The invention provides a method for directly and quickly rooting tissue culture seedlings. According to the rooting method, the basal stem of a tissue culture seedling is beveled, rooting hormone-containing gelatinoids is uniformly applied to the bevel of the basal stem, and the tissue culture seedling is inserted into a substrate to induce the tissue culture seedling to root in a culturing mode. The method has high rooting rate and high survival rate in late transplanting, is simple and convenient to operate, does not need cultivation on a rooting culture medium or preparation of massive rooting culture medium so as to greatly reduce cost, and does not need to clean the culture medium at the root part of the tissue culture seedling in the process of replacing a culture medium or subsequent transplanting so as to save time and labor. The variety and dosage of rooting hormone are optimized by the method, the optimized method is very suitable for ligneous plants, especially Chinese chestnuts and cherry, and the rooting rates of Chinese chestnuts and cherry are increased to 81-99 percent from 55 percent and 70 percent of a traditional rooting method.