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Optimization of Genetic Transformation Conditions in Pogonatherum paniceum Mediated by Agrobactrium tumefaciens |
LI Mei-yu, LI Rui, YU Min, WANG Sheng-hua, CHEN Fang |
Key Laboratory of Bio-resources and Eco-environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu 610064, China |
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Abstract A perennial herbaceous plant, Pogonatherum paniceum, plays an important role inecological restoration and landscape construction. To establish highly efficient transformation system, factors which influenced transformation including Agrobacterium concentration, infection time, concentration of acetosyringone and glucose, and co-culture time were studied with the mature embryo callus of Pogonatherum paniceum based on the variance of GUS (β-glucuronidase) trsnsient expression rate. The results showed that the transformation efficiency was improved with Agrobacterium concentration of OD600=0.6, incubation for 10 min, co-culture time for 5d. Acetosyringone with the concentration of 20mg/L and 10g/L glucose could significantly enhance the transformation rate. Utilizing this system, more than 57% kanamycin resistance selection efficiency was obtained. GUS gene has successfully integrated into the plant genome by GUS and PCR detection. The establishment of this transformation system lays a foundation for genetic modified and study of functional genes of Pogonatherum paniceum.
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Received: 28 May 2012
Published: 25 January 2013
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