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Tuan, Pham Anh,Kim, Jae Kwang,Lee, Sanghyun,Chae, Soo Cheon,Park, Sang Un American Chemical Society, Books and Journals Divi 2012 Journal of agricultural and food chemistry Vol.60 No.48
<P>Riboflavin (vitamin B2) is the universal precursor of the coenzymes flavin mononucleotide and flavin adenine dinucleotide--cofactors that are essential for the activity of a wide variety of metabolic enzymes in animals, plants, and microbes. Using the RACE PCR approach, cDNAs encoding lumazine synthase (McLS) and riboflavin synthase (McRS), which catalyze the last two steps in the riboflavin biosynthetic pathway, were cloned from bitter melon (Momordica charantia), a popular vegetable crop in Asia. Amino acid sequence alignments indicated that McLS and McRS share high sequence identity with other orthologous genes and carry an N-terminal extension, which is reported to be a plastid-targeting sequence. Organ expression analysis using quantitative real-time RT PCR showed that McLS and McRS were constitutively expressed in M. charantia, with the strongest expression levels observed during the last stage of fruit ripening (stage 6). This correlated with the highest level of riboflavin content, which was detected during ripening stage 6 by HPLC analysis. McLS and McRS were highly expressed in the young leaves and flowers, whereas roots exhibited the highest accumulation of riboflavin. The cloning and characterization of McLS and McRS from M. charantia may aid the metabolic engineering of vitamin B2 in crops.</P>
Tuan, Pham Anh,Kim, Jae Kwang,Kim, Haeng Hoon,Lee, Sook Young,Park, Nam Il,Park, Sang Un American Chemical Society 2011 Journal of agricultural and food chemistry Vol.59 No.10
<P>Phytoene synthase (PSY) and phytoene desaturase (PDS), which catalyze the first and second steps of the carotenoid biosynthetic pathway, respectively, are key enzymes for the accumulation of carotenoids in many plants. We isolated 2 partial cDNAs encoding PSY (<I>AsPSY-1</I> and <I>AsPSY-2</I>) and a partial cDNA encoding PDS (<I>AsPDS</I>) from <I>Allium sativum</I>. They shared high sequence identity and conserved motifs with other orthologous genes. Quantitative real-time PCR analysis was used to determine the expression levels of AsPSY1, AsPSY2, and AsPDS in the bulbils, scapes, leaves, stems, bulbs, and roots of garlic. High-performance liquid chromatography demonstrated that carotenoids were not biosynthesized in the underground organs (roots and bulbs), but were very abundant in the photosynthetic organs (leaves) of <I>A. sativum</I>. A significantly higher amount of β-carotene (73.44 μg·g<SUP>–1</SUP>) was detected in the leaves of <I>A. sativum</I> than in the other organs.</P><P><B>Graphic Abstract</B> <IMG SRC='http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/jafcau/2011/jafcau.2011.59.issue-10/jf2009827/production/images/medium/jf-2011-009827_0003.gif'></P>
Hairy Root Culture of Platycodon grandiflorum Transformed with Agrobacterium rhizogenes
Pham Anh Tuan,Nam Il Park,Xiaohua Li,Yong Kyoung Kim,Sang Un Park 한국작물학회 2010 한국작물학회 학술발표대회 논문집 Vol.2010 No.04
We developed an efficient protocol for transformation of balloon flower (Platycodon grandiflorum) root cultures by using cotyledon leaf explants that were infected by Agrobacterium rhizogenes. We found that four different strains of A. rhizogenes differed in their ability to transform P. grandiflorum hairy root cultures. We also found correct antibiotics concentration for selection after transformation by Agrobacterium. Our results demonstrate that use of suitable strain of A. rhizogenes and correct level of antibiotics for the hairy root culture, and may allow to study and apply of valuable metabolites like phenolic compounds from P. grandiflorum hairy root culture.
Pham Anh Tuan,Nam Il Park,Xiaohua Li,Hui Xu,Yong Kyoung Kim,Sang Un Park 한국작물학회 2010 한국작물학회 학술발표대회 논문집 Vol.2010 No.04
Allium sativum, belongs to a member of the onion family (Alliaceae) are economically important vegetables because of the culinary value and medicinal purpose. Using PCR strategy with degenerated primers targeted to conserved regions of orthologous phenylalanine ammonia-lyase (PAL) and cinnamate 4-hydroxylase (C4H) sequences available, full-length PAL and C4H from A. sativum. The amino acid sequence of these genes is highly conserved, particularly AsPAL and AsC4H has greater than 70% amino acid identity to other plants. AsPAL and AgC4H were most highly expressed in roots of A. sativum, whereas lowest level of transcript was detected in flower. Phenolic compounds most highly produced in flowers of A. sativum. The presented sequences and expression an alysis of PAL and C4H will provide possible material to enhance the understading of phenolic compounds synthesis in A. sativum.