Improvement<i>of</i>sugarcane for borer resistance using<i>Agrobacterium</i>mediated transformation of<i>cry1Ac</i>gene

Abstract

The sugarcane (<i>Saccharum</i> X <i>officinarum</i>) is one of the most important crops used to produce sugar and raw material for biofuel in the world. One of the main causes for sucrose content and yield losses is the attack by insect. In this investigation, <i>cry1Ac</i> gene was introduced into sugarcane variety GT54-9(C9) using the <i>Agrobacterium tumefaciens</i> transformation method for transgenic sugarcane production presenting insect-resistance. The <i>A. tumefaciens</i> strain GV1303 including pART<i>cry1Ac</i> vector was used for the production of transformed sugarcane. The <i>Bacillus thuringiensis cry</i> gene were successfully used to produce transgenic plants used for the improvement of both agronomic efficiency and product quality by acquiring insect resistance. PCR and Southern hybridization techniques were used to confirm the <i>cry1Ac</i> gene incorporation into sugarcane genome. Transformation percentage was 22.2% using PCR analysis with specific primers for <i>cry1Ac</i> and <i>npt-II</i> (Neomycin phosphotransferase) genes. The expression of <i>cry1Ac</i> gene was determined using reverse transcriptase polymerase chain reaction (RT-PCR), QuickStix test, and insect bioassays. Bioassays for transformed sugarcane plants showed high level of toxicity to <i>Sesamia cretica</i> giving 100% mortality of the larvae. Sugarcane insect resistance was improved significantly by using <i>cry1Ac</i> gene transformation.