iGEM 2021 — Team MTU-CORK

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# Team:MTU-CORK/Notebook

[Home](https://2021.igem.org/Team%3AMTU-CORK)

Overview

[Description](https://2021.igem.org/Team%3AMTU-CORK/Description)
[Literature](https://2021.igem.org/Team%3AMTU-CORK/Literature)

Team

[Collaborations](https://2021.igem.org/Team%3AMTU-CORK/Collaborations)
[Attributions](https://2021.igem.org/Team%3AMTU-CORK/Attributions)
[Team](https://2021.igem.org/Team%3AMTU-CORK/Team)
[Support Team](https://2021.igem.org/Team%3AMTU-CORK/Support_Team)

Human Practices

[Integrated Human Practices](https://2021.igem.org/Team%3AMTU-CORK/Integrated_Human_Practices)
[Human Practices](https://2021.igem.org/Team%3AMTU-CORK/Human_Practices)

Design

[Proof Of Concept](https://2021.igem.org/Team%3AMTU-CORK/Proof_Of_Concept)
[Implementation](https://2021.igem.org/Team%3AMTU-CORK/Implementation)
[Contribution](https://2021.igem.org/Team%3AMTU-CORK/Contribution)
[Biology Design](https://2021.igem.org/Team%3AMTU-CORK/Biology_Design)

Build

[Experiments](https://2021.igem.org/Team%3AMTU-CORK/Experiments)
[Engineering](https://2021.igem.org/Team%3AMTU-CORK/Engineering)

Test

[Results](https://2021.igem.org/Team%3AMTU-CORK/Results)
[Parts](https://2021.igem.org/Team%3AMTU-CORK/Parts)

[Education](https://2021.igem.org/Team%3AMTU-CORK/Communication)

[Safety](https://2021.igem.org/Team%3AMTU-CORK/Safety)

[Hardware Model](https://2021.igem.org/Team%3AMTU-CORK/Model)

☰

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# Notebook

Lorem ipsum dolor sit amet, consectetur adipiscing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat. Duis aute irure dolor in reprehenderit in voluptate velit esse cillum dolore eu fugiat nulla pariatur. Excepteur sint occaecat cupidatat non proident, sunt in culpa qui officia deserunt mollit anim id est laborum.

Lorem ipsum dolor sit amet, consectetur adipiscing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat. Duis aute irure dolor in reprehenderit in voluptate velit esse cillum dolore eu fugiat nulla pariatur. Excepteur sint occaecat cupidatat non proident, sunt in culpa qui officia deserunt mollit anim id est laborum.

Lorem ipsum dolor sit amet, consectetur adipiscing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat. Duis aute irure dolor in reprehenderit in voluptate velit esse cillum dolore eu fugiat nulla pariatur. Excepteur sint occaecat cupidatat non proident, sunt in culpa qui officia deserunt mollit anim id est laborum.

# example 1

Lorem ipsum dolor sit amet, consectetur adipiscing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat. Duis aute irure dolor in reprehenderit in voluptate velit esse cillum dolore eu fugiat nulla pariatur. Excepteur sint occaecat cupidatat non proident, sunt in culpa qui officia deserunt mollit anim id est laborum.

# example 2

Lorem ipsum dolor sit amet, consectetur adipiscing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat. Duis aute irure dolor in reprehenderit in voluptate velit esse cillum dolore eu fugiat nulla pariatur. Excepteur sint occaecat cupidatat non proident, sunt in culpa qui officia deserunt mollit anim id est laborum.

# Notebook

Document the dates you worked on your project. This should be a detailed account of the work done each day for your project.

### What should this page have?

* Chronological notes of what your team is doing.
* Brief descriptions of daily important events.
* Pictures of your progress.
* Mention who participated in what task.

