RNAi-Mediated Silencing of the Flavanone 3-Hydroxylase Gene into Nelumbo nucifera Geartn. Using Particle Bombardment

Abstract

Flavanone 3-hydroxylase (F3H) is an important enzyme in the synthesis anthocyanin pathway
involved in color production in flower plants. In this study transformation of F3H gene in pJA8F3H plasmid
which can stimulate the process of RNA interference (RNAi) and subsequently inhibits the expression of
F3H, into lotus (Nelumbo nucifera Gaertn.) cv. Buntharik by particle bombardment was evaluated. After
bombardment, 105 pieces of shoot clusters of lotus were selected on MS medium supplemented with 5μM
BA and 10 mg/L glufosinate ammonium for 16 weeks. The results found that 27 pieces of shoot clusters
survived (28.35%). And selection mature plants for existence detection gene 5 pieces. The PCR analysis
of F3H gene was determined by primer F3H sense F and F3H sense R which specific to F3H gene of 308
nucleotides and bar gene was determined by primer bar gene F and bar gene R which specific to gene
phosphinothricin-acetyl transferase (bar gene) of 430 nucleotides. The result found that existence of gene

5 pieces total of 100 percent. The expression of F3H gene was detected by RT-PCR technique. The results
showed that the expression of F3H gene reduced in transgenic plants comparing with the lotus
non - transgenic plants.