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MB Sample ID: SA193979
| Local Sample ID: | Hydrated_Lipids_Neg_2 |
| Subject ID: | SU002142 |
| Subject Type: | Plant |
| Subject Species: | Arabidopsis thaliana |
| Taxonomy ID: | 3702 |
| Genotype Strain: | Col-0 |
| Age Or Age Range: | Mature pollen, Pollen hydration Stage (45 min), Pollen germination stage (4 hours) |
| Species Group: | Pollen |
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Collection:
| Collection ID: | CO002135 |
| Collection Summary: | The Arabidopsis Col-0 plants were grown under controlled temperature (22°C) with a 16-h light (100-150 µmol m-2 s-1)/ 8-h dark photoperiod. Mature pollen grains from the fully opened flowers were collected from more than 1000 plants using a vacuum cleaner method (Johnson-Brousseau and McCormick, 2004) at around 5 hours into the light period. For mature pollen samples, collected pollen grains were resuspended in 2 ml of ice-cold Pollen Isolation Buffer (PIB, composed of 100 mM NaPO4, pH 7.5, 1 mM EDTA, and 0.1% (v/v) Triton X-100) right after collection followed by centrifuging at 15,000 g for 1 min (4°C). For hydrated pollen and germinated pollen samples, mature pollen grains were germinated in vitro according to a previously described pollen transcriptome study (Wang et al., 2008). In brief, pollen pellets were washed with 1 ml of liquid Pollen Germination Medium (PGM, composed of 15% (w/v) sucrose, 1.5 mM boric acid, 0.8 mM MgSO4, 1 mM KCl, 5 mM MES, 0.05% (w/v) lactalbumin hydrolysate, 10 µM myo-inositol, 5 mM CaCl2) before they were resuspended in 30 µl of liquid PGM and subsequently cultured in Petri dishes (35 mm in diameter). A 70 µm mesh was used to cover the pollen droplet to create a thin layer for optimal germination for each Petri dish. The Petri dishes were covered and placed in the dark for 45 min or 4 h and collected as hydrated pollen or germinated pollen, respectively. All pollen samples were washed by 1 ml ice-cold ddH2O three times before being stored in a -80 °C freezer. |
| Collection Protocol Filename: | PlantGrowth_PollenCollection.docx |
| Sample Type: | Plant |
| Collection Method: | Flash frozen in Liquid N2 |
| Collection Location: | University of Illinois, Urbana-Champain |
| Storage Conditions: | -80℃ |