### Inspiration

You can see what others teams have done to organize their notes:

* [2018 Munich](https://2018.igem.org/Team%3AMunich/Notebook)
* [2019 Georgia State](https://2019.igem.org/Team%3AGeorgia_State/Notebook)
* [2019 Newcastle](https://2019.igem.org/Team%3ANewcastle/Notebook)
* [2020 IISER Pune India](https://2020.igem.org/Team%3AIISER-Pune-India/Notebook)
* [2020 Lund](https://2020.igem.org/Team%3ALund/Notebook)
* [2020 NOVA LxPortugal](https://2020.igem.org/Team%3ANOVA_LxPortugal/Notebook)
* [2020 RDFZ China](https://2020.igem.org/Team%3ARDFZ-China/NoteBook)

* Email Us : igemmtu2021@gmail.com

* [Home](https://2021.igem.org/Team%3AMTU-CORK)
* [Description](https://2021.igem.org/Team%3AMTU-CORK/Description)
* [Parts](https://2021.igem.org/Team%3AMTU-CORK/Parts)
* [Members](https://2021.igem.org/Team%3AMTU-CORK/Team)
* [Support Team](https://2021.igem.org/Team%3AMTU-CORK/Support_Team)

* ![](https://static.igem.org/mediawiki/2021/a/aa/T--MTU-CORK--shannonabc.jpg)
* ![](https://static.igem.org/mediawiki/2021/1/14/T--MTU-CORK--cappa.jpg)
* ![](https://static.igem.org/mediawiki/2021/8/80/T--MTU-CORK--themtulogo.jpg)
* ![](https://static.igem.org/mediawiki/2021/7/7b/T--MTU-CORK--BCO.jpg)

© MTU-CORK | All Rights Reserved

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# Team:MTU-CORK/Engineering

[Home](https://2021.igem.org/Team%3AMTU-CORK)

Overview

[Description](https://2021.igem.org/Team%3AMTU-CORK/Description)
[Literature](https://2021.igem.org/Team%3AMTU-CORK/Literature)

Team

[Collaborations](https://2021.igem.org/Team%3AMTU-CORK/Collaborations)
[Attributions](https://2021.igem.org/Team%3AMTU-CORK/Attributions)
[Team](https://2021.igem.org/Team%3AMTU-CORK/Team)
[Support Team](https://2021.igem.org/Team%3AMTU-CORK/Support_Team)

Human Practices

[Integrated Human Practices](https://2021.igem.org/Team%3AMTU-CORK/Integrated_Human_Practices)
[Human Practices](https://2021.igem.org/Team%3AMTU-CORK/Human_Practices)

Design

[Proof Of Concept](https://2021.igem.org/Team%3AMTU-CORK/Proof_Of_Concept)
[Implementation](https://2021.igem.org/Team%3AMTU-CORK/Implementation)
[Contribution](https://2021.igem.org/Team%3AMTU-CORK/Contribution)
[Biology Design](https://2021.igem.org/Team%3AMTU-CORK/Biology_Design)

Build

[Experiments](https://2021.igem.org/Team%3AMTU-CORK/Experiments)
[Engineering](https://2021.igem.org/Team%3AMTU-CORK/Engineering)

Test

[Results](https://2021.igem.org/Team%3AMTU-CORK/Results)
[Parts](https://2021.igem.org/Team%3AMTU-CORK/Parts)

[Education](https://2021.igem.org/Team%3AMTU-CORK/Communication)

[Safety](https://2021.igem.org/Team%3AMTU-CORK/Safety)

[Hardware Model](https://2021.igem.org/Team%3AMTU-CORK/Model)

☰

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![](https://static.igem.org/mediawiki/2021/9/9b/T--MTU-CORK--marsgif.gif)

# Engineering

![](https://static.igem.org/mediawiki/2021/7/75/T--MTU-CORK--Red.png)

The Biofrag was designed on openSCAD and the code used to manipulate the different areas and attributes of the 3D mechanism can be found below. Both the inner and outer constructions of the Biofrag used various versions of the same code, with the inner layer being scaled down to allow for it to fit within the outer layer.

//This code below is the syntax for Biofrag v1. The code has gone through several iterations to include screw locking mechanism and multiple layers.
//All units are in mm
//the parameters of the bioballs
$fn=100; //number of faces
outer\_radius=30;
inner\_radius=29;
hole\_radius=2;
n\_holes=12;
n\_angle=360/n\_holes;
//end of defining parameters
difference(){ //You delete group of objects from another group of objects
union() {sphere(r=outer\_radius); //This is Big sphere from which you delete another sphere}
//here we delete group of objects from the above sphere
union() {sphere(r=inner\_radius); //This is small sphere from which you delete another sphere

