C synthetic gene of CryIAb-CryIAc fusion to generate resistant sugarcane to shoot or stem borer

Abstract

The sugar yield lost caused by borer is about 10% or equal to 1000 kg of sugar/ha. There is no resistance trait to stem or shoot borer available in sugarcane germplasm. Genetic engineering by expressing of the Cry1Ab-Cry1Ac gene fusion is efforts to develop resistant variety to borer. A synthetic gene which consists of Rubisco gene promoter, chloroplast specific transit peptide (CTP) and the Cry1Ab-Cry1Ac was designed and assembled for a total size of 4019bp. It was inserted into pU3775CE plasmid cloning, and then into pCAMBIA5300_Ubi-tNOS plasmid vector at HindIII and KpnI sites, producing of pCAMBIA5300_RbcS::Cry1Ab- Cry1Ac. This plasmid was transforming into Agrobacterium tumefaciens and then transformed into Bulu Lawang (BL) sugarcane calli. Research aims were to identify lines produced from plant transformation molecularly and to evaluate their resistances against shoot borer. Methods applied were a DNA isolation and PCR using KAPA 2G ready mix and CryIAc-316F and CryIAc-316R primers, and plant bioassay with larva instar 1. Research progresses were 30 lines had been identified and proved containing of the cryIAc gene. Preliminary results of bioassay showed that there was variation among 10 BL Cry lines on shoots (from healthy to wilt and dry) and also on shoot borer conditions.