The invention relates to a high-efficiency
mutagenesis method of
nicotiana tabacum L.
pollen. The high-efficiency
mutagenesis method comprises the following steps of putting
nicotiana tabacum L.
pollen into a
liquid medium under an aseptic condition to carry out
mutagenesis treatment by 0.1 to 0.2% of
ethyl methanesulfonate (EMS) in a dark place at a temperature of 30 to 32 DEG C, carrying out
chromosome doubling treatment by a NLN-16
liquid medium in a dark environment at a temperature of 30 to 32 DEG C, putting the
nicotiana tabacum L.
pollen obtained by the previous step into a NLN-13
liquid medium to carry out
embryo induction culture in a dark environment at a temperature of 24 to 26 DEG C, carrying out shake culture by a shaking table in a dark environment at a temperature of 24 to 26 DEG C when an immature
embryo appears, and putting the immature
embryo growing a
cotyledon into a
solid medium to carry out subculture, wherein during a
transplanting season, after
seedling hardening, a
seedling obtained by the high-efficiency mutagenesis method can be transplanted in a field for
mutant character identification. The high-efficiency mutagenesis method overcomes dominant and recessive barriers of
gene expression, is beneficial for selection, shortens greatly a
mutant character stabilization period, saves breeding
generation time, and has the characteristics of wide variation range, high mutagenesis frequency and the like.