//The following is the number of balls

for(i = [0:n\_holes]) {rotate([0, 0, n\_angle \* i]) translate([0,7, -9])
rotate([45, 0, 0])
cylinder(r = 4, h = 50);}

for(i = [0:n\_holes]) {rotate([0, 0, n\_angle \* i]) translate([0, 6, 4])
rotate([45, 0, 0])
cylinder(r = 3, h = 50);}

for(i = [0:n\_holes])
{rotate([0, 0, n\_angle \* i]) translate([0, 10, 14])
rotate([45, 0, 0])
cylinder(r = 4, h = 50);}

for(i = [0:n\_holes]) {rotate([0, 0, n\_angle \* i]) translate([0, -1, -15])
rotate([45, 0, 0])
cylinder(r = 4, h = 50);}

// This is to make the half sphere, basically we delete a big cube at hlaf-the sphere
translate([0,0,-20])
cube([outer\_radius\*2,outer\_radius\*2,40], center=true)}}

Figure 1 below shows the final version of the Biofrag and its inner and outer components

![](https://static.igem.org/mediawiki/2021/e/ee/T--MTU-CORK--biofraghardwareengineering.png)

Figure 1. The inner and outer layer of the Biofrag

# The Biofrag Isolation Unit

The Biofrag Isolation Unit (BIU) was also designed using the openSCAD software. The top and bottom were developed using two separate sets of codes.

The code below was manipulated to develop the bottom of the BIU.

//box length
$fn=100;
box\_length = 80;
//box\_breadth
box\_breadth = 50;
//box height
box\_height = 4;
//outer radius for rounded box
radius\_outer = 5;
//radius for screw holes
radius\_hole = 2;
//height of the holes
height\_holes=-20;
//diameter of the banana connector
banana\_diameter=8.5;
module faceplate(l,b,r\_outer,r\_hole) {
minkowski() {
circle(r\_outer);
square([l,b]);}}
module inner\_shape(l,b,r\_outer) {minkowski() {circle(r\_outer-3);
square([l,b]);}
module box(l,b,h,r\_outer,r\_hole)
{linear\_extrude(height = h)faceplate(l,b,r\_outer,r\_hole);}
difference()
{union() {box(box\_length,box\_breadth,box\_height,radius\_outer,radius\_hole);
translate([-2,-2,4])
box(box\_length+4,box\_breadth+4,box\_height,radius\_outer,radius\_hole);
translate([-4,-4,8])
box(box\_length+8,box\_breadth+8,box\_height,radius\_outer,radius\_hole);}

translate([0,0,4])
box(box\_length,box\_breadth,box\_height+5,radius\_outer,radius\_hole);}

The top of the BIU was designed using the code below.
//box length
$fn=100;
box\_length = 80;
//box\_breadth
box\_breadth = 50;
//box height
box\_height = 30;
//outer radius for rounded box
radius\_outer = 5;
//radius for screw holes
radius\_hole = 2;
//height of the holes
height\_holes=-20;
//diameter of the banana connector
banana\_diameter=8.5;
module faceplate(l,b,r\_outer,r\_hole)
{difference() {
minkowski()
{circle(r\_outer);
square([l,b]);}
circle(r\_hole);
translate([l,0,0])circle(r\_hole);
translate([0,b,0])circle(r\_hole);
translate([l,b,0])circle(r\_hole);}}
module inner\_shape(l,b,r\_outer) {difference() {minkowski() {circle(r\_outer-3);
square([l,b]);}

circle(r\_outer);
translate([l,0,0])circle(r\_outer);
translate([0,b,0])circle(r\_outer);
translate([l,b,0])circle(r\_outer);}}
module box(l,b,h,r\_outer,r\_hole) {difference() {union()
{linear\_extrude(height = h)
{difference() {faceplate(l,b,r\_outer,r\_hole);
inner\_shape(l,b,r\_outer);}}
linear\_extrude(height = 2)faceplate(l,b,r\_outer,r\_hole);}

union() {rotate([0,0,0])
translate([box\_length/2,box\_breadth/2,-0.2])
cylinder(d=3, h=8, $fn=100); //Top Vent

rotate([0,90,0])
translate([height\_holes+4,25,box\_length-2])
cylinder(d=banana\_diameter, h=10, $fn=100); //For Raman Probe}}}
box(box\_length,box\_breadth,box\_height,radius\_outer,radius\_hole);
//translate([box\_length+2\*radius\_outer+5,0,0])linear\_extrude(height = 2)faceplate(box\_length,box\_breadth,radius\_outer,radius\_hole);

Figure 2 below depicts a complete render of the Biofrag Isolation Unit

![](https://static.igem.org/mediawiki/2021/1/11/T--MTU-CORK--harwareengineeringpic.png)

Figure 2: Biofrag Isolation Unit (the portholes allowing for insertion of oxygen and chlorite sensors are also depicted), the top will also be transparent.

