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African Journal of Biotechnology

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Optimization of Agrobacterium-mediated transformation parameters for sweet potato embryogenic callus using β-glucuronidase (GUS) as a reporter

Y Xing, Q Yang, Q Ji, Y Luo, Y Zhang, K Gu, D Wang

Abstract


Agrobacterium-mediated transformation factors for sweet potato embryogenic calli were optimized using -glucuronidase (GUS) as a reporter. The binary vector pTCK303 harboring the modified GUS gene
driven by the CaMV 35S promoter was used. Transformation parameters were optimized including bacterial concentration, pre-culture period, co-cultivation period, immersion time, acetosyringone (AS)
concentration and mannitol treated time. Results were obtained based on the percentage of GUS expression. Agrobacterium tumefaciens strain EHA105 at concentration OD600 nm = 0.8 showed the highest virulence on sweet potato embryogenic callus. Four days of pre-culture, four days of
co-cultivation, 10 min of immersion, 200 M acetosyringone and 60 min of mannitol-treated embryogenic callus gave the highest percentage of GUS positive transformants.



http://dx.doi.org/10.5897/AJB2007.000-2411
AJOL African Journals Online