EXPERIMENT “BIOPHOBOS/ANABYOSIS” IN PROJECT “PHOBOS-GRUNT”

EXPERIMENT “BIOPHOBOS/ANABYOSIS” IN PROJECT “PHOBOS-GRUNT”

О.I.Orlov1

V.R.Alekseev2

Е.А.Vorobyeva3 V.F.Galchenko4

Е.А.Deshevaya1 А.V.Zakharov5 М.А.Levinskikh1

О.V.Muravenko6

N.D.Novikova1

S.V.Poddubko1

N.А.Polykarpov1

V.N.Sychev1

1 - SSC RF – Institute of Biomedical Problems RAS, Moscow2 - Zoological Institute RAS, St.Petersburg 3 - Soil Science Faculty, Moscow State University, Moscow4 - Institute of Microbiology RAS, Moscow5 - Space Research Institute RAS, Moscow6 - Institute of Molecular Biology RAS, Moscow

LONG-TERM ASTROBIOLOGICAL RESEARCH PROGRAM

Пути достижения поставленной цели

Long-term program of applied science research and planned experiments onboard RS ISS

Experiments : Biorisk-MSV; Biorisk-MSN; Expose-R; Aquarium; Plants-

Seeds

Project “PHOBOS-GRUNT”Experiment

“Biophobos/Anabiosis”

MARS-500 Program

Satellite experiments

Elucidation of viability of resting forms of

organisms of different taxonomical groups

under conditions of spaceflight and continuous

interplanetary flights for solving problems of

planetary quarantine, astrobiology and bio-

medical problems of manned cosmonautics.

AIMS OF THE ASTROBIOLOGICAL RESEARCH

INSIDE: RASTENIYA-2 AQUARIUM BRADOS BIORISK - KM BIORISC - MSV OUTSIDE: BIORISK – MSNEXPOSE-R

To expose dormant forms of various organisms on the external surface of the ISS, which include not only bacterial and fungal spores but also seeds and dormant embryos of lower crustaceans, as well as multi-cellular organism in the cryptobiotic state.

THE STUDY OF DORMANT FORMS OF VARIOUS ORGANISMS UNDER SPACE FLIGHT CONDITIONS

Cooperation with:Moscow State University, Moscow, Russia;Zoological Institute of RAS, St. Petersburg, Russia;Institute of microbiology of RAS, Moscow, Russia;St. Petersburg St. State University, St. Petersburg, Russia;Space Dynamic Laboratory, Utah State University, Utah, USA;Research Institute for Bioresources, Okayama University, Okayama, Japan;National Institute of Agrobiological Sciences, Ibaraki, Japan.

HIGHER PLANTSAir-dry seeds of mustard Brassica rapa, Improved Basic lineAir-dry seeds of mustard Brassica rapa, High Anthocyanin lineAir-dry seeds of mustard Brassica rapa, Anthocyaninless lineAir-dry seeds of mustard Brassica rapa, Rosette lineAir-dry seeds of mustard Brassica rapa, AstroPlants lineAir-dry seeds of mustard Brassica rapa, Somatic Variegation lineAir-dry seeds of mustard Brassica juncea, RGAir-dry seeds of Arabidopsis thaliana, K-310 lineAir-dry seeds of Arabidopsis thaliana, Columbia lineAir-dry seeds of weed Nicandra physaloidesAir-dry seeds of tomatoes Lycopersicum esculentium, Micro-Tom var.Air-dry seeds of radishes Raphanus sativus, Cherry Bomb var.Air-dry seeds of barley Hordeum vulgare, Haruna Nijo cvAir-dry seeds of barley Hordeum vulgare, K-305 cvAir-dry seeds of rice Oryza sativa, Rsp-5Air-dry seeds of rice Oryza sativa, Rsp-7VERTEBRATESDry eggs of fish Nothobranchius guenteriINSECTSCryptobiotic larvae of chyronamide Polypedium vanderplankiiLOWER CRUSTACEANSDormant eggs of Daphnia magnaDormant eggs of Daphnia pullicariaDormant eggs of Artemia salinaDormant eggs of Streptocephalus torvicornisDormant eggs of Eucypris ornateMOLDSSpores of Aspergillus sedowii # 9-6Spores of Aspergillus versicolor # 4-3-4Spores of Penicillium aurantiogresium # 9-9Spores of Penicillium expansum # 4-3-3BACTERIASpores of Bacillus subtilis # 20Spores of Bacillus licheniformis # 24Spores of Bacillus subtilis # 25Spores of Bacillus subtilis # 2335\105

