The invention relates to a molecular marking method for carrying out genetic relationship and clustering analysis on citrus fruit flies and belongs to the technical field of molecular marking. The molecular marking method for carrying out genetic relationship and clustering analysis on the citrus fruit flies comprises the following steps: carrying out PCR (polymerase chain reaction) amplification on DNA of citrus fruit flies by utilizing seven RAPD (random amplification polymorphism DNA) primers and 10 pairs of SRAP (sequence-related amplified polymorphism) primer combinations, carrying out agarose gel electrophoresis on amplification products, taking pictures, recording results, and carrying out statistics on an amplification zone of a detected material; and analyzing with genetic analysis software to obtain genetic distance between individuals, and drawing a dendrogram of citrus fruit flies in different geographical populations for distinguishing different class groups. The molecular marking method for carrying out genetic relationship and clustering analysis on the citrus fruit flies has the advantages of stable reaction, clear amplified fragments and high polymorphism, a molecular reaction system for population genetic diversity of the citrus fruit flies is constructed, genetic diversity among populations can be shown, clustering analysis can be rapidly and accurately carried out on the citrus fruit flies, different populations can be distinguished, and the molecular marking method for carrying out genetic relationship and clustering analysis on the citrus fruit flies has an important significance on prevention and control of the citrus fruit flies.