* Email Us : igemmtu2021@gmail.com

* [Home](https://2021.igem.org/Team%3AMTU-CORK)
* [Description](https://2021.igem.org/Team%3AMTU-CORK/Description)
* [Parts](https://2021.igem.org/Team%3AMTU-CORK/Parts)
* [Members](https://2021.igem.org/Team%3AMTU-CORK/Team)
* [Support Team](https://2021.igem.org/Team%3AMTU-CORK/Support_Team)

* ![](https://static.igem.org/mediawiki/2021/a/aa/T--MTU-CORK--shannonabc.jpg)
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* ![](https://static.igem.org/mediawiki/2021/7/7b/T--MTU-CORK--BCO.jpg)

© MTU-CORK | All Rights Reserved

[![](https://static.igem.org/mediawiki/2021/8/8b/HQ_page_logo.jpg)](https://2021.igem.org)

# Team:MTU-CORK/Experiments

[Home](https://2021.igem.org/Team%3AMTU-CORK)

Overview

[Description](https://2021.igem.org/Team%3AMTU-CORK/Description)
[Literature](https://2021.igem.org/Team%3AMTU-CORK/Literature)

Team

[Collaborations](https://2021.igem.org/Team%3AMTU-CORK/Collaborations)
[Attributions](https://2021.igem.org/Team%3AMTU-CORK/Attributions)
[Team](https://2021.igem.org/Team%3AMTU-CORK/Team)
[Support Team](https://2021.igem.org/Team%3AMTU-CORK/Support_Team)

Human Practices

[Integrated Human Practices](https://2021.igem.org/Team%3AMTU-CORK/Integrated_Human_Practices)
[Human Practices](https://2021.igem.org/Team%3AMTU-CORK/Human_Practices)

Design

[Proof Of Concept](https://2021.igem.org/Team%3AMTU-CORK/Proof_Of_Concept)
[Implementation](https://2021.igem.org/Team%3AMTU-CORK/Implementation)
[Contribution](https://2021.igem.org/Team%3AMTU-CORK/Contribution)
[Biology Design](https://2021.igem.org/Team%3AMTU-CORK/Biology_Design)

Build

[Experiments](https://2021.igem.org/Team%3AMTU-CORK/Experiments)
[Engineering](https://2021.igem.org/Team%3AMTU-CORK/Engineering)

Test

[Results](https://2021.igem.org/Team%3AMTU-CORK/Results)
[Parts](https://2021.igem.org/Team%3AMTU-CORK/Parts)

[Education](https://2021.igem.org/Team%3AMTU-CORK/Communication)

[Safety](https://2021.igem.org/Team%3AMTU-CORK/Safety)

[Hardware Model](https://2021.igem.org/Team%3AMTU-CORK/Model)

☰

![](https://static.igem.org/mediawiki/2021/7/75/T--MTU-CORK--Red.png)

![](https://static.igem.org/mediawiki/2021/9/9b/T--MTU-CORK--marsgif.gif)

# Experiments

![](https://static.igem.org/mediawiki/2021/7/75/T--MTU-CORK--Red.png)

# Plant Choice Materials and Method

* A quick google search suggested radish and lettuce could be suitable as are
  observed to be fast growing plants.
* Seeds were bought from a local garden centre along with growing boxes, and
  smaller growing containers (all with clear Perspex lids allowing for ventilation).
* Soil was taken from the garden and distributed into the large growing box to a depth
  of 150mm.
* Radish seeds were planted in 3 rows with approximately 80-90mm between
  them.
* Lettuce seeds were planted in 3 rows with approximately 80-90mm between
  them.
* 30mL of water was added to each seeded area initially and this was
  supplemented every 2nd day with an additional 20mL of water.
* The grow box was positioned in the same area continuously to both observe
  identical sunlight exposure and to minimise temperature fluctuations from drafts,
  shadows etc.
* Perspex lid was maintained in place with ventilation slots left open (except for
  days when additional rainfall would affect water volume to seeds) and seeds
  were inspected daily.
* Growth was allowed to continue for 21 days post seeding.

