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

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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.



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