EXPERIMENT “BIORISK-MSN-2”

Duration of the exposure: The 1-st container – 13 monthsThe 2-nd container - 18 months The 3-rd container – 31 months

EXPERIMENT “BIORISK-MSN” ONBOARD OF RUSSIAN SEGMENT OF THE INTERNATIONAL SPACE STATION

Reactivation of “space” and control resting eggs of Artemia salina

Growth of Bacillus subtilis # 24 strain after 13-months exposure on the outer side of ISS

Larvae of chironomid Polypedilum vanderplanki revived after 13, 18 and 31 months of exposition on the outer side of RS of ISS

“Biorisk-MSN” hardware and its location on “Pirs” Docking Compartment

Germinated “space” seeds of barley Hordeum vulgare cv. Haruna Nijo and plants of radish Raphanus sativus strain «Cherry Bomb», grown from “space” seeds

BACTERIAGroup 18, subgroup Bacillaceae, genera Bacillus, species:Bacillus subtilis-2335/105 ;Bacillus subtilis-20 ;Bacillus pumilus-25 ;Bacillus licheniformis-24.Gram-positive rods, 0,5-2,5 x 1,2-10 mkm, spore - 6 mkmFUNGITaxon - Mitosporic fungi, genera Aspergillus, Penicillium, Geomyces species:Aspergillus sydowi - 9-6; Geomyces pannorum – 2241;Aspergillus versicolor - 4-3-4;Penicillium aurantiogresium – 9-9;Penicillium expansum – 4-3-3.Spore 3-10 mkmANIMALSDried encysted embryos (diameter of a single egg is 200 micron) Daphnia magna

belongs to family Daphniidae, order Daphniiformes, class Crustacea.Dried encysted embryos (diameter of a single egg is 160 micron) Streptocephalus

torvicornis belongs to family Streptocephalidae, order Anostraca, class Crustacea.

Dried encysted embryos (diameter of a single egg is 180-200 micron) Artemia salina belongs to family Artemiidae, order Anostraca, class Branchiopoda, subphylum: Crustacea.

Dried encysted embryos (diameter of a single egg is 180-200 micron) Eucypris species belongs to family Cyprididae, order Ostracoda, class Crustacea.

Cryptobiotic dried larvae (length 5 mm 0,8 mm) of Polypedilum vanderplanki belongs to family Chironomidae, order Diptera, class Insecta.

PLANTS Seeds of Lycopersicum esculentum (Tomato Micro-Tom) Seeds of Raphanus sativus (Radish Cherry Bomb)Seeds of Arabidopsis thaliana line WT–2.

EXPERIMENT “EXPOSE-R” (Start March 10, 2009)

At present Russian scientists participate ESA experiment EXPOSE-R onboard ISS. In contrast to Biorsik-MSN experiment, a part of biological samples in EXPOSE-R is being exposed to solar UV.

EXPERIMENT “EXPOSE-R” (Start March 10, 2009)

The biological objects in “EXPOSE-R” experiment are placed in plastic film bags 7x7 mm. The bags are fixed in the cells of special sets assembled in trays. EXPOSE-R hardware contains three trays composed by 4 sets of the cells with biological samples. The biological samples arranged in three levels, and those of the top level are exposed to solar UV.

