Extraction of DNA from Phyllosphere Microbiota
  1. First, 6.0 g of rubber leaves was transferred into a sterile conical flask containing 100 mL 0.1 mol/L phosphate buffer solution (pH=7.0), and then 100 μL Tween 80 was added.
  2. Each sample was shaken with ultrasonic oscillation for 8 minutes, and then shaken at a speed of 200 rpm for 30 minutes at 25°C.
  3. After 2 minutes of ultrasonic oscillation, sterilized forceps were used to remove the leaves on the super-clean worktable.
  4. The microorganisms in the conical flask were collected using suction filtration with a 0.45-μm microporous membrane.
  5. DNA was extracted from each sample using the MP Biomedicals™ FastDNA® Spin Kit for Soil (CA, USA).
  6. The concentration and purity of DNA were verified using the NanoDrop 2000C (Thermo Scientific, USA) to A260/A280 = 1.8–2.0, and stored at −20°C until use.