Team:Tokyo Tech/Iron-oxidizing bacteria

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==Iron-oxidizing bacteria==
 
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===Role in the story===
 
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Why do we need to use Iron-oxidizing bacteria for terraforming of Mars?
 
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Heterotrophic bacteria cannot live in native Martian environment because there are no organic matters on Mars, however autotrophic bacteria can live without organic matters.
 
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Iron-oxidizing bacteria is autotrophic bacteria.
 
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Iron-oxidizing bacteria gets energy by oxidizing Fe(Ⅱ).
 
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To grow on Mars, iron-oxidizing bacteria require only energy injection of us because of composition of the Martian surface, crust and atmosphere. (リンク)
 
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We propose culturing iron-oxidizing bacteria on Mars and producing organic matters.
 
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Accumulation of organic matters provides habitable environment for heterotrophic bacteria (for instance, E.coli).
 
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===About iron-oxidizing bacteria===
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==Achievement==
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Iron-oxidizing bacterium is a kind of autotrophic bacteria.
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Iron-oxidizing bacteria reduce NAD+ to NADH by transfer of hydride.
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In laboratory, iron-oxidizing bacteria are cultured in 9K medium (リンク) which contains Fe(Ⅱ) and no organic matters.
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===Experimental result===
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*We cultured iron-oxidizing bacteria (''Acidithiobacillus ferrooidans'').
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We cultured Acidithiobacillus ferrooxidans (iron-oxidizing bacteria).
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The medium for A.ferrooxidans was blue before cultivation owing to Fe(Ⅱ).
+
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The presence of A.ferrooxidans changed the medium into brown one after culturing for two days.
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This change was observed owing to the production of Fe(OH)3.
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We confirmed the growth by using microscope of 200 magnifications.
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 +
[[Image:Tokyo_tech Iron bacteria.jpg|300px|right|change of the color of 9K medium]]
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==Introduction ~Why do we need to use Iron-oxidizing bacteria for terraforming of Mars?~ ==
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===Protocols===
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Heterotrophic bacteria cannot live in native Martian environment because there are no organic matters on the Martian surface, however autotrophic bacteria can live without organic matters.<br\>
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====Preparation of 9K medium (for Acidithiobacillus ferrooxidans)====
+
Iron-oxidizing bacteria is a kind of autotrophic bacteria.<br\>
-
*Prepare following A&B solution.
+
Iron-oxidizing bacteria get energy by oxidizing Fe(Ⅱ) and grow with CO<sub>2</sub>.
 +
To grow on Mars, iron-oxidizing bacteria require only energy injection by human beings because of composition of the Martian surface, crust and atmosphere. <br\>
 +
Martian surface and crust contains carbonate(Bandfield,2003 ''et al.'') and FeO(Fe(Ⅱ)). In addition, Martian atmosphere contains much CO<sub>2</sub>.(For detail, see [[Team:Tokyo_Tech/Project#The Most Earth-Like Planet “Mars”|here]]) <br\>
 +
We propose culturing iron-oxidizing bacteria on Mars to produce organic matters.<br\>
 +
Accumulation of organic matters provides habitable environment for heterotrophic bacteria (for instance, ''E.coli'').<br\>
 +
We plan to culture iron-oxidizing bacteria on the Martian surface in early stage of terraforming to accumulate enough organic matters for heterotrophic bacteria to live.
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*Sterilize A solution by autoclaving.
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==Material and method==
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*Sterilize B solution by using filter.
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*Mix solution A and B before use.
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*A solution can be stored for 2 weeks.
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====Culturing method====
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===Iron-oxidizing bacteria===
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Culture in 9K medium at 30℃ with vigorous shaking.
+
Iron-oxidizing bacterium is a kind of autotrophic bacteria.<br\>
-
Use the microscope when confirm the growth.
+
Iron-oxidizing bacteria reduce NAD<sup>+</sup> to NADH by oxidizing Fe(Ⅱ).<br\>
 +
Iron-oxidizing bacteria grow with dissolved CO<sub>2</sub> as carbon source, not organic matters.<br\>
 +
In laboratory, iron-oxidizing bacteria are cultured in 9K medium which contains Fe(Ⅱ) and no organic matters.
 +
 