# Biofrag 1.0 Seed Growth Ability Testing

* Both internal and external Biofrag 1.0 were filled with dense sponge.
* Seeds were added into the Biofrag via a small tunnel cut into the foam centre.
  This tunnel centred on holes within the inner Biofrag (which would hope to allow
  both stem and root growth of seed to be unimpeded as it exited the Biofrag
  structure).
* Biofrag was planted within a glass jar (see pic) so upper surface was level with
  soil and 30mL of water was gently administered directly over it (to not potentially
  disturb its position).
* An additional 20mL of water was applied every 2nd day.
* The glass jar was positioned in the same area continuously to both observe
  identical sunlight exposure and to minimise temperature fluctuations from drafts,
  shadows etc.
* Growth was allowed to continue for 21 days post seeding.

# Alternate Biofrag 1.0 Seed Growth Ability Testing

* See *Biofrag 1.0 Seed Growth Ability Testing* for initial steps of Biofrag setup.
* Biofrag was then planted in the same manner but within the grow box and 30mL
  of water was gently administered directly over it (to not potentially disturb its
  position).
* An additional 20mL of water was applied every 2nd day.
* The grow box was positioned in the same area continuously to both observe
  identical sunlight exposure and to minimise temperature fluctuations from drafts,
  shadows etc.
* Perspex lid was maintained in place with ventilation slots left open (except for
  days when additional rainfall would affect water volume to seeds) and seeds
  were inspected daily.
* Growth was allowed to continue for 21 days post seeding.

# Biofrag 2.0 Seed Growth Ability Testing

* The inner frag was filled with soil surrounded by a thin layer of dense foam. Foam
  was arranged as to not interfere with the newly incorporated portals on both poles
  of the inner Biofrag This was accomplished by placing a hose pipe section
  through the portals creating space within the foam matrix that could be filled with
  soil afterwards.
* The outer Biofrag halves were lined with dense foam and the inner frag was
  placed into one half with the protruding hose section lined up with the larger
  portal. The adjoining half was then secured to it with the other end of the hose
  section lined up with the adjoining portal.
* The hose was filled with earth and the Biofrag was buried into soil (within a grow
  box) where the hose was removed allowing the earth within to fill the previous
  aperture made. Additional soil was added post hose removal to fill the Biofrag to
  the portal level.
* One radish seed was placed into the portal opening to a depth of approximately
  20mm and had 30mL of water added onto it.
* The grow box was positioned in the same area continuously to both observe
  identical sunlight exposure and to minimise temperature fluctuations from drafts,
  shadows etc.
* Perspex lid was maintained in place with ventilation slots left open (except for
  days when additional rainfall would affect water volume to seeds) and seeds
  were inspected daily.
* 20mL of additional water was added in the same way every 2nd day until
  completion of test.
* Controls involving radish seeds being planted as normal around the Biofrag were
  used to compare potential growth differences.
* This whole assay was repeated in a second grow box and growth was allowed to continue for 21 days post seeding.

---

# Seed Growth in Varying Perchlorate (ClO4¯) Concentrations.

### Soil Preparation

* Oven tray filled with soil approximately 15mm in depth.
* Oven pre heated to 130ºC and tray inserted on middle shelf for 60mins.
* Tray was removed and allowed to cool before soil transferred to blender and
  blended at high speed for 60 seconds.

### Falcon tube filling and Autoclaving

* Five Falcon tubes were found to be 12.5g (mean) when weighed.
* Each tube was filled with 8g of prepared soil (approx 25mL).
* All weighing performed on laboratory scales (total weight:20.5g) and this was
  repeated for all 24 tubes needed for testing.
* All tubes had the cap loosely screwed on and were placed inside an autoclaving
  bag and autoclaved for 15mins at 121°C and 15psi.
* Once autoclaved, bag was placed into a biosafety cabinet before all tubes had
  their caps tightened to prevent potential contamination.

| ClO4¯ Concentration | RODI | DMM |
| --- | --- | --- |
| 1% | 3 | 3 |
| 0.5% | 3 | 3 |
| 0.25% | 3 | 3 |
| 0% (control) | 3 | 3 |
| Total tubes | 12 | 12 |

Table 1: Falcon Tube Sample Numbers

### Soil: Fluid Ratio (for ClO4¯ testing)

* Two of the filled falcon tubes were set aside and 5mL of water was added to the
  first, the tube was vortexed for 20 seconds and inspected for soil/moisture
  homogenisation.
* This was repeated until desired consistency was produced (20mL water).
* The second Falcon tube had 20mL of water added in one go, was vortexed for 20
  seconds, and inspected for soil/water homogenisation (same result).

