The invention discloses an induction method of poplar tetraploid plants, belonging to the field of plant inheritance breeding. The method of the invention comprises the following steps: pollination is carried out on female inflorescence of poplar female parent by powder of poplar male parent to form infructescence, when flocculent fiber is formed and wraps ovule in the infructescence ovary, physicochemical treatment is carried out on the infructescence, seed selection is carried out on tetraploid plants; the inventor of the invention discovers by tests that the effective period for carrying out physicochemical treatment to induce the zygote chromosome to be reduplicated is when most zygotes of the poplar is at the first mitosis process and the flocculent fiber starts to wrap but does not completely covet the ovule, thus the induction efficiency of tetraploid plants is improved. The invention avoids verbose links such as fixing, dehydrating, waxing, embedding, slicing, pigmentation, microexamination of the ovule and the like, thus enhancing the timeliness of physicochemical treatment, saving induction reagent and experimental material and improving the precision and stability of the reduplication of zygote chromosome.