Global Proteomics of the Extremophile Black Fungus <i>Cryomyces antarcticus</i>Using 2D-Electrophoresis

The microcolonial black fungus Cryomyces antarcticus is an extremophile organism growing on and in rock in the Antarctic desert. Ecological plasticity and stress tolerance make it a perfect model organism for astrobiology. 2D-gel electrophoresis and MALDI-TOF/TOF mass spectrometry were performed to explore the protein repertoire, which allows the fungus to survive in the harsh environment. Only a limited number of proteins could be identified by using sequence homologies in public databases. Due to the rather low identification rate by sequence homology, this study reveals that a major part of the proteome of C. antarcticus varies significantly from other fungal species.

Extraterrestrial Life in the Thermosphere: Plasmas, UAP, Pre-Life, Fourth State of Matter

“Plasmas” up to a kilometer in size and behaving similarly to multicellular organisms have been filmed on 10 separate NASA space shuttle missions, over 200 miles above Earth within the thermosphere. These self-illuminated “plasmas” are attracted to and may “feed on” electromagnetic radiation. They have different morphologies: 1) cone, 2) cloud, 3) donut, 4) spherical-cylindrical;and have been filmed flying towards and descending from the thermosphere into thunderstorms;congregating by the hundreds and interacting with satellites generating electromagnetic activity;approaching the Space Shuttles. Computerized analysis of flight path trajectories documents these plasmas travel at different velocities from different directions and change their angle of trajectory making 45°, 90°, and 180° shifts and follow each other. They’ve been filmed accelerating, slowing down, stopping, congregating, engaging in “hunter-predatory” behavior and intersecting plasmas leaving a plasma dust trail in their wake. Similar life-like behaviors have been demonstrated by plasmas created experimentally. “Plasmas” may have been photographed in the 1940s by WWII pilots (identified as “Foo fighters”);repeatedly observed and filmed by astronauts and military pilots and classified as Unidentified Aerial—Anomalous Phenomenon. Plasmas are not biological but may represent a form of pre-life that via the incorporation of elements common in space, could result in the synthesis of RNA. Plasmas constitute a fourth state of matter, are attracted to electromagnetic activity, and when observed in the lower atmosphere likely account for many of the UFO-UAP sightings over the centuries.

A Review beyond the borders: Proteomics of microclonial black fungi and black yeasts

Black microcolonial fungi and black yeasts are inhabitants of extreme environments like vulcanic, desert and polar regions, where they are exposed to enhanced temperature alterations and desiccation. They have developed, therefore, extraordinary biologic characteristics which are mainly based on the expression of proteins, however, these are rarely studied and known. The review article presented here is focused on the obstacles and solutions for the proteomic analyses of this very particular fungal species.

Progress of Research on Origins of Life in China

The development of Chinese space science and technology plays a great role in promoting the researches in the field of the origin of life.With the multidisciplinary cooperation,there are fruitful achievements in this research field obtained over the past two years.This report summarizes the major progress of the basic researches about the origin of life in China during 2018–2020.

A Novel Halotolerant Bacterium Isolated from El Golea Lake in Algeria and Antimicrobial Potential of this Strain

A halotolerant, gram positive, motile and rod, was isolated from the E1 Gol6a lake of Ghardaia, Alegria and screened for its antimicrobial potential. The strain showed an inhibitory effect to gram positive and gram negative bacteria against Cladosporium spp. and a slight inhibitory effect against C. albicans using chloroform as the extraction solvent and the nutrient broth as production medium. 16S rRNA sequencing showed that the isolate named LMB3981 is a new taxon in the family of Bacillaceae （with 96% similarity）. The strain is close to Filobacillus milosensis and Bacillus haloalkaliphilum with 95% similarity. The phenotypic study showed differences between LMB3981 and two strains that are near and have confirmed the results of 16S rRNA sequencing by specific metabolic properties of the strain.

Cyanobacteria from Extreme Deserts to Space

The development of space technology makes the exposure of organisms and molecules to the space environment possible by using the ESA Biopan and Expose facilities and NASA nanosatellites;the aim is to decipher the origin, evolution and distribution of life on Earth and in the Universe. The study of microbial communities thriving in lithic habitats in cold and hot deserts is gathering appreciation when dealing with the limits of life as we know it, the identification of biosignatures for searching life beyond Earth and the validation of the (litho)-Panspermia theory. Cyanobacteria of the genus Chroococcidiopsis dominate rock-dwelling communities in extreme deserts that are considered terrestrial analogues of Mars, like the Dry Valleys in Antarctica, the Atacama Desert in Chile or the Mojave Desert in California. The extraordinary tolerance of these cyanobacteria towards desiccation, ionizing and UV radiation makes them suitable experimental strains which have been already used in astrobiological experiments and already selected for future space missions. Evidence gained so far supports the use of desert cyanobacteria to develop life support systems and insitu resource utilization for the human space exploration and settlement on the Moon or Mars.

