Fuente: WIPO "tobacco agriculture" The invention discloses a semi-in-vitro culture method of tobacco pollen tubes, which comprises the following specific steps of: taking a pistil in the initial blooming stage of tobacco, and pre-culturing the pistil in a 28 DEG C incubator after pollination; cutting half or three quarters of a pre-cultured stylus by using a razor blade, placing the pre-cultured stylus on a glass slide containing aculture medium after cutting, and continuously culturing for 24h at 28 DEG C in a moisturizing box to ensure that after the culture, pollen tubes grow out of the cut part of the stylus into bundles;and cutting a pollen tube by ophthalmic forceps under a microscope, suspending the pollen tube in a centrifuge tube containing a culture medium, and carrying out centrifugation at a speed of 5000 rpmfor 10min to obtain semi-in-vitro cultured tobacco pollen tubes. The culture medium provided by the invention has simple and effective components and is high in repeatability; and different culture times and culture media are arranged at different cutting positions of a stylus, so that the method is easy to operate and provides an effective way for efficiently collecting the semi-in-vitro culturedtobacco pollen tubes. The method is suitable for selfing pollination and hybrid pollination combination without stylus incompatibility barrier, and provides a stable system for studying the regulation mechanism of interaction of tobacco pollen tubes and pistils.