Establishment of an Agrobacterium mediated genetic transformation system for Eremochloa ophiuroides

Abstract

Abstract: Agrobacterium tumefaciens strain LBA4404 harboring the plasmid pCAMBIA1301 which contains stress tolerance-related transcription factor ZjDREB1 gene and reporter gene HPT was used to transform axillary bud-derived embryogenic calluses of Eremochloa ophiuroides ‘E126’. To systematically optimize the conditions of centipedegrass transformation, several factors which could influence Agrobacterium-mediated DNA transfer were examined, including concentration of selective agent and bacteriostat, bacterial concentration, duration of infection and co-cultivation. Consequently, a transformation system of transgenic centipedegrass plants was established. It was found that the efficiency of transformation highly relied on the following optimal conditions: The concentration of hygromycin for selection was 30 mg/L; the cefotaxime concentration for restraining the Agrobacterium was 400 mg/L; OD₆₀₀ as 0.3, 30 min of infection time and 3 days. Co-cultivation time was the optimized condition for genetic transformation system in E. ophiuroides. Ten transgenic plants were identified by PCR detection from initial 44 regenerated plants.

CLC Number:

ZHANG Fang, WANG Zhou, ZONG Jun-qin, LIU Jian-xiu, SHE Jian-ming. Establishment of an Agrobacterium mediated genetic transformation system for Eremochloa ophiuroides[J]. Acta Prataculturae Sinica, 2011, 20(2): 184-192.

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