genetic modification of the soybean to enhance the β-carotene content through seed-specific expression转基因大豆的提高通过seed-specificβ-carotene内容表达.pdfVIP

Genetic Modification of the Soybean to Enhance the b-Carotene Content through Seed-Specific Expression 1. 2. 1 1 1 1 Mi-Jin Kim , Jae Kwang Kim , Hye Jeong Kim , Jung Hun Pak , Jai-Heon Lee , Doh-Hoon Kim , Hong 1 1 3 1 2 Kyu Choi , Ho Won Jung , Jeong-Dong Lee , Young-Soo Chung *, Sun-Hwa Ha * 1 Department of Genetic Engineering, Dong-A University, Busan, Republic of Korea, 2 National Academy of Agricultural Science, Rural Development Administration, Suwon, Republic of Korea, 3 School of Applied Biosciences, Kyungpook National University, Daegu, Republic of Korea Abstract The carotenoid biosynthetic pathway was genetically manipulated using the recombinant PAC (Phytoene synthase-2A- Carotene desaturase) gene in Korean soybean (Glycine max L. cv. Kwangan). The PAC gene was linked to either the b- conglycinin (b) or CaMV-35S (35S) promoter to generate b-PAC and 35S-PAC constructs, respectively. A total of 37 transgenic lines (19 for b-PAC and 18 for 35S-PAC) were obtained through Agrobacterium-mediated transformation using the modified half-seed method. The multi-copy insertion of the transgene was determined by genomic Southern blot analysis. Four lines for b-PAC were selected by visual inspection to confirm an orange endosperm, which was not found in the seeds of the 35S- PAC lines. The strong expression of PAC gene was detected in the seeds of the b-PAC lines and in the leaves of the 35S-PAC lines by RT-PCR and qRT-PCR analyses, suggesting that these two different promoters function distinctively. HPLC analysis of the seeds and leaves of the T2 gen