paper are the influence of ultraviolet radiation on biogeography, photosynthesis, and desiccation resistance.

NASA Center ARC↗

The effects of UV radiation A and B on diurnal variation in photosynthesis in three taxonomically and ecologically diverse microbial mats

Photosynthetic primary production, the basis of most global food chains, is inhibited by UV radiation. Evaluating UV inhibition is therefore important for assessing the role of natural levels of UV radiation in regulating ecosystem behavior as well as the potential impact of stratospheric ozone depletion on global ecosystems. As both photosynthesis and UV fluxes are subject to diurnal variations, we examined the diurnal variability of the effect of UV radiation on photosynthesis in three diverse algal mats. In one of the mats (Cyanidium caldarium) a small mean decrease in primary productivity over the whole day occurred when both UVA and UVB were screened out. In two of the mats (Lyngbya aestuarii and Zygogonium sp.) we found a mean increase in the total primary productivity over the day when UVB alone was screened and a further increase when UVA and UVB were both screened out. Variations in the effects of UV radiation were found at different times of the day. This diurnal variability may be because even under the same solar radiation flux, there are different factors that may control photosynthetic rate, including nutritional status and other physiological processes in the cell. The results show the importance of assessing the complete diurnal productivity. For some of the time points the increase in the mean was still within the standard deviations in primary productivity, illustrating the difficulty in dissecting UV effects from other natural variations.

Ultraviolet Rays↗

A 'crytic' microbial mat: A new model ecosystem for extant life on Mars

If life were present on Mars today, it would face potentially lethal environmental conditions such as a lack of water, frigid temperatures, ultraviolet radiation, and soil oxidants. In addition, the Viking missions did not detect near-surface organic carbon available for assimilation. Autotrophic organisms that lived under a protective layer of sand or gravel would be able to circumvent the ultraviolet radiation and lack of fixed carbon. Two terrestrial photosynthetic near-surface microbial communities have been identified, one in the inter- and supertidal of Laguna Ojo de Liebere (Baja California Sur, Mexico) and one in the acidic gravel near several small geysers in Yellowstone National Park (Wyoming, U.S.A.). Both communities have been studied with respect to their ability to fix carbon under different conditions, including elevated levels of inorganic carbon. Although these sand communities have not been exposed to the entire suite of Martian environmental conditions simultaneously, such communities can provide a useful model ecosystem for a potential extant Martian biota.

Rothschild, L. J.↗

Elevated CO2: Impact on diurnal patterns of photosynthesis in natural microbial ecosystems

Algae, including blue-green algae (cyanobacteria), are the major source of fixed carbon in many aquatic ecosystems. Previous work has shown that photosynthetic carbon fixation is often enhanced in the presence of additional carbon dioxide (CO2). This study was undertaken to determine if this CO2 fertilization effect extended to microbial mats, and, if so, at what times during the day might the addition of CO2 affect carbon fixation. Four microbial mats from diverse environments were selected, including mats from a hypersaline pond (area 5, Exportadora de Sal, Mexico), the marine intertidal (Lyngbya, Laguna Ojo de Liebre, Mexico), an acidic hotspring (Cyanidium, Nymph Creek, Yellowstone National Park), and an acidic stream at ambient temperature (Zygogonium, Yellowstone National Park). Carbon fixation in the absence of additional CO2 essentially followed the rising and falling sunlight levels, except that during the middle of the day there was a short dip in carbon fixation rates. The addition of CO2 profoundly enhanced carbon fixation rates during the daylight hours, including during the midday dip. Therefore, it is unlikely that the midday dip was due to photoinhibition. Surprisingly, enhancement of carbon fixation was often greatest in the early morning or late afternoon, times when carbon fixation would be most likely to be light limited.

Rothschild, L. J.↗

Metabolic activity of microorganisms in evaporites

Crystalline salt is generally considered so hostile to most forms of life that it has been used for centuries as a preservative. Here, we present evidence that prokaryotes inhabiting a natural evaporite crust of halite and gypsum are metabolically active while inside the evaporite for at least 10 months. In situ measurements demonstrated that some of these "endoevaporitic" microorganisms (probably the cyanobacterium Synechococcus Nageli) fixed carbon and nitrogen. Denitrification was not observed. Our results quantified the slow microbial activity that can occur in salt crystals. Implications of this study include the possibility that microorganisms found in ancient evaporite deposits may have been part of an evaporite community.

NASA Discipline Exobiology↗

Paleobiomarkers and defining exobiology experiments for future Mars experiments

Mars is a cold, dry planet with an oxidizing surface bombarded by ultraviolet and ionizing radiation, making prospects for an extant Mars biota bleak. Yet, it is suggested that early Earth and early Mars were similar enought that life may have evoled on Mars. If life did evolve on Mars, what evidence for its existence might we find? What constitutes a Martian paleobiomarker, and how can we distinguish such a marker from abiotically produced substances? The topics studied to answer this question include carbon and nitrogen cycling, as well as the stability and relative abundance of their intermediates in microbially dominated ecosystems. The microbially dominated ecosystems studied are the crytoendolithic microbial community living within sand rocks, the endoevaporite microbial community living inside salt crystals, and the microbial communities living beneath perennially ice-covered lakes and hypersaline ponds. The nitrogen cycle of these communities ranges from simple, where only assimilation occurs, to the more complex, where a complete cycle occurs. The carbon cycle of these communities appears to be complete.

Mancinelli, R. L.↗

A model for diurnal patterns of carbon fixation in a Precambrian microbial mat based on a modern analog

Microbial mat communities are one of the first and most prevalent biological communities known from the Precambrian fossil record. These fossil mat communities are found as laminated sedimentary rock structures called stromatolites. Using a modern microbial mat as an analog for Precambrian stromatolites, a study of carbon fixation during a diurnal cycle under ambient conditions was undertaken. The rate of carbon fixation depends primarily on the availability of light (consistent with photosynthetic carbon fixation) and inorganic carbon, and not nitrogen or phosphorus. Atmospheric PCO2 is thought to have decreased from 10 bars at 4 Ga (10(9) years before present) to approximately 10(-4) bars today, implying a change in the availability of inorganic carbon for carbon fixation. Experimental manipulation of levels of inorganic carbon to levels that may have been available to Precambrian mat communities resulted in increased levels of carbon fixation during daylight hours. Combining these data with models of daylength during the Precambrian, models are derived for diurnal patterns of photosynthetic carbon fixation in a Precambrian microbial mat community. The models suggest that, even in the face of shorter daylengths during the Precambrian, total daily carbon fixation has been declining over geological time, with most of the decrease having occurred during the Precambrian.

Review↗

Stable carbon isotope fractionation in the search for life on early Mars

The utility of measurements of C-13/C-12 ratios in organic vs inorganic deposits for searching for signs of life on early Mars is considered. It is suggested that three assumptions are necessary. First, if there was life on Mars, it caused the fractionation of carbon isotopes in analogy with past biological activity on earth. Second, the fractionation would be detectable. Third, if a fractionation would be observed, there exist no abiotic explanations for the observed fractionation pattern.

Rothschild, L. J.↗