Sep 22, 2024
Version 2
Arabidopsis-Microbe interaction seed treatment V.2
- 1Plant Health Care Program, Academy of Circular Economy, National Chung Hsing University, Taichung
Protocol Citation: Tao-Ho Chang 2024. Arabidopsis-Microbe interaction seed treatment. protocols.io https://dx.doi.org/10.17504/protocols.io.kqdg394z1g25/v2Version created by Tao-Ho Chang
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: December 06, 2023
Last Modified: September 22, 2024
Protocol Integer ID: 91876
Keywords: Arabidopsis thaliana, Seed priming, Beneficial microbe, Plant stress, Root structure architecture
Funders Acknowledgement:
National Sciecne and Technology Council, Taiwan
Grant ID: 112-2313-B-005 -008 -
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Abstract
The treated seeds can grow in medium or bulk soil to determine the impact of novel materials on plants.
Guidelines
Seed treatment of arabidopsis seeds is a suitable, simple method for many plant-microbe interactions.
Materials
Arabidopsis thaliana seeds (Col-0)
50% Bleach
Distilled water
1/2 MS medium (pH = 5.7)
Safety warnings
The only harmful chemical is 50% bleach (2% sodium hypochlorite). Please make sure ware gloves when in the seed sterile steps.
Before start
Seed priming is an important method that increases the health of the plant.
Seeds sterilisation
Seeds sterilisation
15m
15m
The arabidopsis seeds are immersed with 50% bleach (2% Sodium hypochlorite) in 1.5 µL tubes.
100 rpm, 28°C, 00:10:00
10m
100 rpm, 28°C, 00:00:10
10s
Gently remove the supernatant and leave the seeds in the tube.
The sterile seeds are immersed in distilled water.
Repeat the process of step #3 to #5 for 10 times
Seeds treatment
Seeds treatment
1d
1d
The seeds were immersed in the specific concentration of treatment
1. Bacteria treatment: OD value to 0.1 and diluted 10 times.
2. Fungus treatment: The final concentration of spore suspension is 100 spores mL-1.
100 rpm, 28°C, 01:00:00
1h
150 rpm, 4°C, 23:00:00
23h
Wash seeds
Wash seeds
10s
10s
The treated seeds are moved in a filter mesh (Favorgen) with a collection tube.
100 rpm, 28°C, 00:00:10
10s
Remove the filter through in collection tube
Add 500 µL of distilled water to the filter column and resuspend the seeds.
Repeat the process of step #11 to #13 for 10 times