Abstract:

This paper researched the preparation and transformation technologies of sugarcane pokkah boeng disease pathogen——Fusarium moniliformis's protoplast through observing the precipitation's compactness, regeneration rate and the OD valve. In the paper, the effect of liquid height, centrifugal inertia force, the centrifuge tube size, the rate of deceleration to the deposition's were researched. The types of lyases and the concentration of lysing enzyme from Trichoderma harzianum’s effect to generating protoplasts were introduced in this paper too. This paper also introduced one balance method to the centrifuge. The research showed using 10 mL 0.1 g/mL lysing enzyme from Trichoderma harzianum to digest 0.5 g hypha could get lots of protoplasts; using lysing enzyme from Trichoderma harzianum mixed with driselase could get more protoplasts. Using 1 mol/L sorbital or 1.2 mol/L KCl to rinse the precipitation and then centrifuge, using the small bottom centrifuge tubes and little amount solution and slow down the centrifuge speed slowly could avoid the precipitation loosen and get more precipitation.