Accesso libero

Arabidopsis thaliana for Spaceflight Applications–Preparing Dormant Biology for Passive Stowage and On-Orbit Activation

Natasha J. Sng
Natasha J. Sng
, 
Jordan Callaham
Jordan Callaham
, 
Robert J. Ferl
Robert J. Ferl
 e 
Anna-Lisa Paul
Anna-Lisa Paul
   | 02 mar 2022
Gravitational and Space Research's Cover Image
INFORMAZIONI SU QUESTO ARTICOLO
Articolo precedente
Articolo Successivo
Cita
Article Category: Methods Article
Pubblicato online: 02 mar 2022
Pagine: 81 - 89
DOI: https://doi.org/10.2478/gsr-2014-0018
Parole chiave
© 2014 Natasha J. Sng et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Figure 1

Dry sterilization and planting for dormant plates. A) Sterilized seeds drying on filter paper in a laminar flow hood. B) Collection of sterilized seeds into sterile, screw-top microcentrifuge tubes, which will be stored at 4°C until use. C) Plating sterilized seeds suspended in water across the top of a single plate with a sterile, disposable pipette. D) Plates are sealed with porous, surgical Micropore® tape and labeled.
Dry sterilization and planting for dormant plates. A) Sterilized seeds drying on filter paper in a laminar flow hood. B) Collection of sterilized seeds into sterile, screw-top microcentrifuge tubes, which will be stored at 4°C until use. C) Plating sterilized seeds suspended in water across the top of a single plate with a sterile, disposable pipette. D) Plates are sealed with porous, surgical Micropore® tape and labeled.

Figure 2

Securing plates in a light-tight configuration with Duvetyne Black-Out fabric. A) A survey photograph is taken to show that the configuration of the outer label (top left of image) matches the genotype label on the plate. The plate is then wrapped by first aligning to the bottom of the cloth as shown in (B), then the plate is rolled up in the cloth (C and D), and then the sides are folded in and secured with the outer label (E). The grid shown supporting the plate and cloth in the figure is divided into one inch squares.
Securing plates in a light-tight configuration with Duvetyne Black-Out fabric. A) A survey photograph is taken to show that the configuration of the outer label (top left of image) matches the genotype label on the plate. The plate is then wrapped by first aligning to the bottom of the cloth as shown in (B), then the plate is rolled up in the cloth (C and D), and then the sides are folded in and secured with the outer label (E). The grid shown supporting the plate and cloth in the figure is divided into one inch squares.

Figure 3

Dormancy test using Wassilewskija (WS) ecotype seeds. A) Whole plate view of dormant WS seeds after they were stored for 2, 4, and 6 weeks. All seeds remained dormant. B) Close-up view of the selected seeds corresponding to those boxed in red, in 3A. C) Exact plates from 3A that have been exposed to the light and allowed to germinate to show seed viability.
Dormancy test using Wassilewskija (WS) ecotype seeds. A) Whole plate view of dormant WS seeds after they were stored for 2, 4, and 6 weeks. All seeds remained dormant. B) Close-up view of the selected seeds corresponding to those boxed in red, in 3A. C) Exact plates from 3A that have been exposed to the light and allowed to germinate to show seed viability.

Figure 4

Dormancy test using Colombia-0 (Col-0) ecotype seeds. A) Whole plate view of dormant Col-0 seeds after they were stored for 2, 4, and 6 weeks. All seeds remained dormant. B) Close-up view of the selected seeds corresponding to those boxed in red, in 3A. C) Exact plates from 3A that have been exposed to the light and allowed to germinate to show seed viability.
Dormancy test using Colombia-0 (Col-0) ecotype seeds. A) Whole plate view of dormant Col-0 seeds after they were stored for 2, 4, and 6 weeks. All seeds remained dormant. B) Close-up view of the selected seeds corresponding to those boxed in red, in 3A. C) Exact plates from 3A that have been exposed to the light and allowed to germinate to show seed viability.