Unidentified Anomalous Phenomena, Extraterrestrial Life, Plasmoids, Shape Shifters, Replicons, Thunderstorms, Lightning, Hallucinations, Aircraft Disasters, Ocean Sightings

As documented by NASA space shuttle films and detailed in this report, self-illuminating, pulsating, plasma-like UAP/UFO (“plasmoids”) have multiple shapes and sizes, are attracted to electromagnetic activity, and travel at different velocities from different directions, making 90 to 180-degree turns, as well as colliding, intersecting and piercing other plasma;and have been filmed by U.S. Navy personnel and a U.S. Customs and Border Protection DHC-8 flying above and diving/sinking beneath the ocean;and by NASA following, circling, and hovering near the space shuttles, satellites, and the MIR International Space Station and congregating above and descending into thunderstorms and the lower atmosphere, which is the air corridor favored by commercial and military aircraft;and this may account for reports of UAPs following, harassing, chasing, and “toying with” aircraft. Plasmas also have explosive properties, negatively affect electronics and mental activity (possibly inducing hallucinations of “alien abductions”), pass through glass, plastic, metal, and enter the cockpits of airplanes and have been observed by astronauts inside spacecraft, the MIR and ISS. It is hypothesized that given their propensity to collide, plasmoids may be responsible for at least some unexplained, inexplicable aircraft disasters. Thunder-lightning-storms are the main drivers of Earth’s GEC and direct positive currents into the ionosphere, which attract plasmas. The troposphere also has a positive charge, and the ocean surface under white water and turbulent conditions develops a positive charge, and we hypothesize that this accounts for sightings of UAP in the lower atmosphere and soaring above and diving into the oceans, including, as reported here, shape-shifting UAP replicons that split into or generate additional shape-shifting UAPs as filmed by NASA and U.S. Customs. Plasmoids appear to purposefully interact and engage in complex behaviors, and it is suspected they are sentient and represent a fourth domain of life. Although plasmas in the lower atmosphere may be responsible for UAP sightings over the centuries, including those that appear to “battle” over cities or follow and harass military ships and planes, plasmoids cannot account for all UAPs, which may include extraterrestrial spacecraft from other worlds.

Efficient lignin biodegradation triggered by alkali-tolerant ligninolytic bacteria through improving lignin solubility in alkaline solution

Low lignin solubility in aqueous solution is one of the major bottlenecks for lignin biodegradation and bioconversion.Alkaline solution contributes to improving lignin solubility,whereas most microbes can not survive in alkaline conditions.Herein,lignin dissolution behaviors in different pH solutions were systematically investigated,which indicated that solution pH above 10.5 contributed to high solubility of alkali lignin.To match with alkaline lignin aqueous system,several alkali-tolerant ligninolytic bacteria were isolated,most of which are distinct to previously reported ones.Then,the ligninolytic capabilities of these isolates were assessed in different pH conditions by determining their assimilation on alkali lignin,lignin-derived monomers and dimers,their decolorization capabilities,and their lignin peroxidase activities.Thereafter,the underlying ligninolytic and alkali-tolerant mechanisms of Sutcliffiella sp.NC1,an alkalophilic bacterium,was analyzed on the basis of its genome information.The results not only provide valuable information for lignin biodegradation and lignin valorization,but also expand knowledge on alkali-tolerant bacteria.

Life in extreme environments: Approaches to study life-environment co-evolutionary strategies

The term ＂extreme environments＂ describes the conditions that deviate from what mesophilic cells can tolerate. These condi- tions are ＂extreme＂ in the eye of mankind, but they may be suitable or even essential living conditions for most microorgan- isms. Hypertherrnophilic microorganisms form a branch at the root of the phylogenetic tree, indicating that early life originated from extreme environments similar to that of modern deep-sea hydrothermal vents, which are characterized by high-tempera- ture and oxygen-limiting conditions. During the inevitable cooling and gradual oxidation process on Earth, microorganisms developed similar mechanisms of adaptation. By studying modem extremophiles, we may be able to decode the mysterious history of their genomic evolution and to reconstruct e~,rly life. Because life itself is a process of energy uptake to maintain a dissipative structure that is not in thermodynamic equilibrium, the energy metabolism of microorganisms determines the path- way of evolution, the structure of an ecosystem, and the physiology of cells. ＂Following energy＂ is an essential approach to understand the boundaries of life and to search for life beyond Earth.