### Davis Minimal Media Broth (DMM) Preparation

* 2.9g of DMM powder was weighed out on a plastic weigh boat and transferred
  into a 500mL conical flask. 250mL of RODI was added via a volumetric flask and then placed into a microwave. The media was boiled for 1 minute and allowed to cool before use.

### Soil ClO4¯ Concentrations

There was 60mL of each perchlorate concentration (1%, 0.5%, and 0.25%) needed
to be made up in RODI and DMM.

* The 1% ClO4¯ Soil Concentration needed 0.08g ClO4 ̄/8g soil.
* 20mL fluid (RODI or DMM) x 3 tubes = 60mL.
* 0.08g ClO4¯ x 3 tubes = 0.24g ClO4¯ .
* 0.24g ClO4¯ + 60mL DMM = 1% ClO4¯ solution.
* 0.5% solution = 0.12g ClO4¯ /60mL DMM.
* 0.25% solution = 0.06g ClO4¯ /60mL DMM.
* All concentrations were duplicated in RODI.

RODI= Reverse Osmosis Deionised Water

| ClO4¯ Concentration | ClO4¯ (g) | RODI (mL) | DMM (mL) |
| --- | --- | --- | --- |
| 1% ClO4¯ (DMM) | 0.24 | N/A | 60 |
| 0.5% ClO4¯ (DMM) | 0.12 | N/A | 60 |
| 0.25% ClO4¯ (DMM) | 0.06 | N/A | 60 |
| 1% ClO4¯ (RODI) | 0.24 | 60 | N/A |
| 0.5% ClO4¯ (RODI) | 0.12 | 60 | N/A |
| 0.25% ClO4¯ (RODI) | 0.06 | 60 | N/A |
| Ctrl: 0% ClO4¯ (DMM) | 0 | N/A | 60 |
| Ctrl: 0% ClO4¯ (RODI) | 0 | 60 | N/A |

Table 2: Medium Perchlorate concentrations for soil testing

### Falcon tube seed planting

Each tube was labelled with the date, ClO4¯ concentration, and either RODI or
DMM. Using a pipette, 20mL of each liquid medium was transferred into its
corresponding tube. The tube was vortexed for 20 seconds ensuring complete
homogenisation of perchlorate and soil. Using a disposable loop, a small furrow was
made into the soil and one seed was inserted at a depth of approximately 15mm.
The cap was reattached, and this was repeated for each tube. Tubes were placed upright into a grow box and allowed to grow for 10 days.

* Email Us : igemmtu2021@gmail.com

* [Home](https://2021.igem.org/Team%3AMTU-CORK)
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* [Parts](https://2021.igem.org/Team%3AMTU-CORK/Parts)
* [Members](https://2021.igem.org/Team%3AMTU-CORK/Team)
* [Support Team](https://2021.igem.org/Team%3AMTU-CORK/Support_Team)

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# Team:MTU-CORK/Results

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Test

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## Results

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# Plant Choice Results

Fig 1

Fig 2

Fig 3

Fig 4

Fig 5

Fig 6

Grow box seeded with both radish and lettuce seeds (Fig 1.). Initial sprouting of radish seeds (Fig 2.) and lettuce seeds (Fig 3.). Growth at 11-day post seeding for both radish (Fig 4.) and lettuce (Fig 5.). Growth of radish and lettuce seeds at 28-days post initial seeding.

# Plant Choice Discussion

As expected from seed growth information provided with the seeds, and from online
research, the radish seeds not only sprouted first but they were close to maturity by the
21-day timepoint. Each seed observed the same growth pattern with all growth being
initially observed at 5 days +/- 12 hours.

The lettuce was initially slower to sprout (8 days approx for the first observed
growth) and was not consistent across those that were planted (some seeds initially
took 24-96 hours longer to sprout). However, by day 21 there seemed to be no clear
difference in sizes of lettuces and an even growth was observed across that section of
the grow box.

