Efficient method of seed transformation via Agrobacterium tumefaciens for obtaining transgenic plants of Hibiscus cannabinus L.

Abstract

Kenaf (Hibiscus cannabinus L.) is an economic and ecological fiber crop but suffers severe losses in fiber yield and quality under the stressful conditions of excess salinity and drought. Therefore, in order to obtain new cultivars of kenaf that could face and overcome abiotic stress, it is crucial to have a suitable protocol of genetic transformation. Therein, experiments were carried out on transformation of kenaf mature seeds using a co-culture of Agrobacterium tumefaciens. In this study, a A. tumefaciens-mediated transformation of kenaf seed have been developed by tissue-culture-independent procedure. The GV3010 Agrobacterium strain harboring the pGreenII binary vector that carries the neomymycin phosphotransferase II (npt II) gene for selection was used for transformation. The presence of the transgene and its stable expression were confirmed by PCR. In addition, the transgenic character of the selected transgenic T0 and T1 plants has been confirmed by germination test in the presence of kanamycin. Molecular analysis of F0 plants of three transgenic lines revealed the real integration of VvWRKY2 gene into the kenaf genome. Thus, our described method was an efficient, fast, and reliable procedure by which stable transgenic flowering plants were obtained within a short period of 3 months with 6% transformation efficiency.