Transformasi Gen Pengendali Flavanone 3-Hydroxylase (F3H) untuk Meningkatkan Kandungan Sakuranetin Pada Tanaman Padi

Abstract

Rice (Oryza sativa L.) is one of the staple food sources for most of the world's population, such as in Asia and Indonesia, which are among the countries with high rice consumption rates. The challenges faced in rice cultivation are extreme weather changes, starting with a significant increase in temperature during the dry season, which increases the potential for drought, and high rainfall during the rainy season. These conditions make rice plants susceptible to disease. One of the defense mechanisms of rice plants when attacked by disease is to produce a secondary metabolite compound called sakuranetin. The production of sakuranetin is influenced by the activity of several other enzymes such as flavone synthase (FNS), flavanone 3-hydroxylase (F3H), flavonoid 3'-hydroxylase (F3'H), flavonoid 3',5'- hydroxylase (F3'5'H) to uridine diphosphate (UDP)-glycosyltransferases (UGTs). Increased production of sakuranetin in plants can be achieved by knocking out the biosynthesis of naringenin into dihydrokaempferol by the flavanone 3-hydroxylase (F3H) enzyme using genome editing methods. This study has successfully designed an sgRNA construct with an efficiency of 53.53%. The suitability of the sgRNA construct has been confirmed by sequencing analysis, as evidenced by the same pattern. The success of plasmid transformation into Agrobacterium was confirmed by colony growth on selective media and detection of the hptII gene. In the rice plant transformation stage, the Ketan Hitam and Koshihikari varieties showed the best results, producing one and two putative transformants, respectively. All planlets were confirmed as transformant plants, as evidenced by the presence of the hptII transgene integrated into the plants.