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MB Sample ID: SA316641
| Local Sample ID: | Mock(2-6) |
| Subject ID: | SU003030 |
| Subject Type: | Plant |
| Subject Species: | Arabidopsis thaliana |
| Taxonomy ID: | 3702 |
| Age Or Age Range: | 4-5 weeks |
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Collection:
| Collection ID: | CO003023 |
| Collection Summary: | Apoplastic wash fluid (AWF) was isolated from the leaves of 5- to 6-week-old Arabidopsis plants treated with flg22 or a mock control. To initiate PAMP responses, a needle-less syringe was used to infiltrate the leaves with a solution of 100 nM flg22 in water, or with DMSO in water as a negative control. Six to eight leaves were infiltrated on each plant, and a total of six plants were infiltrated for each treatment condition. Individual plants were infiltrated with flg22 or the DMSO only control, never both. After eight hours, AWF was isolated by syringe-infiltrating the mock- and flg22-treated leaves with sterile H2O containing 164 µM ribitol as an internal standard. Immediately after infiltration, the aerial portion of the plant was removed by cutting the primary stem and briefly washed with H2O to remove surface contaminants. The infiltrated leaves were detached from the rosette and stacked between layers of parafilm, with 2 to 4 leaves in each layer. The parafilm booklet of leaves was wrapped with tape and suspended inside a 15 mL conical centrifuge tube. The tube was centrifuged at 750 x g for seven minutes. The AWF that collected at the bottom of the conical tube was transferred to a clean 1.7 mL microcentrifuge tube, then centrifuged at 21,000 x g for 10 minutes at 4°C. The resulting supernatant was transferred to a clean 1.7 mL microcentrifuge tube. After addition of 50 µL of chloroform, the samples were vortexed for 10 seconds and centrifuged at 21,000 x g for 10 minutes at 4°C. The upper aqueous phase was transferred to a clean 1.7 mL microcentrifuge tube, and the volume recovered was measured with a pipette. The AWF samples were then lyophilized to dryness and stored at -80°C until further use. |
| Sample Type: | Apoplastic washing fluid |
