Twenty two common millet (Panicum miliaceum L.) varieties collected from Korea, China and Russia were investigated for their phylogenetic relationship using 5S ribosomal DNA sequences with a hope to provide the basic information on their exact origin. Sequences of 5S rDNA were isolated by PCR. The primers, 5s-rRNA1 and 5s-rRNA2, were designed to isolate the complete NTS. Genomic DNA amplification produced two fragments with different length, 900 bp and 400 bp fragments, confirming the presence of two types of 5S rDNA repeats that differed from each other in the length of the NTS region. Amplified DNAs of 400 bp fragment were subcloned and used for further investigation. The obtained NTS sequences ranged from 200 to 300 bp and homology of sequences among plant materials was much higher than long repeat. CLUSTALW multiple aligment of 5S rDNA sequences from 22 different common millets revealed the clear difference by their origin. And critically different areas with insert or deletion were also confirmed. Those sequence difference seems to be used for discrimination of cultivars from different origin and use as molecular markers for origin identification. In phylogenic tree construction, the clear classification was shown where the genotypes from China and Russia is positioned together and stay away from domestic genotypes.

Tocopherol belongs to the Vitamin E class of lipid soluble antioxidants that are essential for human nutrition. Tocopherols consist of a chromanol ring and a 15-carbon tail derived from homogentisate (HGA) and phytyl diphosphate, respectively. Condensation of HGA and phytyl diphosphate, the committed step in tocopherol biosynthesis, is catalyzed by HGA phytyltransferase (HPT). To increase tocopherol level in cucumber fruit, we performed Agrobacterium-mediated transformation with mdhpt gene, encoding HPT, isolated from apple (Malus domestica). We obtained two independent T0 cucumber plants from transforming 2,142 explants. Southern blot analysis of T0 showed that mdhpt was inserted in the genome. In order to test the tocopherol level, T1 cucumber fruits were screened by GC analysis. From 39 T1 fruits tested, some fruits showed 1.5-2 fold in α- and β-tocopherol content comparing to control. However, most of fruits contained lower levels of γ- and δ- tocopherol indicating a partition of metabolites in tocopherol biosynthesis pathway. We selected these T1 line and made a self-cross to obtain T2 seeds.