For realization of biological research in “Phobos-Grunt” project, a Workgroup chaired by academician RAS O.I. Orlov was organized by decision of the section “Space biology and physiology” of RAS Council on March 19, 2007. The workgroup consists of representatives of GSC IMBP RAS, ISS RAS, ZIN RAS, MSU, IMB RAS, FSCP “SAO after S.A. Lavochkin”. The workgroup has defined the main aims of biological experiments (Experiment “BioPhobos/Anabiosis”) in “Phobos-Grunt” project:

• investigation of survival of resting stages of bacteria, fungi, animals and plants under conditions of interplanetary flights for estimation of damaging effect of factors of space environment on cryptobiotic stages of organisms and further development of technology of long-term preservation of organisms in conditions of biological resting;

• investigation of biological, biochemical and molecular-genetic changes in organisms after long-term spaceflight;

EXPERIMENT “BIOPHOBOS/ANABYOSIS”

• estimation of risk related to possibility of transformation of microorganisms inhabiting the space object after their exposure to outer space with further return inside of the space object.

• comparison of survival under conditions of the spaceflight of resting stages with different level of survivability, including that of artificially induced be mean of cultivation with biologically active substances, for developing new biotechnological methods of investigations of resting stages for continuous space transportation.

• comparison of survival of resting stages of organisms under space flight conditions (BioPhobos/Anabiosis) and an orbital flight in control experiment on low Earth orbit on the outer side of RS of ISS (experiment “Biorisk-MSN”).



Род, вид Штамм

Leptolyngbia sp. Dp 6606

Methanosarcina lacustris sp. nov. ZS (= DSM13486T), VKM B-2268

Methylosinus trichosporium № 34

Methylosinus sporium № 29

Methylocystis minimus № 28

Methylobacter chroococcum № 72

Deinococcus radiodurans № NP902

Artrobacter oxydens № Prm1

Pseudomonas aurefaciens № В1393

Bacillus subtilis # 20

Bacillus licheniformis #24

Bacillus pumilus # 25

Bacillus subtilis # 2335\105

Ferroplasma acidiphilum gen. nov., sp. nov.

Acinetobacter sp. К30А

Rhodococcus sp. K-25

Sphingomonas sp. К42

Acetobacterium tundrae sp. nov. Z-4493 (=DSM1 9173T)

Micrococcus luteus 6/90-111

Aquaspirillum sp. 6/90-111/С505

Rudobacter spheroides 323

Streptomyces sp. 315

Spores of the following prokaryotes (bacteria) are included in the list of experiment “BioPhobos/ Anabiosis”


Spores of the following eukaryotes (fungi) are included in the list of experiment “BioPhobos/ Anabiosis”

Род, вид Штамм

Ulocladium botrytis 16-12

Cladosporium cladosporioides 2-3

Aspergillus sydowii 9-6

Aspergillus versicolor 4-3-4

Penicillium aurantiogresium 9-9

Penicillium expansum 4-3-3

Aspergillus sydowii ВКМ

Penicillium aurantiogresium ВКМ

Aspergillus versicolor ВКМ

Penicillium expansum ВКМ

Aureobasidium pullulans Viala&Boyer КБП-3847

Ulocladium botrytis Preuss M-54 ВКМ F-4032D

Saccharomyces cerevisiae Strain W303. ATCC 200060


Resting forms of the following higher eukaryotes are included in the list of experiment “BioPhobos/Anabiosis”

Род, вид Порода

Artemia salina Видовая форма

Streptocephalus torvicornis Видовая форма

Eucypris sp. Видовая форма

Daphnia magna Видовая форма

Hemidiaptomus ingens Видовая форма

Polypedilum vanderplankii Видовая форма

Animals

Род, вид Сорт

Arabidopsis thaliana Видовая форма

Zingeria beibersteiniana Видовая форма

Mesembryanthemum crystallinum Видовая форма

Nicandra physaloides Видовая форма

Plants

Capsule with Phobos’ grunt and biological samples to be returned to the Earth

2 “Phobos-capsule” with 122 (1010 mm) packs with different biological objects

For placing of the biological objects in two titanium “Phobos-capsules” with the size of 11 35 mm, technology similar to that of “EXPOSE-R” experiment will be used.

Thank you for your attention

Documents

EXPERIMENT “BIOPHOBOS/ANABYOSIS” IN PROJECT “PHOBOS-GRUNT”