Agrobacterium tumefaciens-Mediated Transformation of Maize Embryos Using a Standard Binary Vector System

Oxford University Press (OUP) - Tập 129 Số 1 - Trang 13-22 - 2002
Bronwyn Frame1, Huixia Shou1, Rachel Chikwamba1, Zhanyuan Zhang1, Cheng‐Bin Xiang2, Tina M. Fonger1, Sue Ellen Pegg1, Baochun Li1, Dan Nettleton3, Deqing Pei3, Kan Wang1
1Plant Transformation Facility, Departments of Agronomy (B.R.F., H.S., R.K.C., Z.Z., T.M.F., S.E.K.P., B.L., K.W.),
2Botany (C.X.), and
3Statistics (D.S.N., D.P.), Iowa State University, Ames, Iowa 50011

Tóm tắt

Abstract We have achieved routine transformation of maize (Zea mays) using an Agrobacterium tumefaciensstandard binary (non-super binary) vector system. Immature zygotic embryos of the hybrid line Hi II were infected with A. tumefaciens strain EHA101 harboring a standard binary vector and cocultivated in the presence of 400 mg L−1  l-cysteine. Inclusion of l-cysteine in cocultivation medium lead to an improvement in transient β-glucuronidase expression observed in targeted cells and a significant increase in stable transformation efficiency, but was associated with a decrease in embryo response after cocultivation. The average stable transformation efficiency (no. of bialaphos-resistant events recovered per 100 embryos infected) of the present protocol was 5.5%. Southern-blot and progeny analyses confirmed the integration, expression, and inheritance of the bar andgus transgenes in R0, R1, and R2 generations of transgenic events. To our knowledge, this represents the first report in which fertile, stable transgenic maize has been routinely produced using an A. tumefaciensstandard binary vector system.

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Oxford University Press (OUP)

Tập 129 Số 1

13-22

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