Agrobacterium—Mediated Transformation of Embryogenic Calli of Anliucheng and Regeneration of Plants Containing the Chimaeric Ribonuclease Gene

Anliucheng(Cltrus sinensis Osbeck), a very seedy and widely spread acidless sweet orange culutivar in south of China，was transformed by the strain of Agrobacterium Tumefaciens EHA105 carrying pTA29-barnase gene，which will induce pollen sterility in transgenic plants． The embryogenic calli of Anliucheng were co-cultivated with Agrobacterium tumefaciens for 3 days，and then transferred to selective medium containing 50 mg/L basta(a kind of herbicide) for 5 weeks．The resistant calli were recovered and regenerated 118 embryoids．A total of 13 entire plants were obtained after micro-grafted on trifoliate orange． These regenerated plants were verified by PCR am plification and confirmed by PCR-Southern blotting analysis.