Agrobacterium-mediated transformation of Jatropha curcas young leaf explants with lateral shoot-inducing factor (LIF).

Abstract

Limited branching is considered as one of the major factors limiting oil-bearing seed yield in Jatropha curcas L. Seeking for promoting branches, Lateral shoot-Inducing Factor (LIF) was going to be transferred into the J. curcas genome. For this transgenic approach to be successful, we developed a stable and efficient transformation protocol using Agrobacterium tumefaciens strain LBA4404 for young leaf explants of J. curcas. Parameters optimized were bacterial strain, bacterial concentration, co-cultivation period, immersion time and concentration of cefotaxime and kanamycin. Transformation with LIF gene was achieved subsequently. PCR amplification confirmed that the transformation efficiency was 23.91±5.78%. Southern blot hybridization was performed to reveal that single and multiple copy events and further confirm that LIF was stably integrated into the J. curcas genome. © 2010 Friends Science Publishers