The invention discloses application of a tobacco C2H2-type zinc finger protein gene Nt540. A gene fragment with a nucleotide sequence shown as SEQ ID NO.1 is provided. The invention further discloses a recombinant vector comprising the gene fragment, a recombinant strain and application of the recombinant vector and the recombinant strain. Through transfer of the gene Nt540 and the recombinant strain, the tobacco quality is improved effectively, novel materials and novel approaches are provided for tobacco variety improvement, and accordingly such application has a promising application prospect.