Engineering bacteria for enhanced polyhydroxyalkanoates(PHA)biosynthesis

Polyhydroxyalkanoates(PHA)have been produced by some bacteria as bioplastics for many years.Yet their commercialization is still on the way.A few issues are related to the difficulty of PHA commercialization:namely,high cost and instabilities on molecular weights(Mw)and structures,thus instability on thermo-mechanical properties.The high cost is the result of complicated bioprocessing associated with sterilization,low conversion of carbon substrates to PHA products,and slow growth of microorganisms as well as difficulty of downstream separation.Future engineering on PHA producing microorganisms should be focused on contamination resistant bacteria especially extremophiles,developments of engineering approaches for the extremophiles,increase on carbon substrates to PHA conversion and controlling Mw of PHA.The concept proof studies could still be conducted on E.coli or Pseudomonas spp.that are easily used for molecular manipulations.In this review,we will use E.coli and halophiles as examples to show how to engineer bacteria for enhanced PHA biosynthesis and for increasing PHA competitiveness.

Solutions:how adaptive changes in cellular fluids enable marine life to cope with abiotic stressors

The seas confront organisms with a suite of abiotic stressors that pose challenges for physiological activity.Variations in temperature,hydrostatic pressure,and salinity have potential to disrupt structures,and functions of all molecular systems on which life depends.During evolution,sequences of nucleic acids and proteins are adaptively modified to“fit”these macromolecules for function under the particular abiotic conditions of the habitat.Complementing these macromolecular adaptations are alterations in compositions of solutions that bathe macromolecules and affect stabilities of their higher order structures.A primary result of these“micromolecular”adaptations is preservation of optimal balances between conformational rigidity and flexibility of macromolecules.Micromolecular adaptations involve several families of organic osmolytes,with varying effects on macromolecular stability.A given type of osmolyte generally has similar effects on DNA,RNA,proteins and membranes;thus,adaptive regulation of cellular osmolyte pools has a global effect on macromolecules.These effects are mediated largely through influences of osmolytes and macromolecules on water structure and activity.Acclimatory micromolecular responses are often critical in enabling organisms to cope with environmental changes during their lifetimes,for example,during vertical migration in the water column.A species’breadth of environmental tolerance may depend on how effectively it can vary the osmolyte composition of its cellular fluids in the face of stress.Micromolecular adaptations remain an under-appreciated aspect of evolution and acclimatization.Further study can lead to a better understanding of determinants of environmental tolerance ranges and to biotechnological advances in designing improved stabilizers for biological materials.

Intrasexual competition enhances reproductive isolation between locally adapted populations

During adaptation to different habitat types, both morphological and behavioral traits can undergo divergent selection. Males often fight for status in dominance hierarchies and rank positions predict reproductive success. Ecotypes with reduced fighting abilities should have low reproductive success when migrating into habitats that harbor ecotypes with superior fighting abilities. Livebearing fishes in the Poecilia mexicana-species complex inhabit not only regular freshwater environments, but also independently colonized sulfidic （H2S-containing） habitats in three river drainages. In the current study, we found fighting intensities in staged contests to be considerably lower in some but not all sulfidic surface ecotypes and the sulfidic cave ecotype compared with populations from non-sulfidic surface sites. This is perhaps due to selection imposed by H2S, which hampers oxygen uptake and transport, as well as cellular respiration. Furthermore, migrants from sulfidic habitats may lose fights even if they do not show overall reduced aggressiveness, as phys- iological performance is likely to be challenged in the non-sulfidic environment to which they are not adapted. To test this hypothesis, we simulated migration of H2S-adapted males into H2S-free waters, as well as H2S-adapted cave-dwelling males into sulfidic surface waters. We found that intruders established dominance less often than resident males, independent of whether or not they showed reduced aggressiveness overall. Our study shows that divergent evolution of male aggressive behavior may also contribute to the maintenance of genetic differentiation in this system and we call for more careful evaluation of male fighting abilities in studies on ecological speciation.