 +
===Preparation of 9K medium (for iron-oxidizing bacteria)===
 +
We Prepared following A and B solution.
 +
{|
 +
|-
 +
|[[Image:Tokyo_Tech_A_Solution.png|200px|thumb|A Solution]]
 +
|[[Image:Tokyo_Tech_B_Solution.png|200px|thumb|B Solution]]
 +
|}
 +
We sterilized A solution by autoclaving.
 +
We sterilized B solution by using filter.<br\>
 +
We Mixed A and B solution before use.
 +
 
 +
===Cultivation of ''Acidithiobacillus ferrooxidans''===
 +
We cultured in 9K medium at 30℃ with vigorous shaking.
 +
We used the microscope when confirmed the growth.
 +
 
 +
==Result==
 +
===Cultivation of ''A.ferrooxidans''===
 +
[[Image:Tokyo_tech Iron bacteria.jpg|300px|right|change of the color of 9K medium]]
 +
We succeeded in culturing ''Acidithiobacillus ferrooxidans'' (iron-oxidizing bacteria).<br\>
 +
The medium for ''A.ferrooxidans'' was blue before cultivation owing to Fe(Ⅱ).<br\>
 +
The presence of ''A.ferrooxidans'' changed the medium into brown one after culturing for two days.<br\>
 +
We confirmed the growth by using microscope of 200 magnifications.<br\>
 +
<br\>This experiment was conducted by Shinya Tahara (student member).<br\><br\><br\><br\><br\><br\>
 +
 
 +
==Discussion==
 +
We cultured iron-oxidizing bacteria in 9K medium.
 +
This experimental result means that iron-oxidizing bacteria require only dissolved CO<sub>2</sub> as carbon source.
 +
Therefore, iron-oxidizing bacteria is useful to terraform Mars, where lacks organic matters and contains Fe(Ⅱ).
 +
Iron-oxidizing bacteria require us only to heat the surface to control the surface temperature.<br\><br\>
 +
To apply to terraform Mars, we have to introduce genes to iron-oxidizing bacteria.
 +
However, there are few methods for selecting iron-oxidizing bacteria with introduced genes, because antibiotics are deactivated in culture conditions of iron-oxidizing bacteria.
 +
Some researchers succeeded in introducing genes to iron-oxidizing bacteria in their previous researches.
 +
They introduced mercury-resistance gene to iron-oxidizing bacteria and selected in the medium that contains mercury ion.
 +
 
 +
==Reference==
 +
*Joshua L. Bandfield,* Timothy D. Glotch, Philip R. Christensen (2003) Spectroscopic Identification of Carbonate Minerals in the Martian Dust, science 22 August 2003 Vol. 301. no. 5636, pp. 1084 - 1087
 +
*TOMONOBU KUSANO, KAZUYUKI SUGAWARA, CHIHIRO INOUE, TOSHIYUKI TAKESHIMA, MASAHIKO NUMATA, AND TOSHIKAZU SHIRATORI2 (1992) Electrotransformation of Thiobacillus ferrooxidans with Plasmids Containing a mer Determinant, JOURNAL OF BACTERIOLOGY, Oct. 1992, p. 6617-6623
 +
*MEDIA FOORMULATIONS http://wdcm.nig.ac.jp/catalogue/ncim/document/Ncim_media.pdf <br\>
 +
 
 +
==Acknowledgement==
 +
*Chihiro Inoue (Tohoku university,  Graduate School of Environmental Studies Department of Environmental Studies )

Latest revision as of 01:54, 22 October 2009

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Contents

Achievement

  • We cultured iron-oxidizing bacteria (Acidithiobacillus ferrooidans).
change of the color of 9K medium

Introduction ~Why do we need to use Iron-oxidizing bacteria for terraforming of Mars?~

