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.

Buckwheat (Fagopyrum esculentum Moench.) is an alternative crop belonging to the Polygonaceae family. Buckwheat has been well known as a plant source of rutin, quercetin, kaempferol-3-rutinoside and other phenolic compounds. Buckwheat contains more rutin than most of the other plants, which exhibits antioxidative, antihemorrhagic and blood vessel protecting properties. Phenylalanin ammonia-lyase (PAL), chalcone synthase (CHS), flavone 3-hydroxylase (F3H), and anthocyanidin synthase (ANS) were important enzymes involved in the flavonoid biosynthesis in buckwheat. In this study, we have purified full-length PAL, CHS, F3H and ANS from buckwheat using RACE PCR with degenerated primers targeted to conserved regions of orthologous sequences PAL, CHS, F3H and ANS available. The open reading frame of FePAL was 2112 nucleotides long, encodes for 704 amino acids. FePAL gene shares 80-81% identity and 88-90% similarity with Arabidopsis thaliana PAL1, Capsicum annuum PAL, Lactuca sativa PAL, and Populus trichocarpa PAL2. The cDNA of FeCHS has 1179 nucleotides open reading frame, encoding for 393 amino acids. FeCHS has identities and similarities ranging 84-86% and 92-95%, respectively, with other CHSs from P. trichocarpa, Citrus sinensis, Gerbera hybrid, and Vitis rotundifolia, whereas this gene shares 93% identity and 98% similarity with Polygonum cuspidatum. The cDNA of FeF3H has 1101 nucleotides open reading frame, encoding for 367 amino acids. FeF3H has identities and similarities ranging 76-81% and 92-97%, respectively, with other F3Hs from Gossypium hirsutum, Dimocarpus longan, Garcinia mangostana, Camellia sinensis, Vitis vinifera, Rubus coreanus, Pyrus communis, Glycine max, and Actinidia chinensis. The open reading frame of FeANS was 1074 nucleotides long, encodes for 358 amino acids. FeANS gene shares 74-76% identity and 94-96% similarity with Gypsophila elegans, C. sinensis and G. hirsutum. These result provieded the theoretical basis for the metabolic engineering on buckwheat.

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.