Four novel 2-methoxyimino phenylacetate derivatives containing 1,3,4-oxadiazole ring were designed and synthesized from the key intermediate of Trifloxystrobin or Azoxystrobin via intermediate derivatization and active structure splicing. The chemical structures of the target compounds were confirmed by ¹H NMR, ¹³C NMR and elemental analysis. The crystal structure of methyl (E)-2-(methoxyimino)-2-(2-(((5-((4-methoxyphenoxy)methyl)- 1,3,4-oxadiazol-2-yl)thio)methyl)phenyl)acetate (A1) was determined by single-crystal X-ray diffraction. Compound A1 belongs to triclinic system, space group P with two molecules in each unit cell. The benzene ring plane C(2)–C(3)–C(4)–C(5)–C(6)–C(7) and oxazole ring plane are nearly parallel with the dihedral angle of 6.4°. The benzene ring plane C(12)–C(13)–C(14)–C(15)–C(16)–C(17) and oxazole ring plane are not perpendicular with the dihedral angle of 49.4°. The crystal of compound A1 is stabilized by π-π stacking interactions. The fungicidal activities of the target compounds against four plant pathogenic fungi in vitro were tested, and some of them had good activities. The DFT calculation was carried out to study the structure-activity relationship of the title derivatives using Gasian 09 and Multiwfn 3.6.