Construction of the CRISPR/Cas9 module and genetic transformation of aromatic rice cv. Mentik Wangi for developing bacterial leaf blight resistance

NURUL FARIDAH RIFHANI (Graduate Program of Bioscience, Faculty of Mathematics and Natural Sciences, Universitas Sebelas Maret. Jl. Ir. Sutami 36A, Surakarta 57126, Central Java, Indonesia)
ANIVERSARI APRIANA (Research Center for Food Crops, Research Organization for Agriculture and Food, National Research and Innovation Agency. Jl. Raya Jakarta-Bogor Km. 46, Cibinong, Bogor 16911, West Java, Indonesia)
ATMITRI SISHARMINI (Research Center for Food Crops, Research Organization for Agriculture and Food, National Research and Innovation Agency. Jl. Raya Jakarta-Bogor Km. 46, Cibinong, Bogor 16911, West Java, Indonesia)
TRI JOKO SANTOSO (Research Center for Horticulture and Estate Crops, Research Organization for Agriculture and Food, National Research and Innovation Agency. Jl. Raya Jakarta-Bogor Km. 46, Cibinong, Bogor 16915, West Java, Indonesia)
KURNIAWAN RUDI TRIJATMIKO (International Rice Research Institute. 10th Floor Suite 1009, Security Bank Center 6776 Ayala Avenue, Makati Metro Manila 1226, Philippines)
INEZ H. SLAMET-LOEDIN (International Rice Research Institute. 10th Floor Suite 1009, Security Bank Center 6776 Ayala Avenue, Makati Metro Manila 1226, Philippines)
AHMAD YUNUS (Department of Agrotechnology, Faculty of Agriculture, Universitas Sebelas Maret. Jl. Ir. Sutami 36A, Surakarta 57126, Central Java, Indonesia)

Biodiversitas Journal of Biological Diversity
Vol. 24 No. 6 (2023)

Abstract. Rifhani NF, Apriana A, Sisharmini A, Santoso TJ, Trijatmiko KR, Slamet-Loedin IH, Yunus A. 2023. Construction of the CRISPR/Cas9 module and genetic transformation of aromatic rice cv. Mentik Wangi for developing bacterial leaf blight resistance. Biodiversitas 24: 3258-3268.Rice cv. Mentik Wangi is a local aromatic rice that is susceptible to pests and diseases, such as bacterial leaf blight (BLB) caused by Xanthomonas oryzae(Xoo). This bacteria can cause damage to plants thereby reducing crop yields. This study aimed to obtain the CRISPR/Cas9 module construct as well as introducing the construct into rice cv. Mentik Wangi for developing BLB resistance. The manufacture of the CRISPR/Cas9 module was carried out using the Golden Gate method, and the introduction of that construct into rice cv. Mentik Wangi was carried out using Agrobacterium tumefaciens. Constructing a CRISPR/Cas9 module with multiple gRNA of OsSWEET11 and OsSWEET14 genes has succeeded, 129 putative transformant lines of T0 generation produced with the regeneration and transformation efficiency values ??are respectively 9.4% and 9.8%. The results showed that 36 lines were positive for the hpt gene, indicating that the CRISPR/Cas9-gRNAOsSWEET module construct was successfully entered into rice cv. Mentik Wangi. Further analysis is needed to identify mutagenesis in the target gene region of TI generation transgenic plants as well as phenotypic tests for BLB resistance.