Transgenic Wheat Plants Derived from Agrobacterium-mediated Transformation of Mature Embryo Tissues
Although significant progress has been made on Agrobacterium-mediated wheat transformation, the current methodologies use immature embryos as recipient tissues, a process which is labor intensive, time consuming and expensive. In this study, we have managed to develop an Agrobacterium-based transformation scheme using explants derived from mature embryos. Based on transient expression of β-glucuronidase (GUS) marker, mature embryo halves prepared from freshly imbibed seeds were generally most susceptible to Agrobacterium-mediated T-DNA transfer. According to the results of callus induction and shoot production, Yumai 66 and Lunxuan 208 showed higher selection and regeneration efficiency than Bobwhite. In line with this finding, fertile T 0 transgenic plants were most readily obtained for both spring and winter wheat when mature embryo halves were used for co-inoculation by Agrobacterium cells. The presence of the antibiotic selection marker (nptII, encoding neomycin phosphotransferase II) in the T 0 plants was revealed by both genomic PCR amplification and the enzyme-linked immunosorbent assay (ELISA). Additional analysis showed that the transgene was stably inherited from the two different generations and segregated normally among the T 1 progenies. Further development along this line will raise the efficiency of wheat transformation and increase the use of this approach in the molecular breeding of wheat crop.
KeywordsAgrobacterium tumefaciens genetic transformation mature embryo wheat transgenic plant
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