3 years ago

Developing a flexible, High-efficiency Agrobacterium-mediated Sorghum Transformation System with Broad Application

Jeffry D. Sander, Ajith Anand, Marissa K. Simon, Gina Zastrow-Hayes, Shai J. Lawit, Michael Miller, Amy L. Sigmund, Ping Che, Todd J. Jones, Zuo-Yu Zhao, Donglong Liu, Emily Wu, Marc C. Albertsen, Weiwei Zhu
Sorghum is the fifth most widely planted cereal crop in the world and is commonly cultivated in arid and semi-arid regions such as Africa. Despite its importance as a food source, sorghum genetic improvement through transgenic approaches has been limited because of an inefficient transformation system. Here we report a ternary vector (also known as co-habitating vector) system using a recently described pVIR accessory plasmid that facilitates efficient Agrobacterium-mediated transformation of sorghum. We report regeneration frequencies ranging from 6-29% in Tx430 using different selectable markers and single copy, backbone free “quality events” ranging from 45-66% of the total events produced. Furthermore, we successfully applied this ternary system to develop transformation protocols for popular but recalcitrant African varieties including Macia, Malisor 84-7 and Tegemeo. In addition, we report the use of this technology to develop the first stable CRISPR/Cas9 mediated gene knockouts in Tx430. This article is protected by copyright. All rights reserved.

Publisher URL: http://onlinelibrary.wiley.com/resolve/doi

DOI: 10.1111/pbi.12879

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