Whilst both radish and lettuce seeds produced post 21-day growth (Fig 6.) that could be
used throughout this project to easily quantify potential growth-inhibiting/promoting
factors in further testing, radish seeds were chosen for further testing due to
their efficient and replicable growth patterns observed during this test.

# Biofrag 1.0 Seed Growth Ability Results

Fig 7

Fig 8

Fig 9

Position of Biofrag in glass jar allowing for unimpeded sunlight (Fig 7.) and with soil filled around it and up to its apex (Fig 8). No growth was observed 21-days post seeding (Fig 9).

# Biofrag 1.0 Seed Growth Ability Discussion

The lack of any growth may have been due to several reasons. The compact nature
of the sponge and lack of room may not have been conducive to seed germination.
The obvious reason may be just that the removal of growth medium (soil) from
around the seed did not allow for germination and may be a deciding factor upon
choice of Biofrag design going forwards. Although the choice of housing was
different from initial growth experiments, it was though the material choice (glass)
would not negatively influence assay outcome. However, the increased light and
potentially differential temperature from that within the Perspex contained grow box
may have been contributing factors to null growth observed. It was decided to repeat
the assay using the exact criteria previously used in the initial growth studies.

# Alternate Biofrag 1.0 Seed Growth Ability Results

Fig 10

Fig 11

Fig 12

Biofrag replanted within the grow box (Fig 10.) and growth of seeds through both the outer Biofrag (Fig 11) and inner Biofrag (Fig 12).

# Alternate Biofrag 1.0 Seed Growth Ability Discussion

Growth was observed from two seeds overall, but the growth rate was much slower than
expected with observable growth not until approx 7-days post seeding. Whilst
observable growth proceeded there was a halt after approximately a further 7-days
after which there was no increase in plant size. However, plant health remained until
end of 21-day timeframe, and though no additional growth was observed, the plant maintained colour and rigidity comparable with live plants.
From these results, it was deemed an alternative Biofrag design was needed and
possibly an alternative to the dense sponge used at this time.

# Biofrag 2.0 Seed Growth Ability Testing Results

![](https://static.igem.org/mediawiki/2021/3/37/T--MTU-CORK--improvedbiofragpicture.png)

Figure 13

Fig 14

Fig 15

Fig 16

Fig 17

Fig 18

The 3D printed Biofrag 2.0 model (Fig 13) and the hose adaption for filling (Fig 14 and Fig 15). Buried Biofrag 2.0 with portal facing upwards for unimpeded growth (Fig 16) and observed growth at days 11 and 21 (Fig 17 and Fig 18).

# Biofrag 2.0 Seed Growth Ability Discussion

Although both Biofrags were seeded 2-days apart, initial growth was much faster in
one compared to the other. Whilst one Biofrag produced growth like the control
seeds around it (sprouting 5-days +/- 12hrs post planting) the other did not have
observable growth for an additional 4-days approximately (post-seeding) though the
controls were as expected. In grow box 1, growth between the Biofrag seed and the
control seeds followed a very similar pattern with no clear difference between any of
the plants. Although slower to initiate, the seed growth in grow box 2, within the
Biofrag, continued at the same rate of growth as all other controls and the other
Biofrag.

# Seed Growth in Varying Perchlorate Concentration Results

Fig 19

Fig 20

Fig 21

Fig 22

The controls (0% Perchlorate) had similar growth in both the RODI (Left) and DMM (Right) tubes (Fig 19). Both the RODI (Left) and DMM (Right) 0.25% concentrations produced growth with slightly more observed in the DMM (Fig 20). Neither the 0.5% or 1% Perchlorate concentrations produced growth in either the RODI or DMM (Fig 21 and Fig 22 respectively).

# Seed Growth in Varying Perchlorate Concentrations Discussion

From the growth observed, it is clear there is little difference in growing these seeds in either RODI or DMM. Any small differences seen may be down to natural seed composition or growth patterns. The lowest perchlorate concentration (0.25%) allowed growth but at a slower rate than the controls and all concentrations above this (0.5% and 1.0%) did not provide an environment suitable for seed growth. These results highlight the need for remediation of Martian soil, as introducing plant life, even as a potential oxygen source, would require soil perchlorate concentrations to be much lower than those currently experienced.

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