The invention discloses a sandy plant tissue culture medium, a preparation method thereof and an application thereof. The sandy plant tissue culture medium consists of river sand, nutrient solution, plant growth regulator, ascorbic acid, polyvinylpyrrolidone and fungicide, wherein with 1 liter as the standard, the nutrient solution accounts for 100-200 milliliters per liter, the various plant growth regulator such as 6-BA (6-Benzylaminopurine), NAA (1-naphthlcetic acid), IBA (indole-3-butytric acid) and IAA (indole-3-acetic acid) each account for 0.01-10.0 milliliters, the ascorbic acid accounts for 0.1-5.0 grams per liter, the polyvinylpyrrolidone accounts for 1-5 grams per liter, the fungicide accounts for 3-15 grams per liter, and the river sand accounts for the remaining volume, and the nutrient solution includes MS nutrient solution, sucrose, Zt, 6-BA and NAA. The preparation method of the sandy plant tissue culture medium includes various steps such as pretreatment, culture medium preparation, and post-formulation treatment. The sandy plant tissue culture medium can be widely applied to the tissue culture of of lianas, herbaceous plants and woody plants. The sandy plant tissue culture medium, the preparation method thereof and the application thereof have the advantages that with fine clean river sand as solid medium instead of agar, the cost of the culture medium is reduced; the river sand can be recycled for multiple times; accurately adjusting the pH of the nutrient solution, plant growth hormone and plant organic nutrient solution in plant tissue culture medium isunnecessary.