Optimized Agrobacterium -mediated sorghum transformation protocol and molecular data of transgenic sorghum plants

In Vitro Cellular & Developmental Biology. Plant : Journal of the Tissue Culture Association
Emily WuZuo-Yu Zhao

Abstract

Agrobacterium-mediated sorghum transformation frequency has been enhanced significantly via medium optimization using immature embryos from sorghum variety TX430 as the target tissue. The new transformation protocol includes the addition of elevated copper sulfate and 6-benzylaminopurine in the resting and selection media. Using Agrobacterium strain LBA4404, the transformation frequency reached over 10% using either of two different selection marker genes, moPAT or PMI, and any of three different vectors in large-scale transformation experiments. With Agrobacterium strain AGL1, the transformation frequencies were as high as 33%. Using quantitative PCR analyses of 1,182 T0 transgenic plants representing 675 independent transgenic events, data was collected for T-DNA copy number, intact or truncated T-DNA integration, and vector backbone integration into the sorghum genome. A comparison of the transformation frequencies and molecular data characterizing T-DNA integration patterns in the transgenic plants derived from LBA4404 versus AGL1 transformation revealed that twice as many transgenic high-quality events were generated when AGL1 was used compared to LBA4404. This is the first report providing molecular data for T-DNA integra...Continue Reading

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Citations

Jan 1, 2020·Biotechnology for Biofuels·Christine A Ondzighi-AssoumeC Neal Stewart
Jun 27, 2017·Biotechnology for Biofuels·Tesfamichael H KebromJohn E Mullet
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Methods Mentioned

BETA
transgenic
PCR
fluorescence microscopy

Software Mentioned

SAS
LAPLACE

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