User:David Johnston Monje/Notebook/Maize Endophyte Biofertilizers/2011/08/01

Maize Endophyte Biofertilizers

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The Daily Grind Took more measurements of light in the greenhouse, today being a particularly cloudy day. With dense cloud cover, the light hitting the bench from one side to the other varied from 31/37/39 to 39/46/48 umol m-2 s-1. Outside the greenhouse, there was from 222/230/255/268 umol m-2 s-1 of light. Phyllosphere and rhizoplane bacterial sampling method will be this: Bacteria isolation. Using axenic procedures, 12 g of leaves (fresh weight) from individual trees were placed into 50 mL polypropylene tubes containing washing solution (0.1 M potassium phosphate buffer, pH 7.0) and sonicated for 10 min at 22.5 kHz in an ultrasonic cell disrupter (Misonix Inc., Atlantic Beach, NY, USA ) (6). The resulting bacterial suspension was centrifuged at 30,000 g for 15 min at 4oC. The pellet was resuspended in 1.5 mL of washing solution and further concentrated by centrifugation at 30,000 g for 15 min at 4oC. Bacteria were resuspended in 200 μL of washing solution and frozen at -20oC until processing for DNA extraction. Its from Lambais et al (http://www.sciencemag.org/content/312/5782/1917.full.pdf). I plan to use 30 mL of liquid per 50 mL tube, and add 5 drops of Tween 20 per litre. Will also sample plants using R2A culture to visualize differences in culturable microbes. One plant per genotype per treatment will be plated out to see visualize differences.