Abstract: Suitable control content of culture medium can benefit growth, proliferation rate of Williams B6 seedling, save spending of production material. The content of 30ml per bottle show the highest proliferation rate and the higher usage of culture medium, the contrary result will appear when preponderate over the content of 30ml per bottle, but the increase weight of single seedling. For the losing of medium by sterilization, seedling SO2 resistance and other effects, it’s considered the content of 25~30ml per bottle is fit for production of tissue culture seedling. The varieties of Williams B6 and Brazil banana show the higher SO2 sensitivity compare with the powder plantain and plantain when the Na2SO3 concentration reaches 15g/100ml. The result will benefit standard production and planting management of tissue culture seedling of banana against SO2 adversity.