Developing a flexible, high‐efficiency Agrobacterium‐mediated sorghum transformation system with broad application

• DOI: 10.1111/pbi.12879
• Publication Date: 2018-01-12T01:22:39Z
• Authors: Ping Che; Ajith Anand; Emily Wu; Jeffry D. Sander; Marissa K. Simon; Weiwei Zhu; Amy L. Sigmund; Gina Zastrow‐Hayes; Michael Miller; Donglong Liu; Shai J. Lawit; Zuo‐Yu Zhao; Marc C. Albertsen; Todd J. Jones
• Keywords: Genetic Markers; 2. Zero hunger; 0303 health sciences; Genetic Vectors; Gene Transfer Techniques; Agrobacterium; 15. Life on land; Plants, Genetically Modified; 03 medical and health sciences; Transformation, Genetic; CRISPR-Cas Systems; Genetic Engineering; Research Articles; Sorghum

SummarySorghum 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 cohabitating vector) system using a recently described pVIR accessory plasmid that facilitates efficient Agrobacterium‐mediated transformation of sorghum. We report regeneration frequencies ranging from 6% to 29% in Tx430 using different selectable markers and single copy, backbone free ‘quality events’ ranging from 45% to 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.