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  • 1 Chinese Academy of Agricultural Sciences National Key Facility of Crop Gene Resources and Genetic Improvement, Key Laboratory of Crop Genetics and Breeding, Ministry of Agriculture, Institute of Crop Science Beijing 100081 China
  • 2 Chinese Academy of Sciences The State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology Beijing 100101 China
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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.

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