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Agrobacterium-mediated transformation of shoot apices of Kentucky bluegrass (Poa pratensis L.) and production of transgenic plants carrying a betA gene

Zhang, Kewei, Wang, Juan, Hu, Xiaorui, Yang, Aifang, Zhang, Juren
Plant cell, tissue, and organ culture 2010 v.102 no.2 pp. 135-143
Poa pratensis, transgenic plants, Agrobacterium tumefaciens, genetic transformation, shoot meristems, apical meristems, herbicide resistance, acetolactate synthase, inheritance (genetics), in vitro regeneration, micropropagation
Apical meristems of multiple shoots produced from axenic seedlings of Kentucky bluegrass (Poa pratensis L.) were used for Agrobacterium tumefaciens-mediated transformation. Transformation parameters were optimized for concentration of bacterial cells, duration of infection, and vacuum infiltration. The highest transformation frequency (1.42%) was obtained by infection with Agrobacterium suspension of OD₆₀₀ = 0.6 for 5 min, under a negative pressure of 0.5 × 10⁵ Pa. After co-cultivation, the herbicide-resistant plants were rooted and transplanted into flowerpots. Transgenic plants were confirmed by polymerase chain reaction (PCR) assay and Southern blot analysis. Using this transformation system, the betA gene encoding choline dehydrogenase and mutant als gene encoding the enzyme acetolactate synthase were introduced into three Kentucky bluegrass cultivars.