Plant regeneration and Agrobacterium-mediated transformation of Achyranthes bidentata using cotton EREBP gene

ABSTRACT

The aim of this work was to study the plant regeneration and Agrobacterium-mediated transformation of Achyranthes bidentata using cotton EREBP gene. Results showed that the callus induction rate of stems from A. bidentata was the highest (100%) and bud was in approximately 70% of calli from stems. However bud differentiation rate of the callus from leaves and petioles was very low. Compared with ceftriaxone, 200mg/L cefotaxime could completely control Agrobacterium tumefaciens and had relatively less toxic action on the stems of A. bidentata. In addition, the induction rate of callus resistant to hygromycin was the highest when infected for 3 min and co-cultivated for 3 d. Six positive transgenic plants transformed with pCAMBIA1304-GhEREB2 expression vector were obtained and confirmed by PCR. The expression of target gene GhEREB2 was detected in five transgenic plants by RT-PCR. In brief, an efficient system of genetic transformation and plant regeneration was established for A. bidentata.