The invention provides a molecular biological differentiation method aiming at genders of
asparagus dioecian plants and application of the molecular biological differentiation method. According to the technical scheme, four functional STS (Sequence-tagged Site) molecular markers of the
gene are developed based on a CDS (Coding Sequence) of a female inhibitor
gene SOFF on a
Y chromosome of
asparagus. On the basis, corresponding primers and universal PCR (
Polymerase Chain Reaction) amplification conditions are designed for self-characteristics of the four STS molecular markers; furthermore, the length of a basic group of a characteristic strip of a male
plant is definitely oriented. An experiment proves that the four molecular markers provided by the invention are all located in a
coding region of the female inhibitor
gene SOFF of the
asparagus, and are co-separated from phenotypes of the female and male genders in the aspect of molecular
heredity, and the differentiation accuracy reaches 100 percent; the four molecular markers can be widely used for the differentiation of the female and male genders of the asparagus and
molecular marker assisted selection of all-male or all-female varieties. Moreover, the experiment shows that the method can still be used for gender differentiation of the asparagus dioecian plants including asparagus dauricus (A.dauricus), asparagus longiflorus (A.longiflorus), A.maritumus and the like.