Heterotrophic bacteria cannot live in native Martian environment because there are no organic matters on the Martian surface, however autotrophic bacteria can live without organic matters.
Iron-oxidizing bacteria is a kind of autotrophic bacteria.
Iron-oxidizing bacteria get energy by oxidizing Fe(Ⅱ) and grow with CO2. To grow on Mars, iron-oxidizing bacteria require only energy injection by human beings because of composition of the Martian surface, crust and atmosphere.
Martian surface and crust contains carbonate(Bandfield,2003 et al.) and FeO(Fe(Ⅱ)). In addition, Martian atmosphere contains much CO2.(For detail, see here)
We propose culturing iron-oxidizing bacteria on Mars to produce organic matters.
Accumulation of organic matters provides habitable environment for heterotrophic bacteria (for instance, E.coli).
We plan to culture iron-oxidizing bacteria on the Martian surface in early stage of terraforming to accumulate enough organic matters for heterotrophic bacteria to live.

Material and method

Iron-oxidizing bacteria

Iron-oxidizing bacterium is a kind of autotrophic bacteria.
Iron-oxidizing bacteria reduce NAD+ to NADH by oxidizing Fe(Ⅱ).
Iron-oxidizing bacteria grow with dissolved CO2 as carbon source, not organic matters.
In laboratory, iron-oxidizing bacteria are cultured in 9K medium which contains Fe(Ⅱ) and no organic matters.

Preparation of 9K medium (for iron-oxidizing bacteria)

We Prepared following A and B solution.

A Solution
B Solution

We sterilized A solution by autoclaving. We sterilized B solution by using filter.
We Mixed A and B solution before use.

Cultivation of Acidithiobacillus ferrooxidans

We cultured in 9K medium at 30℃ with vigorous shaking. We used the microscope when confirmed the growth.

Result

Cultivation of A.ferrooxidans

change of the color of 9K medium

We succeeded in culturing Acidithiobacillus ferrooxidans (iron-oxidizing bacteria).
The medium for A.ferrooxidans was blue before cultivation owing to Fe(Ⅱ).
The presence of A.ferrooxidans changed the medium into brown one after culturing for two days.
We confirmed the growth by using microscope of 200 magnifications.

This experiment was conducted by Shinya Tahara (student member).





Discussion

We cultured iron-oxidizing bacteria in 9K medium. This experimental result means that iron-oxidizing bacteria require only dissolved CO2 as carbon source. Therefore, iron-oxidizing bacteria is useful to terraform Mars, where lacks organic matters and contains Fe(Ⅱ). Iron-oxidizing bacteria require us only to heat the surface to control the surface temperature.

To apply to terraform Mars, we have to introduce genes to iron-oxidizing bacteria. However, there are few methods for selecting iron-oxidizing bacteria with introduced genes, because antibiotics are deactivated in culture conditions of iron-oxidizing bacteria. Some researchers succeeded in introducing genes to iron-oxidizing bacteria in their previous researches. They introduced mercury-resistance gene to iron-oxidizing bacteria and selected in the medium that contains mercury ion.

Reference

  • Joshua L. Bandfield,* Timothy D. Glotch, Philip R. Christensen (2003) Spectroscopic Identification of Carbonate Minerals in the Martian Dust, science 22 August 2003 Vol. 301. no. 5636, pp. 1084 - 1087
  • TOMONOBU KUSANO, KAZUYUKI SUGAWARA, CHIHIRO INOUE, TOSHIYUKI TAKESHIMA, MASAHIKO NUMATA, AND TOSHIKAZU SHIRATORI2 (1992) Electrotransformation of Thiobacillus ferrooxidans with Plasmids Containing a mer Determinant, JOURNAL OF BACTERIOLOGY, Oct. 1992, p. 6617-6623
  • MEDIA FOORMULATIONS http://wdcm.nig.ac.jp/catalogue/ncim/document/Ncim_media.pdf

Acknowledgement

  • Chihiro Inoue (Tohoku university, Graduate School of Environmental Studies Department of Environmental Studies )
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