Tocopherols, with antioxidant properties, are synthesized by photosynthetic organisms andplay important roles in human and animal nutrition.In the major oilseed crops, γ-tocopherol, andthe biosynthetic precursor to α-tocopherol, is the predominant form found on the leaves.It wassuggested that the final step of the α-tocopherol biosynthetic pathway was catalyzed by γ-tocopherol methyltransferase.　　The full-length γ-PfTMT was obtained from the total RNA of Perilla frutescens leaves byRT-PCR.Sequence analysis indicates that γ-PfTMT consisted the open reading frame of 894nucleotides encoding the protein of 34 kD polypeptide.Our results demonstrated that the E.ColiBL21(DE3) expression of the γ-PfTMT resulted in the α-tocopherol contents (and γ-tocopherolconversion yield) from 18％ in the reaction products.Transgenic Trichoderma reesei Rut-C30strains, over-expressing the γ-PfTMT was also generated by Agrobacterium tumefaciens-mediated transformation.The presence of hph and γ-PfTMT genes in the transformants wereconfirmed by PCR analysis.The expression of the γ-PfTMT gene of the transgenes wasdemonstrated by SDS-PAGE.Furthermore, we demonstrated that the Trichoderma reesei Rut-C30 expression of the γ-PfTMT gene resulted in the tocopherol composition 5.9-fold increase inα-tocopherol content by using high-performance liquid chromatographic (HPLC) method.Theincrease in the α-tocopherol content indicates that a regulatory function of the γ-PfTMT proteinconverts γ-tocopherol to α-tocopherol.　　The full-length γ-BoTMT was obtained from the total RNA of Brassica oleracea leaves byRT-PCR.Sequence analysis indicates that γ-BoTMT consisted the open reading frame of 1041nucleotides encoding the protein of 39 kD polypeptide.Our results demonstrated that the E.ColiBL21(DE3) expression of the γ-BoTMT resulted in the α-tocopherol contents (andγ-tocopherolconversion yield) from 23％ of the reaction products by using HPLC method.TransgenicTrichoderma reesei Rut-C30 strains, over-expressing the γ-BoTMT gene was also generated byAgrobacterium tumefaciens-mediated transformation.The presence of hph and γ-BoTMT gene inthe transformants was confirmed by PCR analysis.The expression of the γ-BoTMT gene of thetransgenes was demonstrated by SDS-PAGE.　　Fatty acids are the main groups of components of plant membrane lipid and seed storagelipid, and the major source of energy in plant.According to bioinformation analysis of the cDNAsequence, the specific fragment of FAD2 from immature maize embryos was isolated by RT-PCR.Results of sequence analysis indicate that FAD2 fragment contains the open reading frameof 1,236 bp long coding for the 46 kD polypeptide.Transgenic Trichoderma reesei Rut-C30strains, over-expressing the FAD2 gene from maize were generated by Agrobacteriumtumefaciens-mediated transformation.The presence of hph and FAD2 gene in the transformantswere confirmed by polymerase chain reaction (PCR) analysis.The expression of the FAD2 geneof the transgenes from Trichoderma reesei and E.coli BL21 was demonstrated by SDS-PAGE.　　In this study, we developed novel plasmids containing three plasmids designated pBI121-TMT, pCAMBIA 1301 S-FAD2 and pCAMBIA 1301S-FAD2-TMT that incorporate modified andimproved expression omega-3 and vitamin E content in seeds of the plant transformation.TheFAD2 and γ-PfTMT genes of each plasmid were driven by the constitutive CaMV 35S promoterwhich are mostly used for driving trangene expressions in both monocot and dicot planttransformation.The binary vector pCAMBIA1301S-FAD2 and vector pBI121-TMT containsFAD2, γ-PfTMT genes respectively, whereby the binary vector pCAMBIA1301S-FAD2-TMTcontain both FAD2 and γ-PfTMT gene.All three plasmid vectors were introduced into A.tumefaciens EHA105 by electroporation.