On the diversity of fungi from soda soils

The diversity of filamentous fungi that can grow at high ambient pH values(i.e.,8–11)remains largely understudied.Here we study 100 alkalitolerant and alkaliphilic isolates from the soils around the basin of soda lakes in Asia and Africa to assess the major evolutionary lineages and morphologies pertinent to the alkaliphilic trait in filamentous fungi.The Emericellopsis lineage(Hypocreales,Hypocreomycetidae),along with Plectosphaerellaceae(Hypocreomycetidae),Pleosporaceae(Dothideomycetes),Chaetomiaceae(Sordariomycetidae)families appeared to be overrepresented with strong alkalitolerants and effective alkaliphiles.In particular,Sodiomyces species(Plectosphaerellaceae),Acrostalagmus luteoalbus(Plectosphaerellaceae),Emericellopsis alkalina(Hypocreales),Thielavia sp.(Chaetomiaceae),and Alternaria sect.Soda(Pleosporaceae)grew best at high ambient pH.The pH tolerance of Chordomyces antarcticum,Acrostalagmus luteoalbus and some other species was largely affected by the presence of extra Na+in the growth medium.Moderate alkalitolerants included Scopulariopsis members(Microascales),Fusarium,Cladosporium,and many asexual acremonium-like species from Bionectriaceae.Weak alkalitolerants were represented by sporadic isolates of Penicillium,Purpureocillium lilacinum,and Alternaria alternata species,with the growth optimum at neutral or acidic pH.Weak alkalitolerants develop loose dry chains of spores easily dispersed by air.Their presence at low frequency with the growth optimum at neutral or acidic pH leads us to treat them as transient species in the alkaline soils,as those are also ubiquitous saprobes in normal soils.Our phylogenetic analyses show that the alkaliphilic trait in filamentous fungi has evolved several times.Several lineages harboring strong alkalitolerants derived from the known marine-borne fungi(Emericellopsis,Alternaria sect.Phragmosporae),or fall within the fungi associated with halophytic grasses(Pleosporaceae).Soda soils contain a diversity of fungi that range from weak alkalitolerant to alkaliphilic,which in few cases is associated with darkly pigmented mycelium and formation of microsclerotia.The alkaliphilic trait is spread throughout the Ascomycota,and usually juxtaposes with slime-covered polyphyletic acremonium-,verticillium-,gliocladium-types of asexual morphology,hyphae aggregating in chords,and enclosed fruit bodies.

Xerotolerant foliar endophytic fungi of Populus euphratica from the Tarim River basin,Central China are conspecific to endophytic ITS phylotypes of Populus tremula from temperate Europe

Fungal research on non-lichenized taxa in xeric and other unusual habitats for fungi is comparatively scarce but constantly reveal an enormous ecological and functional adaptiveness of these organisms to extremes.This holds true in the topmost forest canopies as well as in marine,arctic,antarctic or desert habitats.In the present study dried leaves of Populus euphratica from a continental desert area in Central China were investigated for endophytic fungi 6 weeks after sampling by cultivation of small leaf fragments.Thirty-five species,many of them well-known from extreme environments,were delimitated from as few as 58 isolates by a combined analysis of morphotypes,micromorphology and ITS sequences.Meta-analyses revealed pronounced conspecificity to endophytes from Populus tremula leaves,but significant differences to foliar endophytic assemblages in Pinus monticola and Fagus sylvatica.The deduced high similarity of endophytic communities on both Populus species compared with that on Pinus and Fagus suggests that many fungal phyllosphere species show a well-pronounced host preference,at least for their endophytic stages.In addition,it demonstrates that leaving leaves as a microhabitat might provide similar conditions such as constantly high relative humidity,regardless of the surrounding climatic conditions.The evolution of extremotolerance might be of particular importance for retaining competitiveness of foliar endophytes in deciduous host trees after loss of pathogenicity.

Dishwashers provide a selective extreme environment for human-opportunistic yeast-like fungi

Urban life has led to the creation of human-made environments that,from a microbiological perspective,provide extreme life conditions.Certain non-ubiquitous microorganisms such as thermophilic members of the black yeast genus Exophiala are enriched within these habitats for which no counterpart is known in nature.Dishwashers consistently accumulate a number of specific black,white and red yeasts on the rubber seals of doors and in stagnant water at the interior.Several of these yeasts are primarily known as agents of human opportunistic infections.In this review,the literature data are supported by a screening study involving 937 households in 15 cities in Turkey.Fungi were detected in 230 samples(24.5%).Using rDNA sequencing,the prevalent species were identified as Exophiala dermatitidis(n=116),Candida parapsilosis(n=44),E.phaeomuriformis(n=35),Magnusiomyces capitatus(n=22),Rhodotorula mucilaginosa(n=15),and C.lusitaniae(n=14).The possible role of dishwashers in transmitting disease is discussed.