Figure 5

Dormancy and germination (viability) rates of WS and Col-0 and ecotype seeds. A) Dormancy (red) and germination (blue) rates of WS seeds after they were stored for 2, 4, and 6 weeks. In WS ecotype, dormancy rates were > 98% and germination rates were > 94%. B) Dormancy (red) and germination (blue) rates of Col-0 seeds after they were stored for 2, 4, and 6 weeks. Dormancy rates for Col-0 were > 95% and germination rates were > 93%. For WS, there were between three and six replicates of each time point. Error bars are provided for each set, all standard deviations were less than 0.07. For Col-0, there were four replicates of each time point. Error bars are provided for each set, all standard deviations were less than 0.06.
Dormancy and germination (viability) rates of WS and Col-0 and ecotype seeds. A) Dormancy (red) and germination (blue) rates of WS seeds after they were stored for 2, 4, and 6 weeks. In WS ecotype, dormancy rates were > 98% and germination rates were > 94%. B) Dormancy (red) and germination (blue) rates of Col-0 seeds after they were stored for 2, 4, and 6 weeks. Dormancy rates for Col-0 were > 95% and germination rates were > 93%. For WS, there were between three and six replicates of each time point. Error bars are provided for each set, all standard deviations were less than 0.07. For Col-0, there were four replicates of each time point. Error bars are provided for each set, all standard deviations were less than 0.06.

Figure 6

Visual inspection of dormant, Duvetyne-wrapped plates: A,B,C) Plates with WS seeds were treated with Far-red light and stored at 4°C for 1 week before being unwrapped and exposed to 30 seconds of ambient room lighting, and then re-wrapped for storage at 4°C for 4 weeks. The full view of the plate three weeks after visual inspection is shown in (A), and an enlargement of the seed row is shown in (B). C) Exact plate from 5A that has been exposed to the light and allowed to germinate to show seed viability. D,E,F) Plates with WS seeds were kept at room temperature for 1 week before being unwrapped and exposed to 30 seconds of ambient room lighting, and re-wrapped for storage at room temperature for 4 weeks. The full view of the plate three weeks after visual inspection is shown in (D), and an enlargement of the seed row is shown in (E). D) Exact plate from 5A that has been exposed to the light and allowed to germinate to show seed viability.
Visual inspection of dormant, Duvetyne-wrapped plates: A,B,C) Plates with WS seeds were treated with Far-red light and stored at 4°C for 1 week before being unwrapped and exposed to 30 seconds of ambient room lighting, and then re-wrapped for storage at 4°C for 4 weeks. The full view of the plate three weeks after visual inspection is shown in (A), and an enlargement of the seed row is shown in (B). C) Exact plate from 5A that has been exposed to the light and allowed to germinate to show seed viability. D,E,F) Plates with WS seeds were kept at room temperature for 1 week before being unwrapped and exposed to 30 seconds of ambient room lighting, and re-wrapped for storage at room temperature for 4 weeks. The full view of the plate three weeks after visual inspection is shown in (D), and an enlargement of the seed row is shown in (E). D) Exact plate from 5A that has been exposed to the light and allowed to germinate to show seed viability.

Figure 7

Actual TAGES flight experiment on-board the ISS, documenting the successful application of this technique in delivering dormant arabidopsis seeds on-orbit. A) Visual inspection of dormant plate 4 (1B49137TK) run 1B showing dormant seeds, and no contamination. B) Identical plate 4 in Figure 6B being harvested on ISS after 12 days of growth. C) Wrapped TAGES plate (3A19363TK). D) Identical plate in Figure 6C shows dormant seeds, unwrapped and light-stimulated for germination onboard the ISS. The image was taken with the ABRS/GIS hardware. E,F) Gradual growth of the plant from once dormant seed shows the viability of the seed.
Actual TAGES flight experiment on-board the ISS, documenting the successful application of this technique in delivering dormant arabidopsis seeds on-orbit. A) Visual inspection of dormant plate 4 (1B49137TK) run 1B showing dormant seeds, and no contamination. B) Identical plate 4 in Figure 6B being harvested on ISS after 12 days of growth. C) Wrapped TAGES plate (3A19363TK). D) Identical plate in Figure 6C shows dormant seeds, unwrapped and light-stimulated for germination onboard the ISS. The image was taken with the ABRS/GIS hardware. E,F) Gradual growth of the plant from once dormant seed shows the viability of the seed.
eISSN:
2332-7774
Lingua:
Inglese
Frequenza di pubblicazione:
2 volte all'anno
Argomenti della rivista:
Life Sciences, other, Materials Sciences, Physics
Feed RSS della rivista